From ernestinemiddleton <@t> yahoo.ca Sun May 1 14:54:30 2005 From: ernestinemiddleton <@t> yahoo.ca (Ernestine Middleton) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] fungus control blocks Message-ID: <20050501195430.97482.qmail@web51502.mail.yahoo.com> Hi; Sorry, my e-mail address is ernestinemiddleton@yahoo.ca. Ernestine. --------------------------------- Post your free ad now! Yahoo! Canada Personals From christian <@t> eisbo.dk Mon May 2 03:31:07 2005 From: christian <@t> eisbo.dk (christian@eisbo.dk) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Ki67 - kidney. Message-ID: Hey users of histonet. I am trying to stain rat kidneys for Ki67! I have bought the recomended antibody. But this one is for human! Do anyone know if this can be used since i dont have a outcome! Do anyone have a protocol that works? Christian Eisbo From TillRenee <@t> uams.edu Mon May 2 07:55:02 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] antigen retrieval/enzyme digestion Message-ID: When trying both on the same slides, which would go first, the antigen retrieval or the enzyme digestion? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From kelly.mcqueeney <@t> bms.com Mon May 2 08:26:49 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Ki67 - kidney. In-Reply-To: References: Message-ID: <42762A99.7010505@bms.com> Hi Christian, I have used rabbit polyclonal human Ki67 antibody on rat kidney and it worked beautifully. Make sure you use a wash buffer with tween-20. Simply follow the protocol recommended by Abcam (ab833). I used heat antigen retrieval in citrate buffer (Dako), 3% BSA (only) as a block and Envision+ rabbit kit (Dako) rather than ABC. The incubation time is O/N at 4C. Let me know if you need more detailed information. You can also use Ki67 antibody from Santa Cruz. Good luck, Kelly christian@eisbo.dk wrote: >Hey users of histonet. >I am trying to stain rat kidneys for Ki67! >I have bought the recomended antibody. But this one is for human! >Do anyone know if this can be used since i dont have a outcome! >Do anyone have a protocol that works? >Christian Eisbo > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From pex0220 <@t> yahoo.com.cn Mon May 2 08:30:08 2005 From: pex0220 <@t> yahoo.com.cn (=?gb2312?q?=CC=EC=20=D0=C1?=) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Normal donkey serum block solution Message-ID: <20050502133008.26004.qmail@web15507.mail.cnb.yahoo.com> Hello,all, I am doing double-immunofluorescence. Both the hosts of my secondary antibodies are donkey, so I plan to use normal donkey serum for blocking, but I do not know how to make up this solution. I have ordered donkey serum from sigma company, but the specification doesnot show how to make up it. so if you have done it, I hope that you can do me a favor. In addition, if the hosts of my secondary antiboies are different, one is horse, the other is donkey, which solution do I use for blocking? Thank you! Best wishes! Guofeng Giessen --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From timothy.m.coskran <@t> pfizer.com Mon May 2 12:11:54 2005 From: timothy.m.coskran <@t> pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Ki67 - kidney Message-ID: <599089EF29304C44AF1D84B1C223AC63019D8E28@groamrexm03.amer.pfizer.com> Christian, We've been using an anti-rat Ki-67, clone MIB5, from DakoCytomation with good results on formalin fixed paraffin embedded tissues. We use a standard ABC detection and anti mouse rat adsorbed secondary, after pre-treatment using pH citrate buffer in a pressure cooker. For Ki-67 antibodies, we've had better success using a pressure cooker rather than a steamer for antigen retrieval. If you have any questions let me know, Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From drsvsrini <@t> yahoo.com Mon May 2 13:15:24 2005 From: drsvsrini <@t> yahoo.com (Mr srinivas sv) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] i need some technical help! In-Reply-To: 6667 Message-ID: <20050502181524.84357.qmail@web31503.mail.mud.yahoo.com> Hello Vinnie and all users of histonet, Thanks for the help.I think i was not clear in my mail. I am looking at the expression of steroidogenic enzymes in sheep ovary at different physiological status. my primary antibodies are polyclonal rabbit anti-mouse and my seconadary is Goat anti rabbit conjugated with HRP. one primary antibody is home grown but from different lab and the other is commercial from Research diagnostics. My seconadary is not biotilynated and i am using VIP ( has H2O2 as a substrate) as a color reagent. I am using 1% H2O2 to block the endogenous peroxides and Normal goat serum with 1%BSA for blocking the nonspecific antigens (Serum blocking). I am using Normal Rabbit sera for negative control as my primary antibody is raised in rabbit. I have reached the dilution of primary antibody to work with by looking at the response obtained with different dilution and 1:200 dilutions works good for the primary and 1:100 dilution works good for the secondary antibody. my problem is that i am getting the color reaction in both positive ( ACTUAL PRIMARY Ab) as well my negative (NORMAL RABBIT SERA) controls. so the problem is with my normal rabbit sera (source from SIGMA) . i beleive that there is less problem with the background staining. I am facing problem with both the home grown as well the commercial source of antibodies. so my tissue sections are cryosections and are from sheep. but i am using the primary antibody raised in rabbit. i am planning to use the liver acetone powder with the idea of blocking non specific sites in my tissues or blocking non specific antibodies in my polyclonal primary antibody raised in rabbit and the normal rabbit sera(NRS) which i am using as a negative control. i am not sure about the source of the liver acetone powder i need to use. unfortunately sheep liver powder is not available commercially. so i was planning to go for the calf one. i have not yet ordered and waiting for your suggestions. but as you suggested, i have got the sheep liver with me and it would be of great help if you let me know how to prepare the sheep liver acetone powder by my own. whether it helps to correct my problem??? Also, for your information, i am not left with 1ml of primary antibody but have sufficient NRS. or would you suggest me any alteration in my protocol??? Thanks in advance for any kind of help. This is my first trial of IHC and am planning to do lots in future. I enjoy doing it and learning from the mistakes and trouble shooting the problem. but it will be fun if antibodies, time and the cost is not the constraint. Thanks once again. waiting for your reply Srinivas Srinivas Seekallu PhD student Dept of Vet Biomed.Sci. Western College of Veterinary Medicine, University of Saskatchewan. Saskatoon, Canada. Ph: 306-477-0849 (Home) 306-966-7373 (Lab) 306-966-7382 (Office) __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From ahenn <@t> vaccinex.com Mon May 2 13:29:56 2005 From: ahenn <@t> vaccinex.com (Alicia Henn Ph.D.) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] mycoplasma staining References: <200505021716.j42HGXD08762@nexserverpro.vaccinex.com> Message-ID: <000c01c54f44$f0b28950$2c01a8c0@alicia> Hi All, I use a bisbenzimide stain for routine mycoplasma detection of cell cultures. Lately, I've had problems with a bright blue haze covering dimly-stained cells. I've tried changing every step of the stain, but still get blue fog. I fix with methanol/acetone (I've tried formalin as well) stain the cells in chamberslides or in solution (for non-adherent cells) for 30 minutes, wash, air-dry chamberslides, mount with anti-fade mounting medium (I've tried several). I can visualize fluorescent beads with exquisite detail, so my microscope is OK. Anyone had any experience with this? I don't want to have to go back to PCR-based detection as that was a nightmare. Alicia Henn, Ph.D. Vaccinex, Inc. From drsvsrini <@t> yahoo.com Mon May 2 23:20:07 2005 From: drsvsrini <@t> yahoo.com (Mr srinivas sv) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Liver acetone powder In-Reply-To: 6667 Message-ID: <20050503042007.23475.qmail@web31512.mail.mud.yahoo.com> Thanks Ms. Vinnie Della Speranza and Mr. kharazia for the reply. Can anyone tell me the protocol for preparing the liver acetone powder, which i am planning to use as an adsorbent to non specific agents in a polyclonal antibody. I am looking at the expression of steroidogenic enzymes in sheep ovary. I think most have heard of my problem in my previous mails. To tell briefly, I am getting same color reaction in both positive and negative controls. so i am planning to try the adsorbtion technique using liver acetone powder. There is no commercial preparation of liver acetone powder from sheep. only liver powder from calf, rat are available. it would help me, if anyone tell me which one i can use or the protocol for preparation of sheep liver acetone powder as i have got sheep liver with me. Also i need to know the concentration of the liver powder i should use with the antibodies. Thanks waiting for the replies.. Srinivas Srinivas Seekallu PhD student Dept of Vet Biomed.Sci. Western College of Veterinary Medicine, University of Saskatchewan. Saskatoon, Canada. Ph: 306-477-0849 (Home) 306-966-7373 (Lab) 306-966-7382 (Office) --------------------------------- Do you Yahoo!? Yahoo! Small Business - Try our new resources site! From jkiernan <@t> uwo.ca Tue May 3 00:51:20 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Liver acetone powder References: <20050503042007.23475.qmail@web31512.mail.mud.yahoo.com> Message-ID: <42771158.232B0B9F@uwo.ca> Dear Srinivas Seekallu, A Few words of advice about abbreviated titles beginning with M. Mr = Mister (any man, married or single) Miss = Miss (an unmarried lady) Mrs (pronounced Missiz) = a married lady Ms = a female person unwilling or ashamed to disclose her marital status. Pronounced umzzz or mizz. This advice won't help with steroidogenic enzymes in sheep, but it could be useful generally in Saskatoon and other Canadian cities. John Kiernan London, Canada. __________________________________ Mr srinivas sv wrote: > > Thanks Ms. Vinnie Della Speranza and Mr. kharazia for the reply. > > Can anyone tell me the protocol for preparing the liver acetone powder, which i am planning to use as an adsorbent to non specific agents in a polyclonal antibody. > > I am looking at the expression of steroidogenic enzymes in sheep ovary. I think most have heard of my problem in my previous mails. To tell briefly, I am getting same color reaction in both positive and negative controls. so i am planning to try the adsorbtion technique using liver acetone powder. There is no commercial preparation of liver acetone powder from sheep. only liver powder from calf, rat are available. it would help me, if anyone tell me which one i can use or the protocol for preparation of sheep liver acetone powder as i have got sheep liver with me. > > Also i need to know the concentration of the liver powder i should use with the antibodies. > > Thanks > > waiting for the replies.. > > > > Srinivas Seekallu > PhD student > Dept of Vet Biomed.Sci. > Western College of Veterinary Medicine, > University of Saskatchewan. > Saskatoon, Canada. > > Ph: 306-477-0849 (Home) > 306-966-7373 (Lab) > 306-966-7382 (Office) > --------------------------------- From c.m.vanderloos <@t> amc.uva.nl Tue May 3 01:34:28 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] RE: Ki67 - kidney Message-ID: <261bb425ec61.25ec61261bb4@amc.uva.nl> Christian, You didn't tell us what rabbit Ki67antibody you purchased, but just in case it isn't working: we used rabbit monoclonal anti-Ki67, clone SP6 from LabVision/Neomarkers successfully on mouse, rat and human FFPE tissue sections. Just a citrate6.0 or Tris-EDTA9.0 pretreatment and 2-step EnVision detection. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "christian@eisbo.dk" Date Mon, 02 May 2005 10:31:07 +0200 To "histonet@lists.utsouthwestern.edu" Subject [Histonet] Ki67 - kidney. Hey users of histonet. I am trying to stain rat kidneys for Ki67! I have bought the recomended antibody. But this one is for human! Do anyone know if this can be used since i dont have a outcome! Do anyone have a protocol that works? Christian Eisbo References 1. mailto:c.m.vanderloos@amc.uva.nl From c.m.vanderloos <@t> amc.uva.nl Tue May 3 01:50:23 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] RE: Normal donkey serum block solution Message-ID: <261ce425ef62.25ef62261ce4@amc.uva.nl> Guofeng, Indeed if you have two secondary antibodies raised in donkey, normal donkey serum should be used for blocking. Just prepare a 1:10-20 dilution in PBS or TBS and incubate 15 min at RT. Do not wash, but blot off the normal serum and apply your primary. In case of two secondaries raised in different species you have to be aware first there will be no cross-reaction between those species. Two secondaries from the same species is always the best. However, in the situation of horse and donkey secondaries, cross-reaction seems highly unlikely. Just prepare a mixture for blocking composed of normal horse 1:20 and normal donkey 1:20 in PBS or TBS. Otherwise use the DakoCyto Protein Block, serum-free (X0909). Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From ? ? Date Mon, 02 May 2005 21:30:08 +0800 (CST) To histonet@pathology.swmed.edu Subject [Histonet] Normal donkey serum block solution Hello,all, I am doing double-immunofluorescence. Both the hosts of my secondary antibodies are donkey, so I plan to use normal donkey serum for blocking, but I do not know how to make up this solution. I have ordered donkey serum from sigma company, but the specification doesnot show how to make up it. so if you have done it, I hope that you can do me a favor. In addition, if the hosts of my secondary antiboies are different, one is horse, the other is donkey, which solution do I use for blocking? Thank you! Best wishes! Guofeng References 1. mailto:c.m.vanderloos@amc.uva.nl From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue May 3 04:03:09 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] RE: Histonet Digest, Vol 18, Issue 2 Message-ID: Hi Enzyme digestion is an antigen retrieval system too, so guess you mean heat or enzyme first - on bone marrow trephine biopsies for Igs, we used heat retrieval followed by a brief enzyme. Haven't heard of it the other way round. Cheers -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 02 May 2005 18:04 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 18, Issue 2 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. fungus control blocks (Ernestine Middleton) 2. Ki67 - kidney. (christian@eisbo.dk) 3. antigen retrieval/enzyme digestion (Till, Renee) 4. Re: Ki67 - kidney. (Kelly D Mcqueeney) 5. Normal donkey serum block solution (=?gb2312?q?=CC=EC=20=D0=C1?=) ---------------------------------------------------------------------- Message: 1 Date: Sun, 1 May 2005 15:54:30 -0400 (EDT) From: Ernestine Middleton Subject: [Histonet] fungus control blocks To: histonet@pathology.swmed.edu Message-ID: <20050501195430.97482.qmail@web51502.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi; Sorry, my e-mail address is ernestinemiddleton@yahoo.ca. Ernestine. --------------------------------- Post your free ad now! Yahoo! Canada Personals ------------------------------ Message: 2 Date: Mon, 02 May 2005 10:31:07 +0200 From: "christian@eisbo.dk" Subject: [Histonet] Ki67 - kidney. To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=windows-1252 Hey users of histonet. I am trying to stain rat kidneys for Ki67! I have bought the recomended antibody. But this one is for human! Do anyone know if this can be used since i dont have a outcome! Do anyone have a protocol that works? Christian Eisbo ------------------------------ Message: 3 Date: Mon, 2 May 2005 07:55:02 -0500 From: "Till, Renee" Subject: [Histonet] antigen retrieval/enzyme digestion To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii When trying both on the same slides, which would go first, the antigen retrieval or the enzyme digestion? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 4 Date: Mon, 02 May 2005 09:26:49 -0400 From: Kelly D Mcqueeney Subject: Re: [Histonet] Ki67 - kidney. To: "christian@eisbo.dk" Cc: "histonet@lists.utsouthwestern.edu" Message-ID: <42762A99.7010505@bms.com> Content-Type: text/plain; format=flowed; charset=windows-1252 Hi Christian, I have used rabbit polyclonal human Ki67 antibody on rat kidney and it worked beautifully. Make sure you use a wash buffer with tween-20. Simply follow the protocol recommended by Abcam (ab833). I used heat antigen retrieval in citrate buffer (Dako), 3% BSA (only) as a block and Envision+ rabbit kit (Dako) rather than ABC. The incubation time is O/N at 4C. Let me know if you need more detailed information. You can also use Ki67 antibody from Santa Cruz. Good luck, Kelly christian@eisbo.dk wrote: >Hey users of histonet. >I am trying to stain rat kidneys for Ki67! >I have bought the recomended antibody. But this one is for human! >Do anyone know if this can be used since i dont have a outcome! >Do anyone have a protocol that works? >Christian Eisbo > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 5 Date: Mon, 2 May 2005 21:30:08 +0800 (CST) From: =?gb2312?q?=CC=EC=20=D0=C1?= Subject: [Histonet] Normal donkey serum block solution To: histonet@pathology.swmed.edu Message-ID: <20050502133008.26004.qmail@web15507.mail.cnb.yahoo.com> Content-Type: text/plain; charset=gb2312 Hello,all, I am doing double-immunofluorescence. Both the hosts of my secondary antibodies are donkey, so I plan to use normal donkey serum for blocking, but I do not know how to make up this solution. I have ordered donkey serum from sigma company, but the specification doesnot show how to make up it. so if you have done it, I hope that you can do me a favor. In addition, if the hosts of my secondary antiboies are different, one is horse, the other is donkey, which solution do I use for blocking? Thank you! Best wishes! Guofeng Giessen --------------------------------- Do You Yahoo!? ?????????????????????????????? ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 2 *************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From ASelf <@t> gmhsc.com Tue May 3 04:54:53 2005 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] test Message-ID: <39836CD6DB61654E8F95A35898C92186D70E4A@exchange.gmhpost.com> From ctsblack <@t> capeheart.uct.ac.za Tue May 3 05:59:27 2005 From: ctsblack <@t> capeheart.uct.ac.za (Melanie Black) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Alk Phos Crystals Message-ID: Hi There Can anybody shed some light onto the subject of crystals forming after the use of BCIP/NBT on alk phos staied tissue. At first when I mount the slides, you do not see the precipitate, btu within hours, it deveopes. I must add that the tissue is ZINC fixed. As zinc is a precippitant, I am not sure where to look for the problem. DAKO has no idea!! This problem has been discussed before, but I cannot recall if it was ever resolved. Many Thanks Melanie Black -- Cardiovascular Research Unit Div. of Cardiothoracic Surgery Chris Barnard Building University of Cape Town Anzio Road Observatory 7925 Republic of South Africa Tel +27 21 406-6589 Cel +27 82 469-3352 Fax +27 21 448-5935 From Wanda.Smith <@t> HCAhealthcare.com Tue May 3 08:33:33 2005 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:25:01 2005 Subject: **SPAM** [Histonet] slide storage Message-ID: Cynthia, Cardinal Health has the cabinet you are talking about. The order number is M6350-2L and the metal tray order number is M6349-1. The cabinet holds 100 metal trays. Catalog price is $1570.00 for the cabinet and $19.77 ea for the metal trays. You may be able to get a discount through your organization. Hope this helps!!! Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Favara, Cynthia (NIH/NIAID) Sent: Friday, April 29, 2005 6:08 PM To: histonet@pathology.swmed.edu Subject: **SPAM** [Histonet] slide storage Many years ago -like in the 60's-70's I remember seeing a slide storage system that allowed on to leave slides on metal trays and then put them in a cabinet. Does anyone out there know of such a system? The reason I ask is that I have an investigator that wants all the slides in trays until he figures things out. This sometimes takes a year or more. Needless to say my counter space and book shelves are being over run. Please help if you can. Good weekend to all. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abee <@t> pml.ac.uk Tue May 3 08:41:17 2005 From: abee <@t> pml.ac.uk (abee@pml.ac.uk) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Pecten maximus Message-ID: <000001c54fe5$cad68a50$2ca2abc0@npm.ac.uk> Dear all, I am working on the structure of the gill of Pectin maximus and am having problems identifying one of the structures. The samples have been fixed in Davidsons and are in wax blocks. These structures are visible with a light microscope and are in whole sections. It is present on the descending lamella starting at the gill filament and runs about halfway down the lamella. It has the appearance of a sinusoidal shape. It is not on the ascending lamella at all. Can anyone help me identify this, or at least point me in the direction of a good reference please? Many Thanks, Amanda Beesley From mmmlyons <@t> hotmail.com Tue May 3 09:05:00 2005 From: mmmlyons <@t> hotmail.com (Molly Lyons) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Pecten maximus In-Reply-To: <000001c54fe5$cad68a50$2ca2abc0@npm.ac.uk> Message-ID: Hello, Here is a reference for bivalve mollusc histology - it has a photograph of the gill of Pecten maximus (compared to gills of an oyster, mussel, and clam). Editor: Grizel, H. 2003. An atlas of histology and cytology of marine bivavle molluscs. By Ifremer ISBN 2-84433-111-4. Hope that helps. Maille Lyons Department of Marine Sciences University of Connecticut 1080 Shennecossett Road Groton, CT 06340 From ROrr <@t> enh.org Tue May 3 09:12:43 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] RE: Histonet Digest, Vol 18, Issue 2 Message-ID: Hi Renee, As much as you could do this really any way you want, In my experience, I have done the antigen retrieval first then used the enzyme digestion. In this case, the antigen retrieval can beat up the tissue and a full enzyme digestion incubation might wipe out what you are looking for...or even wipe out the tissue! I would recommend using a highly diluted digestion or least less time... For example, a Citrate AR process and a 30 second incubation (at RT) for Pepsin... Becky Becky Orr, CLA HT (ASCP) IHC Lead Evanston Northwestern Healthcare ph: 847-570-2771 From cgfields <@t> lexhealth.org Tue May 3 09:53:27 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Histotechnologist Needed in South Carolina Message-ID: I am posting this for Spartanburg Regional Healthcare System, Spartanburg, S.C....(no access HistoNet). Histotechnologist Needed Spartanburg Regional Healthcare System offers an environment that combines state-of-the-art technology, a wealth of expertise and a staff that cares for what matters. We invite you to consider joining our special team. We have a full-time dayshift position (college graduate preferred; 2-5 years experience, HT or HTL certification preferred or eligible). We offer competitive salaries, a sign-on-bonus, relocation assistance up to $2500, exceptional programs and opportunities for professional growth. If you are interested in a career with Spartanburg Regional, please visit our website at www.spartanburgregional.com, call Jeff Fields at 800-288-7762 or forward your resume to jfields@srhs.com. Equal Opportunity Employer Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From godsgirlnow <@t> msn.com Tue May 3 10:45:10 2005 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] cutting angle Message-ID: What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne From mprice26 <@t> juno.com Tue May 3 11:37:50 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] RE: USED GLX LINEAR STAINER Message-ID: <20050503.093854.24587.70994@webmail31.nyc.untd.com> We have an GLX Linear Stainer for sale if anyone is interested, contact me @ 903-791-1777 or via e-mail. It is $1000 or Best Offer. Thank you. Marsha Price From dellav <@t> musc.edu Tue May 3 11:31:05 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Re: Liver acetone powder Message-ID: Srinivas sorry for the delay in getting back to you. I will outline the technique listed in Fluorescent Protein Tracing by Nairn that I cited in last message to you. before doing so, I am not convinced from the message you posted yesterday that the problem is in your primary antibody. I recommend that you omit the primary antibody on a known positive control slide, substituting the buffer you use for rinsing your slides and incubate along with another slide that receives the primary antibody. If you are getting positive staining on the slide that had buffer instead of the primary antibody, you can conclude that the secondary antibody (goat anti-rabbit) may be the problem also, are you making certain that your sections never dry out once the method has begun? this will be essential to avoiding any non-specific staining since you observe the problem with two different primary antibodies from two different sources (one home made and the other purchase commercially) I am inclined to think that your problem is not caused by the primaries so I recommend that you look elsewhere before embarking on the absorption. as you will see below, the method of preparing the sheep liver is quite involved and time consuming. if you are using an avidin-biotin detection system it is critical that you perform a biotin blocking step to avoid non-specific staining. the method for liver is as follows: wash the organ or tissue clear of blood with physiologic saline, dice with scissors or knife and wash again with saline. the diced material is then mixed with an equal volume of cold saline and homogenized without allowing excessive warming. the product is frozen at -76 degrees C and thawed to facilitate centrifuging and washing. centrifuge at 4000 g for 10 minutes and wash the cellular material twice with phosphate buffered saline this suspension is centrifuged at 10,000g for 15 minutes in separate amounts designed to give 0.5 ml or 1.0 ml volumes of packed tissue in each centrifuge tube, which is drained, stoppered and frozen at -20 degrees C. absoprtion is carried out by stirring each ml of antibody with 0.5 ml of thawed homogenate, incubating at 37 degrees C for 30-60 minutes and leaving to stand overnight at 0- -2 degrees C. alternately, antibody and homogenate may be shaken for 2 hours at room temperature. the antibody is recovered by centrifuging at 10,000 g for 20 minutes the powders are prepared either by lyophilization of homogenates or by acetone drying. for the latter, wash the diced liver and gind with acetone in a mortar and pestle or low speed homogenizer. the product is filtered through coarse filter paper. the deposit on the paper is washed several times with acetone to complete the dehydration and then dried at 37 degrees C or overnight at room temperature. the dried tissue is ground to powder in a mortar or mechanical grinder and sieved through a 1 mm wire mesh to remove coarse debris. the dehydrated powder may be stored at room temperature until required. as stated in my last message, for absoption of your antibody, the ratio of liver powder to antibody does not need to be precise, but more is better than less to ensure complete absorption. the ratio of 100 mg of liver to 1 ml of antibody is given in the reference. if you have 0.5 ml of antibody you would use 50 mg of liver powder etc. good luck Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> Mr srinivas sv 05/03/05 12:20AM >>> Thanks Ms. Vinnie Della Speranza and Mr. kharazia for the reply. Can anyone tell me the protocol for preparing the liver acetone powder, which i am planning to use as an adsorbent to non specific agents in a polyclonal antibody. I am looking at the expression of steroidogenic enzymes in sheep ovary. I think most have heard of my problem in my previous mails. To tell briefly, I am getting same color reaction in both positive and negative controls. so i am planning to try the adsorbtion technique using liver acetone powder. There is no commercial preparation of liver acetone powder from sheep. only liver powder from calf, rat are available. it would help me, if anyone tell me which one i can use or the protocol for preparation of sheep liver acetone powder as i have got sheep liver with me. Also i need to know the concentration of the liver powder i should use with the antibodies. Thanks waiting for the replies.. Srinivas Srinivas Seekallu PhD student Dept of Vet Biomed.Sci. Western College of Veterinary Medicine, University of Saskatchewan. Saskatoon, Canada. Ph: 306-477-0849 (Home) 306-966-7373 (Lab) 306-966-7382 (Office) --------------------------------- Do you Yahoo!? Yahoo! Small Business - Try our new resources site! From pruegg <@t> ihctech.net Tue May 3 11:46:14 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Need help finding some antibodies Message-ID: <200505031646.j43Gk5KB020663@chip.viawest.net> I am looking for cytokeratin 3 and collagen VII. If anyone knows where I can purchase these and if you have experience with using them please let me know. Thank you, Patsy From nienhuis <@t> ucla.edu Tue May 3 12:27:28 2005 From: nienhuis <@t> ucla.edu (NIENHUIS,ROBERT ) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] antigen retrieval/enzyme digestion Message-ID: <1115141248.4277b48005b64@mail.ucla.edu> Has anyone any experience with using gelatin (or something else) to embed tissue to hold it together during antigen retrieval? Some have said that it can inerfere with immuno staining, others say no. We sometines deal with poorly fixed or over fixed post-mortem tissue in various species. Bob Nienhuis UCLA Neurobiology Research Quoting "Till, Renee" : > When trying both on the same slides, which would go first, the antigen > retrieval or the enzyme digestion? > > > > Renee' Till, HT > > > > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > > > (501) 364-2774 > > > > > Confidentiality Notice: This e-mail message, including any attachments, > is for the sole use of the intended recipient(s) and may contain > confidential and privileged information. Any unauthorized review, use, > disclosure or distribution is prohibited. If you are not the intended > recipient, please contact the sender by reply e-mail and destroy all > copies of the original message. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ----- End forwarded message ----- From JNocito <@t> Pathreflab.com Tue May 3 13:35:52 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Which RPMI media to use Message-ID: Greetings Histoland, once, again, I need to tap into your vast knowledge. We will be sending out bone marrow, lymph node and placenta cases for flow cytometry and cytogenetic studies. Looking through the catalogs, there are about 15 different formulas for RPMI. Can any one tell me which formula is the best one. I'm at a loss. thanks in advance Joe Nocito From BlazekL <@t> childrensdayton.org Tue May 3 13:57:20 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Which RPMI media to use Message-ID: Joe, The best one to uses is the one that the facility you are sending them to wants you to use. The facility that we send ours to even furnishes it to us. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Joe Nocito" 05/03/2005 2:35:52 PM >>> Greetings Histoland, once, again, I need to tap into your vast knowledge. We will be sending out bone marrow, lymph node and placenta cases for flow cytometry and cytogenetic studies. Looking through the catalogs, there are about 15 different formulas for RPMI. Can any one tell me which formula is the best one. I'm at a loss. thanks in advance Joe Nocito _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pzeitlow <@t> bbpllab.com Tue May 3 14:05:36 2005 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Which RPMI media to use Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEBEC@bbplsrv1.bbpl> We use RPMI from Fisher (#MT-15-041-CV). We add Pen-Strep prior to use to inhibit bacterial growth. Pat -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Tuesday, May 03, 2005 1:36 PM To: Histonet Subject: [Histonet] Which RPMI media to use Greetings Histoland, once, again, I need to tap into your vast knowledge. We will be sending out bone marrow, lymph node and placenta cases for flow cytometry and cytogenetic studies. Looking through the catalogs, there are about 15 different formulas for RPMI. Can any one tell me which formula is the best one. I'm at a loss. thanks in advance Joe Nocito _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vsailes <@t> nd.edu Tue May 3 14:26:52 2005 From: vsailes <@t> nd.edu (vsailes@nd.edu) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] slide storage cabinets Message-ID: <1115148412.4277d07c18790@webmail.nd.edu> Hi Cynthia, VWR International has two types of the wood slide storage cabinets that comes with the aluminum trays. Ordering information: Micro Slide storage cabinet,wood cat#48466-006 $410.00 each (comes with 25 trays) Micro slide storage cabinet, wood cat # 48467-009 $1432.00 (comes with 100 trays) Hopes this information helps. Valerie From jhabecke <@t> seattlecca.org Tue May 3 14:34:52 2005 From: jhabecke <@t> seattlecca.org (Randolph-Habecker, Julie) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] Canine Herpes Virus Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B9484EB08E@wala01.seattlecca.org> Histonetters, I am looking for an antibody to detect canine herpes virus in either FFPE or frozen tissue. Any experience in this area? Thanks! Julie Julie Randolph-Habecker, Ph.D. Experimental Histopathology Shared Resources Fred Hutchinson Cancer Research Center 1100 Fairview Ave. N, G1-300 PO Box 19023 Seattle, WA 98109-1024 Tel: (206) 288-1187 FAX: (206) 288-1345 jhabecke@fhcrc.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From JWEEMS <@t> sjha.org Tue May 3 14:48:13 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] cutting angle Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45BDB@sjhaexc02.sjha.org> I think you should be allowed to cut at whatever angle you produce the best sections! My 2 cents....Joyce:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Roxanne Soto Sent: Tuesday, May 03, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From dellav <@t> musc.edu Tue May 3 16:22:22 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:01 2005 Subject: [Histonet] cutting angle Message-ID: Of course, Joyce's remark is right on target. when I first saw your post Roxanne, I wasn't quite sure if I understood entirely what you were asking. for example, do you use the same blades in your new job as you used in the previous lab? if the answer is yes, then it is very unlikely that the angles used could be so different and both yield acceptable results. the angle set on the microtome is directly related to the facet angle on the blade and of course this can vary between manufacturers. in essence, you must find the correct angle to a degree through trial and error until your sections have minimal compression. if the sections are rolling off the blade smoothly and with little compression, you are at the correct angle for that brand and model blade. I am also not convinced that the angle setting between microtomes, especially between different brands of microtomes, are all that precise. what I mean is that if you used an angle of 10 on a Leica for example, you might need an angle of 8 or 12 on a microm. I don't know what the actual angle settings should be but you understand my point. even with a number of microtomes from the same manufacturer I've seen it occasionally necessary where the optimal setting on one might a degree or two away from another. I believe these settings are intended as general guides. the ultimate proof is in the ease of sectioning and the quality of the sections you are getting from a particular machine. back to Joyce's remark. as long as you are able to attain the quality required by your new employer, it matters little what angle setting you are using. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Weems, Joyce" 05/03/05 03:48PM >>> I think you should be allowed to cut at whatever angle you produce the best sections! My 2 cents....Joyce:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Roxanne Soto Sent: Tuesday, May 03, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From GODSGIRLNOW <@t> msn.com Tue May 3 17:28:10 2005 From: GODSGIRLNOW <@t> msn.com (Roxanne Soto) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle References: <1115143433.4277bd09b4fca@imp.vet.upenn.edu> Message-ID: The microtome is an olympus. They use an infiltrating paraffin and a different one for embedding. The blades are accuedge. ----- Original Message ----- From: pmarcum@vet.upenn.edu To: Roxanne Soto Sent: Tuesday, May 03, 2005 2:03 PM Subject: Re: [Histonet] cutting angle Roxanne, Did the model of the microtome change? I find I had to change the angle when I used a different brand of microtome. I also have to decide for myself what knife angle is best for me on a given microtome. I dont think there is one absolute knife angle for all microtomes or blades. I am currently using a Leica 2255 and use 3 to 5 for tungsten carbide D profile and 5 to 6 for low profile knife holder and blades. I am using Thermo Shandon Premier S35 low profile knives for paraffin. They are working well for me. Pam Marcum Quoting Roxanne Soto >: > What angles does everyone out there cut at and what kind of blades do you > use? I have always cut at 10 with high or low profile blades, but I have > recently changed jobs and they cut at an angle of 21. They were told that > with high profile blades they should use an angle of 21. I am having a hard > > time cutting at this angle....... > any thoughts? > Roxanne > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From bills <@t> icpmr.wsahs.nsw.gov.au Tue May 3 17:31:23 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle In-Reply-To: Message-ID: <000001c5502f$d682aba0$0ecd080a@wsahs.nsw.gov.au> Vinnie, I agree with you. We have several diferent vintage Leicas and the markings for the "angle" are in fact reversed. One reads 0-20 top to bottom the other 20-0 top to bottom. Like you I have told the microtomists to use the angle which for them produces the best ribbon. I have spoken to several mictotome manufacturers over the years and they agree these marks do not mark "angles" as such merely a guide to where you should set your knife. We have several Cryostats (different brands) and there is no consistency amongst these either, two have no numbers on the knife angle guide. As I have been a histotechnologist for over forty years and have cut many sections in wax and other substances (including resin, celloidin etc) using many types of knife (Ralph, Tungsten carbide, carbon steel and disposable) I have found that for most conditions the "perfect angle is one where the block only just clears the back of the knife. Even using a sledge microtome for the celloidin sections (whole lobes of lung and hemisheres of brain) the same rule applies. There is some great material written about knife clearance angles and the facet angles of knives. Baker and Silverton in Introduction to Medical Laboratory Technology suggest 1-6 degress clearance. Bancroft and Stevens in Theory and Practice of Histological Techniques recommend a clearance angle of 3-4 degrees. When teaching students microtomy we begin by setting all the mictotomes at 7 on the scale then do minor adjustments according to the tissue being cut. However, the choice of "angle" is the users if the sections come off the knife well. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vinnie Della Speranza Sent: Wednesday, 04 May 2005 7:22 AM To: histonet@lists.utsouthwestern.edu; godsgirlnow@msn.com; JWEEMS@sjha.org Subject: RE: [Histonet] cutting angle Of course, Joyce's remark is right on target. when I first saw your post Roxanne, I wasn't quite sure if I understood entirely what you were asking. for example, do you use the same blades in your new job as you used in the previous lab? if the answer is yes, then it is very unlikely that the angles used could be so different and both yield acceptable results. the angle set on the microtome is directly related to the facet angle on the blade and of course this can vary between manufacturers. in essence, you must find the correct angle to a degree through trial and error until your sections have minimal compression. if the sections are rolling off the blade smoothly and with little compression, you are at the correct angle for that brand and model blade. I am also not convinced that the angle setting between microtomes, especially between different brands of microtomes, are all that precise. what I mean is that if you used an angle of 10 on a Leica for example, you might need an angle of 8 or 12 on a microm. I don't know what the actual angle settings should be but you understand my point. even with a number of microtomes from the same manufacturer I've seen it occasionally necessary where the optimal setting on one might a degree or two away from another. I believe these settings are intended as general guides. the ultimate proof is in the ease of sectioning and the quality of the sections you are getting from a particular machine. back to Joyce's remark. as long as you are able to attain the quality required by your new employer, it matters little what angle setting you are using. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Weems, Joyce" 05/03/05 03:48PM >>> I think you should be allowed to cut at whatever angle you produce the best sections! My 2 cents....Joyce:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Roxanne Soto Sent: Tuesday, May 03, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From GODSGIRLNOW <@t> msn.com Tue May 3 17:32:21 2005 From: GODSGIRLNOW <@t> msn.com (Roxanne Soto) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle References: Message-ID: Anyone from TBS out there want to chime in on this? As the microtome is an Olympus.......... ----- Original Message ----- From: Vinnie Della Speranza To: histonet@lists.utsouthwestern.edu ; godsgirlnow@msn.com ; JWEEMS@sjha.org Sent: Tuesday, May 03, 2005 5:22 PM Subject: RE: [Histonet] cutting angle Of course, Joyce's remark is right on target. when I first saw your post Roxanne, I wasn't quite sure if I understood entirely what you were asking. for example, do you use the same blades in your new job as you used in the previous lab? if the answer is yes, then it is very unlikely that the angles used could be so different and both yield acceptable results. the angle set on the microtome is directly related to the facet angle on the blade and of course this can vary between manufacturers. in essence, you must find the correct angle to a degree through trial and error until your sections have minimal compression. if the sections are rolling off the blade smoothly and with little compression, you are at the correct angle for that brand and model blade. I am also not convinced that the angle setting between microtomes, especially between different brands of microtomes, are all that precise. what I mean is that if you used an angle of 10 on a Leica for example, you might need an angle of 8 or 12 on a microm. I don't know what the actual angle settings should be but you understand my point. even with a number of microtomes from the same manufacturer I've seen it occasionally necessary where the optimal setting on one might a degree or two away from another. I believe these settings are intended as general guides. the ultimate proof is in the ease of sectioning and the quality of the sections you are getting from a particular machine. back to Joyce's remark. as long as you are able to attain the quality required by your new employer, it matters little what angle setting you are using. Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Weems, Joyce" > 05/03/05 03:48PM >>> I think you should be allowed to cut at whatever angle you produce the best sections! My 2 cents....Joyce:>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Roxanne Soto Sent: Tuesday, May 03, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Megan.Clarke <@t> hnehealth.nsw.gov.au Tue May 3 17:39:30 2005 From: Megan.Clarke <@t> hnehealth.nsw.gov.au (Megan Clarke) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] BAF47 Message-ID: Thank you for the information on BAF47. Megan Clarke HAPS Newcastle Australia From lpwenk <@t> sbcglobal.net Tue May 3 17:26:57 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle In-Reply-To: Message-ID: Try contacting the company that makes the microtome, and tell them the brand of knife you are using. They will tell you the best angle for that microtome for that knife. Each brand of microtome is a little bit different. And each brand of blade is a little bit different. So as you switch from one blade to another (or same blade but different microtome), the angles have to be adjusted. There is not one "true" setting, as in "one size fits all". Doesn't work with microtomes and blades. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roxanne Soto Sent: Tuesday, May 03, 2005 11:45 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Tue May 3 18:37:49 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Which RPMI media to use References: Message-ID: <007c01c55039$1e2a3e10$7929f318@yourxhtr8hvc4p> Linda, that would be great, if we were dealing with only one lab, but, as things would have it here at PRL, we send specimens to at least two different labs (don't even go there. I know what you're going to say.) The labs currently send us RPMI media, but since we are going to be increasing our specimen load, I'd like to have some available just in case something happens like people dropping open tubes and stuff. Thanks for the info. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Linda Blazek" To: ; Sent: Tuesday, May 03, 2005 1:57 PM Subject: Re: [Histonet] Which RPMI media to use Joe, The best one to uses is the one that the facility you are sending them to wants you to use. The facility that we send ours to even furnishes it to us. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Joe Nocito" 05/03/2005 2:35:52 PM >>> Greetings Histoland, once, again, I need to tap into your vast knowledge. We will be sending out bone marrow, lymph node and placenta cases for flow cytometry and cytogenetic studies. Looking through the catalogs, there are about 15 different formulas for RPMI. Can any one tell me which formula is the best one. I'm at a loss. thanks in advance Joe Nocito _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Anti-Virus. Version: 7.0.308 / Virus Database: 266.11.2 - Release Date: 5/2/2005 From jnocito <@t> satx.rr.com Tue May 3 18:38:43 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Which RPMI media to use References: <813FB33DA405334F947F8BFC6EBD0B2A0BEBEC@bbplsrv1.bbpl> Message-ID: <008801c55039$3ec54200$7929f318@yourxhtr8hvc4p> Thanks Pat. How much pen-strep do you use? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Pat Zeitlow" To: "Joe Nocito" ; "Histonet" Sent: Tuesday, May 03, 2005 2:05 PM Subject: RE: [Histonet] Which RPMI media to use We use RPMI from Fisher (#MT-15-041-CV). We add Pen-Strep prior to use to inhibit bacterial growth. Pat -----Original Message----- From: Joe Nocito [mailto:JNocito@Pathreflab.com] Sent: Tuesday, May 03, 2005 1:36 PM To: Histonet Subject: [Histonet] Which RPMI media to use Greetings Histoland, once, again, I need to tap into your vast knowledge. We will be sending out bone marrow, lymph node and placenta cases for flow cytometry and cytogenetic studies. Looking through the catalogs, there are about 15 different formulas for RPMI. Can any one tell me which formula is the best one. I'm at a loss. thanks in advance Joe Nocito _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Anti-Virus. Version: 7.0.308 / Virus Database: 266.11.2 - Release Date: 5/2/2005 From jnocito <@t> satx.rr.com Tue May 3 18:43:19 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Cold acetone References: Message-ID: <00cd01c55039$e2e5dc00$7929f318@yourxhtr8hvc4p> Steve, the only "cold" acetone I used was just placing a coplin jar with acetone in a refrigerator, which brought the temp down to + 4,. I've never used it at -20. What application are you using the cold acetone for? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Steven Coakley" To: Sent: Wednesday, April 27, 2005 9:12 AM Subject: [Histonet] Cold acetone > > > I hope this doesn't sound to lame. 13 years in this field and I've never > used "cold acetone". The only ref. I have is to use -20 cold acetone. So > how do I get my acetone to -20. Do I place a coplin jar of covered > actenone > in the cyostat for a period of time?????. \\Steve > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.308 / Virus Database: 266.11.2 - Release Date: 5/2/2005 > > From bruyntjes <@t> voeding.tno.nl Wed May 4 04:39:11 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Cold acetone Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DAE@ntexch1.voeding.tno.nl> Steve I've used cold acetone (-20) to fix tissues. First of all the acetone was cooled down to 4 degrees in the refrigerator, the tissues were collected and placed in the cold acetone, 15 minutes later the jar was placed in a freezer (+ - 20 degrees) for another 2 hours. I used it to fix and embed rat mammary glands using the AMEX method about 15 years ago in order to do some immunostaining with all kind of different monoclonal keratins (with good results). Joost -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: woensdag 4 mei 2005 1:43 To: steven.coakley@mirusbio.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cold acetone Steve, the only "cold" acetone I used was just placing a coplin jar with acetone in a refrigerator, which brought the temp down to + 4,. I've never used it at -20. What application are you using the cold acetone for? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Steven Coakley" To: Sent: Wednesday, April 27, 2005 9:12 AM Subject: [Histonet] Cold acetone > > > I hope this doesn't sound to lame. 13 years in this field and I've never > used "cold acetone". The only ref. I have is to use -20 cold acetone. So > how do I get my acetone to -20. Do I place a coplin jar of covered > actenone > in the cyostat for a period of time?????. \\Steve > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.308 / Virus Database: 266.11.2 - Release Date: 5/2/2005 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From juan.gutierrez <@t> christushealth.org Wed May 4 05:06:50 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Cold acetone Message-ID: Just make sure your freezer is explosion proof!!! There was an article in HISTOLOGIC about a freezer blowing up in the middle of the night because of flammable fumes accumulating inside. The pictures in the article were pretty scary. Contact SAKURA for a copy of the article. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bruijntjes, J.P. Sent: Wednesday, May 04, 2005 4:39 AM To: Joe Nocito; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cold acetone Steve I've used cold acetone (-20) to fix tissues. First of all the acetone was cooled down to 4 degrees in the refrigerator, the tissues were collected and placed in the cold acetone, 15 minutes later the jar was placed in a freezer (+ - 20 degrees) for another 2 hours. I used it to fix and embed rat mammary glands using the AMEX method about 15 years ago in order to do some immunostaining with all kind of different monoclonal keratins (with good results). Joost -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: woensdag 4 mei 2005 1:43 To: steven.coakley@mirusbio.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Cold acetone Steve, the only "cold" acetone I used was just placing a coplin jar with acetone in a refrigerator, which brought the temp down to + 4,. I've never used it at -20. What application are you using the cold acetone for? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Steven Coakley" To: Sent: Wednesday, April 27, 2005 9:12 AM Subject: [Histonet] Cold acetone > > > I hope this doesn't sound to lame. 13 years in this field and I've never > used "cold acetone". The only ref. I have is to use -20 cold acetone. So > how do I get my acetone to -20. Do I place a coplin jar of covered > actenone > in the cyostat for a period of time?????. \\Steve > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Anti-Virus. > Version: 7.0.308 / Virus Database: 266.11.2 - Release Date: 5/2/2005 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abright <@t> brightinstruments.com Wed May 4 06:11:25 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle Message-ID: Dear Roxanne, I hope this is going to be of assistance to you, we manufacture all our microtomes & cryostats from a true 0? to 25? angle on the knife or blade holders. In this way we have set a standard for all our users. I cannot see why others choose arbitrary calibrations and vary them too. You will need to place a place a flat sample onto the cassette or object holder and adjust the knife or blade close to the face of the sample and view from the side of the edge of the knife or blades facet to make sure you have a 1 to 3? clearance. If this is not a clear description please come back to me and I will email you a sketch or try to place it on "list server images" of the Histonet.org website if someone can inform me how to place it there. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Roxanne Soto [mailto:godsgirlnow@msn.com] Sent: 03 May 2005 16:45 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Wed May 4 07:45:32 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Canine Herpes Virus Message-ID: <42788BAB.31307.B2E5B@localhost> ------- Forwarded message follows ------- From: Greg Dobbin To: "Randolph-Habecker, Julie" Subject: Re: [Histonet] Canine Herpes Virus Date sent: Wed, 4 May 2005 08:42:09 -0400 Check VMRD first, then Serotec. Then if you still aren't satisfied- search on Abcam. Good luck. Greg From: "Randolph-Habecker, Julie" To: "'histonet@lists.utsouthwestern.edu'" Date sent: Tue, 3 May 2005 12:34:52 -0700 Subject: [Histonet] Canine Herpes Virus > Histonetters, > > I am looking for an antibody to detect canine herpes virus in either > FFPE or frozen tissue. Any experience in this area? > > Thanks! > > Julie > > Julie Randolph-Habecker, Ph.D. > Experimental Histopathology Shared Resources > Fred Hutchinson Cancer Research Center > 1100 Fairview Ave. N, G1-300 > PO Box 19023 > Seattle, WA 98109-1024 > Tel: (206) 288-1187 > FAX: (206) 288-1345 > jhabecke@fhcrc.org > > > > This electronic message transmission contains information which may > be > confidential or privileged. The information is intended to be for > the use of the individual or entity named above. If you are not the > intended recipient, be aware that any disclosure, copying, > distribution or use of the contents of this information is > prohibited. If you have received this electronic transmission in > error, please leave a message via telephone at (206) 288-6266, > notify me by electronic reply, and delete this message. Opinions and > ideas in this message that do not relate to official business are > understood as neither given nor endorsed by the Seattle Cancer Care > Alliance. To view our complete Notice of Privacy Practices, visit > our web site at www.seattlecca.org. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------- End of forwarded message ------- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From NSEARCY <@t> swmail.sw.org Wed May 4 08:11:37 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Automated Microtome Message-ID: Can I have the latest and greatest regarding automated microtomes? I have an employee with carpal tunnel- anyone have experience with such employees using this instrument to lessen their problems? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Nita Searcy TEL;WORK:4-2438 ORG:;Anatomic Pathology EMAIL;WORK;PREF;NGW:NSEARCY@swmail.sw.org N:Searcy;Nita TITLE:Manager, Pathology Division END:VCARD From kelly.mcqueeney <@t> bms.com Wed May 4 08:15:17 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Cold acetone In-Reply-To: References: Message-ID: <4278CAE5.8040901@bms.com> Alcohol does not freeze at -20C. Cold acetone refers to -20C acetone. Place a jar in the freezer! Kelly Steven Coakley wrote: >I hope this doesn't sound to lame. 13 years in this field and I've never >used "cold acetone". The only ref. I have is to use -20 cold acetone. So >how do I get my acetone to -20. Do I place a coplin jar of covered actenone >in the cyostat for a period of time?????. \\Steve > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jqb7 <@t> cdc.gov Wed May 4 08:24:39 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Automated Microtome Message-ID: I had carpal tunnel surgery on my left hand 5 years ago. While my right hand needed it as well, it was not as badly damaged so I put it off and am now having the surgery tomorrow. Although it is not a truly automated microtome, the Microm Ergostar has been a life-saver for me. I highly recommend it. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nita Searcy Sent: Wednesday, May 04, 2005 9:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated Microtome Can I have the latest and greatest regarding automated microtomes? I have an employee with carpal tunnel- anyone have experience with such employees using this instrument to lessen their problems? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 From cwscouten <@t> myneurolab.com Wed May 4 11:06:17 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle Message-ID: <5784D843593D874C93E9BADCB87342AB44F90F@tpiserver03.Coretech-holdings.com> To post a histology picture or drawing to histonet.org, send it as an attachment to pictures@histonet.org. When you post your message to the Histonet listserver, you can tell the Histonet community that you have posted a picture to www.histonet.org and give the histonet community the name of the picture file you sent to pictures@histonet.org. Please allow 24 hours for posting your picture to www.histonet.org. They are reviewed to ensure that they meet histonet guidelines. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: Alan Bright [mailto:abright@brightinstruments.com] Sent: Wednesday, May 04, 2005 6:11 AM To: Roxanne Soto; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cutting angle Dear Roxanne, I hope this is going to be of assistance to you, we manufacture all our microtomes & cryostats from a true 0? to 25? angle on the knife or blade holders. In this way we have set a standard for all our users. I cannot see why others choose arbitrary calibrations and vary them too. You will need to place a place a flat sample onto the cassette or object holder and adjust the knife or blade close to the face of the sample and view from the side of the edge of the knife or blades facet to make sure you have a 1 to 3? clearance. If this is not a clear description please come back to me and I will email you a sketch or try to place it on "list server images" of the Histonet.org website if someone can inform me how to place it there. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Roxanne Soto [mailto:godsgirlnow@msn.com] Sent: 03 May 2005 16:45 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cutting angle What angles does everyone out there cut at and what kind of blades do you use? I have always cut at 10 with high or low profile blades, but I have recently changed jobs and they cut at an angle of 21. They were told that with high profile blades they should use an angle of 21. I am having a hard time cutting at this angle....... any thoughts? Roxanne _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From julien_lambreydesouza <@t> uqar.qc.ca Wed May 4 12:30:38 2005 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien LambreyDeSouza) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Masson's trichrome Message-ID: <6.1.1.1.1.20050504131027.019835c0@pop3.uqar.qc.ca> Hello, I am trying out Masson's trichrome for the first time. I used the Goldner modification from Gabe 1976, with Fuschine-Ponceau, orang-G phosphomolybdic and Light green. We are staining 7 micron sections of entire fish larvae heads. When following the protocol (5 minutes in each color followed by a 1% acetic water rinse) I get a general green staining of all tissues. The only way to differentiate cartilage from the rest in our section is by recognizing cellular topography, same as with Hematox-eosine staining, therefore loosing the whole point in using Masson's trichrome. I seem to be doing something wrong. Would anybody familiar with the technique be inclined to share their expertise or protocol? I would be happy to send a pic of our staining result by email to have input on the possible problem. Thanks, Julien. Julien Lambrey de Souza Assistant de recherche, Biologiste M.Sc. Biologie ?volutive. Universit? du Qu?bec ? Rimouski D?partement de Biologie, Chimie et Sciences sant? 300, all?e des Ursulines Rimouski (Qu?bec) Canada G5L 3A1 T?l.: (418) 723-1986 ext. 1714 Fax.: (418) 724-1849 Courriel: julien_lambreydesouza@uqar.qc.ca From cormier <@t> MIT.EDU Wed May 4 12:50:31 2005 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Veterinary version of Ackermans' surgical pathology Message-ID: <5.2.1.1.2.20050504134537.015c63d0@po14.mit.edu> Hello All! Question, is there an vet version out there that is similar to Ackermans Surgical Pathology? I am looking more for "average" ranges of tissues for rodents such as "average size of mouse kidney varies between x and x cm. Any one know if such a thing/guide exists? Published papers? Thanks for guidance! Kathy From steven.coakley <@t> mirusbio.com Wed May 4 13:17:16 2005 From: steven.coakley <@t> mirusbio.com (Steven Coakley) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Herovici's picropolychrome Message-ID: I'm, looking for this procedure. It is specific to differenciate collagen types Steve From Thomas.Crowell <@t> biogenidec.com Wed May 4 13:34:14 2005 From: Thomas.Crowell <@t> biogenidec.com (Thomas Crowell) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Mouse alveolar macrophages In-Reply-To: Message-ID: Hi, Can anyone suggest an antibody that labels mouse lung macrophages in FFPE tissue? As confirmed with Serotec, F4/80 labels weakly or not at all. Thanks Tom Crowell BiogenIdec Cambridge, MA From bjrenquist <@t> ucdavis.edu Wed May 4 13:57:08 2005 From: bjrenquist <@t> ucdavis.edu (Ben) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Vector Red and True Blue Message-ID: <200505041857.j44Iv8sm003193@pop19.ucdavis.edu> Hello All, I have been attempting some double labeling for fos and estrogen receptor for about a month with little luck. I am using Vector Red (fos) and True Blue (ER) for my staining. I am having no problems with the ER labeling, but the fos labeling is not that clear. I don't appear to be getting a strong reaction product from the Vector Red. Has anyone else had this problem? How strong should the staining be (light pink, dark pink, red)? I have been careful to only place in xylene for a few moments before coverslipping and therefore don't think fading is my problem. I have been thinking that my antigen retrieval may be the problem. I am using triton X to improve antigen retrieval, but was thinking about microwaving. These are free floating sections. First is this possible to do with free floating sections and second how long do I microwave? Any help for this lost soul is greatly appreciated. Ben From nmuvarak <@t> facstaff.wisc.edu Wed May 4 14:06:37 2005 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Masson's trichrome Message-ID: I also did MTS for the first time on human saphenous vein. Got intense blue staining throughout the vessel and some red in the middle, but no nuclear staining. Here's the protocol I followed (if you're interested, Julien): Overnight in Bouin's solution Wash in tap water for 1 min Weigert's hematoxylin for 10 min Wash in tap water for 1 min, blue in 37mM ammonia water Beibrich scarlet for 5 min Wash in distilled water for 1 min Phosphotungstic/phosphomolybdic acid for 10 min Discard and directly add aniline blue for 5 minutes Wash in distilled water for 1 min Wash in acetic acid for 1 min, then again with distilled water for 1 min Dehydrate, clear, coverslip Anything wrong with this protocol? ----- Original Message ----- From: Julien LambreyDeSouza Date: Wednesday, May 4, 2005 12:30 pm Subject: [Histonet] Masson's trichrome > > Hello, > > I am trying out Masson's trichrome for the first time. I used the > Goldner > modification from Gabe 1976, with Fuschine-Ponceau, orang-G > phosphomolybdic > and Light green. We are staining 7 micron sections of entire fish > larvae > heads. When following the protocol (5 minutes in each color > followed by a > 1% acetic water rinse) I get a general green staining of all > tissues. The > only way to differentiate cartilage from the rest in our section > is by > recognizing cellular topography, same as with Hematox-eosine > staining, > therefore loosing the whole point in using Masson's trichrome. I > seem to be > doing something wrong. > > Would anybody familiar with the technique be inclined to share > their > expertise or protocol? I would be happy to send a pic of our > staining > result by email to have input on the possible problem. > > Thanks, > > Julien. > Julien Lambrey de Souza > Assistant de recherche, Biologiste M.Sc. > Biologie ?volutive. > Universit? du Qu?bec ? Rimouski > D?partement de Biologie, Chimie et Sciences sant? > 300, all?e des Ursulines > Rimouski (Qu?bec) Canada G5L 3A1 > T?l.: (418) 723-1986 ext. 1714 > Fax.: (418) 724-1849 > Courriel: julien_lambreydesouza@uqar.qc.ca > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From dobbin <@t> upei.ca Wed May 4 14:52:09 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Veterinary version of Ackermans' surgical pathology In-Reply-To: <5.2.1.1.2.20050504134537.015c63d0@po14.mit.edu> Message-ID: <4278EFAA.22116.191CBF9@localhost> Hi Kathy, There is at least one text dedicated to rodent pathology and I've seen another for pathology of laboratory animals. You'll have to do some digging on your own though (or wait to hear from someone else on the list), because I don't have access to the afore mentioned volumes, it's just that I know I have come across them at one time or another over the years. Sorry I couldn't be more helpful than that! Greg Date sent: Wed, 04 May 2005 13:50:31 -0400 To: histonet@lists.utsouthwestern.edu From: Kathy Cormier Subject: [Histonet] Veterinary version of Ackermans' surgical pathology > Hello All! > > Question, is there an vet version out there that is similar to > Ackermans Surgical Pathology? I am looking more for "average" ranges > of tissues for rodents such as "average size of mouse kidney varies > between x and x cm. Any one know if such a thing/guide exists? > Published papers? Thanks for guidance! > > Kathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From jwatson <@t> gnf.org Wed May 4 14:57:20 2005 From: jwatson <@t> gnf.org (James Watson) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Masson's trichrome Message-ID: I have done the Masson Trichrome on human and all kinds of animal tissue with success, I follow the protocol in the AFIP "Laboratory methods in Histotechnology" with a couple of modifications: Deparaffinize and hydrate to distilled water Bouins overnight or 60 degrees for 1 hour (If heated seal staining jar and open in hood after cooling) Wash in running water until sections are clear Weigerts hematoxylin 10 min. Running water 10 min Rinse in distilled water Biebrich Scarlet-acid fuchsin 15 min Rinse in distilled water Differentiate in Phosphotungstic-phosphomolybdic acid for 10-15 minutes (check under microscope if collagen tissue is clear, if not differentiate for 5-10 more minutes until collagen is clear.) Counterstain in aniline blue for 2 minutes (or light green 10 min.) Rinse in distilled water Dehydrate, clear and mount. Checking the differentiation is very important to ensure that all the red in not removed from the muscle and the collagen is clear and can pick up the counterstain. Shortening the aniline blue time decreases the possibility of overstaining and eliminates the need for differentiating in acetic water which can also remove your hematoxylin. Hope this helps. James Watson HT, ASCP Facilities Manager of Histology GNF, Genomics Institute of the Novartis Research Foundation Room C015 858-332-4647 jwatson@gnf.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of NIDAL E MUVARAK Sent: Wednesday, May 04, 2005 12:07 PM To: Julien LambreyDeSouza Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Masson's trichrome I also did MTS for the first time on human saphenous vein. Got intense blue staining throughout the vessel and some red in the middle, but no nuclear staining. Here's the protocol I followed (if you're interested, Julien): Overnight in Bouin's solution Wash in tap water for 1 min Weigert's hematoxylin for 10 min Wash in tap water for 1 min, blue in 37mM ammonia water Beibrich scarlet for 5 min Wash in distilled water for 1 min Phosphotungstic/phosphomolybdic acid for 10 min Discard and directly add aniline blue for 5 minutes Wash in distilled water for 1 min Wash in acetic acid for 1 min, then again with distilled water for 1 min Dehydrate, clear, coverslip Anything wrong with this protocol? ----- Original Message ----- From: Julien LambreyDeSouza Date: Wednesday, May 4, 2005 12:30 pm Subject: [Histonet] Masson's trichrome > > Hello, > > I am trying out Masson's trichrome for the first time. I used the > Goldner > modification from Gabe 1976, with Fuschine-Ponceau, orang-G > phosphomolybdic > and Light green. We are staining 7 micron sections of entire fish > larvae > heads. When following the protocol (5 minutes in each color > followed by a > 1% acetic water rinse) I get a general green staining of all > tissues. The > only way to differentiate cartilage from the rest in our section > is by > recognizing cellular topography, same as with Hematox-eosine > staining, > therefore loosing the whole point in using Masson's trichrome. I > seem to be > doing something wrong. > > Would anybody familiar with the technique be inclined to share > their > expertise or protocol? I would be happy to send a pic of our > staining > result by email to have input on the possible problem. > > Thanks, > > Julien. > Julien Lambrey de Souza > Assistant de recherche, Biologiste M.Sc. > Biologie ?volutive. > Universit? du Qu?bec ? Rimouski > D?partement de Biologie, Chimie et Sciences sant? > 300, all?e des Ursulines > Rimouski (Qu?bec) Canada G5L 3A1 > T?l.: (418) 723-1986 ext. 1714 > Fax.: (418) 724-1849 > Courriel: julien_lambreydesouza@uqar.qc.ca > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AGrobe2555 <@t> aol.com Wed May 4 15:29:40 2005 From: AGrobe2555 <@t> aol.com (AGrobe2555@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Sheep p47 phox Message-ID: <1c9.28084dbd.2faa8ab4@aol.com> Hello folks, I am having a difficult time detecting p47 phox from sheep with several different antibodies available. Can anyone recommend an antibody that works for IHC in cryosections and especially for western blot detection of protein from tissue lysates. Thanks in advance, Albert Albert C. Grobe, PhD Vascular Biology Lab, Rm 3300 Saint Patrick Hospital 554 W. Broadway Missoula, MT 59802 (406) 327-1676 Lab (406) 327-1679 Fax From BDUE <@t> PARTNERS.ORG Wed May 4 15:55:34 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Herovici's picropolychrome Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB50279F1@PHSXMB7.partners.org> Try PubMed. The original paper and a derivative are listed in the literature. You will likely have to pull the journals. -brice "THE INFORMATION TRANSMITTED IN THIS EMAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON, THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVE THIS EMAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER." HIPPA Policy, 2003, Brigham & Women's Hospital, Boston, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: Wednesday, May 04, 2005 2:17 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Herovici's picropolychrome I'm, looking for this procedure. It is specific to differenciate collagen types Steve _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Traczyk7 <@t> aol.com Wed May 4 16:07:26 2005 From: Traczyk7 <@t> aol.com (Traczyk7@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle Message-ID: <82.277be099.2faa938e@aol.com> Hi Alan, Help me out here, are you saying that the knife gauge angle and clearance angle are the same? If not, then does it really matter that the gauge markings are accurate? Setting a 1 to 3 degree clearance angle is still a problem. Can't very well stick my head in the cryostat chamber to view the block/knife interface from the end. And of course there is then the problem of the bevel angle of the blade varying between manufacturers. Also, you mentioned "flat" specimen face but wouldn't having the block parallel to the knife edge be important also? I have successfully set up many cryostats, in this life and in my past lives as both a rep and a tech. I have never been able to determine a universal method for adjusting knife angle but welcome any advice that will make me look brilliant to my customers. Thanks, Dorothy From Barb.Richmond <@t> mckennan.org Wed May 4 16:42:18 2005 From: Barb.Richmond <@t> mckennan.org (Barb Richmond) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] VIP PROCESSORS Message-ID: We currently are processing 1100 blocks per week on one of our processors and on the other we process small biopsies that total around 250/week. Currently we change both processors twice a week and are considering cutting it back to once a week. I need some support before I go ahead and change this procedure. Any comments?? ____________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From Kathy.Paton <@t> waitematadhb.govt.nz Wed May 4 18:54:10 2005 From: Kathy.Paton <@t> waitematadhb.govt.nz (Kathy Paton (WDHB)) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Automated Microtome Message-ID: We have a staff member with early degenerative shoulder arthritis. The manual microtome did aggravate her condition badly, effecting her ability to work efficiently and without pain. I purchased the Leica RM2155 motorized microtome and very very rapidly indeed her condition improved. Now two years later ( I have just spoken to her) she has absolutely no pain at all. I must mention tho' the other staff are very reluctant to use this "new-fangled-technology". Just habit I guess. Kathy Paton Auckland New Zealand -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Nita Searcy Sent: Thursday, 5 May 2005 01:12 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated Microtome Can I have the latest and greatest regarding automated microtomes? I have an employee with carpal tunnel- anyone have experience with such employees using this instrument to lessen their problems? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 From llewllew <@t> shaw.ca Wed May 4 19:26:20 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Masson's trichrome References: <6.1.1.1.1.20050504131027.019835c0@pop3.uqar.qc.ca> Message-ID: <002c01c55109$0fb73e70$7e034246@yourlk4rlmsu> Go to http://stainsfile.info and read up the principles on which trichrome stains work. I suggest you then start with the standard Masson method listed there. It is one of the easiest to control. The commonest reason for getting all green (or blue) staining is the fixation. Formalin fixation is the likely cause. Either modify the staining protocol to 10-2-2 minutes with plasma-polyacid-fibre stains, or refix sections in Bouin's fluid for an hour at 56C. The nuclei are not usually black, regardless of what the books say they should be. They are usually a muddy brown. Bryan Llewellyn ----- Original Message ----- From: "Julien LambreyDeSouza" To: Sent: Wednesday, May 04, 2005 10:30 AM Subject: [Histonet] Masson's trichrome Hello, I am trying out Masson's trichrome for the first time. I used the Goldner modification from Gabe 1976, with Fuschine-Ponceau, orang-G phosphomolybdic and Light green. We are staining 7 micron sections of entire fish larvae heads. When following the protocol (5 minutes in each color followed by a 1% acetic water rinse) I get a general green staining of all tissues. The only way to differentiate cartilage from the rest in our section is by recognizing cellular topography, same as with Hematox-eosine staining, therefore loosing the whole point in using Masson's trichrome. I seem to be doing something wrong. Would anybody familiar with the technique be inclined to share their expertise or protocol? I would be happy to send a pic of our staining result by email to have input on the possible problem. Thanks, Julien. Julien Lambrey de Souza Assistant de recherche, Biologiste M.Sc. Biologie ?volutive. Universit? du Qu?bec ? Rimouski D?partement de Biologie, Chimie et Sciences sant? 300, all?e des Ursulines Rimouski (Qu?bec) Canada G5L 3A1 T?l.: (418) 723-1986 ext. 1714 Fax.: (418) 724-1849 Courriel: julien_lambreydesouza@uqar.qc.ca _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bucana <@t> audumla.mdacc.tmc.edu Wed May 4 22:15:29 2005 From: bucana <@t> audumla.mdacc.tmc.edu (Corazon D. Bucana) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Mouse alveolar macrophages In-Reply-To: References: Message-ID: <5.1.1.6.0.20050504221207.0369b218@audumla.mdacc.tmc.edu> You might want to test anti-scavenger macrophage antibody. It works on frozen sections very well but Serotec might be able to tell you if it works on paraffin sections. At 02:34 PM 5/4/2005 -0400, you wrote: >Hi, > >Can anyone suggest an antibody that labels mouse lung macrophages in FFPE >tissue? As confirmed with Serotec, F4/80 labels weakly or not at all. > >Thanks >Tom Crowell >BiogenIdec >Cambridge, MA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Thu May 5 03:38:02 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] VIP PROCESSORS In-Reply-To: Message-ID: Buy an alcohol hydrometer, and test the last absolute alcohol. If you have never used one, you can buy them from Fisher for $20-50 dollars. Looks like a large thermometer, with markings in percent. Pour some of the last absolute alcohol in a large (1000 mL) graduate cylinder. Place the hydrometer in the alcohol. It will float. Read the marking at the meniscus of the alcohol (you may have to adjust for room temperature - usually comes with a chart). If the final absolute alcohol is still 100% - then go ahead and extend the number of days processing before changing. If it's under about 98%, don't extend. Has too much carry over water in it. By the way, do you change all solutions at once, or do you rotate (change just the first solutions of each percent alcohol, first clearant, first paraffin)? Might be cheaper to rotate. Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barb Richmond Sent: Wednesday, May 04, 2005 5:42 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] VIP PROCESSORS We currently are processing 1100 blocks per week on one of our processors and on the other we process small biopsies that total around 250/week. Currently we change both processors twice a week and are considering cutting it back to once a week. I need some support before I go ahead and change this procedure. Any comments?? ____________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cormier <@t> MIT.EDU Thu May 5 04:38:59 2005 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Mouse alveolar macrophages In-Reply-To: References: Message-ID: <5.2.1.1.2.20050505053719.016149b8@po14.mit.edu> Hi Tom, We use Caltag F4/80 (rm2915) (rat mono) at 1:150 on FFPE mouse tissues with much success. Kathy Cormier DCM MIT At 02:34 PM 5/4/2005 -0400, Thomas Crowell wrote: >Hi, > >Can anyone suggest an antibody that labels mouse lung macrophages in FFPE >tissue? As confirmed with Serotec, F4/80 labels weakly or not at all. > >Thanks >Tom Crowell >BiogenIdec >Cambridge, MA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cormier <@t> MIT.EDU Thu May 5 05:05:44 2005 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Veterinary version of Ackermans' surgical pathology In-Reply-To: <4278EFAA.22116.191CBF9@localhost> References: <5.2.1.1.2.20050504134537.015c63d0@po14.mit.edu> Message-ID: <5.2.1.1.2.20050505055148.01621970@po14.mit.edu> > >Hey Greg and the rest of the Histonet Gang, I suppose that I should mention that this is for training a med tech (snatched one from the dark side!!) in mouse surveillance, we are trying to get a guideline in place so that this tech will have a better idea of what is normal and what is outside of normal limits. Spontaneous tumors and lesions are pretty clear and easy to see, but "slightly enlarged kidney" is a bit harder to define and to learn. We know that this is just one of those learning curves and it the more mice this tech sees the better he will be, but we were hoping to make it a bit easier for him. We do have a nice atlas for him to help ID organs in rodents, so we are not completely out in the cold! Thanks for all the help! Kathy DCM -MIT >Hi Kathy, >There is at least one text dedicated to rodent pathology and I've >seen another for pathology of laboratory animals. You'll have to do >some digging on your own though (or wait to hear from someone >else on the list), because I don't have access to the afore >mentioned volumes, it's just that I know I have come across them at >one time or another over the years. Sorry I couldn't be more helpful >than that! >Greg > >Date sent: Wed, 04 May 2005 13:50:31 -0400 >To: histonet@lists.utsouthwestern.edu >From: Kathy Cormier >Subject: [Histonet] Veterinary version of Ackermans' >surgical pathology > > > Hello All! > > > > Question, is there an vet version out there that is similar to > > Ackermans Surgical Pathology? I am looking more for "average" ranges > > of tissues for rodents such as "average size of mouse kidney varies > > between x and x cm. Any one know if such a thing/guide exists? > > Published papers? Thanks for guidance! > > > > Kathy > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Greg Dobbin >Research Technologist, >Pathology Lab, >Atlantic Veterinary College, U.P.E.I. >550 University Ave. >Charlottetown, P.E.I. >Canada, C1A 4P3 >Phone: (902)566-0744 >Fax: (902)566-0851 >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Happiness is a journey, not a destination. From Andrew.Prior <@t> Smith-Nephew.com Thu May 5 08:03:09 2005 From: Andrew.Prior <@t> Smith-Nephew.com (Prior, Andrew) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Large slides and Exakt resin Message-ID: Our lab is now doing a lot of work on resin embedded samples that are too large for standard slides (76x25 mm) so we are using over-size slides. These are 100 x 50 x 1.5 mm. These are great for fitting the samples on BUT the slides don't fit in any racks that we have, or in staining pots. Does anyone out there use these sized slides and know of a supplier of suitable slide boxes, racks and staining jars? I have had a look through some of the main suppliers and couldn't see anything. Any help greatly appreciated. As for the Exakt resin. We were told that we could recycle the Technovit 7200VLC resin by using it for the previous dilution i.e. the 100% becomes 70:30 which becomes 50:50 etc. System is still new to us so haven't tried it out yet. Don't know if this would work with 7100 resin. Can't see why not as long as the last solution is fresh. Would be very interested to hear how it turns out >>Date: Thu, 28 Apr 2005 14:27:46 +0200 >>From: Jorge Tornero >>Subject: [Histonet] Recycling Technovit 7100 >>To: histonet@lists.utsouthwestern.edu >>Message-ID: <8c964a790504280527384ce43c@mail.gmail.com> >>Content-Type: text/plain; charset=ISO-8859-1 >>Hello, >> >>does anybody have experience about re-using technovit 7100 solutions? >> >>For instance, preparing the 1:1 absolute alcohol-resin solution with >>used preinfiltration solution. >> >>Cheers >> >>Jorge Tornero >>IEO-C?diz (Spain) Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. From abright <@t> brightinstruments.com Thu May 5 08:30:26 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Large slides and Exakt resin Message-ID: Surgipath should be able to supply this size Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Prior, Andrew [mailto:Andrew.Prior@Smith-Nephew.com] Sent: 05 May 2005 14:03 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Large slides and Exakt resin Our lab is now doing a lot of work on resin embedded samples that are too large for standard slides (76x25 mm) so we are using over-size slides. These are 100 x 50 x 1.5 mm. These are great for fitting the samples on BUT the slides don't fit in any racks that we have, or in staining pots. Does anyone out there use these sized slides and know of a supplier of suitable slide boxes, racks and staining jars? I have had a look through some of the main suppliers and couldn't see anything. Any help greatly appreciated. As for the Exakt resin. We were told that we could recycle the Technovit 7200VLC resin by using it for the previous dilution i.e. the 100% becomes 70:30 which becomes 50:50 etc. System is still new to us so haven't tried it out yet. Don't know if this would work with 7100 resin. Can't see why not as long as the last solution is fresh. Would be very interested to hear how it turns out >>Date: Thu, 28 Apr 2005 14:27:46 +0200 >>From: Jorge Tornero >>Subject: [Histonet] Recycling Technovit 7100 >>To: histonet@lists.utsouthwestern.edu >>Message-ID: <8c964a790504280527384ce43c@mail.gmail.com> >>Content-Type: text/plain; charset=ISO-8859-1 >>Hello, >> >>does anybody have experience about re-using technovit 7100 solutions? >> >>For instance, preparing the 1:1 absolute alcohol-resin solution with >>used preinfiltration solution. >> >>Cheers >> >>Jorge Tornero >>IEO-C?diz (Spain) Andrew Andrew Prior Histologist Smith & Nephew Research Centre York Science Park Heslington York Confidentiality. This electronic transmission is strictly confidential to Smith & Nephew and intended solely for the addressee. It may contain information which is covered by legal, professional or other privilege. If you are not the intended addressee, or someone authorised by the intended addressee to receive transmissions on behalf of the addressee, you must not retain, disclose in any form, copy or take any action in reliance on this transmission. If you have received this transmission in error, please notify the sender as soon as possible and destroy this message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mario <@t> bu.edu Thu May 5 08:48:58 2005 From: mario <@t> bu.edu (Mario Muscedere) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] [IHC] Whole-mount tissue permeablization Message-ID: <22ae6b2aaea41352004430a46474e9f4@bu.edu> Hello, I'm relatively new to immunohistochemistry, and am trying to optimize a protocol for staining whole ant brains. The brains are relatively small pieces of tissue (about 300 microns x 300 microns) that should be (if I understand correctly) well within the size range of tissue pieces that can be successfully processed whole. I have been using the standard 4% paraformaldehyde in PBS at 4 C fix, and have been incubating in primary antibody 1:50 for 3 days at RT, and secondary 1:200 for 2 days at RT. Despite these long incubations I'm having trouble with antibody penetration, and am starting to look into permeabliziation methods. I see several techniques in the literature: enzyme digestion, adding Triton-X 100 and/or DMSO to the washing and incubation buffers, and incubations in acetone or methanol after fixation, but there's no explanation (that I can find) about how these techniques work. Does anyone have experience with these issues? What does DMSO or Triton-X or acetone "do" to the tissues? What kinds of concentrations have you found useful or not useful? There is some indication of a protective, very very thin "sheath" of tissue around the actual neurons of the brain. If this is the case, is enzymatic digestion of the tissue the only option? Pros or cons of the different treatments? I know this is a long and detailed question. Thanks so much for any help you might provide. Mario Muscedere??? ? ? Biology Department Boston University 5 Cummington Street Boston, MA, 02215? Office: 409 BRB Office #: 617-353-6977 From cfavara <@t> niaid.nih.gov Thu May 5 08:57:23 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Mouse alveolar macrophages Message-ID: I am not familiar with this antibody can you give more information? Is it from Serotec? c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Corazon D. Bucana [mailto:bucana@audumla.mdacc.tmc.edu] Sent: Wednesday, May 04, 2005 8:15 PM To: Thomas Crowell Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Mouse alveolar macrophages You might want to test anti-scavenger macrophage antibody. It works on frozen sections very well but Serotec might be able to tell you if it works on paraffin sections. At 02:34 PM 5/4/2005 -0400, you wrote: >Hi, > >Can anyone suggest an antibody that labels mouse lung macrophages in FFPE >tissue? As confirmed with Serotec, F4/80 labels weakly or not at all. > >Thanks >Tom Crowell >BiogenIdec >Cambridge, MA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abright <@t> brightinstruments.com Thu May 5 09:24:12 2005 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] cutting angle Message-ID: Dorothy, I will try. I am saying when the angle indicator on the knife holder is at 0? on the specimen side of a standard 'c' profile knife, the knife is at 0?. As most knives are sharpened with a 5? to 7 1/2? facet angle would then say on a 5? facet angle that you could section with a clearance angle of 3? with the knife set at 8? using the angle indicator on the knife holder. If the knife has been stropped which would increase the knife facet angle then this would have to be allowed for by increasing the angle indicator on the knife holder by another 2-3?. Disposable blade holders are manufactured with an offset angle built in and will section with the angle indicator on the knife holder set at 3? to 7 1/2?. This is the main point, it would be very beneficial if all manufacturers used the same angle indicator on their knife holders, as inexperience users having an issue with knife angle settings would be able to indicate when asking for assistance what true angles they are using. Disposable blades could have a fixed angle set as there is no variation between blades purchased from a single source, but experienced users would argue that being able to adjust the angle for different types of specimens and perfection is desirable and I tend to agree. The main results to look for is if the angle is to steep then you will get no sections for awhile and then a very thick section as the specimen is hitting the base of the facet and compressing the specimen to a point where eventually the specimens compression is so great it will spring forward and be forced into cutting a thick section or forcing the specimen from the object holder. If the angle it to wide you will get chatter on the specimen face as the knife edge is just scrapping the specimen face. Taking into account that the knife or blade is sharp. A cold mirror could be used to view the specimen/knife interface in a cryostat. Orientation of the specimen face is very important but not an issue I wish to cover in this email. The last part of your email is the hardest to answer, as I would have thought by now you should have had no problems setting up knife angles, but hopefully what I have written above may help. Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Traczyk7@aol.com [mailto:Traczyk7@aol.com] Sent: 04 May 2005 22:07 To: Alan Bright Cc: HACKERLAB@aol.com; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] cutting angle Hi Alan, Help me out here, are you saying that the knife gauge angle and clearance angle are the same? If not, then does it really matter that the gauge markings are accurate? Setting a 1 to 3 degree clearance angle is still a problem. Can't very well stick my head in the cryostat chamber to view the block/knife interface from the end. And of course there is then the problem of the bevel angle of the blade varying between manufacturers. Also, you mentioned "flat" specimen face but wouldn't having the block parallel to the knife edge be important also? I have successfully set up many cryostats, in this life and in my past lives as both a rep and a tech. I have never been able to determine a universal method for adjusting knife angle but welcome any advice that will make me look brilliant to my customers. Thanks, Dorothy From JNocito <@t> Pathreflab.com Thu May 5 10:09:18 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Which RPMI media to use In-Reply-To: Message-ID: I want to thank everyone for their responses. Since we deal with two labs and only one lab sends us RPMI media, I wanted to have enough on hand. I guess I'll have to nicely ask the other lab to either send us some, or tell me which formula to use. This ought to rock the boat a little. (like I haven't done that before). Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Linda Blazek Sent: Tuesday, May 03, 2005 1:57 PM To: histonet@pathology.swmed.edu; JNocito@Pathreflab.com Subject: Re: [Histonet] Which RPMI media to use Joe, The best one to uses is the one that the facility you are sending them to wants you to use. The facility that we send ours to even furnishes it to us. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Joe Nocito" 05/03/2005 2:35:52 PM >>> Greetings Histoland, once, again, I need to tap into your vast knowledge. We will be sending out bone marrow, lymph node and placenta cases for flow cytometry and cytogenetic studies. Looking through the catalogs, there are about 15 different formulas for RPMI. Can any one tell me which formula is the best one. I'm at a loss. thanks in advance Joe Nocito _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Thu May 5 10:12:41 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] VIP PROCESSORS In-Reply-To: Message-ID: Barb, we change out our solutions on Monday and then rotate the solutions on Wednesday and Friday. It may be a little extreme, but we never have to worry about poor processing. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Barb Richmond Sent: Wednesday, May 04, 2005 4:42 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] VIP PROCESSORS We currently are processing 1100 blocks per week on one of our processors and on the other we process small biopsies that total around 250/week. Currently we change both processors twice a week and are considering cutting it back to once a week. I need some support before I go ahead and change this procedure. Any comments?? ____________________ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mward <@t> wfubmc.edu Thu May 5 10:13:42 2005 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] PCR laboratory Message-ID: <61135F0455D33347B5AAE209B903A3040DA5B803@EXCHVS2.medctr.ad.wfubmc.edu> We are in the process of designing new lab space. In addition to doing immunos, we also do PCR for various infectious disease PCR (herpes, CMV, enterovirus, etc.) I would be interesting in talking with anyone out there who performs PCR. If you would pass this message along to anyone in your various institutions I would really appreciate it. I am interested in square footage, etc. Martha Ward Wake Forest University Baptist Medical Center Winston-Salem, NC 27157 1-336-716-2756 From cekallsen <@t> ucdavis.edu Thu May 5 11:16:40 2005 From: cekallsen <@t> ucdavis.edu (Craig Kallsen) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Substitutes for picric acid using crystal violet stains Message-ID: <427A46E8.7000105@ucdavis.edu> Dear List, I am new to this and would like to stain phloem, xylem and cambial cells in citrus tree-trunk tissue in paraplast embedded tissue. I have a protocol for using crystal violet stain that requires ethnolic picric acid in the procedure. Is there a substitute for picric acid? I am having trouble obtaining picric acid in alcohol (as well as finding the price intimidating). From GDawson <@t> dynacaremilwaukee.com Thu May 5 12:06:29 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] titanium microtome blades Message-ID: All, I have a pathologist who is running into problems with metal contamination due to various microtome knives running into prostatic concretions. He asked me to inquire as to whether or not anyone out there in histo-land has heard of a manufacturer of titanium microtome blades. Non disposable or disposable will do. Thanx In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From Melissa.Gonzalez <@t> cellgenesys.com Thu May 5 12:19:31 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Pericyte, smooth muscle cell IHC problems Message-ID: Dear all, I have been trying unsuccessfully to stain for pericytes/ smooth muscle cells in sub cutaneous gliomas in a mouse model. (Tissues are either 4% PFA perfused or 10% FFPE) I have tried several products, including: * NG2 (Chemicon): works nicely in normal mouse organs, but binds to much of the tissue in the gliomas, making it impossible to interpret. *PDGFRB (Santa Cruz): resulted in a few crazy-appearing vascular areas, very weird * ASMA (DAKO, Sigma, US Bio, Neomarkers, Abcam) *Desmin (DAKO, Santa Cruz) --in both monoclonals (using a mouse on mouse kit which did not work) and polyclonals, for which I only see staining in the attached skin and a very rare area in the tissue. I have tried both immunofluorescence on the perfused, and chromogenic detection on paraffin sections, using Citrate, high pH, pepsin, and proteinase K retrievals. Is it possible that smooth muscle cells would not be well developed around blood vessels in a sub cutaneous glioma, and that is the reason I only see staining in the attached skin with a rare occurrence in the tumor? (Tumors are very vascular, blood vessels are definitely present) Also, what is everyone using for blood vessel markers? We used to use BD Pharm. Pecam-1, which doesn't work anymore on anything except frozens, Santa Cruz goat Pecam-1 was working great but the most recent lots I ordered now appear completely negative using any detection. I have now switched to using CD105. Any help would be greatly appreciated as I have tried all the tricks I know, and am left completely puzzled. Thanks so much, Melissa From pmarcum <@t> vet.upenn.edu Thu May 5 12:34:55 2005 From: pmarcum <@t> vet.upenn.edu (pmarcum@vet.upenn.edu) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] titanium microtome blades In-Reply-To: References: Message-ID: <1115314495.427a593f7f339@imp.vet.upenn.edu> Hi Glen, I know I use the tungsten carbide blades and have not heard of titanium. If you find some would you let us all know about them. Pam Marcum Quoting "Dawson, Glen" : > All, > > I have a pathologist who is running into problems with metal contamination > due to various microtome knives running into prostatic concretions. He > asked me to inquire as to whether or not anyone out there in histo-land has > heard of a manufacturer of titanium microtome blades. Non disposable or > disposable will do. > > Thanx In Advance, > > Glen Dawson BS, HT & QIHC (ASCP) > IHC Manager > Milwaukee, WI > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ssalesky <@t> LowellGeneral.org Thu May 5 12:38:22 2005 From: ssalesky <@t> LowellGeneral.org (Shawn Salesky) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] IHC Billing CPT question. Message-ID: <9E49B4E3A6200C4B91E5FB99FB1F7EDB571FC0@NTMAIL> Hi All, >From the discussion a few days ago I had some questions concerning IHC Billing. Patient: B. Smith Has a Left and Right Breast Core Bx submitted as parts A and B. Both of which have ER, PR and Her2 done on them. The Her2 is negative so there is no reflex to F.I.S.H. >From the discussion on Histo net I gather that the majority of facilities would be charging for Quantitative/Semi-quantitative IHC x 6. Is there any literature or references that anyone is using to determine that it is legal to charge for 6 instances? At my current facility that same scenario would only be billed as 3 instances. Any help would be appreciated. This e-mail and any attachments is intended only for the person or entity to which it is addressed and may contain confidential and privileged information. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this e-mail and destroy any copies. Any dissemination or use of this information by a person other than the intended recipient is unauthorized and may be illegal. From cfranci <@t> rigel.com Thu May 5 12:48:57 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] really basic question Message-ID: <25e84fccbf5224c4155829ddc523675d@rigel.com> I'm doing staining of mouse tissues and need to get vascular tissue from different areas. What places/organs are the best bets? Kidneys come to mind but, where else? Thanks a bunch, Chris From Jackie.O'Connor <@t> abbott.com Thu May 5 13:02:45 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Pericyte, smooth muscle cell IHC problems Message-ID: HEY! I've had the same problem with the Santa Cruz CD31! How utterly frustrating! I had a lot from 11/04 that didn't work, and when I requested a replacement this month, that didn't work either!! I've really come to depend on that antibody for microvessel density in my xenografts. I haven't been able to find a comparable CD31 to work in FFPE mouse. The BD will work on zinc fixed, tho, without antigen retrieval. Anyone else have any tidbits for FFPE murine tissue? Jackie O' Jacqueline M. O'Connor HT(ASCP) Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Melissa Gonzalez" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/05/2005 12:19 PM To: cc: Subject: [Histonet] Pericyte, smooth muscle cell IHC problems Dear all, I have been trying unsuccessfully to stain for pericytes/ smooth muscle cells in sub cutaneous gliomas in a mouse model. (Tissues are either 4% PFA perfused or 10% FFPE) I have tried several products, including: * NG2 (Chemicon): works nicely in normal mouse organs, but binds to much of the tissue in the gliomas, making it impossible to interpret. *PDGFRB (Santa Cruz): resulted in a few crazy-appearing vascular areas, very weird * ASMA (DAKO, Sigma, US Bio, Neomarkers, Abcam) *Desmin (DAKO, Santa Cruz) --in both monoclonals (using a mouse on mouse kit which did not work) and polyclonals, for which I only see staining in the attached skin and a very rare area in the tissue. I have tried both immunofluorescence on the perfused, and chromogenic detection on paraffin sections, using Citrate, high pH, pepsin, and proteinase K retrievals. Is it possible that smooth muscle cells would not be well developed around blood vessels in a sub cutaneous glioma, and that is the reason I only see staining in the attached skin with a rare occurrence in the tumor? (Tumors are very vascular, blood vessels are definitely present) Also, what is everyone using for blood vessel markers? We used to use BD Pharm. Pecam-1, which doesn't work anymore on anything except frozens, Santa Cruz goat Pecam-1 was working great but the most recent lots I ordered now appear completely negative using any detection. I have now switched to using CD105. Any help would be greatly appreciated as I have tried all the tricks I know, and am left completely puzzled. Thanks so much, Melissa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Thu May 5 13:11:26 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] IHC Billing CPT question. In-Reply-To: <9E49B4E3A6200C4B91E5FB99FB1F7EDB571FC0@NTMAIL> Message-ID: Shawn, some one there should have a current CPT book. Look under the immuno code, there it will tell you that you may charge for each antibody. If there are two specimens from two distinct sites, we charge twice. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Shawn Salesky Sent: Thursday, May 05, 2005 12:38 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] IHC Billing CPT question. Hi All, >From the discussion a few days ago I had some questions concerning IHC Billing. Patient: B. Smith Has a Left and Right Breast Core Bx submitted as parts A and B. Both of which have ER, PR and Her2 done on them. The Her2 is negative so there is no reflex to F.I.S.H. >From the discussion on Histo net I gather that the majority of facilities would be charging for Quantitative/Semi-quantitative IHC x 6. Is there any literature or references that anyone is using to determine that it is legal to charge for 6 instances? At my current facility that same scenario would only be billed as 3 instances. Any help would be appreciated. This e-mail and any attachments is intended only for the person or entity to which it is addressed and may contain confidential and privileged information. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this e-mail and destroy any copies. Any dissemination or use of this information by a person other than the intended recipient is unauthorized and may be illegal. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Melissa.Gonzalez <@t> cellgenesys.com Thu May 5 13:09:53 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Pericyte, smooth muscle cell IHC problems Message-ID: Thank God! I thought I was going crazy! When I found Santa Cruz's I was thrilled because it worked on everything and looked great. Then it suddenly went dead no matter what I try it on. I hate it when these things happen. FYI- R&D systems rat x mouse CD105 works really well, as does Serotec's CD34. But I know some people insist on Pecam-1 as being the most specific and reliable for blood vessels, and now I have no product to use. Melissa -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: Thursday, May 05, 2005 11:03 AM To: Melissa Gonzalez Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Pericyte, smooth muscle cell IHC problems HEY! I've had the same problem with the Santa Cruz CD31! How utterly frustrating! I had a lot from 11/04 that didn't work, and when I requested a replacement this month, that didn't work either!! I've really come to depend on that antibody for microvessel density in my xenografts. I haven't been able to find a comparable CD31 to work in FFPE mouse. The BD will work on zinc fixed, tho, without antigen retrieval. Anyone else have any tidbits for FFPE murine tissue? Jackie O' Jacqueline M. O'Connor HT(ASCP) Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Melissa Gonzalez" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/05/2005 12:19 PM To: cc: Subject: [Histonet] Pericyte, smooth muscle cell IHC problems Dear all, I have been trying unsuccessfully to stain for pericytes/ smooth muscle cells in sub cutaneous gliomas in a mouse model. (Tissues are either 4% PFA perfused or 10% FFPE) I have tried several products, including: * NG2 (Chemicon): works nicely in normal mouse organs, but binds to much of the tissue in the gliomas, making it impossible to interpret. *PDGFRB (Santa Cruz): resulted in a few crazy-appearing vascular areas, very weird * ASMA (DAKO, Sigma, US Bio, Neomarkers, Abcam) *Desmin (DAKO, Santa Cruz) --in both monoclonals (using a mouse on mouse kit which did not work) and polyclonals, for which I only see staining in the attached skin and a very rare area in the tissue. I have tried both immunofluorescence on the perfused, and chromogenic detection on paraffin sections, using Citrate, high pH, pepsin, and proteinase K retrievals. Is it possible that smooth muscle cells would not be well developed around blood vessels in a sub cutaneous glioma, and that is the reason I only see staining in the attached skin with a rare occurrence in the tumor? (Tumors are very vascular, blood vessels are definitely present) Also, what is everyone using for blood vessel markers? We used to use BD Pharm. Pecam-1, which doesn't work anymore on anything except frozens, Santa Cruz goat Pecam-1 was working great but the most recent lots I ordered now appear completely negative using any detection. I have now switched to using CD105. Any help would be greatly appreciated as I have tried all the tricks I know, and am left completely puzzled. Thanks so much, Melissa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dharclerode <@t> macropore.com Thu May 5 13:47:16 2005 From: dharclerode <@t> macropore.com (Donna Harclerode) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Over sized slide racks and free floating tissue handling Message-ID: Brain Research Laboratories have wonderful items for both oversized specimens (slides, coverslips, racks etc.) and free floating tissues. They are a wonderful small company to deal with and I have used their products for a long time. They have also proven to be less expensive for the few things I can find elsewhere. They even have "plus" oversized slides 38 x 75mm (1-1/2" x 3") 1.0mm thickness, adhesion superfrost plus. Good luck Donna Harclerode, HT, (ASCP), HTL, QIHC Immunohistochemist MacroPore Biosurgery, Inc 6740 Top Gun St. San Diego, CA 92121 858-458-0900 xt 322 dharclerode@macropore.com Brain Research Laboratories Division of Cambridge Intelligent Systems, Inc. Waban P.O. Box 88 Newton, MA 02468 Please use this for mailing orders as well as remittance Telephone: (617)965-5544 Toll-Free: (888)BRL-5544 Fax: (617)965-6220 E-mail: brl@brainresearchlab.com From Don.Birgerson <@t> leica-microsystems.com Thu May 5 13:48:14 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] titanium microtome blades Message-ID: Hi Glen, Could the contamination problem be solved by using Ralph glass knives( 25 or 38mm widths)?? We have both triangular and Ralph glass knife holders for our microtomes. Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Dawson, Glen" cc: Sent by: Subject: [Histonet] titanium microtome blades histonet-bounces@lists.utsouth western.edu 05/05/2005 12:06 PM All, I have a pathologist who is running into problems with metal contamination due to various microtome knives running into prostatic concretions. He asked me to inquire as to whether or not anyone out there in histo-land has heard of a manufacturer of titanium microtome blades. Non disposable or disposable will do. Thanx In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From RCHIOVETTI <@t> aol.com Thu May 5 13:49:05 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] titanium microtome blades Message-ID: <1c7.27fef74d.2fabc4a1@aol.com> In a message dated 5/5/2005 10:09:02 AM US Mountain Standard Time, GDawson@dynacaremilwaukee.com writes: > He > asked me to inquire as to whether or not anyone out there in histo-land has > heard of a manufacturer of titanium microtome blades. Glen, I looked at the Sturkey line of premium blades a while back, and I tried the "gold" version. The gold color is a titanium nitride coating. I don't think anyone makes a blade out of solid titanium. Sturkey now also has a blade with a diamond coating. I haven't tried it yet, but I'd think the diamond coated blade would also do the trick for you. you can go to their website here, then click on the link for the "Extremus" knives for more info. Bob Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From RCHIOVETTI <@t> aol.com Thu May 5 13:54:45 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] really basic question Message-ID: <14.44bb663a.2fabc5f5@aol.com> In a message dated 5/5/2005 10:49:52 AM US Mountain Standard Time, cfranci@rigel.com writes: > I'm doing staining of mouse tissues and need to get vascular tissue > from different areas. > Chris, I'd add heart, lungs, liver and spleen to the list. All are highly vascular. Cheers, Bob Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From RCHIOVETTI <@t> aol.com Thu May 5 13:57:36 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] titanium microtome blades Message-ID: <1a0.333a5d63.2fabc6a0@aol.com> All, Sorry, the hyperlink didn't show up on my posting. For info on Sturkey blades, the website is: Bob Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From mcauliff <@t> umdnj.edu Thu May 5 14:00:14 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] really basic question/blood vessels In-Reply-To: <25e84fccbf5224c4155829ddc523675d@rigel.com> References: <25e84fccbf5224c4155829ddc523675d@rigel.com> Message-ID: <427A6D3E.6010604@umdnj.edu> Spleen will have vessles of different sizes and the red pulp is a veritable swamp, liver has lots of small vessels, connective tissue of salivary glands might be good, the brain is highly vascular as is the heart. Geoff Christian Franci wrote: > I'm doing staining of mouse tissues and need to get vascular tissue > from different areas. > What places/organs are the best bets? Kidneys come to mind but, where > else? > > Thanks a bunch, > Chris > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From bucana <@t> audumla.mdacc.tmc.edu Thu May 5 14:05:36 2005 From: bucana <@t> audumla.mdacc.tmc.edu (Corazon D. Bucana) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Mouse alveolar macrophages In-Reply-To: Message-ID: <5.1.1.6.0.20050505134854.01e350e0@audumla.mdacc.tmc.edu> This antibody came from the lab of S. Gordon (the same lab that developed F4/80 antibody). The antibody reacts to mouse macrophage scavenger receptor types I and II. It stains tissue macrophages and is known to be present in abundance in atherosclerotic lesions. My personal experience with this antibody is that macrophages found in wound healing reacts very strongly to this antibody and at a certain stage of wound healing it is possible to see macrophages stained by both antibodies but in the later stages of wound healing (eg. 10 days),F4/80 reactivity becomes weak but the anti-scavenger receptor is still very strong. Strong reaction is also seen in several tumors as well. since alveolar macrophages are essentially scavengers it seems reasonable that a strong reaction will also be seen in these cells. I hope this helps. At 09:57 AM 5/5/2005 -0400, you wrote: >I am not familiar with this antibody can you give more information? Is it >from Serotec? >c > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > >-----Original Message----- >From: Corazon D. Bucana [mailto:bucana@audumla.mdacc.tmc.edu] >Sent: Wednesday, May 04, 2005 8:15 PM >To: Thomas Crowell >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Mouse alveolar macrophages > >You might want to test anti-scavenger macrophage antibody. It works on >frozen sections very well but Serotec might be able to tell you if it works >on paraffin sections. > >At 02:34 PM 5/4/2005 -0400, you wrote: > > >Hi, > > > >Can anyone suggest an antibody that labels mouse lung macrophages in FFPE > >tissue? As confirmed with Serotec, F4/80 labels weakly or not at all. > > > >Thanks > >Tom Crowell > >BiogenIdec > >Cambridge, MA > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Thu May 5 14:15:01 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] [IHC] Whole-mount tissue permeablization In-Reply-To: <22ae6b2aaea41352004430a46474e9f4@bu.edu> References: <22ae6b2aaea41352004430a46474e9f4@bu.edu> Message-ID: <427A70B5.4010801@umdnj.edu> Hi Mario: \ Just because the tissue can be processed whole does not mean that antibody molecules, which are huge, will penetrate all the way to interesting places. The various permeablization methods make cell membranes more permeable by removing lipids. Do put some TritonX (try 0.1 to 0.3%) in your primary and secondary Ab solutions. Also 1. cut the "brain" in half or quarters to improve penetration, or cut Vibratome or frozen sections. 2. are you sure your Ab gives good results with buffered paraformaldehyde fixation for whatever time you are fixing? 3. have you tried an acid -alcohol fixative (which works differently than formlin)? 4. a freeze-thaw cycle, after appropriate cryoprotection, also improves penetration by punching holes in cell membranes. As far as which is the best method for your tissue+fixation+antibody combination, you should do a literature search (PubMed) to see what has been done and what their results look like. Geoff Mario Muscedere wrote: > Hello, > I'm relatively new to immunohistochemistry, and am trying to > optimize a protocol for staining whole ant brains. The brains are > relatively small pieces of tissue (about 300 microns x 300 microns) > that should be (if I understand correctly) well within the size range > of tissue pieces that can be successfully processed whole. I have > been using the standard 4% paraformaldehyde in PBS at 4 C fix, and > have been incubating in primary antibody 1:50 for 3 days at RT, and > secondary 1:200 for 2 days at RT. Despite these long incubations I'm > having trouble with antibody penetration, and am starting to look into > permeabliziation methods. I see several techniques in the literature: > enzyme digestion, adding Triton-X 100 and/or DMSO to the washing and > incubation buffers, and incubations in acetone or methanol after > fixation, but there's no explanation (that I can find) about how these > techniques work. Does anyone have experience with these issues? What > does DMSO or Triton-X or acetone "do" to the tissues? What kinds of > concentrations have you found useful or not useful? There is some > indication of a protective, very very thin "sheath" of tissue around > the actual neurons of the brain. If this is the case, is enzymatic > digestion of the tissue the only option? Pros or cons of the > different treatments? > > I know this is a long and detailed question. Thanks so much for any > help you might provide. > > Mario Muscedere > Biology Department > Boston University > 5 Cummington Street > Boston, MA, 02215 > Office: 409 BRB > Office #: 617-353-6977 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From sharon.osborn <@t> dnax.org Thu May 5 14:17:14 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] RE: automated microtomes Message-ID: <29B25753F6B1D51196110002A589D44402397F84@PALMSG30.us.schp.com> Nita, I have used both the Microm and Leica automated microtomes. They both greatly ease the pains/problems of repetitive motion (carpal tunnel) problems. In addition, a correct sitting height, well designed bench and good chair are important. I suggest you have the reps bring in each microtome for you and the tech try out. Have them leave for at least one week so you have sustained use to assist in making the decision. Currently, I am using the Leica RM 2155 and love it. When I used the Microm, I loved it. Both are manufactured in Germany, basically, across the street from each other. Some engineers from Leica formed their own company and produced the Microm. Microm and Leica seem to have the smoothest, easiest functioning of the automated microtomes. Both have their positives...it may be a matter of personal preference. If you use a manual rotary, the ErgoStar that another tech mentioned, has been designed specifically for ergonomic motion and for use with either left or right hand. As a manual, it is easy and easy on the joints to use. sharon osborn DNAX Palo Alto, CA Message: 13 Date: Thu, 5 May 2005 11:54:10 +1200 From: "Kathy Paton \(WDHB\)" Subject: RE: [Histonet] Automated Microtome To: "Nita Searcy" , Message-ID: Content-Type: text/plain; charset="us-ascii" We have a staff member with early degenerative shoulder arthritis. The manual microtome did aggravate her condition badly, effecting her ability to work efficiently and without pain. I purchased the Leica RM2155 motorized microtome and very very rapidly indeed her condition improved. Now two years later ( I have just spoken to her) she has absolutely no pain at all. I must mention tho' the other staff are very reluctant to use this "new-fangled-technology". Just habit I guess. Kathy Paton Auckland New Zealand -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu ] On Behalf Of Nita Searcy Sent: Thursday, 5 May 2005 01:12 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated Microtome Can I have the latest and greatest regarding automated microtomes? I have an employee with carpal tunnel- anyone have experience with such employees using this instrument to lessen their problems? Thanks Nita Searcy, HT/HTL (ASCP) Scott and White Hospital Temple, Texas Division Manager, Anatomic Pathology 254-724-2438 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From RCHIOVETTI <@t> aol.com Thu May 5 14:32:03 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] titanium microtome blades Message-ID: <12e.5d5e797a.2fabceb3@aol.com> I give up...time for a drink, I guess...other than coffee! Link is: sturkey.com Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From gcallis <@t> montana.edu Thu May 5 14:50:37 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Scavenger receptor antibody info Message-ID: <6.0.0.22.1.20050505134850.01b37a30@gemini.msu.montana.edu> SEROTEC rat anti-mouse CD204 for scavenger recepetor type I/II Clone 2F8, Rat IgG2b Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From liz <@t> premierlab.com Thu May 5 14:56:32 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] F4/80 and alveloar macrophages Message-ID: <000001c551ac$8d535280$a7d48a80@AMY> I needed to chime in on this discussion. I have used serotec's F4/80 antibody a lot and in a wide variety of mouse tissues and different disease states. I have run this antibody on mouse lungs that have been exposed to tuberculosis and from my experiences it does not stain all alveolar macrophages. I have seen variations in staining patterns and intensity in different lesions in one lung and differences from animal to animal. So if you are looking for a marker that will detect all alvelolar macrophages this antibody will not. I consider it more of an activated macrophage marker and expression will vary. The literature states that this particular antibody is expressed at low levels on avelolar macrophages, but expressed in all mouse macrophages, but the expression of the Ag varies greatly depending upon the state of cell activation. We use it mostly in detection macrophages in disease states, since macrophages are effected by the release of growth factors, cytokines etc during the disease process. 30% of normal mouse bone marrow will be positive and it does stain kupffer cells quite well. I'll post an image to the histonet of F4/80 of a mouse lung lesion. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From gcallis <@t> montana.edu Thu May 5 15:13:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Oversize slides for EXAKT system Message-ID: <6.0.0.22.1.20050505135322.01b5ac70@gemini.msu.montana.edu> You wrote: Our lab is now doing a lot of work on resin embedded samples that are too large for standard slides (76x25 mm) so we are using over-size slides. These are 100 x 50 x 1.5 mm. These are great for fitting the samples on BUT the slides don't fit in any racks that we have, or in staining pots. Does anyone out there use these sized slides and know of a supplier of suitable slide boxes, racks and staining jars? I have had a look through some of the main suppliers and couldn't see anything. Any help greatly appreciated. ********************************************************************** Try Arthur Thomas, they have a website, and they have some huge oversized coverslips - we also had some custom made - these were VERY pricey, from Erie. Also, Brain Research Laboratories, www.brainresearchlab.com the latter has more to offer for large slides, storages, folders, probably coverslips. Erie Scientific will make custom slides for people in any size you want, we have done that in the past and I think they will make smaller sized lots now. As for staining racks, containers, we used our staining dishes but carried the slides through with forceps and allowing slides to be slanted byresting slide on a forceps angled in the staining solution. A bit tedious, but it worked. We also used cardboard slide flats, tore out some of the slots to make them bigger. Photo-negative storage drawers also worked for large slides on edge. If you are using plastic slides, you can have them custom made by a company in Illinois to your specifications, slide thickness, etc. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From billk1ibz <@t> yahoo.com Thu May 5 15:24:07 2005 From: billk1ibz <@t> yahoo.com (Bill) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Alveolar macrophages Message-ID: <20050505202407.88951.qmail@web53602.mail.yahoo.com> CD68 clone FA11 labels alveolar macrophages very strongly. Serotec has it- cat. #MCA1957. In my opinion, it is a much better marker of mouse macrophages than F4/80. --------------------------------- Do you Yahoo!? Yahoo! Small Business - Try our new resources site! From Charlene.Henry <@t> STJUDE.ORG Thu May 5 15:27:41 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Microwave Tissue Processing Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C756F7@SJMEMXMB02.stjude.sjcrh.local> I have been ask to check into Microwave Tissue Processing and was wondering what kind of results others are getting with this method of processing. Do you fix with NBF before processing? Do IHC and ISH work on these tissues? What kind do you use? Any information will be appreciated. Vendors: Please let me know what type of microwave tissue processors you have. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 From BMolinari <@t> heart.thi.tmc.edu Thu May 5 16:04:15 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] unsubscribe Message-ID: From lmezo <@t> statlab.com Thu May 5 16:30:07 2005 From: lmezo <@t> statlab.com (Laura Mezo) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Microwave Tissue Processing References: <5CB39BCA5724F349BCB748675C6CA1A202C756F7@SJMEMXMB02.stjude.sjcrh.local> Message-ID: <00d201c551b9$9c785aa0$4203a8c0@statlab.com> Here at StatLab Medical Products we have the Energy Beam Sciences Microwave H2850. If you would like a reference and or information packet you may contat me, Laura M. Mezo at (800) 442-3573 X 289, or lmezo@statlab.com. ----- Original Message ----- From: "Henry, Charlene" To: Sent: Thursday, May 05, 2005 3:27 PM Subject: [Histonet] Microwave Tissue Processing I have been ask to check into Microwave Tissue Processing and was wondering what kind of results others are getting with this method of processing. Do you fix with NBF before processing? Do IHC and ISH work on these tissues? What kind do you use? Any information will be appreciated. Vendors: Please let me know what type of microwave tissue processors you have. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bwhitaker <@t> brownpathology.com Thu May 5 17:35:03 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] billing question Message-ID: <000201c551c2$ae3ac6c0$3601a8c0@brownpathology.net> Hi All, Many of you have a lot more CPT coding/billing experience than I do, and maybe you can help me out. We are in the midst of a debate about billing for determining specimen adequacy on a liver bx, or similar u/s guided needle bx. On at least one occaision the pathologist has done a touch prep to determine if the target lesion had been sampled. How should these be billed? My opinion is as an 88329, even though the procedure is in radiology, not the OR. Everyone does not agree with this. What do you all think? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From jmahoney <@t> alegent.org Thu May 5 17:47:18 2005 From: jmahoney <@t> alegent.org (Janice A Mahoney) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] billing question Message-ID: Actually I think the apropriate CPT code for this is 88172 or 88173 Cytopathology, evaluation of fine needle; immediate cytohistological study to determine adequacy of specimen (88173 for interp and report). These FNA codes can get very confusing. Jan Omaha >>> "Bonnie Whitaker" 05/05/2005 5:35:03 PM >>> Hi All, Many of you have a lot more CPT coding/billing experience than I do, and maybe you can help me out. We are in the midst of a debate about billing for determining specimen adequacy on a liver bx, or similar u/s guided needle bx. On at least one occaision the pathologist has done a touch prep to determine if the target lesion had been sampled. How should these be billed? My opinion is as an 88329, even though the procedure is in radiology, not the OR. Everyone does not agree with this. What do you all think? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tony_Reilly <@t> health.qld.gov.au Thu May 5 20:44:36 2005 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Substitutes for picric acid using crystal violet stains Message-ID: Picric acid is usually purchased under water as it is explosive when dry. A procedure I have used in the past is to get picric acid in water, put some into a flask and give it a few washes with alcohol, allowing it to settle each time and then decanting the excess. Finally add some more alcohol with enough of the picric acid to produce a saturated alcoholoic solution. With picric acid having a solubility of 8g/100ml you now have an 8% stock solution which can be diluted to your working concentration. A precaution I take with the storage of the stock is to use a plastic stopper as the friction of glass on glass could ignite any dried picric acid at the mouth of the storage bottle. I am not aware of the price of alcoholoic picric acid but this may prove cheaper even though some of the picric acid will be lost during the wash process. regards Tony Reilly Chief Scientist Anatomical Pathology Northside Pathology Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au >>> Craig Kallsen 05/06/05 02:16am >>> Dear List, I am new to this and would like to stain phloem, xylem and cambial cells in citrus tree-trunk tissue in paraplast embedded tissue. I have a protocol for using crystal violet stain that requires ethnolic picric acid in the procedure. Is there a substitute for picric acid? I am having trouble obtaining picric acid in alcohol (as well as finding the price intimidating). _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet *********************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. *********************************************************************************** From Jason.PALMER <@t> svhm.org.au Fri May 6 00:49:04 2005 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] re: problems with BD PECAM-1 / CD31 Message-ID: Hi. In response to Jackie O's and Melissa Gonzalez's comments on BD CD31 IHC problems on FFPE tissues, I have been getting very nice staining recently using a protocol developed with tips from Carrie Kyle Byrne, after i made a similar query to histonet some time ago. Mostly use it on our Dako autostainer, but works fine on the bench too. Tissues fixed O/N in 4% PFA (Zn fixation not necessary) processed to paraffin. H2O2 block 5 min, Dako proteinase k 8min RT, Dako protein block 30 min, primary 1:100 in Dako diluent 60 mins RT, secondary Dako rabbit anti rat biotin 1:300 30 min RT, vector ABC elite 30 min RT then DAB 5 min. Gives consistently good vascular staining for me, in connective and adipose rich mouse tissues. I think the ABC elite is the key. And i'm going to give the Santa Cruz PECAM Ab that has been talked about recently a try too. Has been sitting in our fridge for a while as a second option, and though don't really need it at the moment, thought i'd take it out for a spin to see how it performs... Hope this helps, Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 email: palmerj@unimelb.edu.au -------------- next part -------------- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From Kemlo.Rogerson <@t> elht.nhs.uk Fri May 6 02:15:10 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] really basic question Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F27B@bhrv-nt-11.bhrv.nwest.nhs.uk> Spleen? Liver? -----Original Message----- From: Christian Franci [mailto:cfranci@rigel.com] Sent: 05 May 2005 18:49 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] really basic question I'm doing staining of mouse tissues and need to get vascular tissue from different areas. What places/organs are the best bets? Kidneys come to mind but, where else? Thanks a bunch, Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mplhisto <@t> aol.com Fri May 6 06:02:23 2005 From: mplhisto <@t> aol.com (mplhisto@aol.com) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Dallas Ft. Worth Texas Wages Information In-Reply-To: <00d201c551b9$9c785aa0$4203a8c0@statlab.com> References: <5CB39BCA5724F349BCB748675C6CA1A202C756F7@SJMEMXMB02.stjude.sjcrh.local> <00d201c551b9$9c785aa0$4203a8c0@statlab.com> Message-ID: <8C72042375E1BEE-74C-2687@FWM-R39.sysops.aol.com> Can anyone tell me if a current wage survey for the Dallas/FT. Worth area exsists or if anyone is willing to share Salaries? We are currently looking to make adjustments to our technicians salaries and any information would be greatly appreciated. We would like both hospital and private lab figures. Thanks in advance!!!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KPercival <@t> wyeth.com Fri May 6 06:50:46 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] CYTOKINES FOR MOUSE TISSUE Message-ID: Hi Everyone, I am new to the Histonet, but looking over previous communications, you all have a lot of information to share. Anyway, I am trying to optimize TNFa, IL6, and IL1a on mouse tissue. Nothing I have tried works so far. Any suggestions? Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com From KPercival <@t> wyeth.com Fri May 6 07:17:47 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Hi again, Message-ID: Hi again, I think I forgot to mention that the mouse tissue for TNF, IL6 and IL1 is FFPE. we may have to resort to frozen tissue, so protocols for that would be helpful, too. Thanks! Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com From cfockler <@t> mail1.vcu.edu Fri May 6 07:25:08 2005 From: cfockler <@t> mail1.vcu.edu (cfockler@mail1.vcu.edu) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Dallas Ft. Worth Texas Wages Information In-Reply-To: <8C72042375E1BEE-74C-2687@FWM-R39.sysops.aol.com> Message-ID: <200505061225.IAA04543@arrakis.vcu.edu> We are trying to do the same for the southeast region (VA, NC, SC, TN, GA). So if there is any information to be given on these regions we'd greatly appreciate the info as well. Thanks in advance Candyce Fockler Histotechnologist VCUHS ------------------- > > > > > Can anyone tell me if a current wage survey for the Dallas/FT. Worth area exsists or if anyone is willing to share Salaries? We are currently looking to make adjustments to our technicians salaries and any information would be greatly appreciated. We would like both hospital and private lab figures. Thanks in advance!!!! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From LINDA.MARGRAF <@t> childrens.com Fri May 6 08:27:55 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] billing question Message-ID: Our practice would be to charge the intraoperative consultation 88329 and a touch preparation 88161. As this is a biopsy and not an aspirate, I don't think that the fine needle aspirate charges would be appropriate. Linda Margraf MD Histonet administrator Director of Anatomic Pathology Children's Medical Center of Dallas >>> "Janice A Mahoney" 5/5/2005 5:47:18 PM >>> Actually I think the apropriate CPT code for this is 88172 or 88173 Cytopathology, evaluation of fine needle; immediate cytohistological study to determine adequacy of specimen (88173 for interp and report). These FNA codes can get very confusing. Jan Omaha >>> "Bonnie Whitaker" 05/05/2005 5:35:03 PM >>> Hi All, Many of you have a lot more CPT coding/billing experience than I do, and maybe you can help me out. We are in the midst of a debate about billing for determining specimen adequacy on a liver bx, or similar u/s guided needle bx. On at least one occaision the pathologist has done a touch prep to determine if the target lesion had been sampled. How should these be billed? My opinion is as an 88329, even though the procedure is in radiology, not the OR. Everyone does not agree with this. What do you all think? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Fri May 6 09:09:32 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Hi again, In-Reply-To: Message-ID: <001501c55245$3ce99810$a7d48a80@AMY> Karen Abcam has an IL-6 antibody that works in mouse tissue, it's a rabbit anti-human that works in rat and mouse with proteinase K digestion on FFPE tissue. It even works on formic acid decalcifed tissue. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Karen Percival Sent: Friday, May 06, 2005 5:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hi again, Hi again, I think I forgot to mention that the mouse tissue for TNF, IL6 and IL1 is FFPE. we may have to resort to frozen tissue, so protocols for that would be helpful, too. Thanks! Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cfranci <@t> rigel.com Fri May 6 10:42:09 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] lots of responses to my basic question Message-ID: <84a37fa4ccb588f519232af83b0e719a@rigel.com> Thanks everybody. cheers Chris From dholmes <@t> anatomy.umsmed.edu Fri May 6 10:51:02 2005 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] tracer & diluent dilemma Message-ID: What diluent and at what pH should be used to prepare Floro-Ruby? Has anyone had problems with it not going into solution? From Dixon.Leslie <@t> mayo.edu Fri May 6 11:05:00 2005 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] TRAP on bones Message-ID: <115ED45FBA6D3D4B89E7F40A9EB5339D0361C303@excsrv60.mayo.edu> Good morning all, I have been trying to stain formic acid/EDTA decalcified formalin fixed mouse femurs for osteoclasts with the TRAP stain. I have had little success. Does anyone have a protocol that I might be able to try? Any suggestions would be greatly appreciated. I have been using the Sigma kit #387A thus far. Thanks, Leslie Dixon Mayo Clinic Scottsdale Research Histology From pex0220 <@t> yahoo.com.cn Fri May 6 11:32:49 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Background! Message-ID: <20050506163249.84259.qmail@web15502.mail.cnb.yahoo.com> Hello, all, I am doing double Immunofluorescence in bone sections. But I find there is so much backgound in sections, I have used the normal serum matched to the host of secondary antibodies for blocking, moreover I have also add times for washing, so I do not really know how to reduce the background. Can anybody do me a favor?Thank you! Best wishes! Guofeng Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From gcallis <@t> montana.edu Fri May 6 12:07:46 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] TRAP on bones In-Reply-To: <115ED45FBA6D3D4B89E7F40A9EB5339D0361C303@excsrv60.mayo.edu > References: <115ED45FBA6D3D4B89E7F40A9EB5339D0361C303@excsrv60.mayo.edu> Message-ID: <6.0.0.22.1.20050506110342.01b3ce48@gemini.msu.montana.edu> EDTA decalcification is preferred decalcifier. Formic acid is more than likely killing the enzyme, rendering it unstainable. I have a method from Hermina Borgerink, if you want it. At 10:05 AM 5/6/2005, you wrote: >Good morning all, > >I have been trying to stain formic acid/EDTA decalcified formalin fixed >mouse femurs for osteoclasts with the TRAP stain. I have had little >success. Does anyone have a protocol that I might be able to try? Any >suggestions would be greatly appreciated. I have been using the Sigma kit >#387A thus far. > >Thanks, >Leslie Dixon >Mayo Clinic Scottsdale Research Histology > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From NSEARCY <@t> swmail.sw.org Fri May 6 12:48:20 2005 From: NSEARCY <@t> swmail.sw.org (Nita Searcy) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] IHC Billing CPT question. Message-ID: This is definitely a Beth Sheppard question for final clarification but billing is done per specimen- so the IHC x 6 is correct. Only one specimen would be x 3- this scenario has 2 specimens. >>> Shawn Salesky 05/05/05 12:38 PM >>> Hi All, >From the discussion a few days ago I had some questions concerning IHC Billing. Patient: B. Smith Has a Left and Right Breast Core Bx submitted as parts A and B. Both of which have ER, PR and Her2 done on them. The Her2 is negative so there is no reflex to F.I.S.H. >From the discussion on Histo net I gather that the majority of facilities would be charging for Quantitative/Semi-quantitative IHC x 6. Is there any literature or references that anyone is using to determine that it is legal to charge for 6 instances? At my current facility that same scenario would only be billed as 3 instances. Any help would be appreciated. This e-mail and any attachments is intended only for the person or entity to which it is addressed and may contain confidential and privileged information. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this e-mail and destroy any copies. Any dissemination or use of this information by a person other than the intended recipient is unauthorized and may be illegal. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri May 6 12:59:14 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:02 2005 Subject: [Histonet] Bone, fluorescent background, TRAP staining - triple info In-Reply-To: <20050506163249.84259.qmail@web15502.mail.cnb.yahoo.com> References: <20050506163249.84259.qmail@web15502.mail.cnb.yahoo.com> Message-ID: <6.0.0.22.1.20050506111431.01b65668@gemini.msu.montana.edu> You need to access this article in J Histochemistry Cytochemistry titled Histological Analysis of GFP expression in murine bone, JHC 53(5):593-602,2005. Although GFP is used, they dealt with autofluorescence very nicely using dual band pass filters on their microscope. This is an elegant paper, one of the finest I've seen on fluorescence with GFP along with other staining methods. Autofluorescence is a problem for those working with either double immunofluorescent staining or with GFP work. If you use formalin or paraformaldehyde fixation, you NEVER get rid of autofluorescence, these fixatives usually increase autofluorescence levels. For this very reason our work on bone or tissues attached to bone are done on undecalcified bone frozen sections, using the Cryojane tape transfer device. Washing in general, does not decrease autofluorescence. I read your previous message and reply from Chris van der Loos. You need to use two different species hosted secondaries as one way to reduce nonspecific binding and background fluorescence to avoid nonspecific binding background fluorescence, this is NOT autofluorescence - that means it is in the tissue before you ever start - and is the result of tissue components or induced by formalin or paraformaldehyde fixatives. Both are problems, with autofluorescence either not easy to remove or impossible!! Nonspecific binding by antibodies to tissues or other proteins introduced in your staining method is another story including dilutions, blocking, choice of secondaries, etc and is avoidable by how you chose to set up your double IFA staining. With our double fluorescent staining we have done the following, as examples: 1. Goat antiGFP 2. Donkey antiGoat-FITC 3. Armenian hamster anti dendritic cell-biotinylated 4. Strepavidin-Alexa 555 Or one could do two rat antimouse primaries: 1. Normal serum block is cocktail of goat and donkey serums, 2. Rat antiMouse CD marker 3. Donkey antiRat-FITC 4. Rat serum block to ensure the next secondary does NOT detect the rat again 5. Rat antiMouse 6. Goat antiRat-RRX (rhodamine) Please remember we do NOT use any aldehyde fixed bone samples. Not sure of what you are doing exactly in your staining method, you only talked about secondaries. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From wprichett <@t> ohri.ca Fri May 6 13:41:07 2005 From: wprichett <@t> ohri.ca (Prichett, Wendy) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Growing PC12 cells onto coverslips, their differentiation and subsequent immunohistochemistry Message-ID: <0133EAC2EDC7D74D944267D639E49EF246771F@ohexch04.civic1.ottawahospital.on.ca> Hi, I have mainly been doing immunohistocchemistry on tissues for years but our lab is branching out to manipulate cell cultures. We want to start immunohistochemistry on the PC12 cell line. I have done tissue culture in the past however I need some advice on these cells. We could like to seed them onto glass coverslips if possible and then get them to differentiate into neurons with NGF. I have read some literature on this but need some advice along with clarification. If anyone can give me their protocols for seeding and differentiation on coverslips (either matrix coated or using poly-L-lysine? Not sure which to use?) it would be much appreciated. These neurons are going to be stained with immunofluorescence with a regular fixing, permeabilization, and staining protocol, which I have already optimized. Thanks in advance for any help. Wendy Pejic Research Technician OHRI, Loeb Building Ottawa, Ontario From pruegg <@t> ihctech.net Fri May 6 14:10:38 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] TRAP on bones In-Reply-To: <6.0.0.22.1.20050506110342.01b3ce48@gemini.msu.montana.edu> Message-ID: <200505061910.j46JAYBO000171@chip.viawest.net> Decalcification with formic acid and formalin fixation will destroy the TRAP enzyme. I always use undecalcified bone fixed in cold methanol and embedded in GMA to demostrate osteoclasts. Cold Methanol will also best preserve fluorochrome labels which may be soluable in aqueous fixatives. Leslie, I do contract GMA work. You can visit my website at www.ihctech.net Patsy Ruegg -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Friday, May 06, 2005 10:08 AM To: Dixon, Leslie E.; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] TRAP on bones EDTA decalcification is preferred decalcifier. Formic acid is more than likely killing the enzyme, rendering it unstainable. I have a method from Hermina Borgerink, if you want it. At 10:05 AM 5/6/2005, you wrote: >Good morning all, > >I have been trying to stain formic acid/EDTA decalcified formalin fixed >mouse femurs for osteoclasts with the TRAP stain. I have had little >success. Does anyone have a protocol that I might be able to try? Any >suggestions would be greatly appreciated. I have been using the Sigma >kit #387A thus far. > >Thanks, >Leslie Dixon >Mayo Clinic Scottsdale Research Histology > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri May 6 14:17:23 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] re: problems with BD PECAM-1 / CD31 In-Reply-To: Message-ID: <200505061917.j46JHKBO002885@chip.viawest.net> CD 31 in my hands requires enhanced detection systems as Jason uses with the ABC elite I have also gotten good results with the DAKO CSA II Tyramide detection system, not much luck with non-enhanced systems such as LSAB or even Labelled Polymer. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of PALMER Jason (SVHM) Sent: Thursday, May 05, 2005 10:49 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re: problems with BD PECAM-1 / CD31 Hi. In response to Jackie O's and Melissa Gonzalez's comments on BD CD31 IHC problems on FFPE tissues, I have been getting very nice staining recently using a protocol developed with tips from Carrie Kyle Byrne, after i made a similar query to histonet some time ago. Mostly use it on our Dako autostainer, but works fine on the bench too. Tissues fixed O/N in 4% PFA (Zn fixation not necessary) processed to paraffin. H2O2 block 5 min, Dako proteinase k 8min RT, Dako protein block 30 min, primary 1:100 in Dako diluent 60 mins RT, secondary Dako rabbit anti rat biotin 1:300 30 min RT, vector ABC elite 30 min RT then DAB 5 min. Gives consistently good vascular staining for me, in connective and adipose rich mouse tissues. I think the ABC elite is the key. And i'm going to give the Santa Cruz PECAM Ab that has been talked about recently a try too. Has been sitting in our fridge for a while as a second option, and though don't really need it at the moment, thought i'd take it out for a spin to see how it performs... Hope this helps, Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 email: palmerj@unimelb.edu.au From gcallis <@t> montana.edu Fri May 6 14:32:48 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Growing PC12 cells onto coverslips, their differentiation and subsequent immunohistochemistry In-Reply-To: <0133EAC2EDC7D74D944267D639E49EF246771F@ohexch04.civic1.ott awahospital.on.ca> References: <0133EAC2EDC7D74D944267D639E49EF246771F@ohexch04.civic1.ottawahospital.on.ca> Message-ID: <6.0.0.22.1.20050506132244.01b45158@gemini.msu.montana.edu> Look into buying chamber slides from NUNC designed for this use. They are mini culture chambers and you do not have to manipulate coverslips, sometimes a tedious, delicate proposition. They come in all sizes, large or multichambered. After doing cell cultures, you can do all l the staining right on the slide from fixation to final product. When finished, you mount coverslip and view. Are you using an inverted microscope by any chance to see fluorescent staining? If so there are petri dishes designed for confocal use with inverted where the bottom of dish IS a coverslip. They come sterile and in different coverslip thicknesses and sizes, and are made of glass. Very tidy little items as you can coat them or whatever, grow cells, fix, stain and do inverted fluorescent scope or confocal laster scanning with inverted setup. At 12:41 PM 5/6/2005, you wrote: >Hi, > >I have mainly been doing immunohistocchemistry on tissues for years but >our lab is branching out to manipulate cell cultures. We want to start >immunohistochemistry on the PC12 cell line. I have done tissue culture >in the past however I need some advice on these cells. We could like to >seed them onto glass coverslips if possible and then get them to >differentiate into neurons with NGF. I have read some literature on >this but need some advice along with clarification. If anyone can give >me their protocols for seeding and differentiation on coverslips (either >matrix coated or using poly-L-lysine? Not sure which to use?) it would >be much appreciated. These neurons are going to be stained with >immunofluorescence with a regular fixing, permeabilization, and staining >protocol, which I have already optimized. > >Thanks in advance for any help. > >Wendy Pejic >Research Technician >OHRI, Loeb Building >Ottawa, Ontario > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From cfranci <@t> rigel.com Fri May 6 15:37:13 2005 From: cfranci <@t> rigel.com (Christian Franci) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] follow-up questions... Message-ID: <7cd9b7602729d3dde42b16d865a67193@rigel.com> Since I'll be doing IHC on paraffin sections from these highly-vascular mouse organs, any issues that might arise? (for example, will RBC's be a problem?) Any tricks to dealing with vascular tissues? BTW, my primary Ab's are rat and rabbit. Again, thanks for all your help. C From DRG <@t> Stowers-Institute.org Fri May 6 15:49:55 2005 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] unsubscribe Message-ID: <1096710316-115989344@pathology.swmed.edu> Debby Grant Histology Specialist I Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From linhines <@t> yahoo.com Fri May 6 17:30:16 2005 From: linhines <@t> yahoo.com (Linda Hines) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] histologist to fill in Message-ID: <20050506223021.87346.qmail@web31315.mail.mud.yahoo.com> Hi! Everyone, I am looking for a histotech to fill in on May 20, 2005 in the afternoon appx. 4:00pm in the afternoon and work until appx. 9:00 pm. We are located central Phoenix, AZ if interested please contact Linda Hines @ 602-241-5134. Thank-you Linda Hines HT Specialty Diagnostics --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From histo20 <@t> hotmail.com Fri May 6 19:48:59 2005 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] (no subject) Message-ID: Hello everyone! My department will inherit immunohistochemistry staining from another lab in the very near future. I was just wondering if anyone knew if performing these procedures required a registered tech, and if so, is this considered high, medium or low complexity testing? Thanks so much! Paula Wilder St. Joseph Medical Center Towson, MD 21204 From CCollins <@t> propathlab.com Sat May 7 09:31:19 2005 From: CCollins <@t> propathlab.com (Cory Collins) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] IHC position in Dallas Message-ID: > We have an opening in the IHC Lab at ProPath in Dallas. ProPath is a > reference lab with it's own IHC department. I'm looking for a histotech > that is interested in learning new, cutting edge IHC techniques. We do a > high volume of IHC antibodies (to see a copy of our requistion listing > which antibodies we offer click here: > http://www.propathlab.com/pdf/requisition_immuno.pdf) as well as UroVysion, > PathVysion, InSitu hybridization for HPV and EBER, IF, and we are looking > into doing CISH and some other molecular probes both brightfield and > fluorescent. We are a 24 hour 5 days a week lab with four techs and a > clerical aide. The positon we are looking for would be from 8AM-4:30PM. > We are located at I-35 and Mockingbird. For more information about the > position or applying, please contact Angie Viancourt in our HR department > at aviancourt@propathlab.com or e-mail me at ccollins@propathlab.com if you > have any questions about the lab. > > Thanks, > > Cory > > > > Cory Collins, HT (ASCP) QIHC > Immunohistochemistry Supervisor > ProPath > 8267 Elmbrook Drive Suite 100 > Dallas, TX 75247 > 214-638-2000 ext 2027 > 214-237-1730 Fax > To learn more about ProPath, please visit http://www.ProPathLab.com > > ______________________________________________________________________________ This e-mail may contain confidential or privileged information. If you think you have received this e-mail in error, please advise the sender by reply e-mail and then delete this e-mail immediately. From pruegg <@t> ihctech.net Sun May 8 10:42:06 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] antigen retrieval/enzyme digestion In-Reply-To: Message-ID: <200505081542.j48FgR75001351@pro12.abac.com> I have heard of it being done both ways depending on the epitope of interest. In general I do HIER first and then EIER. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Till, Renee Sent: Monday, May 02, 2005 5:55 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] antigen retrieval/enzyme digestion When trying both on the same slides, which would go first, the antigen retrieval or the enzyme digestion? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Mon May 9 02:43:52 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] RE: Vector Red and True Blue Message-ID: <774aad776fb4.776fb4774aad@amc.uva.nl> Hi Ben, It's good to read that you're at least successful with the application of True Blue since this isn't the most easy chromogen to deal with. Furthermore, your choice for this turquoise-red color combination is a very good one, because of it's very high contrast. Vector Red however, isn't that strong in my hands either. You may try to look with a fluorescence microscope as the Vector Red reaction product shows a very strong (red) fluorescence upon green light excitation (rhodamine-filter pack). Furthermore, I have very good results for both light (and fluorescence microscopy) using the new Dako Permanent Red kit (K0640). The staining intensity is much higher than with Vector Red. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Ben" Date Wed, 4 May 2005 11:57:08 -0700 To Subject [Histonet] Vector Red and True Blue Hello All, I have been attempting some double labeling for fos and estrogen receptor for about a month with little luck. I am using Vector Red (fos) and True Blue (ER) for my staining. I am having no problems with the ER labeling, but the fos labeling is not that clear. I don't appear to be getting a strong reaction product from the Vector Red. Has anyone else had this problem? How strong should the staining be (light pink, dark pink, red)? I have been careful to only place in xylene for a few moments before coverslipping and therefore don't think fading is my problem. I have been thinking that my antigen retrieval may be the problem. I am using triton X to improve antigen retrieval, but was thinking about microwaving. These are free floating sections. First is this possible to do with free floati! ng sectio References 1. mailto:c.m.vanderloos@amc.uva.nl From pex0220 <@t> yahoo.com.cn Mon May 9 04:26:47 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] the skeletal staining Message-ID: <20050509092647.69523.qmail@web15510.mail.cnb.yahoo.com> Hello, all, Recently I do Alizarin Red and Alcian Blue staining of mouse embryo skeletons, but I find that my results are not better due to a wrong color. After staining, bone should be red, and cartilage should be blue, but my results are that bone is stained purple, and cartilage is stained green, then I change my protocol, but I get the similar results, I do not know the reasons, I guess that bioreagents could not be good, because I make them up, the colors of the solutions are purple and green respectively, not red and blue.Our bioreagents are from sigma. If you have some experience about it, can you do me a favor? Best wishes! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From manuelle.debock <@t> UGent.be Mon May 9 07:22:59 2005 From: manuelle.debock <@t> UGent.be (Manuelle De Bock) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] methasol fast blue Message-ID: <000601c55491$d6e7e540$ed54bec1@ugent.be> I am trying to stain collected gastric tissue cells with Maxwell's staining procedure (1963) but we don't have methasol fast blue in our lab. I also can't find any links on the web where I can buy this dye. can someone help me with this? Is it the same dye as Luxol fast blue? Kind regards Manuelle Manuelle De Bock, dierenarts (DVM) Laboratory of Veterinary Pathology Department of Pathology, Bacteriology and Avian Diseases Faculty of Veterinary Medicine Ghent University Salisburylaan 133 B9820 Merelbeke - Belgium Tel 0032(0)9 264 7745 Fax 0032(0)9 264 7789 From t-sherman <@t> comcast.net Mon May 9 08:58:31 2005 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] RE: antigen retrieval/enzyme digestion Message-ID: On Sun, 08 May 2005 12:00:26 -0500, wrote: > Today's Topics: > > 1. RE: antigen retrieval/enzyme digestion (Patsy Ruegg) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sun, 8 May 2005 09:42:06 -0600 > From: "Patsy Ruegg" > Subject: RE: [Histonet] antigen retrieval/enzyme digestion > To: "'Till, Renee'" , > > Message-ID: <200505081542.j48FgR75001351@pro12.abac.com> > Content-Type: text/plain; charset="us-ascii" > > I have heard of it being done both ways depending on the epitope of > interest. In general I do HIER first and then EIER. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Till, > Renee > Sent: Monday, May 02, 2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] antigen retrieval/enzyme digestion > > When trying both on the same slides, which would go first, the antigen > retrieval or the enzyme digestion? > > > Renee' Till, HT > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > (501) 364-2774 Renee, I'd tend to agree with Patsy as far as a sequence best determined empirically. You'll probably need to compare each protocol for each antigen of interest. From a theoretical perspective, both techniques of "unmasking" involve the breaking of strong molecular bonds. HIER is typically designed to cleave the basic side-chains of amino acids on proteins previously fixed (or bound to each other) with x-aldehyde. Enzyme digestion is similarly designed to cleave proteins but at well defined amino acids which are not necessarily basic - it depends on the digesting enzyme and its substrate. My guess would be that progressing from a more basic (read simple/generic) to a more specific cleaving sequence would be more efficient; consequently, treat with HIER first and enzyme digestion second. The HIER would open up the tissue to further access and the enzyme digestion could then more specifically cleave regions hindering an even more localized access to your target antigen. To a great extent, these success derived from these sequences will be greatly influenced by the local milieu of your target antigen. The relative content of various amino acids in a given milieu may or may not be known. To that end empirical observations are "best" which is why mandating one particular protocol over another may be a bit dubious. Good luck. Todd President HistoSoft Corporation -- >>>>>>>> www.histosoft.com <<<<<<<< <<<<<<<< Biology In A New Form (c) >>>>>>> From DELONG_CYNTHIA_A <@t> LILLY.COM Mon May 9 09:13:04 2005 From: DELONG_CYNTHIA_A <@t> LILLY.COM (Cynthia A Delong) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Book- Immunoenzyme Multiple Staining Methods Message-ID: Hello All, I am looking for Chris Van de Loos's book"Immunoenzyme Multiple Staining Methods". Does anyone know of where I can get a copy of this book? I had one and someone walked off with it. Thank you Cindy Cynthia A DeLong LRL - Corporate Center DC0533 - 48B/3042 355 E Merrill St Indianapolis, IN 46225 phone: 317-276-7635 fax: 317-277-6146 From Tbarnhart <@t> primecare.org Mon May 9 09:25:54 2005 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: <1779904B5E82D511914C00D0B793339205BFD930@exchangent> This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From juan.gutierrez <@t> christushealth.org Mon May 9 09:35:39 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: We are located one floor above the surgical suites, but we do maintain a frozen/gross room in the surgical suite area. At the end of the day the gross tech brings the specimens upstairs to be loaded in the processors. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barnhart, Tammy Sent: Monday, May 09, 2005 9:26 AM To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon May 9 09:41:10 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Book- Immunoenzyme Multiple Staining Methods In-Reply-To: References: Message-ID: <6.0.0.22.1.20050509083836.01b613a8@gemini.msu.montana.edu> I replaced my copy from Amazon.com. The book is now 8.99/copy, and the book is no longer in print, so don't hesitate too long to replace. I was so worried this would happen to me i.e. walking off, I now keep a personal copy at home, my first one is worn out from use! At 08:13 AM 5/9/2005, you wrote: >Hello All, >I am looking for Chris Van de Loos's book"Immunoenzyme Multiple Staining >Methods". Does anyone know of where I can get a copy of this book? >I had one and someone walked off with it. >Thank you >Cindy > >Cynthia A DeLong >LRL - Corporate Center >DC0533 - 48B/3042 >355 E Merrill St >Indianapolis, IN 46225 >phone: 317-276-7635 >fax: 317-277-6146 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pmarcum <@t> vet.upenn.edu Mon May 9 09:46:49 2005 From: pmarcum <@t> vet.upenn.edu (pmarcum@vet.upenn.edu) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Book- Immunoenzyme Multiple Staining Methods In-Reply-To: <6.0.0.22.1.20050509083836.01b613a8@gemini.msu.montana.edu> References: <6.0.0.22.1.20050509083836.01b613a8@gemini.msu.montana.edu> Message-ID: <1115650008.427f77d900571@imp.vet.upenn.edu> Hi Gayle and All, I just purchased one from Amazon and only two were still listed at $8.95 p;us shipping and handling it was about $12.50 for the copy. Thanks, Pam Marcum Quoting Gayle Callis : > I replaced my copy from Amazon.com. The book is now 8.99/copy, and the > book is no longer in print, so don't hesitate too long to replace. I was > so worried this would happen to me i.e. walking off, I now keep a personal > copy at home, my first one is worn out from use! > > At 08:13 AM 5/9/2005, you wrote: > >Hello All, > >I am looking for Chris Van de Loos's book"Immunoenzyme Multiple Staining > >Methods". Does anyone know of where I can get a copy of this book? > >I had one and someone walked off with it. > >Thank you > >Cindy > > > >Cynthia A DeLong > >LRL - Corporate Center > >DC0533 - 48B/3042 > >355 E Merrill St > >Indianapolis, IN 46225 > >phone: 317-276-7635 > >fax: 317-277-6146 > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From la.sebree <@t> hosp.wisc.edu Mon May 9 09:49:46 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: Several years ago our surgical pathology, IHC/ISH, and renal labs were moved to rented space in a VA hospital that's physically connected to our University hospital. The grossing and frozen section lab remained in space adjacent to the ORs. The cassetted specimens are brought over from there every day on carts. It seems to work okay. If the surgeons want to talk with a pathologist the next day about a specimen they have to walk over to the VA to do so. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barnhart, Tammy Sent: Monday, May 09, 2005 9:26 AM To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon May 9 09:59:15 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Autoflourescence Eliminator Reagent for lipofuscin in CNS, other cells Message-ID: <6.0.0.22.1.20050509084844.01b22e70@gemini.msu.montana.edu> Dear all For those looking for a way to eliminate or reduce lipofuscin autofluorescence in CNS tissues and other cells, there is a commercial product available. Science magazine just listed this new reagent, and it may be worth a try. Chemicon International at www.chemicon.com is the vendor. They mentioned monkey, human and rat neural and other tissues as well. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From rebecca.riesen <@t> dsilabs.com Mon May 9 10:11:13 2005 From: rebecca.riesen <@t> dsilabs.com (Riesen, Rebecca) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Phoenix fill in Message-ID: <05844092236E7749A1BBBCB1077C34A2DE9FDD@dsi-ex01.gateway.dom> PICK ME! PICK ME! Fly me to Phoenix for the weekend! I'm in Florida. Message: 10 Date: Fri, 6 May 2005 15:30:16 -0700 (PDT) From: Linda Hines Subject: [Histonet] histologist to fill in To: Histonet@lists.utsouthwestern.edu Message-ID: <20050506223021.87346.qmail@web31315.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi! Everyone, I am looking for a histotech to fill in on May 20, 2005 in the afternoon appx. 4:00pm in the afternoon and work until appx. 9:00 pm. We are located central Phoenix, AZ if interested please contact Linda Hines @ 602-241-5134. Thank-you Linda Hines HT Specialty Diagnostics --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour ------------------------------ DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From Kemlo.Rogerson <@t> elht.nhs.uk Mon May 9 10:12:05 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F28D@bhrv-nt-11.bhrv.nwest.nhs.uk> Very interesting question and very pertinent for Laboratory Modernisation in the UK. Traditionally it has always been supposed that Histology Labs must be on Site for the very reasons you suggest. My experience in London and now in the North leads me to question that. I accept that frozen sections may need a local presence, but they are relatively rare nowadays, but a motorbike Courier service may be quicker than a porter! I think a happy intermediary is for there to be a Core Lab, that deals with the bulk of the processing and staining and this will reduce the duplication of tests and increase the critical number of Staff; you should see costs driven down. ICC and such like can be consolidated on one Site again with cost savings. A satellite laboratory may be needed to deal with frozen sections and urgent rapid H&E's, but I wonder if that necessity, once the Core Lab became organised may 'wither on the vine'. The bigger problems are MDT's and alienation of Pathologists from their Clinical colleagues; Telepathology and the use of video cameras are a solution, but not a complete one; the spontaneous ways we humans do business makes that an unacceptable means of communicating. In order to release resources consolidation must occur and if used wisely may result in extension of roles with the positive impact on recruitment and retention of Staff. Do you get Black Pudding in America? -----Original Message----- From: Barnhart, Tammy [mailto:Tbarnhart@primecare.org] Sent: 09 May 2005 15:26 To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Mon May 9 10:19:03 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] microscope companies Message-ID: <18444780.1115651943878.JavaMail.root@web4.mail.adelphia.net> Fellow tech's, Does anyone know of a company in Florida that services microscopes? I have already contacted JL Optical and Southern Microscope. Thanks, Ron Martin From John.Sheppard <@t> Health-Partners.org Mon May 9 10:22:49 2005 From: John.Sheppard <@t> Health-Partners.org (John.Sheppard@Health-Partners.org) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] RE: Hospital based Histology Labs Message-ID: We have the surgical suites located one floor above us. The O.R. sends a runner down with the frozens. We keep the cryostats here in histology. We do get a copy of the surgury schedule, so we have an idea when about half the frozens are comming. It's not perfect, but it works, as long as the runner hand delivers the frozen to someone and does not just leave it on the counter. John Sheppard HT(ASCP) The Community Hospital Springfield, Ohio CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From juan.gutierrez <@t> christushealth.org Mon May 9 10:31:14 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: On a previous life in the alien visited town of Roswell, NM, we had an offsite lab. The frozen sections were handled by the pathologists at the hospital, but the rest of the specimens were picked up by a courier twice a day and brought to the lab for gross. The courier will also ensure that there was enough materials at the frozen room daily. And the doctors were always kind enough to let us know when their mess was too much for us to go and clean it up. Ever try to clean up a weeks worth of shavings in a cryostat? Needles to say we stop waiting for word and visited the hospital often to keep the room nice and tidy;0) Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GUTIERREZ, JUAN Sent: Monday, May 09, 2005 9:36 AM To: Barnhart, Tammy; Histonet (E-mail) Subject: RE: [Histonet] Hospital based Histology Labs We are located one floor above the surgical suites, but we do maintain a frozen/gross room in the surgical suite area. At the end of the day the gross tech brings the specimens upstairs to be loaded in the processors. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barnhart, Tammy Sent: Monday, May 09, 2005 9:26 AM To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RCHIOVETTI <@t> aol.com Mon May 9 10:31:13 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] microscope companies Message-ID: <86.27bbaf54.2fb0dc41@aol.com> In a message dated 5/9/2005 8:21:22 AM US Mountain Standard Time, pathrm35@adelphia.net writes: > Does anyone know of a company in Florida that services microscopes? I have > already contacted JL Optical and Southern Microscope. > Ron, Try Micro Optics of Florida, 3941 S.W. 47th Ave., Davie, FL, 954-791-0082, < www.microopticsfl.com>. They are Leica dealers, but I believe their service shop also maintains and repairs other brands of scopes. Cheers, Bob Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From DELONG_CYNTHIA_A <@t> LILLY.COM Mon May 9 10:39:27 2005 From: DELONG_CYNTHIA_A <@t> LILLY.COM (Cynthia A Delong) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Thanks Book ordered Message-ID: Thanks to all who replied to my request. The book is ordered. I guess I will have to keep this one under lock and key. Cindy Cynthia A DeLong LRL - Corporate Center DC0533 - 48B/3042 355 E Merrill St Indianapolis, IN 46225 phone: 317-276-7635 fax: 317-277-6146 From gu.lang <@t> gmx.at Mon May 9 11:40:15 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:03 2005 Subject: AW: [Histonet] Hospital based Histology Labs In-Reply-To: <1779904B5E82D511914C00D0B793339205BFD930@exchangent> Message-ID: Our lab is on the hospital area, but in another building as the surgery area. We are connected with the surgery room with a pneumatic delivery, that shots the tissue to us. That takes less than 30 sec. We do the frozens in an integrated part of our lab. The pathologists give the results through an intercommunication system to the surgeon. Gudrun Lang Linz, Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Barnhart, Tammy Gesendet: Montag, 09. Mai 2005 16:26 An: Histonet (E-mail) Betreff: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Mon May 9 12:30:51 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] histology lab within hospital Message-ID: <001701c554bc$d8e49110$1d2a14ac@wchsys.org> Our hospital system has two labs, one at the hospital and one about a mile away. The hospital based lab handles all the stat work. The hospital histology side handles grossing the hospital cases and the frozen sections. The other lab has all departments but blood bank. The histology department handles all processing, cutting and staining of all tissue received from the hospital, the surgical center and the outlying doctor offices. We have a several couriers that run to all locations to pick up all specimens for every laboratory department. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From dholmes <@t> anatomy.umsmed.edu Mon May 9 13:26:41 2005 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] tracer & diluent dilemma Message-ID: What diluent and at what pH should be used to prepare Floro-Ruby? Has anyone had problems with it not going into solution? From JEllin <@t> yumaregional.org Mon May 9 14:00:40 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Slide Dryers Message-ID: Hello there everyone thought that I would put another amber on the fire, I am in the need of a slide dryer. And not any slide dryer one that people are using for IHC. Amy info would greatly be appreciated. Jesus Ellin Yuma Regional Medical Center From SJones <@t> cvm.tamu.edu Mon May 9 15:45:19 2005 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Florida License Message-ID: I am having trouble finding the histology category on the Florida Dept. of Health web site. Are histologist still being licensed under Clinical Laboratory Personnel? Any help would be appreciated. Thanks, Sarah Sarah Jones, HT(ASCP) Histology Lab Dept. of Veterinary Integrative Biosciences College of Veterinary Medicine Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 From dcrippen <@t> buckinstitute.org Mon May 9 19:04:04 2005 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] 1uM thick Drosophila sections Message-ID: Dear Experts in HistoLand, I need to make 1uM thick sections of full grown Drosophila for Toluidine Blue staining and have some questions about fixation and processing. 1.) What is the best way to get the fixative into the fly? I've read several techniques ranging from careful incision into abdomen to cutting off the head? Secondarily--is the best tool for this a microscalpel, or can other instruments be employed? 2.) What is the best fixative? We're not doing EM, just need thick sections from the ultramicrotome. Does the standard 2% PFA/ 0.2% gluteraldehyde in CB suffice? 3.) I've read that some fix/process under vacuum? How common/necessary is this? And--how long is the fixation time? 4.) I've also read that some put 1%DMSO into all reagents through absolute alcohol. Is this standard? A thousand thanks in advance!! Cheers, Danielle From weneng2004 <@t> yahoo.com Mon May 9 19:10:17 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] hepcidin antibody Message-ID: <20050510001018.86167.qmail@web53407.mail.yahoo.com> Hello, I was asked to do hepcidin IHC on rat FFPE tissue. Does anybody know a good antibody of that? Thanks in advance! Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From llewllew <@t> shaw.ca Mon May 9 19:20:05 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs References: <1030B679AD69D6119C3F00080210DD9D05A3F28D@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <003501c554f6$0400daa0$7e034246@yourlk4rlmsu> I live in Canada and we can sometimes get the small ring type of black pudding in local supermarkets. I have never seen the big type over here. They don't sell it precooked either, and I had a tremendous shock the first time I cut one open. All that granular red stuff! Bryan Llewellyn From jkiernan <@t> uwo.ca Tue May 10 00:24:42 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] tracer & diluent dilemma [FLUORO-RUBY] References: Message-ID: <4280459A.AD254D2E@uwo.ca> Dear Dianna and other neuro-histonetters concerned about "fluoro-ruby", Try: www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve& db=PubMed&list_uids=7678380&dopt=Abstract This was the 2nd hit with a Google search for "fluoro-ruby", and it identifies the substance as fluorescently labelled dextran amine. These compounds have been used as axon tracers for 10+ years, and there is a large body of literature. Much of it is accessible (full text) on the web if you are at a university or other institute with a library. I think "fluoro-ruby" is one of the tracers developed by Dr Larry C Schmued and published in respected journals without declaration of the compound's chemical identity. It's disconcerting that such secrecy can get past the peer-review procedure. "Fluoro-gold" (a 2-hydroxy-4,4'-diaminostilbene salt) is another one, and so is "Fluoro-jade" (the trisodium salt of mixed isomers of carboxyfluorescein, a dye similar to fluorescein and eosin that binds to the basic proteins in the cytoplasm of dead cells). [Thanks to Adam Halberstadt (Univ of Pittsburgh) for telling me about US Patent 6,229,024, which is more informative than the original "fluoro-jade" publications. Adam's information came as a Histonet reply in 2003. I'm quoting it now so that the reference to the patent will remain in the Histosearch archives for another year or two.] The name "fluoro-gold" (for hydroxystilbamidine) quickly entered the vocabulary of neuroscientists following its introduction in 1986 (Schmued & Fallon. Brain Res. 377:147-154). It's an excellent retrograde tracer and it has been used in hundreds of published investigations. A Google search for hydroxystilbamidine brings up about 450 items, with the first few dozen applying to fluoro-gold as a retrograde tracer. The compound has many other uses. Some of these are noted (with refs) by Richard Horobin in Chapter 22 of the 10th edition of Conn's Biological Stains. Fluoro-gold was adopted as the standard informal name of this fluorochrome because that was the name most often found in catalogues and publications. For more information about "fluoro-ruby" you could ask Larry Schmued. lschmued@nctr.fda.gov His current (federal government) email address suggests that trade secrecy may be in the past. Expect a helpful reply to your question. Fluoro-ruby is probably freely soluble in water and isotonic buffered saline. This is a guess, so don't just go ahead. Ask Schmued. John Kiernan London, Canada. ____________________________________ Dianne Holmes wrote: > > What diluent and at what pH should be used to prepare Floro-Ruby? Has > anyone had problems with it not going into solution? > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue May 10 00:26:47 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs References: <1030B679AD69D6119C3F00080210DD9D05A3F28D@bhrv-nt-11.bhrv.nwest.nhs.uk> <003501c554f6$0400daa0$7e034246@yourlk4rlmsu> Message-ID: <42804617.17EC7CB2@uwo.ca> Dear Bryan, Did you really _like_ the British black pudding, as sold in the 1950s? There was also a red version (in Birmingham, and therefore surely also in Wales). John Kiernan (former Brummie, now in Ontario) ________________________________ Bryan Llewellyn wrote: > > I live in Canada and we can sometimes get the small ring type of black > pudding in local supermarkets. I have never seen the big type over here. > They don't sell it precooked either, and I had a tremendous shock the first > time I cut one open. All that granular red stuff! > > Bryan Llewellyn > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From llewllew <@t> shaw.ca Tue May 10 01:05:37 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs References: <1030B679AD69D6119C3F00080210DD9D05A3F28D@bhrv-nt-11.bhrv.nwest.nhs.uk> <003501c554f6$0400daa0$7e034246@yourlk4rlmsu> <42804617.17EC7CB2@uwo.ca> Message-ID: <001401c55526$499ea8a0$7e034246@yourlk4rlmsu> Yes, I did like it quite a bit. I had it quite often when I was preteen. I stopped eating it in my late teens. My wife doesn't like it, so I didn't buy it. I have bought it in Canada now and again, although I find the Canadian one doesn't have as many fat lumps in as the English one and tastes slightly different. Having to cook it is a pain, too. Last time I went to London, a few years ago, I made sure I had the proper fry up for breakfast including the black pudding. The only red sausage I was familiar with was savaloys. They taste like a cross between cheap bologna and cheap hot dogs and I never liked those. I did like faggots and pease pudding, though. Bryan Llewellyn ----- Original Message ----- From: "John Kiernan" To: "Bryan Llewellyn" Cc: Sent: Monday, May 09, 2005 10:26 PM Subject: Re: [Histonet] Hospital based Histology Labs > Dear Bryan, > > Did you really _like_ the British black pudding, as > sold in the 1950s? There was also a red version > (in Birmingham, and therefore surely also in Wales). > > John Kiernan > (former Brummie, now in Ontario) > ________________________________ > Bryan Llewellyn wrote: >> >> I live in Canada and we can sometimes get the small ring type of black >> pudding in local supermarkets. I have never seen the big type over here. >> They don't sell it precooked either, and I had a tremendous shock the >> first >> time I cut one open. All that granular red stuff! >> >> Bryan Llewellyn >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 10 01:31:22 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F290@bhrv-nt-11.bhrv.nwest.nhs.uk> Cooked???? You cook it? That's heresy. -----Original Message----- From: John Kiernan [mailto:jkiernan@uwo.ca] Sent: 10 May 2005 06:27 To: Bryan Llewellyn Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Hospital based Histology Labs Dear Bryan, Did you really _like_ the British black pudding, as sold in the 1950s? There was also a red version (in Birmingham, and therefore surely also in Wales). John Kiernan (former Brummie, now in Ontario) ________________________________ Bryan Llewellyn wrote: > > I live in Canada and we can sometimes get the small ring type of black > pudding in local supermarkets. I have never seen the big type over here. > They don't sell it precooked either, and I had a tremendous shock the first > time I cut one open. All that granular red stuff! > > Bryan Llewellyn > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 10 01:36:16 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Hospital based Histology Labs Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F292@bhrv-nt-11.bhrv.nwest.nhs.uk> Bolton Black Pudding does not have large fatty lumps in it, if I could ship it then you would be taken to Black Pudding heaven; odd that my ramblings concerning Histology level 3 networking received only a few responses, whilst my short sentence on Black Puddings received loads; my faith in human nature has been restored. -----Original Message----- From: Bryan Llewellyn [mailto:llewllew@shaw.ca] Sent: 10 May 2005 07:06 To: Histonet Subject: Re: [Histonet] Hospital based Histology Labs Yes, I did like it quite a bit. I had it quite often when I was preteen. I stopped eating it in my late teens. My wife doesn't like it, so I didn't buy it. I have bought it in Canada now and again, although I find the Canadian one doesn't have as many fat lumps in as the English one and tastes slightly different. Having to cook it is a pain, too. Last time I went to London, a few years ago, I made sure I had the proper fry up for breakfast including the black pudding. The only red sausage I was familiar with was savaloys. They taste like a cross between cheap bologna and cheap hot dogs and I never liked those. I did like faggots and pease pudding, though. Bryan Llewellyn ----- Original Message ----- From: "John Kiernan" To: "Bryan Llewellyn" Cc: Sent: Monday, May 09, 2005 10:26 PM Subject: Re: [Histonet] Hospital based Histology Labs > Dear Bryan, > > Did you really _like_ the British black pudding, as > sold in the 1950s? There was also a red version > (in Birmingham, and therefore surely also in Wales). > > John Kiernan > (former Brummie, now in Ontario) > ________________________________ > Bryan Llewellyn wrote: >> >> I live in Canada and we can sometimes get the small ring type of black >> pudding in local supermarkets. I have never seen the big type over here. >> They don't sell it precooked either, and I had a tremendous shock the >> first >> time I cut one open. All that granular red stuff! >> >> Bryan Llewellyn >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue May 10 02:53:35 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] RE: Histonet Digest, Vol 18, Issue 11-methasol fast blue Message-ID: We used to use it for myelin stain - can't find mention of it being same as Luxol, but a dye called methazol fast blue can be used instead of Luxol fast blue MBSN for myelin - same dye - different spelling? Jacqui Malam Royal Lancaster Infirmary -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: 09 May 2005 18:04 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 18, Issue 11 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. RE: Vector Red and True Blue (C.M. van der Loos) 2. the skeletal staining (pex) 3. methasol fast blue (Manuelle De Bock) 4. RE: antigen retrieval/enzyme digestion (Todd Sherman) 5. Book- Immunoenzyme Multiple Staining Methods (Cynthia A Delong) 6. Hospital based Histology Labs (Barnhart, Tammy) 7. RE: Hospital based Histology Labs (GUTIERREZ, JUAN) 8. Re: Book- Immunoenzyme Multiple Staining Methods (Gayle Callis) 9. Re: Book- Immunoenzyme Multiple Staining Methods (pmarcum@vet.upenn.edu) 10. RE: Hospital based Histology Labs (Sebree Linda A.) 11. Autoflourescence Eliminator Reagent for lipofuscin in CNS, other cells (Gayle Callis) 12. Phoenix fill in (Riesen, Rebecca) 13. RE: Hospital based Histology Labs (Kemlo Rogerson) 14. microscope companies (pathrm35@adelphia.net) 15. RE: Hospital based Histology Labs (John.Sheppard@Health-Partners.org) 16. RE: Hospital based Histology Labs (GUTIERREZ, JUAN) 17. Re: microscope companies (RCHIOVETTI@aol.com) 18. Thanks Book ordered (Cynthia A Delong) 19. AW: [Histonet] Hospital based Histology Labs (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Mon, 09 May 2005 09:43:52 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: Vector Red and True Blue To: histonet@lists.utsouthwestern.edu Cc: bjrenquist@ucdavis.edu Message-ID: <774aad776fb4.776fb4774aad@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Hi Ben, It's good to read that you're at least successful with the application of True Blue since this isn't the most easy chromogen to deal with. Furthermore, your choice for this turquoise-red color combination is a very good one, because of it's very high contrast. Vector Red however, isn't that strong in my hands either. You may try to look with a fluorescence microscope as the Vector Red reaction product shows a very strong (red) fluorescence upon green light excitation (rhodamine-filter pack). Furthermore, I have very good results for both light (and fluorescence microscopy) using the new Dako Permanent Red kit (K0640). The staining intensity is much higher than with Vector Red. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands phone: +31 20 5665631 fax: +31 20 6960389 e-mail: [1]c.m.vanderloos@amc.uva.nl ----- Original Message ----- From "Ben" Date Wed, 4 May 2005 11:57:08 -0700 To Subject [Histonet] Vector Red and True Blue Hello All, I have been attempting some double labeling for fos and estrogen receptor for about a month with little luck. I am using Vector Red (fos) and True Blue (ER) for my staining. I am having no problems with the ER labeling, but the fos labeling is not that clear. I don't appear to be getting a strong reaction product from the Vector Red. Has anyone else had this problem? How strong should the staining be (light pink, dark pink, red)? I have been careful to only place in xylene for a few moments before coverslipping and therefore don't think fading is my problem. I have been thinking that my antigen retrieval may be the problem. I am using triton X to improve antigen retrieval, but was thinking about microwaving. These are free floating sections. First is this possible to do with free floati! ng sectio References 1. mailto:c.m.vanderloos@amc.uva.nl ------------------------------ Message: 2 Date: Mon, 9 May 2005 17:26:47 +0800 (CST) From: pex Subject: [Histonet] the skeletal staining To: histonet@pathology.swmed.edu Message-ID: <20050509092647.69523.qmail@web15510.mail.cnb.yahoo.com> Content-Type: text/plain; charset=gb2312 Hello, all, Recently I do Alizarin Red and Alcian Blue staining of mouse embryo skeletons, but I find that my results are not better due to a wrong color. After staining, bone should be red, and cartilage should be blue, but my results are that bone is stained purple, and cartilage is stained green, then I change my protocol, but I get the similar results, I do not know the reasons, I guess that bioreagents could not be good, because I make them up, the colors of the solutions are purple and green respectively, not red and blue.Our bioreagents are from sigma. If you have some experience about it, can you do me a favor? Best wishes! Guofeng --------------------------------- Do You Yahoo!? ?????????????????????????????? ------------------------------ Message: 3 Date: Mon, 9 May 2005 14:22:59 +0200 From: "Manuelle De Bock" Subject: [Histonet] methasol fast blue To: Message-ID: <000601c55491$d6e7e540$ed54bec1@ugent.be> Content-Type: text/plain; charset="Windows-1252" I am trying to stain collected gastric tissue cells with Maxwell's staining procedure (1963) but we don't have methasol fast blue in our lab. I also can't find any links on the web where I can buy this dye. can someone help me with this? Is it the same dye as Luxol fast blue? Kind regards Manuelle Manuelle De Bock, dierenarts (DVM) Laboratory of Veterinary Pathology Department of Pathology, Bacteriology and Avian Diseases Faculty of Veterinary Medicine Ghent University Salisburylaan 133 B9820 Merelbeke - Belgium Tel 0032(0)9 264 7745 Fax 0032(0)9 264 7789 ------------------------------ Message: 4 Date: Mon, 09 May 2005 08:58:31 -0500 From: "Todd Sherman" Subject: [Histonet] RE: antigen retrieval/enzyme digestion To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed; delsp=yes; charset=iso-8859-1 On Sun, 08 May 2005 12:00:26 -0500, wrote: > Today's Topics: > > 1. RE: antigen retrieval/enzyme digestion (Patsy Ruegg) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Sun, 8 May 2005 09:42:06 -0600 > From: "Patsy Ruegg" > Subject: RE: [Histonet] antigen retrieval/enzyme digestion > To: "'Till, Renee'" , > > Message-ID: <200505081542.j48FgR75001351@pro12.abac.com> > Content-Type: text/plain; charset="us-ascii" > > I have heard of it being done both ways depending on the epitope of > interest. In general I do HIER first and then EIER. > Patsy > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Till, > Renee > Sent: Monday, May 02, 2005 5:55 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] antigen retrieval/enzyme digestion > > When trying both on the same slides, which would go first, the antigen > retrieval or the enzyme digestion? > > > Renee' Till, HT > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > (501) 364-2774 Renee, I'd tend to agree with Patsy as far as a sequence best determined empirically. You'll probably need to compare each protocol for each antigen of interest. From a theoretical perspective, both techniques of "unmasking" involve the breaking of strong molecular bonds. HIER is typically designed to cleave the basic side-chains of amino acids on proteins previously fixed (or bound to each other) with x-aldehyde. Enzyme digestion is similarly designed to cleave proteins but at well defined amino acids which are not necessarily basic - it depends on the digesting enzyme and its substrate. My guess would be that progressing from a more basic (read simple/generic) to a more specific cleaving sequence would be more efficient; consequently, treat with HIER first and enzyme digestion second. The HIER would open up the tissue to further access and the enzyme digestion could then more specifically cleave regions hindering an even more localized access to your target antigen. To a great extent, these success derived from these sequences will be greatly influenced by the local milieu of your target antigen. The relative content of various amino acids in a given milieu may or may not be known. To that end empirical observations are "best" which is why mandating one particular protocol over another may be a bit dubious. Good luck. Todd President HistoSoft Corporation -- >>>>>>>> www.histosoft.com <<<<<<<< <<<<<<<< Biology In A New Form (c) >>>>>>> ------------------------------ Message: 5 Date: Mon, 09 May 2005 09:13:04 -0500 From: Cynthia A Delong Subject: [Histonet] Book- Immunoenzyme Multiple Staining Methods To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Hello All, I am looking for Chris Van de Loos's book"Immunoenzyme Multiple Staining Methods". Does anyone know of where I can get a copy of this book? I had one and someone walked off with it. Thank you Cindy Cynthia A DeLong LRL - Corporate Center DC0533 - 48B/3042 355 E Merrill St Indianapolis, IN 46225 phone: 317-276-7635 fax: 317-277-6146 ------------------------------ Message: 6 Date: Mon, 9 May 2005 09:25:54 -0500 From: "Barnhart, Tammy" Subject: [Histonet] Hospital based Histology Labs To: "Histonet (E-mail)" Message-ID: <1779904B5E82D511914C00D0B793339205BFD930@exchangent> Content-Type: text/plain; charset="iso-8859-1" This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. ------------------------------ Message: 7 Date: Mon, 9 May 2005 09:35:39 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Hospital based Histology Labs To: "Barnhart, Tammy" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" We are located one floor above the surgical suites, but we do maintain a frozen/gross room in the surgical suite area. At the end of the day the gross tech brings the specimens upstairs to be loaded in the processors. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barnhart, Tammy Sent: Monday, May 09, 2005 9:26 AM To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 09 May 2005 08:41:10 -0600 From: Gayle Callis Subject: Re: [Histonet] Book- Immunoenzyme Multiple Staining Methods To: Cynthia A Delong , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050509083836.01b613a8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed I replaced my copy from Amazon.com. The book is now 8.99/copy, and the book is no longer in print, so don't hesitate too long to replace. I was so worried this would happen to me i.e. walking off, I now keep a personal copy at home, my first one is worn out from use! At 08:13 AM 5/9/2005, you wrote: >Hello All, >I am looking for Chris Van de Loos's book"Immunoenzyme Multiple Staining >Methods". Does anyone know of where I can get a copy of this book? >I had one and someone walked off with it. >Thank you >Cindy > >Cynthia A DeLong >LRL - Corporate Center >DC0533 - 48B/3042 >355 E Merrill St >Indianapolis, IN 46225 >phone: 317-276-7635 >fax: 317-277-6146 >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 9 Date: Mon, 9 May 2005 10:46:49 -0400 From: pmarcum@vet.upenn.edu Subject: Re: [Histonet] Book- Immunoenzyme Multiple Staining Methods To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu, Cynthia A Delong Message-ID: <1115650008.427f77d900571@imp.vet.upenn.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi Gayle and All, I just purchased one from Amazon and only two were still listed at $8.95 p;us shipping and handling it was about $12.50 for the copy. Thanks, Pam Marcum Quoting Gayle Callis : > I replaced my copy from Amazon.com. The book is now 8.99/copy, and the > book is no longer in print, so don't hesitate too long to replace. I was > so worried this would happen to me i.e. walking off, I now keep a personal > copy at home, my first one is worn out from use! > > At 08:13 AM 5/9/2005, you wrote: > >Hello All, > >I am looking for Chris Van de Loos's book"Immunoenzyme Multiple Staining > >Methods". Does anyone know of where I can get a copy of this book? > >I had one and someone walked off with it. > >Thank you > >Cindy > > > >Cynthia A DeLong > >LRL - Corporate Center > >DC0533 - 48B/3042 > >355 E Merrill St > >Indianapolis, IN 46225 > >phone: 317-276-7635 > >fax: 317-277-6146 > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 10 Date: Mon, 9 May 2005 09:49:46 -0500 From: "Sebree Linda A." Subject: RE: [Histonet] Hospital based Histology Labs To: "Barnhart, Tammy" , "Histonet \(E-mail\)" Message-ID: Content-Type: text/plain; charset="us-ascii" Several years ago our surgical pathology, IHC/ISH, and renal labs were moved to rented space in a VA hospital that's physically connected to our University hospital. The grossing and frozen section lab remained in space adjacent to the ORs. The cassetted specimens are brought over from there every day on carts. It seems to work okay. If the surgeons want to talk with a pathologist the next day about a specimen they have to walk over to the VA to do so. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barnhart, Tammy Sent: Monday, May 09, 2005 9:26 AM To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Mon, 09 May 2005 08:59:15 -0600 From: Gayle Callis Subject: [Histonet] Autoflourescence Eliminator Reagent for lipofuscin in CNS, other cells To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050509084844.01b22e70@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Dear all For those looking for a way to eliminate or reduce lipofuscin autofluorescence in CNS tissues and other cells, there is a commercial product available. Science magazine just listed this new reagent, and it may be worth a try. Chemicon International at www.chemicon.com is the vendor. They mentioned monkey, human and rat neural and other tissues as well. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 12 Date: Mon, 9 May 2005 11:11:13 -0400 From: "Riesen, Rebecca" Subject: [Histonet] Phoenix fill in To: Message-ID: <05844092236E7749A1BBBCB1077C34A2DE9FDD@dsi-ex01.gateway.dom> Content-Type: text/plain; charset="iso-8859-1" PICK ME! PICK ME! Fly me to Phoenix for the weekend! I'm in Florida. Message: 10 Date: Fri, 6 May 2005 15:30:16 -0700 (PDT) From: Linda Hines Subject: [Histonet] histologist to fill in To: Histonet@lists.utsouthwestern.edu Message-ID: <20050506223021.87346.qmail@web31315.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi! Everyone, I am looking for a histotech to fill in on May 20, 2005 in the afternoon appx. 4:00pm in the afternoon and work until appx. 9:00 pm. We are located central Phoenix, AZ if interested please contact Linda Hines @ 602-241-5134. Thank-you Linda Hines HT Specialty Diagnostics --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour ------------------------------ DSI Labs Email Disclaimer: Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipients(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 13 Date: Mon, 9 May 2005 16:12:05 +0100 From: Kemlo Rogerson Subject: RE: [Histonet] Hospital based Histology Labs To: "'Barnhart, Tammy'" , "Histonet (E-mail)" Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F28D@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain Very interesting question and very pertinent for Laboratory Modernisation in the UK. Traditionally it has always been supposed that Histology Labs must be on Site for the very reasons you suggest. My experience in London and now in the North leads me to question that. I accept that frozen sections may need a local presence, but they are relatively rare nowadays, but a motorbike Courier service may be quicker than a porter! I think a happy intermediary is for there to be a Core Lab, that deals with the bulk of the processing and staining and this will reduce the duplication of tests and increase the critical number of Staff; you should see costs driven down. ICC and such like can be consolidated on one Site again with cost savings. A satellite laboratory may be needed to deal with frozen sections and urgent rapid H&E's, but I wonder if that necessity, once the Core Lab became organised may 'wither on the vine'. The bigger problems are MDT's and alienation of Pathologists from their Clinical colleagues; Telepathology and the use of video cameras are a solution, but not a complete one; the spontaneous ways we humans do business makes that an unacceptable means of communicating. In order to release resources consolidation must occur and if used wisely may result in extension of roles with the positive impact on recruitment and retention of Staff. Do you get Black Pudding in America? -----Original Message----- From: Barnhart, Tammy [mailto:Tbarnhart@primecare.org] Sent: 09 May 2005 15:26 To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Mon, 9 May 2005 11:19:03 -0400 From: Subject: [Histonet] microscope companies To: histonet@lists.utsouthwestern.edu Message-ID: <18444780.1115651943878.JavaMail.root@web4.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 Fellow tech's, Does anyone know of a company in Florida that services microscopes? I have already contacted JL Optical and Southern Microscope. Thanks, Ron Martin ------------------------------ Message: 15 Date: Mon, 9 May 2005 11:22:49 -0400 From: John.Sheppard@Health-Partners.org Subject: [Histonet] RE: Hospital based Histology Labs To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: TEXT/plain; charset="US-ASCII" We have the surgical suites located one floor above us. The O.R. sends a runner down with the frozens. We keep the cryostats here in histology. We do get a copy of the surgury schedule, so we have an idea when about half the frozens are comming. It's not perfect, but it works, as long as the runner hand delivers the frozen to someone and does not just leave it on the counter. John Sheppard HT(ASCP) The Community Hospital Springfield, Ohio CONFIDENTIALITY NOTICE: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ------------------------------ Message: 16 Date: Mon, 9 May 2005 10:31:14 -0500 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Hospital based Histology Labs To: "GUTIERREZ, JUAN" , "Barnhart, Tammy" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" On a previous life in the alien visited town of Roswell, NM, we had an offsite lab. The frozen sections were handled by the pathologists at the hospital, but the rest of the specimens were picked up by a courier twice a day and brought to the lab for gross. The courier will also ensure that there was enough materials at the frozen room daily. And the doctors were always kind enough to let us know when their mess was too much for us to go and clean it up. Ever try to clean up a weeks worth of shavings in a cryostat? Needles to say we stop waiting for word and visited the hospital often to keep the room nice and tidy;0) Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of GUTIERREZ, JUAN Sent: Monday, May 09, 2005 9:36 AM To: Barnhart, Tammy; Histonet (E-mail) Subject: RE: [Histonet] Hospital based Histology Labs We are located one floor above the surgical suites, but we do maintain a frozen/gross room in the surgical suite area. At the end of the day the gross tech brings the specimens upstairs to be loaded in the processors. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barnhart, Tammy Sent: Monday, May 09, 2005 9:26 AM To: Histonet (E-mail) Subject: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Mon, 9 May 2005 11:31:13 EDT From: RCHIOVETTI@aol.com Subject: Re: [Histonet] microscope companies To: pathrm35@adelphia.net, histonet@lists.utsouthwestern.edu Message-ID: <86.27bbaf54.2fb0dc41@aol.com> Content-Type: text/plain; charset="US-ASCII" In a message dated 5/9/2005 8:21:22 AM US Mountain Standard Time, pathrm35@adelphia.net writes: > Does anyone know of a company in Florida that services microscopes? I have > already contacted JL Optical and Southern Microscope. > Ron, Try Micro Optics of Florida, 3941 S.W. 47th Ave., Davie, FL, 954-791-0082, < www.microopticsfl.com>. They are Leica dealers, but I believe their service shop also maintains and repairs other brands of scopes. Cheers, Bob Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA ------------------------------ Message: 18 Date: Mon, 09 May 2005 10:39:27 -0500 From: Cynthia A Delong Subject: [Histonet] Thanks Book ordered To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" Thanks to all who replied to my request. The book is ordered. I guess I will have to keep this one under lock and key. Cindy Cynthia A DeLong LRL - Corporate Center DC0533 - 48B/3042 355 E Merrill St Indianapolis, IN 46225 phone: 317-276-7635 fax: 317-277-6146 ------------------------------ Message: 19 Date: Mon, 9 May 2005 18:40:15 +0200 From: "Gudrun Lang" Subject: AW: [Histonet] Hospital based Histology Labs To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Our lab is on the hospital area, but in another building as the surgery area. We are connected with the surgery room with a pneumatic delivery, that shots the tissue to us. That takes less than 30 sec. We do the frozens in an integrated part of our lab. The pathologists give the results through an intercommunication system to the surgeon. Gudrun Lang Linz, Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Barnhart, Tammy Gesendet: Montag, 09. Mai 2005 16:26 An: Histonet (E-mail) Betreff: [Histonet] Hospital based Histology Labs This is a question for hospital based histo labs. The hospital we are based in has talked of moving our histology laboratory to an area separated from the surgical suites. My question is, how many hospital histology labs are removed from the surgical suites by a significant distance? Of those that are separated, do you have a frozen/gross area located away from the histo lab but close to the OR? As many responses as possible would be appreciated. This will greatly facilitate our decisions for future locations. Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 11 **************************************** DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From funderwood <@t> mcohio.org Tue May 10 07:19:16 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] asbestos fibers Message-ID: Hi All, I asked this question earlier, but am not sure it got posted. I'm looking for a technic to demonstrate asbestos fibers. Possbily by digesting the tissue. Thanks in advance. Fred Underwood HT(ASCP) Montgomery County Coroner Dayton,OH From histosci <@t> shentel.net Tue May 10 08:45:34 2005 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Giemsa Stain for Mast Cells Message-ID: <000301c55566$90d9dfb0$0200a8c0@HSRLMAIN> Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org From cfavara <@t> niaid.nih.gov Tue May 10 09:37:23 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] slide labeller Message-ID: I am interested in an automated slide labeler. I would like the capacity to do multiple slides with the same information as quiet as possible and of course I have limited space. Any suggestions would be appreciated. I also have tops to the bottle for a Ventana special stainer. I am happy to send to anyone interested. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From liz <@t> premierlab.com Tue May 10 09:40:45 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Giemsa Stain for Mast Cells In-Reply-To: <000301c55566$90d9dfb0$0200a8c0@HSRLMAIN> Message-ID: <001601c5556e$431dff10$a7d48a80@AMY> I would use toluidine blue. It's a great stain for mast cells. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of HSRL Sent: Tuesday, May 10, 2005 6:46 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Giemsa Stain for Mast Cells Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue May 10 09:50:51 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Giemsa Stain for Mast Cells In-Reply-To: <000301c55566$90d9dfb0$0200a8c0@HSRLMAIN> References: <000301c55566$90d9dfb0$0200a8c0@HSRLMAIN> Message-ID: <6.0.0.22.1.20050510084909.01b5ca10@gemini.msu.montana.edu> Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is Churukian Shenk method for mast cells. You don't have to worry about so many other colors from Giemsa. If you want these, I will be happy to attach privately. At 07:45 AM 5/10/2005, you wrote: >Netters, > > > >I am looking for a Giemsa stain for Mast Cells that does not have a >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast >Cells pop out at us on low power. Any suggestions/protocols? > > > >Thanks, > > > >Tom > > > >Tom Galati > >Laboratory Director > >HSRL, Inc.- A GLP Compliant Contract Laboratory > >137 South Main Street > >Woodstock, Virginia 22664 > >(540)459-8211 > >Fax: (540)459-8217 > >tomgalati@hsrl.org > >www.hsrl.org > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From bruyntjes <@t> voeding.tno.nl Tue May 10 09:53:59 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Giemsa Stain for Mast Cells Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DB4@ntexch1.voeding.tno.nl> There is an antibody directed against tryptase, this should be an excellent marker for mast cells only. Joost -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: dinsdag 10 mei 2005 16:41 To: histosci@shentel.net; histonet@pathology.swmed.edu Subject: RE: [Histonet] Giemsa Stain for Mast Cells I would use toluidine blue. It's a great stain for mast cells. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of HSRL Sent: Tuesday, May 10, 2005 6:46 AM To: histonet@pathology.swmed.edu Subject: [Histonet] Giemsa Stain for Mast Cells Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From timothy.m.coskran <@t> pfizer.com Tue May 10 09:58:47 2005 From: timothy.m.coskran <@t> pfizer.com (Coskran, Timothy M) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] CCR7 Message-ID: <599089EF29304C44AF1D84B1C223AC63019D8E4B@groamrexm03.amer.pfizer.com> Does anyone know a source for anti-CCR7 that may be useful in frozen section IHC on mouse tissues? Thanks, Tim Coskran Pfizer LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From gcallis <@t> montana.edu Tue May 10 10:30:13 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] FLUORO-RUBY In-Reply-To: <4280459A.AD254D2E@uwo.ca> References: <4280459A.AD254D2E@uwo.ca> Message-ID: <6.0.0.22.1.20050510092449.01b5eaa8@gemini.msu.montana.edu> Fluoro Ruby is as John Kiernan stated, a dextran. Molecular Probes, in Section 14.5 of the handbook explains its use. the handbook is available in pdf form on website. If you need particulars on how to dissolve, solvent, etc they post this information on their website for this product. It would also have been found in the specification sheet sent with the product. If all else fails, tech_service@invitrogen.com will help you. Molecular Probes is now part of Invitrogen. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From mtitford <@t> aol.com Tue May 10 10:52:23 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Asbestos studies Message-ID: <8C7238F63CB20B8-B5C-14BF9@mblk-d42.sysops.aol.com> Fred Underwood asks about asbestos fibers in lung tissues. Asbestosis and mesothelioma studies are quite big business along the Gulf Coast here with the local shipyards and men of the "Golden Generation" passing away. A lot of lawyers have made a lot of money and most recently, a big lawsuit in Corpus Christi Texas accuses screening companies of misdiagnosis. However, onto things histological: asbestos fibers can be easily seen in an H&E section as brown, cylindrical, and beaded. In our laboratory we digest lung tissue in bleach and then filter the remains through a millipore. Once dried, it can be placed on a slide, the filter dissolved in chloroform and mounted with a coverslip. The fibers are then counted. The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of asbestos-associated diseases" 2ed edition Spinger. 2004 Mike Titford USA Pathology Mobile AL USA From stamptrain <@t> yahoo.com Tue May 10 11:04:27 2005 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] slide labeller In-Reply-To: 6667 Message-ID: <20050510160427.18678.qmail@web31110.mail.mud.yahoo.com> We have the Thermo (was ThermoShandon) Microwriter. It is efficient and does an excellent job. We have attached it to external software for data input, but the software it comes with is also very nice. Two caveats: I wouldn't exactly call it 'quiet' although it is 'relatively quiet'; and it is inexpensive. That said, we have had very little in the way of maintenance problems other than routine maintenance. Get a demo. I'm quite sure they would be willing to bring in a unit for you to try out. That's the best way anyhow, so you get a chance to checkout how it will handle your labeling information and data. Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals Ridgefield, CT --- "Favara, Cynthia (NIH/NIAID)" wrote: > I am interested in an automated slide labeler. I > would like the capacity to > do multiple slides with the same information as > quiet as possible and of > course I have limited space. Any suggestions would > be appreciated. > > > > I also have tops to the bottle for a Ventana special > stainer. I am happy to > send to anyone interested. > > > > c > > > > Cynthia Favara > NIAID/NIH/RML/LPVD > 903 South 4th Street > Hamilton, MT 59840 > 406-363-9317 > > Disclaimer: > The information in this e-mail and any of its > attachments is confidential > and may contain sensitive information. It should not > be used by anyone who > is not the original intended recipient. If you have > received this e-mail in > error please inform the sender and delete it from > your mailbox or any other > storage devices. National Institute of Allergy and > Infectious Diseases shall > not accept liability for any statements made that > are sender's own and not > expressly made on behalf of the NIAID by one of its > representatives > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. http://mobile.yahoo.com/learn/mail From stamptrain <@t> yahoo.com Tue May 10 11:07:41 2005 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] slide labeller In-Reply-To: 6667 Message-ID: <20050510160741.39959.qmail@web31114.mail.mud.yahoo.com> Oops!!! That should have read "not inexpensive" Better proofread it better next time.... Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals Ridgefield, CT --- Roger Moretz wrote: > We have the Thermo (was ThermoShandon) Microwriter. > It is efficient and does an excellent job. We have > attached it to external software for data input, but > the software it comes with is also very nice. Two > caveats: I wouldn't exactly call it 'quiet' > although > it is 'relatively quiet'; and it is inexpensive. > That > said, we have had very little in the way of > maintenance problems other than routine maintenance. > > Get a demo. I'm quite sure they would be willing to > bring in a unit for you to try out. That's the best > way anyhow, so you get a chance to checkout how it > will handle your labeling information and data. > > Roger Moretz, Ph.D. > Dept of Toxicology > Boehringer Ingelheim Pharmaceuticals > Ridgefield, CT > > --- "Favara, Cynthia (NIH/NIAID)" > wrote: > > I am interested in an automated slide labeler. I > > would like the capacity to > > do multiple slides with the same information as > > quiet as possible and of > > course I have limited space. Any suggestions would > > be appreciated. > > > > > > > > I also have tops to the bottle for a Ventana > special > > stainer. I am happy to > > send to anyone interested. > > > > > > > > c > > > > > > > > Cynthia Favara > > NIAID/NIH/RML/LPVD > > 903 South 4th Street > > Hamilton, MT 59840 > > 406-363-9317 > > > > Disclaimer: > > The information in this e-mail and any of its > > attachments is confidential > > and may contain sensitive information. It should > not > > be used by anyone who > > is not the original intended recipient. If you > have > > received this e-mail in > > error please inform the sender and delete it from > > your mailbox or any other > > storage devices. National Institute of Allergy and > > Infectious Diseases shall > > not accept liability for any statements made that > > are sender's own and not > > expressly made on behalf of the NIAID by one of > its > > representatives > > > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your > mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. http://mobile.yahoo.com/learn/mail From eca9 <@t> georgetown.edu Tue May 10 10:41:52 2005 From: eca9 <@t> georgetown.edu (Eva C Anderson) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] rookie wanting to learn to cut Message-ID: <4280D640.2020004@georgetown.edu> Hi Everyone, I am totally new to the cutting of paraffin embedded tissue and would like to try this process. Could anyone provide me with must read information on the subject before I start? I would like to read up as much as possible. I have access to a microtome and old blocks to practice on. Would really appreciate any information you can provide me with as well as suggestions on must have equipment. Thank you all in advance, Eva Andersson Research Assistant From Terry.Marshall <@t> rothgen.nhs.uk Tue May 10 11:41:41 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Asbestos studies Message-ID: Mike, What you are describing is asbestos bodies, not asbestos fibres, which are many times more numerous and not visible on light microscopy. Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: mtitford@aol.com [mailto:mtitford@aol.com] Sent: 10 May 2005 16:52 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Asbestos studies Fred Underwood asks about asbestos fibers in lung tissues. Asbestosis and mesothelioma studies are quite big business along the Gulf Coast here with the local shipyards and men of the "Golden Generation" passing away. A lot of lawyers have made a lot of money and most recently, a big lawsuit in Corpus Christi Texas accuses screening companies of misdiagnosis. However, onto things histological: asbestos fibers can be easily seen in an H&E section as brown, cylindrical, and beaded. In our laboratory we digest lung tissue in bleach and then filter the remains through a millipore. Once dried, it can be placed on a slide, the filter dissolved in chloroform and mounted with a coverslip. The fibers are then counted. The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of asbestos-associated diseases" 2ed edition Spinger. 2004 Mike Titford USA Pathology Mobile AL USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Tue May 10 11:55:02 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] mast cell in guinea pig Message-ID: <000001c55581$05065390$a7d48a80@AMY> Hello All I'm looking for a stain that will identify mast cells in guinea pigs. I have run both giemsa (modified dif quick) and 0.4% Toluidine Blue. I normally get very good staining of mast cells with the toluidine blue. I normally run this stain on mouse and rat tissue. This is the first time I have tried this stain in guinea pigs. Upon review of the slides I could not located one mast cell in 11 lung sections. I know they have to be there. Does the metachromatic nature of the t. blue stain have to do with the amount of histamine present in the cells? I have read in the literature that guinea pigs and humans have less histamine present in their mast cells than rats, hamsters and mice. Is anyone aware of a modification that will stain mast cells in guinea pig? Any help would be appreciated. I would prefer to stick with a histochemical method. I'm aware of Immunohistochemical staining for mast cell tryptase, but in researching this I could not find any references for guinea pig tissue. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From sharon.osborn <@t> dnax.org Tue May 10 12:14:25 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Mast cell stain Message-ID: <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> Tom, Luna's Toluidine Blue Method for Mast Cells is a superior demonstration of mast cells using a pink background. These literally pop up under low power. The method is located on pages 311-312 of Luna's Histopathologic Methods and color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a copy of the procedure, private email me and I will send to you. sharon osborn DNAX ScheringPlough BioPharma Palo Alto, CA Subject: [Histonet] Giemsa Stain for Mast Cells Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From sharon.osborn <@t> dnax.org Tue May 10 12:21:48 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Slide labelers Message-ID: <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> Cynthia, Contact your Leica representative and ask for a demonstration of their slide printers. I think you will be pleasantly surprised as to the amount of information and quietness. I have used several different slide labelers including the etch-a-sketch types and ink printer types; this one is the best of all I have used. Its footprint is a little larger than the others and well worth it. Sakura and Leica partnered on developing these; their main difference is in the software that is provided. Sakura uses off the shelf software while Leica specifically built the software on theirs and it is customizable for stand alone or for integration with MIS of an institution. And, the company service will also make a difference. So, I encourage you to demo several different types of slide labelers and find the one that best suits your needs. Sharon Osborn DNAX ScheringPlough BioPharma Palo Alto, CA Date: Tue, 10 May 2005 10:37:23 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: [Histonet] slide labeller To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I am interested in an automated slide labeler. I would like the capacity to do multiple slides with the same information as quiet as possible and of course I have limited space. Any suggestions would be appreciated. I also have tops to the bottle for a Ventana special stainer. I am happy to send to anyone interested. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From gcallis <@t> montana.edu Tue May 10 12:25:43 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Asbestos studies In-Reply-To: <8C7238F63CB20B8-B5C-14BF9@mblk-d42.sysops.aol.com> References: <8C7238F63CB20B8-B5C-14BF9@mblk-d42.sysops.aol.com> Message-ID: <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the Gulf >Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most recently, >a big lawsuit in Corpus Christi Texas accuses screening companies of >misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen in >an H&E section as brown, cylindrical, and beaded. In our laboratory we >digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter dissolved >in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue May 10 12:28:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Control for Mast cells In-Reply-To: <013351202ADA9E42B7B48602D5DC5A8703A0EE80@smg-exchange.exch ange.co.sclhs.net> References: <013351202ADA9E42B7B48602D5DC5A8703A0EE80@smg-exchange.exchange.co.sclhs.net> Message-ID: <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org > > > > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis >Sent: Tuesday, May 10, 2005 8:51 AM >To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Giemsa Stain for Mast Cells > > > >Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is > >Churukian Shenk method for mast cells. You don't have to worry about so > >many other colors from Giemsa. If you want these, I will be happy to > >attach privately. > > > >At 07:45 AM 5/10/2005, you wrote: > > >Netters, > > > > > > > > > > > >I am looking for a Giemsa stain for Mast Cells that does not have a > > >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast > > >Cells pop out at us on low power. Any suggestions/protocols? > > > > > > > > > > > >Thanks, > > > > > > > > > > > >Tom > > > > > > > > > > > >Tom Galati > > > > > >Laboratory Director > > > > > >HSRL, Inc.- A GLP Compliant Contract Laboratory > > > > > >137 South Main Street > > > > > >Woodstock, Virginia 22664 > > > > > >(540)459-8211 > > > > > >Fax: (540)459-8217 > > > > > >tomgalati@hsrl.org > > > > > >www.hsrl.org > > > > > > > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Gayle Callis > >MT,HT,HTL(ASCP) > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 (lab with voice mail) > >406 994-4303 (FAX) > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >This electronic message and all contents contain information from St. >Mary's Hospital which may be attorney-client privileged, confidential or >otherwise protected from disclosure. The information is intended to be >for the addressee only. If you are not the addressee, any disclosure, >copy, distribution or use of the contents of this message is >prohibited. If you have received this electronic message in error, please >notify the sender immediately and destroy the original message and all >copies. Thank you Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From wurochi <@t> hotmail.com Tue May 10 12:30:54 2005 From: wurochi <@t> hotmail.com (Jeffrey Wu) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Picrosirius red stain Message-ID: Hello all, I am trying to quantitate the amount of fibrosis in mouse cardiac tissue. I based my method on Dr Kiernan's excellent tutorial on picrosirius red staining (posted in 2000). Briefly, slides are deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); 75% ethanol (1x, 3 min); and running distilled water (10-15 min). Next, they are stained in saturated picrosirius red for 60 minutes and washed in acetic acid (2x, 3 min). They are dehydrated in 75% ethanol (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 min). Finally, the slides are mounted with Permount. For image processing, I am using circularly polarized light. Under the microscope, the collagen stains from pink to dark red (almost black), depending on its content; however, the background tissue appears green/blue. It is problematic because I quantify the collagen using ImagePro Plus software, in which I select certain colors. Especially with the pink and some dark blues, there is overlap between the collagen and tissue, causing overestimation of collagen. (Sorry for the long explanation.) I guess what I am seeking is the flaw in my procedure, causing the other tissue not to stain yellow. On a side note, I am using the correct picrosirius red stain, and the solution is saturated with crystals at the bottom. I have tried modifying washing times without any change. I used a short (2 min) 0.2% phosphomolybdic acid wash, which is supposed to reduce background, with no success as well. Once again, sorry for the long question. Any help with my problem would be greatly appreciated. Please do not hesitate to email me with any questions or anything that I have not made clear. Thank you in advance. J Wu From jstaruk <@t> masshistology.com Tue May 10 12:52:27 2005 From: jstaruk <@t> masshistology.com (Jim Staruk) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Control for Mast cells In-Reply-To: <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> Anyone in diagnostic veterinary services gets mast cell tumors all of the time. I'll be glad to share some with someone in need (and I won't even charge you). Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 1:28 PM To: Anna Inman; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Control for Mast cells Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org From jkiernan <@t> uwo.ca Tue May 10 12:58:36 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] mast cell in guinea pig Message-ID: <4280F64C.CD1F5085@uwo.ca> John Kiernan wrote: > > Probably you didn't see any mast cells in the > sections of guinea-pig tissues because there > were none to be seen. > > Mouse and rat mast cells are unusual in that their > granules are preserved and rendered insoluble by > aqueous fixatives such as buffered formaldehyde. > In most other species aqueous fixatives dissolve > out the mast cell granules. Alcoholic fixatives (eg > Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre > etc) will preserve and insolubilize mast cell granules > of any species. > > For more on this, see "The Mast Cells" by Hans Selye > (1965) Chapter 3. Also the "Notes & Queries section in > the current issue of Biotechnic & Histochemistry (Vol 80 > No 1, p.43-45). The latter is available at the publisher's > web site: > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c124b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1,1;homemain,1,1; > > The metachromasia is due to heparin, the major > macromolecular anion of the granules. The staining > properties of heparin are similar to those of > cartilage matrix. Both materials carry a lot > of sulphate-ester groups. Unfortunately heparin > (except that of rats and mice) is water-soluble. > I do not know if any tryptase or chymase immunoreactivity > remains in mast cells whose granules have been > extracted by an aqueous fixative. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Elizabeth Chlipala wrote: > > > > Hello All > > > > I'm looking for a stain that will identify mast cells in guinea pigs. I > > have run both giemsa (modified dif quick) and 0.4% Toluidine Blue. I > > normally get very good staining of mast cells with the toluidine blue. > > I normally run this stain on mouse and rat tissue. This is the first > > time I have tried this stain in guinea pigs. Upon review of the slides > > I could not located one mast cell in 11 lung sections. I know they have > > to be there. Does the metachromatic nature of the t. blue stain have to > > do with the amount of histamine present in the cells? I have read in > > the literature that guinea pigs and humans have less histamine present > > in their mast cells than rats, hamsters and mice. Is anyone aware of a > > modification that will stain mast cells in guinea pig? Any help would > > be appreciated. I would prefer to stick with a histochemical method. > > I'm aware of Immunohistochemical staining for mast cell tryptase, but in > > researching this I could not find any references for guinea pig tissue. > > > > Thanks in advance. > > > > Liz > > > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > > Manager > > Premier Laboratory, LLC > > P.O. Box 18592 > > Boulder, Colorado 80308 > > Office: (303) 735-5001 > > Fax: (303) 735-3540 > > liz@premierlab.com > > www.premierlab.com > > > > Ship to Address: > > Premier Laboratory > > University of Colorado > > MCDB, Room A3B40 > > Boulder, Colorado 80309 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From POWELL_SA <@t> Mercer.edu Tue May 10 13:15:25 2005 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Asbestos studies In-Reply-To: <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> Message-ID: <01LO3GS2JNLI8WX3KK@Macon2.Mercer.edu> The histonet archives can be found at www.histosearch.com. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 12:26 PM To: mtitford@aol.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Asbestos studies If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the >Gulf Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most >recently, a big lawsuit in Corpus Christi Texas accuses screening >companies of misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen >in an H&E section as brown, cylindrical, and beaded. In our laboratory >we digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter >dissolved in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Tue May 10 14:06:06 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Final notice: Missouri Society for Histotechnology Spring Symposium Message-ID: The Missouri Society for Histotechnology Spring Symposium When: May 20-21, 2005 Where: The Lodge of the Four Seasons, Lake Ozark, Missouri (800) 843-5253 www.4seasonsresort.com Note: Rooms for 'The Lodge' may be fully booked so here are 2 alternative places to stay: The Country Club (800) 964-6698 (approximately 1.5 miles away) The Resort at Port Arrowhead (800) 532-3575 (approximately 3 miles away) For additional meeting information please contact: Sharon Walsh Phone: (636) 305-9650 email: userwalsh@aol.com or Rosetta Barkley Phone: (913) 588-2737 email: rbarkley2@kumc.edu We invite you to participate in a unique learning experience. The program is outstanding this year and should have something for everyone. In addition there will be social activities and vendor displays. Please Note: All workshops are CEU approved. Friday Schedule: 8:30am "Ergonomics in the Workplace", Jan Minshew, Leica Microsystems 9:15am "Case Studies in Forensic Pathology", Dr. Michael Graham, St. Louis University 10:45am "Histochemistry of Special Stains", Jerry Fredenburgh 1:30pm-4:30pm Workshops #1, #2, or #3 (Your choice) 6:00-8:00pm - Cruise on the "Lake of the Ozarks" Come enjoy the lake and visit with exhibitor's and friends. Saturday Schedule: 8:15am "Creutzfeldt-Jakob Disease: Tissue Handling and Testing in the United Kingdom", Konnie Zietner, Nebraska Medical Center 9:15am "Histological Techniques in Hematopathology", Dr. Sharad Mather 10:45am "Applications for Microwave Decalcification Procedures", Skip Brown 1:30pm-4:30pm Workshops #4, #5, or #6 (Your choice) Registration Fees: Members - $50, Non-Members - $60 Additional Banquet Ticket - $25 2005 WORKSHOP DESCRIPTIONS Workshop#1 Theory of Routine & Microwave Fixation & Processing - Skip Brown & Jerry Fredenburgh The process of tissue fixation and processing will be illustrated; first from a molecular level to demonstrate what is actually happening at a micro level in the tissue; then from a macro level to show the gross effects on the tissue. Accelerated procedures will be introduced as a way to speed up the process without detrimental effects to the tissue. Workshop #2 (Sponsored by Ventana Medical Systems) A Tour Guide to Immunohistochemistry: How to get there and what to do when you arrive. - Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Immunohistochemistry is defined as, "A method of detecting the presence of specific proteins in cells or tissues." If only it was that easy. A better definition may be, "A method of standardizing each and every step in and out of your control working toward a complex chemical construction of molecules to identify specific proteins in a variety of tissues in a variety of stages in order to answer specific questions." This class will not only identify the building blocks of immunohistochemistry, from rabbit monoclonal antibodies to polymer detection kits, but we will also review how each step before the staining can affect those building blocks. This class will also address some basic dilution techniques for concentrated antibodies, the use of positive and negative controls, where to start trouble shooting your new antibody. Finally, we will discuss the utility of IHC in the clinical and research setting and its relevance to drug discovery and disease treatment. Workshop #3 Immunocytochemical and Immunohistochemical assays. A Pathologist's perspective, Lourdes R. Ylagan MD, Washington University. This workshop will show a brief overview of both immunohistochemical and immunocytochemical techniques. It will also show examples of how double immunostaining techniques can be helpful in both histologic and cytologic samples. It will also show the ways in which immunochemistry is used in: (1) the elucidation of the site of origin of a poorly differentiated neoplasm in the setting of a metastasis, (2) detection of antigens present in the surface of tumor cells amenable to antibody-mediated chemotherapeutic agents, (3) detection of tumor markers known to be potential poor prognostic indicators in tumors. Workshop #4 (Sponsored by Leica Microsystems) Quality and Skill in Microtomy Technique - Jan Minshew, Leica MicroSystems This session will present an understanding of the physical dynamics of microtomy, and discuss the effects of external variables such as knife angle, room temperature, inadequate instrumentation, etc. Workshop #5 (Sponsored by Ventana Medical Systems) Special Stains: the Once and Future King? Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Special Stains are an art of using dyes to stain specific tissues and/or cellular structures that were once thought of as a major advancement in the histology lab. Now we have much more complex chemicals to stain proteins and genes within those cells and tissues, yet we still find a vast usage of special stains in the histology lab. In fact, some of the specials stains have become so routine that they are oftentimes considered as routine as the H&E in certain circumstances. Why do we still utilize special stains when we have far less hazardous chemicals that can pinpoint much more specific things? This class will address that question. We will discuss where special stains has been, why they have been around for so long, and where we see them going. This class will break down the most frequently used special stains; discuss their bio-chemical reactions and their utility. We will then compare those special stains to their relevant immunohistochemistry and in-situ hybridization counterparts to illustrate their utility and futility . Workshop #6 >From Bench Tech to Management, Konnie Zietner, Nebraska Medical Center Basic principles and skills you will need when transitioning from technician to management. From sfarley <@t> seattlecca.org Tue May 10 14:34:23 2005 From: sfarley <@t> seattlecca.org (Farley, Sunni R) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Master's of Histology Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948323349@wala01.seattlecca.org> I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From naje1972 <@t> yahoo.com Tue May 10 16:54:16 2005 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] job Message-ID: <20050510215416.51974.qmail@web41710.mail.yahoo.com> Hi! We are looking for people who might be interested in a pool position in the Chicago land area. Please contact me Cynthia Haynes at 1-773-484-4133. Thanks in Advance Cynthia Haynes H.T. __________________________________ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. http://mobile.yahoo.com/learn/mail From MajorFocus <@t> aol.com Tue May 10 17:56:59 2005 From: MajorFocus <@t> aol.com (MajorFocus@aol.com) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Re: Slide Labeller Message-ID: <1f4.978eaa2.2fb2963b@aol.com> I have used the Thermo Microwriter as well which is quite nice. If you are interested in an inexpensive software program for generating slide labels, check out our website for the Histo-Labels program. It will also interface with most slide and cassette printers/etchers as well. _www.majorfocus.com_ (http://www.majorfocus.com) Greg L. Good, HT(ASCP) Major Focus (800) 888-1152 From laurie.reilly <@t> jcu.edu.au Tue May 10 18:36:52 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Mast cell stain In-Reply-To: <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp. com> Message-ID: <5.1.0.14.0.20050511092655.019a6ec0@mail.jcu.edu.au> Hi Histonetters, We routinely stain all dog skin lesions with Toluidine Blue and it highlights the Mast Cells. They stain metachromatically and show up with red-purple granules in the cytoplasm. These biopsies are all fixed in 10% Neutral Buffered Formalin. The method we use is from Gretchen Humason, "Animal tissue Techniques", 3rd edition. It uses a solution of 0.2% Tol Blue in 60% Ethanol for 2 minutes. Regards, Laurie. At 01:14 PM 05/10/05 -0400, Osborn, Sharon wrote: >Tom, > >Luna's Toluidine Blue Method for Mast Cells is a superior demonstration of >mast cells using a pink background. These literally pop up under low power. >The method is located on pages 311-312 of Luna's Histopathologic Methods and >color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a >copy of the procedure, private email me and I will send to you. > >sharon osborn >DNAX ScheringPlough BioPharma >Palo Alto, CA > >Subject: [Histonet] Giemsa Stain for Mast Cells >Netters, >I am looking for a Giemsa stain for Mast Cells that does not have a pinkish >hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out >at us on low power. Any suggestions/protocols? >Thanks, >Tom >Tom Galati >Laboratory Director >HSRL, Inc.- A GLP Compliant Contract Laboratory >137 South Main Street >Woodstock, Virginia 22664 >(540)459-8211 >Fax: (540)459-8217 >tomgalati@hsrl.org >www.hsrl.org > > > >********************************************************************* >This message and any attachments are solely for the intended recipient. If >you are not the intended recipient, disclosure, copying, use or >distribution of the information included in this message is prohibited -- >Please immediately and permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From ploykasek <@t> phenopath.com Tue May 10 18:41:50 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] job In-Reply-To: <20050510215416.51974.qmail@web41710.mail.yahoo.com> Message-ID: You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From histosci <@t> shentel.net Tue May 10 19:32:13 2005 From: histosci <@t> shentel.net (HSRL) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Mast Cell/Giemsa clarification Message-ID: <000601c555c0$e5a31840$0200a8c0@HSRLMAIN> Netters, I agree with all of you about the Tol Blue stain being a good mast cell stain. However, we are specifically looking for a Giemsa stain for mast cell granules. We already do the Tol Blue, and the Pinacynol Erythrosinate stains for mast cells. Thanks again, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org From Kemlo.Rogerson <@t> elht.nhs.uk Wed May 11 01:42:03 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Master's of Histology Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2A7@bhrv-nt-11.bhrv.nwest.nhs.uk> In the UK you can obtain a Master of Science Degree by pursuing a research project. In fact you could always take a Master's or Doctorate if you could find a Uni, a Supervisor and something of interest to research. I feel that postgraduate degrees have become easier to obtain, but then we old people always say that. In the UK NVQ's and other vocational qualifications are becoming more popular as it is really a judgement call, and I'm going to get into trouble, but maybe postgraduate degrees could be thought of as an overkill for many positions in the Laboratory. Do we really have that much to offer MSc's and PHD's? Other than funding them to get the qualification; I fess up to having an MSc, but...... -----Original Message----- From: Farley, Sunni R [mailto:sfarley@seattlecca.org] Sent: 10 May 2005 20:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Master's of Histology I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eswary <@t> carif.com.my Wed May 11 04:07:14 2005 From: eswary <@t> carif.com.my (eswary) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] (no subject) Message-ID: <200505111707.AA254607564@carif.com.my> hi, does anyone know the advantage of using 0.01M sodium citrate over 0.01M citric acid buffer, pH 6, for antigen retrieval on paraffin embedded sections? also, is the sodium constituent important? would it matter if i used trisodium citrate instead of monosodium? thanks. et From bernaweston <@t> hotmail.com Wed May 11 07:31:25 2005 From: bernaweston <@t> hotmail.com (Bernadette Weston) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] EDTA Message-ID: We can no longer supply EDTA for our bone marrow procedures, can some one recommend a substitution for aspirate collection? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton, OH From wasielewski.reinhard.von <@t> mh-hannover.de Wed May 11 07:40:33 2005 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] macrophages in mouse FFPE Message-ID: <42821961.31188.FCDE1BE2@localhost> Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski From kbroomal <@t> NEMOURS.ORG Wed May 11 07:46:42 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Gayle - floating petri dish snap freezing question Message-ID: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours.org> Gayle, I tried out your snap freezing method with the floating petri dish in the liquid nitrogen yesterday for some itsy bitsy GI biopsies. I really like it! I have a question though. When using this method, do you place your sample in the bottom of your mold, then OCT on top, or put a dab (or more) of OCT, then the tissue, ... or does it really matter? Thanks, Kristen Broomall, HT (ASCP) From Evelyn.Flynn <@t> childrens.harvard.edu Wed May 11 08:21:33 2005 From: Evelyn.Flynn <@t> childrens.harvard.edu (Flynn, Evelyn) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] macrophages in mouse FFPE References: <42821961.31188.FCDE1BE2@localhost> Message-ID: Dear Reinhard, I have had good results with rat anti-mouse Mac-3 antibody from PharMingen (Cat. # 550292). Regards, Evelyn Flynn, M.A. Children's Hospital, Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of wasielewski.reinhard.von@mh-hannover.de Sent: Wed 5/11/2005 8:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] macrophages in mouse FFPE Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Wed May 11 09:08:31 2005 From: emerald_lake77 <@t> yahoo.com (- -) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] New rabbit antibody... but what do I use as negative (isotype) control? Message-ID: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> Hi everyone, I have a new antibody that I was given. It is a rabbit polyclonal anti-mouse antibody. Usually I use a specific isotype control, but I do not have info on how this antibody was made other than the epitope(s) that it should adhere to... but I want a negative control and was wondering if I can just use regular rabbit serum (diluted) as a negative seeing that it should contain the isotype IgG I am using??? I think I am a bit confused here.... I need assistance. Can I use rabbit serum as a negative control? What dilution do I use if that is the case? Anything else I should know before I proceed? Thank you all!! --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! From NMargaryan <@t> childrensmemorial.org Wed May 11 09:26:25 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] job Message-ID: <63B8B599DE283148B92E83C78B32C15DB9164E@cmhexbe2.childrensmemorial.org> What do you mean Pool position? Thanks, Naira -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Tuesday, May 10, 2005 6:42 PM To: histonet Subject: Re: [Histonet] job You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. From tflore <@t> lsuhsc.edu Wed May 11 09:28:22 2005 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:25:03 2005 Subject: [Histonet] Unsubscribe Message-ID: Please unsubscribe. From tflore <@t> lsuhsc.edu Wed May 11 09:28:22 2005 From: tflore <@t> lsuhsc.edu (Flores, Teresa) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Unsubscribe Message-ID: Please unsubscribe. From vsailes <@t> nd.edu Wed May 11 09:44:37 2005 From: vsailes <@t> nd.edu (vsailes@nd.edu) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Help!!! Message-ID: <1115822677.42821a55c5ced@webmail.nd.edu> Hello Histonetters, I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues. I'm new to working out conditons for antibody protocols and could use some help.I'm testing the antibody at different dilutions and I appear to have positive staining but I'm also seeing small areas in the negative that shows positive staining. Liver is my control and I use steamer (HIER) for 30 minutes, my steps include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, Secondary antibody(Biotin), Streptavidin, Chromogen, Counterstain My concern is that I'm using buffers that contain BSA and I'm wondering if that is causing staining to appear or should I be using avidin/biotin block along with a peroxide block. Any suggestion would be helpful. Thank you for your help in advance. Valerie From LINDA.MARGRAF <@t> childrens.com Wed May 11 10:05:33 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] job listing Message-ID: Lead and/or Bench Histology Technician Scott & White is seeking a Lead and/or Bench Histology Technician. Responsible for histology laboratory operations including testing, training of support personnel, QA, personnel utilization and regulatory compliance. Selected candidate will also participate in routine histology laboratory functions. Requirements: An Associate's degree; HT(ASCP); 3 to 5 years experience. Just an hour north of Austin, Scott & White offers highly competitive salaries, comprehensive benefits and advancement opportunities. Candidates should send resumes to: ghollie@swmail.sw.org. Phone: 800.527.JOBS. Fax: 254.724.5591. Apply in person or mail resume to: Human Resources Dept., 2401 S. 31st St., Temple, TX 77508. www.sw.org An equal opportunity employer/tobacco-free environment. From katri <@t> cogeco.ca Wed May 11 10:10:05 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Help!!! References: <1115822677.42821a55c5ced@webmail.nd.edu> Message-ID: <000801c5563b$832b4f40$6a9a9618@Katri> Hi Valerie, I would recommend at least trying the biotin block. Certain tissues (liver, kidney) are rich in endogenous biotin and although formalin fixation often masks it, the HIER will bring it back. Katri ----- Original Message ----- From: To: Sent: Wednesday, May 11, 2005 10:44 AM Subject: [Histonet] Help!!! > Hello Histonetters, > I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues. > I'm > new to working out conditons for antibody protocols and could use some > help.I'm > testing the antibody at different dilutions and I appear to have positive > staining but I'm also seeing small areas in the negative that shows > positive > staining. Liver is my control and I use steamer (HIER) for 30 minutes, my > steps > include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, > Secondary > antibody(Biotin), Streptavidin, Chromogen, Counterstain > My concern is that I'm using buffers that contain BSA and I'm wondering if > that > is causing staining to appear or should I be using avidin/biotin block > along > with a peroxide block. Any suggestion would be helpful. Thank you for your > help > in advance. > Valerie > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed May 11 10:32:24 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] antibody used for macrophages in mouse FFPE In-Reply-To: <42821961.31188.FCDE1BE2@localhost> References: <42821961.31188.FCDE1BE2@localhost> Message-ID: <6.0.0.22.1.20050511093052.01b84780@gemini.msu.montana.edu> F4/80 clone is reported to work, and this protocol has been posted on Histonet several times. Go to www.histosearch.org, and search for the info you need. SEROTEC sells this antibody. At 06:40 AM 5/11/2005, you wrote: >Hi Histonetters, >I am looking for a suitable antibody working in FFPE mouse tissue to detect >macrophages (reproducibly and reliable). > >Many thanks in advance >Reinhard > > > > > >PD Dr. med. Reinhard von Wasielewski > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed May 11 10:41:54 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] New rabbit antibody... but what do I use as negative (isotype) control? In-Reply-To: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> References: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20050511093412.01b4b0f8@gemini.msu.montana.edu> Rabbit IgG will work - you can purchase from Jackson. This is a polyclonal, and if there is no isotype listed, whole IgG will suffice. The concentration of the IgG should match the concentration of your primary antibody, and calculate it in terms of ug/ml - a little math. Rather than rely on some dilution, you will dilute the Rb IgG, but its original concentration in mg/ml may be much higher than the primary antibody's mg/ml, meaning you might be using a different dilution for Rb IgG than you use for the rabbit primary antibody. You could use rabbit serum, but it should be heat inactivated - if the polyclonal is affinity purified and the pooled serum is not, you may get more nonspecific binding with serum. Rabbit IgG would be my choice. t 08:08 AM 5/11/2005, you wrote: >Hi everyone, > >I have a new antibody that I was given. It is a rabbit polyclonal >anti-mouse antibody. Usually I use a specific isotype control, but I do >not have info on how this antibody was made other than the epitope(s) that >it should adhere to... but I want a negative control and was wondering if >I can just use regular rabbit serum (diluted) as a negative seeing that it >should contain the isotype IgG I am using??? I think I am a bit confused >here.... I need assistance. > >Can I use rabbit serum as a negative control? >What dilution do I use if that is the case? >Anything else I should know before I proceed? > >Thank you all!! > > > > >--------------------------------- >Discover Yahoo! > Use Yahoo! to plan a weekend, have fun online & more. Check it out! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From anh2006 <@t> med.cornell.edu Wed May 11 10:57:42 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] New rabbit antibody... but what do I use as negative (isotype) control? In-Reply-To: <6.0.0.22.1.20050511093412.01b4b0f8@gemini.msu.montana.edu> References: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> <6.0.0.22.1.20050511093412.01b4b0f8@gemini.msu.montana.edu> Message-ID: Hi Gayle, I couldn't agree more with what you have said below ... but what is the reason for heat inactivation? There is no worry of complement in dead, sectioned, fixed tissues. I don't HI my serum and have no problems with background. Thanks, Andrea At 9:41 AM -0600 5/11/05, Gayle Callis wrote: >Rabbit IgG will work - you can purchase from Jackson. This is a >polyclonal, and if there is no isotype listed, whole IgG will >suffice. > >The concentration of the IgG should match the concentration of your >primary antibody, and calculate it in terms of ug/ml - a little >math. Rather than rely on some dilution, you will dilute the Rb IgG, >but its original concentration in mg/ml may be much higher than the >primary antibody's mg/ml, meaning you might be using a different >dilution for Rb IgG than you use for the rabbit primary antibody. > >You could use rabbit serum, but it should be heat inactivated - if >the polyclonal is affinity purified and the pooled serum is not, you >may get more nonspecific binding with serum. Rabbit IgG would be my >choice. > -- From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed May 11 10:56:55 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Luna's toluidine blue; also Ventana Benchmark XT morphology Message-ID: I would appreciate it if I could also be sent details of Luna's toluidine blue stain for mast cells, and any others that are considered superior so I can compare with our current method. Thanks! Also - do any of you users of Ventana's Benchmark or Benchmark XT autoimmunostainer have a problem with poor morphology after heat retrieval - usually affecting collagen / fibro-fatty tissues, and also the occasional section detachment? We trialled the Benchmark which was fine but the Benchmark XT which we purchased a year ago produces this problem. It does not affect our positive control tissue (or very little) and I have been through every possibility within the department and with Ventana but to no avail. All ideas / clues welcome! Jacqui Malam Royal Lancaster Infirmary DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From KMH.02 <@t> ex.uchs.org Wed May 11 11:07:38 2005 From: KMH.02 <@t> ex.uchs.org (Hopkins, Karen) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] RE: RVU units Message-ID: <640B23A5DC4B234BB065E56F2DB30596D9634B@uchex2ucmc.uchs.org> I need to set up two tests and need to assign RVU units to two CPT codes in order to get the tests I need in our billing system. I understand this may be a touchy subject but if anyone is willing to share how many RVU units they assigned to the CPT codes for 88189 Flow cytometry interpretation and 88358 DNA ploidy Digital image analysis I would appreciate it greatly. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, May 11, 2005 11:10 AM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 18, Issue 14 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Mast cell stain (Osborn, Sharon) 2. Slide labelers (Osborn, Sharon) 3. Re: Asbestos studies (Gayle Callis) 4. Control for Mast cells (Gayle Callis) 5. Picrosirius red stain (Jeffrey Wu) 6. RE: Control for Mast cells (Jim Staruk) 7. Re: mast cell in guinea pig (John Kiernan) 8. RE: Asbestos studies (Shirley Powell) 9. Final notice: Missouri Society for Histotechnology Spring Symposium (Johnson, Teri) 10. Master's of Histology (Farley, Sunni R) 11. job (cynthia haynes) 12. Re: Slide Labeller (MajorFocus@aol.com) 13. Re: Mast cell stain (Laurie Reilly) 14. Re: job (Patti Loykasek) 15. Mast Cell/Giemsa clarification (HSRL) 16. RE: Master's of Histology (Kemlo Rogerson) 17. (no subject) (eswary) 18. EDTA (Bernadette Weston) 19. macrophages in mouse FFPE (wasielewski.reinhard.von@mh-hannover.de) 20. Gayle - floating petri dish snap freezing question (Kristen Broomall) 21. RE: macrophages in mouse FFPE (Flynn, Evelyn) 22. New rabbit antibody... but what do I use as negative (isotype) control? (- -) 23. RE: job (Margaryan, Naira) 24. Unsubscribe (Flores, Teresa) 25. Unsubscribe (Flores, Teresa) 26. Help!!! (vsailes@nd.edu) 27. job listing (LINDA MARGRAF) ---------------------------------------------------------------------- Message: 1 Date: Tue, 10 May 2005 13:14:25 -0400 From: "Osborn, Sharon" Subject: [Histonet] Mast cell stain To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> Content-Type: text/plain Tom, Luna's Toluidine Blue Method for Mast Cells is a superior demonstration of mast cells using a pink background. These literally pop up under low power. The method is located on pages 311-312 of Luna's Histopathologic Methods and color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a copy of the procedure, private email me and I will send to you. sharon osborn DNAX ScheringPlough BioPharma Palo Alto, CA Subject: [Histonet] Giemsa Stain for Mast Cells Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 2 Date: Tue, 10 May 2005 13:21:48 -0400 From: "Osborn, Sharon" Subject: [Histonet] Slide labelers To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> Content-Type: text/plain Cynthia, Contact your Leica representative and ask for a demonstration of their slide printers. I think you will be pleasantly surprised as to the amount of information and quietness. I have used several different slide labelers including the etch-a-sketch types and ink printer types; this one is the best of all I have used. Its footprint is a little larger than the others and well worth it. Sakura and Leica partnered on developing these; their main difference is in the software that is provided. Sakura uses off the shelf software while Leica specifically built the software on theirs and it is customizable for stand alone or for integration with MIS of an institution. And, the company service will also make a difference. So, I encourage you to demo several different types of slide labelers and find the one that best suits your needs. Sharon Osborn DNAX ScheringPlough BioPharma Palo Alto, CA Date: Tue, 10 May 2005 10:37:23 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: [Histonet] slide labeller To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I am interested in an automated slide labeler. I would like the capacity to do multiple slides with the same information as quiet as possible and of course I have limited space. Any suggestions would be appreciated. I also have tops to the bottle for a Ventana special stainer. I am happy to send to anyone interested. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 3 Date: Tue, 10 May 2005 11:25:43 -0600 From: Gayle Callis Subject: Re: [Histonet] Asbestos studies To: mtitford@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the Gulf >Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most recently, >a big lawsuit in Corpus Christi Texas accuses screening companies of >misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen in >an H&E section as brown, cylindrical, and beaded. In our laboratory we >digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter dissolved >in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 4 Date: Tue, 10 May 2005 11:28:28 -0600 From: Gayle Callis Subject: [Histonet] Control for Mast cells To: "Anna Inman" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org > > > > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis >Sent: Tuesday, May 10, 2005 8:51 AM >To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Giemsa Stain for Mast Cells > > > >Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is > >Churukian Shenk method for mast cells. You don't have to worry about so > >many other colors from Giemsa. If you want these, I will be happy to > >attach privately. > > > >At 07:45 AM 5/10/2005, you wrote: > > >Netters, > > > > > > > > > > > >I am looking for a Giemsa stain for Mast Cells that does not have a > > >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast > > >Cells pop out at us on low power. Any suggestions/protocols? > > > > > > > > > > > >Thanks, > > > > > > > > > > > >Tom > > > > > > > > > > > >Tom Galati > > > > > >Laboratory Director > > > > > >HSRL, Inc.- A GLP Compliant Contract Laboratory > > > > > >137 South Main Street > > > > > >Woodstock, Virginia 22664 > > > > > >(540)459-8211 > > > > > >Fax: (540)459-8217 > > > > > >tomgalati@hsrl.org > > > > > >www.hsrl.org > > > > > > > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Gayle Callis > >MT,HT,HTL(ASCP) > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 (lab with voice mail) > >406 994-4303 (FAX) > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >This electronic message and all contents contain information from St. >Mary's Hospital which may be attorney-client privileged, confidential or >otherwise protected from disclosure. The information is intended to be >for the addressee only. If you are not the addressee, any disclosure, >copy, distribution or use of the contents of this message is >prohibited. If you have received this electronic message in error, please >notify the sender immediately and destroy the original message and all >copies. Thank you Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Tue, 10 May 2005 13:30:54 -0400 From: "Jeffrey Wu" Subject: [Histonet] Picrosirius red stain To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" Hello all, I am trying to quantitate the amount of fibrosis in mouse cardiac tissue. I based my method on Dr Kiernan's excellent tutorial on picrosirius red staining (posted in 2000). Briefly, slides are deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); 75% ethanol (1x, 3 min); and running distilled water (10-15 min). Next, they are stained in saturated picrosirius red for 60 minutes and washed in acetic acid (2x, 3 min). They are dehydrated in 75% ethanol (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 min). Finally, the slides are mounted with Permount. For image processing, I am using circularly polarized light. Under the microscope, the collagen stains from pink to dark red (almost black), depending on its content; however, the background tissue appears green/blue. It is problematic because I quantify the collagen using ImagePro Plus software, in which I select certain colors. Especially with the pink and some dark blues, there is overlap between the collagen and tissue, causing overestimation of collagen. (Sorry for the long explanation.) I guess what I am seeking is the flaw in my procedure, causing the other tissue not to stain yellow. On a side note, I am using the correct picrosirius red stain, and the solution is saturated with crystals at the bottom. I have tried modifying washing times without any change. I used a short (2 min) 0.2% phosphomolybdic acid wash, which is supposed to reduce background, with no success as well. Once again, sorry for the long question. Any help with my problem would be greatly appreciated. Please do not hesitate to email me with any questions or anything that I have not made clear. Thank you in advance. J Wu ------------------------------ Message: 6 Date: Tue, 10 May 2005 13:52:27 -0400 From: "Jim Staruk" Subject: RE: [Histonet] Control for Mast cells To: "'Anna Inman'" , Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> Content-Type: text/plain; charset=us-ascii Anyone in diagnostic veterinary services gets mast cell tumors all of the time. I'll be glad to share some with someone in need (and I won't even charge you). Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 1:28 PM To: Anna Inman; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Control for Mast cells Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org ------------------------------ Message: 7 Date: Tue, 10 May 2005 13:58:36 -0400 From: John Kiernan Subject: Re: [Histonet] mast cell in guinea pig To: Histonet Message-ID: <4280F64C.CD1F5085@uwo.ca> Content-Type: text/plain; charset=us-ascii John Kiernan wrote: > > Probably you didn't see any mast cells in the > sections of guinea-pig tissues because there > were none to be seen. > > Mouse and rat mast cells are unusual in that their > granules are preserved and rendered insoluble by > aqueous fixatives such as buffered formaldehyde. > In most other species aqueous fixatives dissolve > out the mast cell granules. Alcoholic fixatives (eg > Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre > etc) will preserve and insolubilize mast cell granules > of any species. > > For more on this, see "The Mast Cells" by Hans Selye > (1965) Chapter 3. Also the "Notes & Queries section in > the current issue of Biotechnic & Histochemistry (Vol 80 > No 1, p.43-45). The latter is available at the publisher's > web site: > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c12 4b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1, 1;homemain,1,1; > > The metachromasia is due to heparin, the major > macromolecular anion of the granules. The staining > properties of heparin are similar to those of > cartilage matrix. Both materials carry a lot > of sulphate-ester groups. Unfortunately heparin > (except that of rats and mice) is water-soluble. > I do not know if any tryptase or chymase immunoreactivity > remains in mast cells whose granules have been > extracted by an aqueous fixative. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Elizabeth Chlipala wrote: > > > > Hello All > > > > I'm looking for a stain that will identify mast cells in guinea pigs. I > > have run both giemsa (modified dif quick) and 0.4% Toluidine Blue. I > > normally get very good staining of mast cells with the toluidine blue. > > I normally run this stain on mouse and rat tissue. This is the first > > time I have tried this stain in guinea pigs. Upon review of the slides > > I could not located one mast cell in 11 lung sections. I know they have > > to be there. Does the metachromatic nature of the t. blue stain have to > > do with the amount of histamine present in the cells? I have read in > > the literature that guinea pigs and humans have less histamine present > > in their mast cells than rats, hamsters and mice. Is anyone aware of a > > modification that will stain mast cells in guinea pig? Any help would > > be appreciated. I would prefer to stick with a histochemical method. > > I'm aware of Immunohistochemical staining for mast cell tryptase, but in > > researching this I could not find any references for guinea pig tissue. > > > > Thanks in advance. > > > > Liz > > > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > > Manager > > Premier Laboratory, LLC > > P.O. Box 18592 > > Boulder, Colorado 80308 > > Office: (303) 735-5001 > > Fax: (303) 735-3540 > > liz@premierlab.com > > www.premierlab.com > > > > Ship to Address: > > Premier Laboratory > > University of Colorado > > MCDB, Room A3B40 > > Boulder, Colorado 80309 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ ------------------------------ Message: 8 Date: Tue, 10 May 2005 14:15:25 -0400 From: Shirley Powell Subject: RE: [Histonet] Asbestos studies To: 'Gayle Callis' , mtitford@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <01LO3GS2JNLI8WX3KK@Macon2.Mercer.edu> Content-Type: text/plain; charset=us-ascii The histonet archives can be found at www.histosearch.com. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 12:26 PM To: mtitford@aol.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Asbestos studies If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the >Gulf Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most >recently, a big lawsuit in Corpus Christi Texas accuses screening >companies of misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen >in an H&E section as brown, cylindrical, and beaded. In our laboratory >we digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter >dissolved in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 10 May 2005 14:06:06 -0500 From: "Johnson, Teri" Subject: [Histonet] Final notice: Missouri Society for Histotechnology Spring Symposium To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" The Missouri Society for Histotechnology Spring Symposium When: May 20-21, 2005 Where: The Lodge of the Four Seasons, Lake Ozark, Missouri (800) 843-5253 www.4seasonsresort.com Note: Rooms for 'The Lodge' may be fully booked so here are 2 alternative places to stay: The Country Club (800) 964-6698 (approximately 1.5 miles away) The Resort at Port Arrowhead (800) 532-3575 (approximately 3 miles away) For additional meeting information please contact: Sharon Walsh Phone: (636) 305-9650 email: userwalsh@aol.com or Rosetta Barkley Phone: (913) 588-2737 email: rbarkley2@kumc.edu We invite you to participate in a unique learning experience. The program is outstanding this year and should have something for everyone. In addition there will be social activities and vendor displays. Please Note: All workshops are CEU approved. Friday Schedule: 8:30am "Ergonomics in the Workplace", Jan Minshew, Leica Microsystems 9:15am "Case Studies in Forensic Pathology", Dr. Michael Graham, St. Louis University 10:45am "Histochemistry of Special Stains", Jerry Fredenburgh 1:30pm-4:30pm Workshops #1, #2, or #3 (Your choice) 6:00-8:00pm - Cruise on the "Lake of the Ozarks" Come enjoy the lake and visit with exhibitor's and friends. Saturday Schedule: 8:15am "Creutzfeldt-Jakob Disease: Tissue Handling and Testing in the United Kingdom", Konnie Zietner, Nebraska Medical Center 9:15am "Histological Techniques in Hematopathology", Dr. Sharad Mather 10:45am "Applications for Microwave Decalcification Procedures", Skip Brown 1:30pm-4:30pm Workshops #4, #5, or #6 (Your choice) Registration Fees: Members - $50, Non-Members - $60 Additional Banquet Ticket - $25 2005 WORKSHOP DESCRIPTIONS Workshop#1 Theory of Routine & Microwave Fixation & Processing - Skip Brown & Jerry Fredenburgh The process of tissue fixation and processing will be illustrated; first from a molecular level to demonstrate what is actually happening at a micro level in the tissue; then from a macro level to show the gross effects on the tissue. Accelerated procedures will be introduced as a way to speed up the process without detrimental effects to the tissue. Workshop #2 (Sponsored by Ventana Medical Systems) A Tour Guide to Immunohistochemistry: How to get there and what to do when you arrive. - Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Immunohistochemistry is defined as, "A method of detecting the presence of specific proteins in cells or tissues." If only it was that easy. A better definition may be, "A method of standardizing each and every step in and out of your control working toward a complex chemical construction of molecules to identify specific proteins in a variety of tissues in a variety of stages in order to answer specific questions." This class will not only identify the building blocks of immunohistochemistry, from rabbit monoclonal antibodies to polymer detection kits, but we will also review how each step before the staining can affect those building blocks. This class will also address some basic dilution techniques for concentrated antibodies, the use of positive and negative controls, where to start trouble shooting your new antibody. Finally, we will discuss the utility of IHC in the clinical and research setting and its relevance to drug discovery and disease treatment. Workshop #3 Immunocytochemical and Immunohistochemical assays. A Pathologist's perspective, Lourdes R. Ylagan MD, Washington University. This workshop will show a brief overview of both immunohistochemical and immunocytochemical techniques. It will also show examples of how double immunostaining techniques can be helpful in both histologic and cytologic samples. It will also show the ways in which immunochemistry is used in: (1) the elucidation of the site of origin of a poorly differentiated neoplasm in the setting of a metastasis, (2) detection of antigens present in the surface of tumor cells amenable to antibody-mediated chemotherapeutic agents, (3) detection of tumor markers known to be potential poor prognostic indicators in tumors. Workshop #4 (Sponsored by Leica Microsystems) Quality and Skill in Microtomy Technique - Jan Minshew, Leica MicroSystems This session will present an understanding of the physical dynamics of microtomy, and discuss the effects of external variables such as knife angle, room temperature, inadequate instrumentation, etc. Workshop #5 (Sponsored by Ventana Medical Systems) Special Stains: the Once and Future King? Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Special Stains are an art of using dyes to stain specific tissues and/or cellular structures that were once thought of as a major advancement in the histology lab. Now we have much more complex chemicals to stain proteins and genes within those cells and tissues, yet we still find a vast usage of special stains in the histology lab. In fact, some of the specials stains have become so routine that they are oftentimes considered as routine as the H&E in certain circumstances. Why do we still utilize special stains when we have far less hazardous chemicals that can pinpoint much more specific things? This class will address that question. We will discuss where special stains has been, why they have been around for so long, and where we see them going. This class will break down the most frequently used special stains; discuss their bio-chemical reactions and their utility. We will then compare those special stains to their relevant immunohistochemistry and in-situ hybridization counterparts to illustrate their utility and futility . Workshop #6 >From Bench Tech to Management, Konnie Zietner, Nebraska Medical Center Basic principles and skills you will need when transitioning from technician to management. ------------------------------ Message: 10 Date: Tue, 10 May 2005 12:34:23 -0700 From: "Farley, Sunni R" Subject: [Histonet] Master's of Histology To: histonet@lists.utsouthwestern.edu Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948323349@wala01.seattlecca.org> Content-Type: text/plain; charset="ISO-8859-1" I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. ------------------------------ Message: 11 Date: Tue, 10 May 2005 14:54:16 -0700 (PDT) From: cynthia haynes Subject: [Histonet] job To: Histonet@lists.utsouthwestern.edu Message-ID: <20050510215416.51974.qmail@web41710.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi! We are looking for people who might be interested in a pool position in the Chicago land area. Please contact me Cynthia Haynes at 1-773-484-4133. Thanks in Advance Cynthia Haynes H.T. __________________________________ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. http://mobile.yahoo.com/learn/mail ------------------------------ Message: 12 Date: Tue, 10 May 2005 18:56:59 EDT From: MajorFocus@aol.com Subject: [Histonet] Re: Slide Labeller To: cfavara@niaid.nih.gov Cc: histonet@lists.utsouthwestern.edu Message-ID: <1f4.978eaa2.2fb2963b@aol.com> Content-Type: text/plain; charset="US-ASCII" I have used the Thermo Microwriter as well which is quite nice. If you are interested in an inexpensive software program for generating slide labels, check out our website for the Histo-Labels program. It will also interface with most slide and cassette printers/etchers as well. _www.majorfocus.com_ (http://www.majorfocus.com) Greg L. Good, HT(ASCP) Major Focus (800) 888-1152 ------------------------------ Message: 13 Date: Wed, 11 May 2005 09:36:52 +1000 From: Laurie Reilly Subject: Re: [Histonet] Mast cell stain To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5.1.0.14.0.20050511092655.019a6ec0@mail.jcu.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi Histonetters, We routinely stain all dog skin lesions with Toluidine Blue and it highlights the Mast Cells. They stain metachromatically and show up with red-purple granules in the cytoplasm. These biopsies are all fixed in 10% Neutral Buffered Formalin. The method we use is from Gretchen Humason, "Animal tissue Techniques", 3rd edition. It uses a solution of 0.2% Tol Blue in 60% Ethanol for 2 minutes. Regards, Laurie. At 01:14 PM 05/10/05 -0400, Osborn, Sharon wrote: >Tom, > >Luna's Toluidine Blue Method for Mast Cells is a superior demonstration of >mast cells using a pink background. These literally pop up under low power. >The method is located on pages 311-312 of Luna's Histopathologic Methods and >color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a >copy of the procedure, private email me and I will send to you. > >sharon osborn >DNAX ScheringPlough BioPharma >Palo Alto, CA > >Subject: [Histonet] Giemsa Stain for Mast Cells >Netters, >I am looking for a Giemsa stain for Mast Cells that does not have a pinkish >hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out >at us on low power. Any suggestions/protocols? >Thanks, >Tom >Tom Galati >Laboratory Director >HSRL, Inc.- A GLP Compliant Contract Laboratory >137 South Main Street >Woodstock, Virginia 22664 >(540)459-8211 >Fax: (540)459-8217 >tomgalati@hsrl.org >www.hsrl.org > > > >********************************************************************* >This message and any attachments are solely for the intended recipient. If >you are not the intended recipient, disclosure, copying, use or >distribution of the information included in this message is prohibited -- >Please immediately and permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. ------------------------------ Message: 14 Date: Tue, 10 May 2005 16:41:50 -0700 From: Patti Loykasek Subject: Re: [Histonet] job To: histonet Message-ID: Content-Type: text/plain; charset="US-ASCII" You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Tue, 10 May 2005 20:32:13 -0400 From: "HSRL" Subject: [Histonet] Mast Cell/Giemsa clarification To: Message-ID: <000601c555c0$e5a31840$0200a8c0@HSRLMAIN> Content-Type: text/plain; charset="US-ASCII" Netters, I agree with all of you about the Tol Blue stain being a good mast cell stain. However, we are specifically looking for a Giemsa stain for mast cell granules. We already do the Tol Blue, and the Pinacynol Erythrosinate stains for mast cells. Thanks again, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ------------------------------ Message: 16 Date: Wed, 11 May 2005 07:42:03 +0100 From: Kemlo Rogerson Subject: RE: [Histonet] Master's of Histology To: "'Farley, Sunni R'" , histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2A7@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain In the UK you can obtain a Master of Science Degree by pursuing a research project. In fact you could always take a Master's or Doctorate if you could find a Uni, a Supervisor and something of interest to research. I feel that postgraduate degrees have become easier to obtain, but then we old people always say that. In the UK NVQ's and other vocational qualifications are becoming more popular as it is really a judgement call, and I'm going to get into trouble, but maybe postgraduate degrees could be thought of as an overkill for many positions in the Laboratory. Do we really have that much to offer MSc's and PHD's? Other than funding them to get the qualification; I fess up to having an MSc, but...... -----Original Message----- From: Farley, Sunni R [mailto:sfarley@seattlecca.org] Sent: 10 May 2005 20:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Master's of Histology I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Wed, 11 May 2005 17:07:14 +0800 From: "eswary" Subject: [Histonet] (no subject) To: Message-ID: <200505111707.AA254607564@carif.com.my> Content-Type: text/plain; charset=us-ascii hi, does anyone know the advantage of using 0.01M sodium citrate over 0.01M citric acid buffer, pH 6, for antigen retrieval on paraffin embedded sections? also, is the sodium constituent important? would it matter if i used trisodium citrate instead of monosodium? thanks. et ------------------------------ Message: 18 Date: Wed, 11 May 2005 08:31:25 -0400 From: "Bernadette Weston" Subject: [Histonet] EDTA To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed We can no longer supply EDTA for our bone marrow procedures, can some one recommend a substitution for aspirate collection? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton, OH ------------------------------ Message: 19 Date: Wed, 11 May 2005 14:40:33 +0200 From: wasielewski.reinhard.von@mh-hannover.de Subject: [Histonet] macrophages in mouse FFPE To: histonet@lists.utsouthwestern.edu Message-ID: <42821961.31188.FCDE1BE2@localhost> Content-Type: text/plain; charset=US-ASCII Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski ------------------------------ Message: 20 Date: Wed, 11 May 2005 08:46:42 -0400 From: "Kristen Broomall" Subject: [Histonet] Gayle - floating petri dish snap freezing question To: Histonet@lists.utsouthwestern.edu Message-ID: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours.org> Content-Type: text/plain; charset=iso-8859-1 Gayle, I tried out your snap freezing method with the floating petri dish in the liquid nitrogen yesterday for some itsy bitsy GI biopsies. I really like it! I have a question though. When using this method, do you place your sample in the bottom of your mold, then OCT on top, or put a dab (or more) of OCT, then the tissue, ... or does it really matter? Thanks, Kristen Broomall, HT (ASCP) ------------------------------ Message: 21 Date: Wed, 11 May 2005 09:21:33 -0400 From: "Flynn, Evelyn" Subject: RE: [Histonet] macrophages in mouse FFPE To: wasielewski.reinhard.von@mh-hannover.de, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Dear Reinhard, I have had good results with rat anti-mouse Mac-3 antibody from PharMingen (Cat. # 550292). Regards, Evelyn Flynn, M.A. Children's Hospital, Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of wasielewski.reinhard.von@mh-hannover.de Sent: Wed 5/11/2005 8:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] macrophages in mouse FFPE Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Wed, 11 May 2005 07:08:31 -0700 (PDT) From: - - Subject: [Histonet] New rabbit antibody... but what do I use as negative (isotype) control? To: histonet@lists.utsouthwestern.edu Message-ID: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi everyone, I have a new antibody that I was given. It is a rabbit polyclonal anti-mouse antibody. Usually I use a specific isotype control, but I do not have info on how this antibody was made other than the epitope(s) that it should adhere to... but I want a negative control and was wondering if I can just use regular rabbit serum (diluted) as a negative seeing that it should contain the isotype IgG I am using??? I think I am a bit confused here.... I need assistance. Can I use rabbit serum as a negative control? What dilution do I use if that is the case? Anything else I should know before I proceed? Thank you all!! --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! ------------------------------ Message: 23 Date: Wed, 11 May 2005 09:26:25 -0500 From: "Margaryan, Naira" Subject: RE: [Histonet] job To: "Patti Loykasek" , "histonet" Message-ID: <63B8B599DE283148B92E83C78B32C15DB9164E@cmhexbe2.childrensmemorial.org> Content-Type: text/plain What do you mean Pool position? Thanks, Naira -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Tuesday, May 10, 2005 6:42 PM To: histonet Subject: Re: [Histonet] job You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. ------------------------------ Message: 24 Date: Wed, 11 May 2005 09:28:22 -0500 From: "Flores, Teresa" Subject: [Histonet] Unsubscribe To: histonet@lists.utsouthwestern.edu Cc: histonet-request@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, "'histonet@pathology.swmed.edu'" Message-ID: Content-Type: text/plain Please unsubscribe. ------------------------------ Message: 25 Date: Wed, 11 May 2005 09:28:22 -0500 From: "Flores, Teresa" Subject: [Histonet] Unsubscribe To: histonet@lists.utsouthwestern.edu Cc: histonet-request@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, "'histonet@pathology.swmed.edu'" Message-ID: Content-Type: text/plain Please unsubscribe. ------------------------------ Message: 26 Date: Wed, 11 May 2005 09:44:37 -0500 From: vsailes@nd.edu Subject: [Histonet] Help!!! To: histonet@lists.utsouthwestern.edu Message-ID: <1115822677.42821a55c5ced@webmail.nd.edu> Content-Type: text/plain; charset=ISO-8859-1 Hello Histonetters, I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues. I'm new to working out conditons for antibody protocols and could use some help.I'm testing the antibody at different dilutions and I appear to have positive staining but I'm also seeing small areas in the negative that shows positive staining. Liver is my control and I use steamer (HIER) for 30 minutes, my steps include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, Secondary antibody(Biotin), Streptavidin, Chromogen, Counterstain My concern is that I'm using buffers that contain BSA and I'm wondering if that is causing staining to appear or should I be using avidin/biotin block along with a peroxide block. Any suggestion would be helpful. Thank you for your help in advance. Valerie ------------------------------ Message: 27 Date: Wed, 11 May 2005 10:05:33 -0500 From: "LINDA MARGRAF" Subject: [Histonet] job listing To: Message-ID: Content-Type: text/plain; charset=US-ASCII Lead and/or Bench Histology Technician Scott & White is seeking a Lead and/or Bench Histology Technician. Responsible for histology laboratory operations including testing, training of support personnel, QA, personnel utilization and regulatory compliance. Selected candidate will also participate in routine histology laboratory functions. Requirements: An Associate's degree; HT(ASCP); 3 to 5 years experience. Just an hour north of Austin, Scott & White offers highly competitive salaries, comprehensive benefits and advancement opportunities. Candidates should send resumes to: ghollie@swmail.sw.org. Phone: 800.527.JOBS. Fax: 254.724.5591. Apply in person or mail resume to: Human Resources Dept., 2401 S. 31st St., Temple, TX 77508. www.sw.org An equal opportunity employer/tobacco-free environment. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 14 **************************************** From gcallis <@t> montana.edu Wed May 11 11:17:30 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Re: floating petri dish snap freezing question In-Reply-To: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours. org> References: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours.org> Message-ID: <6.0.0.22.1.20050511094857.01b86140@gemini.msu.montana.edu> Kristen, I have done it both ways but if you let the tissue float up, it is harder to find in the OCT during sectioning. I often put a very thin OCT layer in mold then add tissue to have it located in bottom - then add OCT. After than you put mold in canoeing petri dish. It is sometime hard to push a delicate or several pieces into OCT and sort of "glues" them to bottom of mold. Another trick: Put tiny drop of OCT on a dark surface, add biopsies, then roll them into a tiny OCT ball, pick up (we like a curved eye forceps for this, center tissue/OCT into center/bottom of mold but when you add more OCT, go AROUND the little ball of tissues. The pressure of adding gooey OCT around tissues tends to keep everything centered. You tend to lose orientation with this method, something you may not like with gastric bx's. You can dip tiny tissue in OCT, add to mold so it stays oriented, then add OCT slowly and gently. Whatever you do, keep tip of OCT bottle (by laying bottle on side) filled to prevent bubbles - the enemy. tip of bottle can be cut off so hole is small and prevent huge flow of goo when dispensing OCT. One can always add OCT during freezing - so watch bottom turn white, but key word here is is during not after freezing, you do not want an interface of two OCT layers, they will snap part during sectioning at times. Been there, done that, and it was disaster. We purchase eye forceps for embedding in OCT - these are fine round tips, with either straight, half curved or full curved tips. Sharp points are NOT used, to easy to poke holes in tiny tissues. Arista in New York has huge selection and cheap. Another thing we are using is safety glasses that fit over prescription glasses, but the safety glasses have bifocal magnification - wonderful to see when handling tiny tissues. They come in 1, 1.5, 2.5 and 5 magnification - maybe other, available from Fisher, Newcomer Supply, and MarketLab. Hopefully this helps, so many ways. Have a good day At 06:46 AM 5/11/2005, you wrote: >Gayle, > >I tried out your snap freezing method with the floating petri dish in the >liquid nitrogen yesterday for some itsy bitsy GI biopsies. I really like it! >I have a question though. When using this method, do you place your sample >in the bottom of your mold, then OCT on top, or put a dab (or more) of OCT, >then the tissue, ... or does it really matter? > >Thanks, > >Kristen Broomall, HT (ASCP) > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Wed May 11 11:41:35 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] RE: RVU units In-Reply-To: <640B23A5DC4B234BB065E56F2DB30596D9634B@uchex2ucmc.uchs.org> Message-ID: I would like to see the answer posted on the histonet, too. Patti Loykasek PhenoPath Laboratories Seattle, WA > I need to set up two tests and need to assign RVU units to two > CPT codes in order to get the tests I need in our billing system. I > understand this may be a touchy subject but if anyone is willing to > share how many RVU units they assigned to the CPT codes for 88189 Flow > cytometry interpretation and 88358 DNA ploidy Digital image analysis I > would appreciate it greatly. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Wednesday, May 11, 2005 11:10 AM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 18, Issue 14 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Mast cell stain (Osborn, Sharon) > 2. Slide labelers (Osborn, Sharon) > 3. Re: Asbestos studies (Gayle Callis) > 4. Control for Mast cells (Gayle Callis) > 5. Picrosirius red stain (Jeffrey Wu) > 6. RE: Control for Mast cells (Jim Staruk) > 7. Re: mast cell in guinea pig (John Kiernan) > 8. RE: Asbestos studies (Shirley Powell) > 9. Final notice: Missouri Society for Histotechnology Spring > Symposium (Johnson, Teri) > 10. Master's of Histology (Farley, Sunni R) > 11. job (cynthia haynes) > 12. Re: Slide Labeller (MajorFocus@aol.com) > 13. Re: Mast cell stain (Laurie Reilly) > 14. Re: job (Patti Loykasek) > 15. Mast Cell/Giemsa clarification (HSRL) > 16. RE: Master's of Histology (Kemlo Rogerson) > 17. (no subject) (eswary) > 18. EDTA (Bernadette Weston) > 19. macrophages in mouse FFPE > (wasielewski.reinhard.von@mh-hannover.de) > 20. Gayle - floating petri dish snap freezing question > (Kristen Broomall) > 21. RE: macrophages in mouse FFPE (Flynn, Evelyn) > 22. New rabbit antibody... but what do I use as negative > (isotype) control? (- -) > 23. RE: job (Margaryan, Naira) > 24. Unsubscribe (Flores, Teresa) > 25. Unsubscribe (Flores, Teresa) > 26. Help!!! (vsailes@nd.edu) > 27. job listing (LINDA MARGRAF) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 10 May 2005 13:14:25 -0400 > From: "Osborn, Sharon" > Subject: [Histonet] Mast cell stain > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> > Content-Type: text/plain > > Tom, > > Luna's Toluidine Blue Method for Mast Cells is a superior demonstration > of > mast cells using a pink background. These literally pop up under low > power. > The method is located on pages 311-312 of Luna's Histopathologic Methods > and > color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a > copy of the procedure, private email me and I will send to you. > > sharon osborn > DNAX ScheringPlough BioPharma > Palo Alto, CA > > Subject: [Histonet] Giemsa Stain for Mast Cells > Netters, > I am looking for a Giemsa stain for Mast Cells that does not have a > pinkish > hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop > out > at us on low power. Any suggestions/protocols? > Thanks, > Tom > Tom Galati > Laboratory Director > HSRL, Inc.- A GLP Compliant Contract Laboratory > 137 South Main Street > Woodstock, Virginia 22664 > (540)459-8211 > Fax: (540)459-8217 > tomgalati@hsrl.org > www.hsrl.org > > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. > If you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited > -- Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 2 > Date: Tue, 10 May 2005 13:21:48 -0400 > From: "Osborn, Sharon" > Subject: [Histonet] Slide labelers > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> > Content-Type: text/plain > > Cynthia, > Contact your Leica representative and ask for a demonstration of > their slide printers. I think you will be pleasantly surprised as to > the > amount of information and quietness. I have used several different > slide > labelers including the etch-a-sketch types and ink printer types; this > one > is the best of all I have used. Its footprint is a little larger than > the > others and well worth it. Sakura and Leica partnered on developing > these; > their main difference is in the software that is provided. Sakura uses > off > the shelf software while Leica specifically built the software on theirs > and > it is customizable for stand alone or for integration with MIS of an > institution. And, the company service will also make a difference. > So, I encourage you to demo several different types of slide > labelers and find the one that best suits your needs. > Sharon Osborn > DNAX > ScheringPlough BioPharma > Palo Alto, CA > > > Date: Tue, 10 May 2005 10:37:23 -0400 > From: "Favara, Cynthia (NIH/NIAID)" > Subject: [Histonet] slide labeller > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain > > I am interested in an automated slide labeler. I would like the capacity > to > do multiple slides with the same information as quiet as possible and of > course I have limited space. Any suggestions would be appreciated. > > > > I also have tops to the bottle for a Ventana special stainer. I am happy > to > send to anyone interested. > > > > c > > > > Cynthia Favara > NIAID/NIH/RML/LPVD > 903 South 4th Street > Hamilton, MT 59840 > 406-363-9317 > > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. > If you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited > -- Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 3 > Date: Tue, 10 May 2005 11:25:43 -0600 > From: Gayle Callis > Subject: Re: [Histonet] Asbestos studies > To: mtitford@aol.com, Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > If I remember correctly, an iron stain will identify asbestos fibers > nicely > in a section. Histonet had discussion on asbestos fibers some time ago, > > may be worth a search in archives at www.histosearch.org > > At 09:52 AM 5/10/2005, you wrote: >> Fred Underwood asks about asbestos fibers in lung tissues. >> >> Asbestosis and mesothelioma studies are quite big business along the > Gulf >> Coast here with the local shipyards and men of the "Golden Generation" >> passing away. A lot of lawyers have made a lot of money and most > recently, >> a big lawsuit in Corpus Christi Texas accuses screening companies of >> misdiagnosis. >> However, onto things histological: asbestos fibers can be easily seen > in >> an H&E section as brown, cylindrical, and beaded. In our laboratory we >> digest lung tissue in bleach and then filter the remains through a >> millipore. Once dried, it can be placed on a slide, the filter > dissolved >> in chloroform and mounted with a coverslip. The fibers are then > counted. >> The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >> asbestos-associated diseases" 2ed edition Spinger. 2004 >> >> Mike Titford >> USA Pathology >> Mobile AL USA >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 4 > Date: Tue, 10 May 2005 11:28:28 -0600 > From: Gayle Callis > Subject: [Histonet] Control for Mast cells > To: "Anna Inman" , > Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Skin contains mast cells in connective tissues and have seen them in > connective tissues surrounding bones. > > At 10:15 AM 5/10/2005, you wrote: >> --> >> >> I am having difficulty getting a control for Mast cells. Any > suggestions? >> >> >> >> Anna Inman B.S., HT (ASCP) >> >> SMH Pathology >> >> Anna.Inman@stmarygj.org >> >> >> >> >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle > Callis >> Sent: Tuesday, May 10, 2005 8:51 AM >> To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >> Subject: Re: [Histonet] Giemsa Stain for Mast Cells >> >> >> >> Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is >> >> Churukian Shenk method for mast cells. You don't have to worry about > so >> >> many other colors from Giemsa. If you want these, I will be happy to >> >> attach privately. >> >> >> >> At 07:45 AM 5/10/2005, you wrote: >> >>> Netters, >> >>> >> >>> >> >>> >> >>> I am looking for a Giemsa stain for Mast Cells that does not have a >> >>> pinkish hue like the Gaffney Giemsa. We are looking to have the Mast >> >>> Cells pop out at us on low power. Any suggestions/protocols? >> >>> >> >>> >> >>> >> >>> Thanks, >> >>> >> >>> >> >>> >> >>> Tom >> >>> >> >>> >> >>> >> >>> Tom Galati >> >>> >> >>> Laboratory Director >> >>> >> >>> HSRL, Inc.- A GLP Compliant Contract Laboratory >> >>> >> >>> 137 South Main Street >> >>> >> >>> Woodstock, Virginia 22664 >> >>> >> >>> (540)459-8211 >> >>> >> >>> Fax: (540)459-8217 >> >>> >> >>> tomgalati@hsrl.org >> >>> >> >>> www.hsrl.org >> >>> >> >>> >> >>> >> >>> _______________________________________________ >> >>> Histonet mailing list >> >>> Histonet@lists.utsouthwestern.edu >> >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> Gayle Callis >> >> MT,HT,HTL(ASCP) >> >> Research Histopathology Supervisor >> >> Veterinary Molecular Biology >> >> Montana State University - Bozeman >> >> PO Box 173610 >> >> Bozeman MT 59717-3610 >> >> 406 994-6367 (lab with voice mail) >> >> 406 994-4303 (FAX) >> >> >> >> >> >> >> >> _______________________________________________ >> >> Histonet mailing list >> >> Histonet@lists.utsouthwestern.edu >> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> This electronic message and all contents contain information from St. >> Mary's Hospital which may be attorney-client privileged, confidential > or >> otherwise protected from disclosure. The information is intended to be > >> for the addressee only. If you are not the addressee, any disclosure, >> copy, distribution or use of the contents of this message is >> prohibited. If you have received this electronic message in error, > please >> notify the sender immediately and destroy the original message and all >> copies. Thank you > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 5 > Date: Tue, 10 May 2005 13:30:54 -0400 > From: "Jeffrey Wu" > Subject: [Histonet] Picrosirius red stain > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format="flowed" > > > Hello all, > > I am trying to quantitate the amount of fibrosis in mouse > cardiac > tissue. I based my method on Dr Kiernan's excellent tutorial > on > picrosirius red staining (posted in 2000). Briefly, slides > are > deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); > 75% > ethanol (1x, 3 min); and running distilled water (10-15 min). > Next, > they are stained in saturated picrosirius red for 60 minutes > and > washed in acetic acid (2x, 3 min). They are dehydrated in 75% > ethanol > (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 > min). > Finally, the slides are mounted with Permount. > > For image processing, I am using circularly polarized light. > Under > the microscope, the collagen stains from pink to dark red > (almost > black), depending on its content; however, the background > tissue > appears green/blue. It is problematic because I quantify > the > collagen using ImagePro Plus software, in which I select > certain > colors. Especially with the pink and some dark blues, there > is > overlap between the collagen and tissue, causing overestimation > of > collagen. (Sorry for the long explanation.) I guess what I > am > seeking is the flaw in my procedure, causing the other tissue not > to > stain yellow. > > On a side note, I am using the correct picrosirius red stain, and > the > solution is saturated with crystals at the bottom. I > have > tried modifying washing times without any change. I used a short > (2 > min) 0.2% phosphomolybdic acid wash, which is supposed to > reduce > background, with no success as well. > > Once again, sorry for the long question. Any help with my > problem > would be greatly appreciated. Please do not hesitate to email me > with > any questions or anything that I have not made clear. Thank you > in > advance. > > J Wu > > > ------------------------------ > > Message: 6 > Date: Tue, 10 May 2005 13:52:27 -0400 > From: "Jim Staruk" > Subject: RE: [Histonet] Control for Mast cells > To: "'Anna Inman'" , > > Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> > Content-Type: text/plain; charset=us-ascii > > Anyone in diagnostic veterinary services gets mast cell tumors all of > the > time. I'll be glad to share some with someone in need (and I won't even > charge you). > > Jim > > ______________________ > Jim Staruk > Mass Histology Service > www.masshistology.com > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle > Callis > Sent: Tuesday, May 10, 2005 1:28 PM > To: Anna Inman; Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Control for Mast cells > > Skin contains mast cells in connective tissues and have seen them in > connective tissues surrounding bones. > > At 10:15 AM 5/10/2005, you wrote: >> --> >> >> I am having difficulty getting a control for Mast cells. Any > suggestions? >> >> >> >> Anna Inman B.S., HT (ASCP) >> >> SMH Pathology >> >> Anna.Inman@stmarygj.org > > > > > > ------------------------------ > > Message: 7 > Date: Tue, 10 May 2005 13:58:36 -0400 > From: John Kiernan > Subject: Re: [Histonet] mast cell in guinea pig > To: Histonet > Message-ID: <4280F64C.CD1F5085@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > > > John Kiernan wrote: >> >> Probably you didn't see any mast cells in the >> sections of guinea-pig tissues because there >> were none to be seen. >> >> Mouse and rat mast cells are unusual in that their >> granules are preserved and rendered insoluble by >> aqueous fixatives such as buffered formaldehyde. >> In most other species aqueous fixatives dissolve >> out the mast cell granules. Alcoholic fixatives (eg >> Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre >> etc) will preserve and insolubilize mast cell granules >> of any species. >> >> For more on this, see "The Mast Cells" by Hans Selye >> (1965) Chapter 3. Also the "Notes & Queries section in >> the current issue of Biotechnic & Histochemistry (Vol 80 >> No 1, p.43-45). The latter is available at the publisher's >> web site: >> > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c12 > 4b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1, > 1;homemain,1,1; >> >> The metachromasia is due to heparin, the major >> macromolecular anion of the granules. The staining >> properties of heparin are similar to those of >> cartilage matrix. Both materials carry a lot >> of sulphate-ester groups. Unfortunately heparin >> (except that of rats and mice) is water-soluble. >> I do not know if any tryptase or chymase immunoreactivity >> remains in mast cells whose granules have been >> extracted by an aqueous fixative. >> -- >> ------------------------------- >> John A. Kiernan >> Department of Anatomy and Cell Biology >> The University of Western Ontario >> London, Canada N6A 5C1 >> kiernan[AT]uwo.ca >> http://publish.uwo.ca/~jkiernan/ >> http://instruct.uwo.ca/anatomy/530/index.htm >> _______________________________ >> Elizabeth Chlipala wrote: >>> >>> Hello All >>> >>> I'm looking for a stain that will identify mast cells in guinea > pigs. I >>> have run both giemsa (modified dif quick) and 0.4% Toluidine Blue. > I >>> normally get very good staining of mast cells with the toluidine > blue. >>> I normally run this stain on mouse and rat tissue. This is the > first >>> time I have tried this stain in guinea pigs. Upon review of the > slides >>> I could not located one mast cell in 11 lung sections. I know they > have >>> to be there. Does the metachromatic nature of the t. blue stain > have to >>> do with the amount of histamine present in the cells? I have read > in >>> the literature that guinea pigs and humans have less histamine > present >>> in their mast cells than rats, hamsters and mice. Is anyone aware > of a >>> modification that will stain mast cells in guinea pig? Any help > would >>> be appreciated. I would prefer to stick with a histochemical > method. >>> I'm aware of Immunohistochemical staining for mast cell tryptase, > but in >>> researching this I could not find any references for guinea pig > tissue. >>> >>> Thanks in advance. >>> >>> Liz >>> >>> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >>> Manager >>> Premier Laboratory, LLC >>> P.O. Box 18592 >>> Boulder, Colorado 80308 >>> Office: (303) 735-5001 >>> Fax: (303) 735-3540 >>> liz@premierlab.com >>> www.premierlab.com >>> >>> Ship to Address: >>> Premier Laboratory >>> University of Colorado >>> MCDB, Room A3B40 >>> Boulder, Colorado 80309 >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Wed May 11 12:47:50 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Re: floating petri dish snap freezing question In-Reply-To: <6.0.0.22.1.20050511094857.01b86140@gemini.msu.montana.edu> References: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours. org> Message-ID: <42820D07.7788.109264E@localhost> Dear Kristen/ Gayle, To see my detailed reply to a similar question on cryotomy, check the following reference: Biotechnic & Histochemistry 2003, 78(5): 279. The "Notes and Queries" section. Cheers! Greg Date sent: Wed, 11 May 2005 10:17:30 -0600 To: "Kristen Broomall" , Histonet@lists.utsouthwestern.edu From: Gayle Callis Copies to: Subject: [Histonet] Re: floating petri dish snap freezing question > Kristen, > > I have done it both ways but if you let the tissue float up, it is > harder to find in the OCT during sectioning. I often put a very thin > OCT layer in mold then add tissue to have it located in bottom - then > add OCT. After than you put mold in canoeing petri dish. It is > sometime hard to push a delicate or several pieces into OCT and sort > of "glues" them to bottom of mold. > > Another trick: Put tiny drop of OCT on a dark surface, add biopsies, > then roll them into a tiny OCT ball, pick up (we like a curved eye > forceps for this, center tissue/OCT into center/bottom of mold but > when you add more OCT, go AROUND the little ball of tissues. The > pressure of adding gooey OCT around tissues tends to keep everything > centered. You tend to lose orientation with this method, something > you may not like with gastric bx's. > > You can dip tiny tissue in OCT, add to mold so it stays oriented, then > add OCT slowly and gently. Whatever you do, keep tip of OCT bottle (by > laying bottle on side) filled to prevent bubbles - the enemy. tip of > bottle can be cut off so hole is small and prevent huge flow of goo > when dispensing OCT. One can always add OCT during freezing - so watch > bottom turn white, but key word here is is during not after freezing, > you do not want an interface of two OCT layers, they will snap part > during sectioning at times. Been there, done that, and it was > disaster. > > We purchase eye forceps for embedding in OCT - these are fine round > tips, with either straight, half curved or full curved tips. Sharp > points are NOT used, to easy to poke holes in tiny tissues. Arista > in New York has huge selection and cheap. > > Another thing we are using is safety glasses that fit over > prescription glasses, but the safety glasses have bifocal > magnification - wonderful to see when handling tiny tissues. They > come in 1, 1.5, 2.5 and 5 magnification - maybe other, available from > Fisher, Newcomer Supply, and MarketLab. > > Hopefully this helps, so many ways. > > Have a good day > > At 06:46 AM 5/11/2005, you wrote: > >Gayle, > > > >I tried out your snap freezing method with the floating petri dish in > >the liquid nitrogen yesterday for some itsy bitsy GI biopsies. I > >really like it! I have a question though. When using this method, do > >you place your sample in the bottom of your mold, then OCT on top, or > >put a dab (or more) of OCT, then the tissue, ... or does it really > >matter? > > > >Thanks, > > > >Kristen Broomall, HT (ASCP) > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From Barry.R.Rittman <@t> uth.tmc.edu Wed May 11 12:02:26 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] RE: RVU units Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0018B1A26@UTHEVS3.mail.uthouston.edu> Patti You have raised an interesting question that I feel should be discussed on Histonet. There seems to be a national tendency to use RVUs to compare different procedures and provide some type of standardization. The same process is being examined in order to compare teaching loads. On the surface it would appear to be desirable. However there are some drawbacks that I have seen in discussions of this topic for teaching. If x number of units are for a lecture given for the first time this does not take into account the skill of the teacher, the availability of resources or the different fields of teaching, biochemistry, histology etc. I would suggest that the same applies to your question. While it is nice to be able to compare the RVU of a specific technique that you are carrying out with that used by other laboratories in the field it may not be a fair comparison. The actual cost to you depends on several factors such as, skill of the individual, efficiency of technique and equipment (which may depend on newness of machine etc), availability of equipment and resources etc., etc. This is a complex problem that I feel would be in laboratories interest to handle at a local level. Just my opinion. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Wednesday, May 11, 2005 11:42 AM To: histonet Subject: Re: [Histonet] RE: RVU units I would like to see the answer posted on the histonet, too. Patti Loykasek PhenoPath Laboratories Seattle, WA > I need to set up two tests and need to assign RVU units to two > CPT codes in order to get the tests I need in our billing system. I > understand this may be a touchy subject but if anyone is willing to > share how many RVU units they assigned to the CPT codes for 88189 Flow > cytometry interpretation and 88358 DNA ploidy Digital image analysis I > would appreciate it greatly. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Wednesday, May 11, 2005 11:10 AM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 18, Issue 14 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Mast cell stain (Osborn, Sharon) > 2. Slide labelers (Osborn, Sharon) > 3. Re: Asbestos studies (Gayle Callis) > 4. Control for Mast cells (Gayle Callis) > 5. Picrosirius red stain (Jeffrey Wu) > 6. RE: Control for Mast cells (Jim Staruk) > 7. Re: mast cell in guinea pig (John Kiernan) > 8. RE: Asbestos studies (Shirley Powell) > 9. Final notice: Missouri Society for Histotechnology Spring > Symposium (Johnson, Teri) > 10. Master's of Histology (Farley, Sunni R) > 11. job (cynthia haynes) > 12. Re: Slide Labeller (MajorFocus@aol.com) > 13. Re: Mast cell stain (Laurie Reilly) > 14. Re: job (Patti Loykasek) > 15. Mast Cell/Giemsa clarification (HSRL) > 16. RE: Master's of Histology (Kemlo Rogerson) > 17. (no subject) (eswary) > 18. EDTA (Bernadette Weston) > 19. macrophages in mouse FFPE > (wasielewski.reinhard.von@mh-hannover.de) > 20. Gayle - floating petri dish snap freezing question > (Kristen Broomall) > 21. RE: macrophages in mouse FFPE (Flynn, Evelyn) > 22. New rabbit antibody... but what do I use as negative > (isotype) control? (- -) > 23. RE: job (Margaryan, Naira) > 24. Unsubscribe (Flores, Teresa) > 25. Unsubscribe (Flores, Teresa) > 26. Help!!! (vsailes@nd.edu) > 27. job listing (LINDA MARGRAF) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 10 May 2005 13:14:25 -0400 > From: "Osborn, Sharon" > Subject: [Histonet] Mast cell stain > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> > Content-Type: text/plain > > Tom, > > Luna's Toluidine Blue Method for Mast Cells is a superior demonstration > of > mast cells using a pink background. These literally pop up under low > power. > The method is located on pages 311-312 of Luna's Histopathologic Methods > and > color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a > copy of the procedure, private email me and I will send to you. > > sharon osborn > DNAX ScheringPlough BioPharma > Palo Alto, CA > > Subject: [Histonet] Giemsa Stain for Mast Cells > Netters, > I am looking for a Giemsa stain for Mast Cells that does not have a > pinkish > hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop > out > at us on low power. Any suggestions/protocols? > Thanks, > Tom > Tom Galati > Laboratory Director > HSRL, Inc.- A GLP Compliant Contract Laboratory > 137 South Main Street > Woodstock, Virginia 22664 > (540)459-8211 > Fax: (540)459-8217 > tomgalati@hsrl.org > www.hsrl.org > > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. > If you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited > -- Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 2 > Date: Tue, 10 May 2005 13:21:48 -0400 > From: "Osborn, Sharon" > Subject: [Histonet] Slide labelers > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> > Content-Type: text/plain > > Cynthia, > Contact your Leica representative and ask for a demonstration of > their slide printers. I think you will be pleasantly surprised as to > the > amount of information and quietness. I have used several different > slide > labelers including the etch-a-sketch types and ink printer types; this > one > is the best of all I have used. Its footprint is a little larger than > the > others and well worth it. Sakura and Leica partnered on developing > these; > their main difference is in the software that is provided. Sakura uses > off > the shelf software while Leica specifically built the software on theirs > and > it is customizable for stand alone or for integration with MIS of an > institution. And, the company service will also make a difference. > So, I encourage you to demo several different types of slide > labelers and find the one that best suits your needs. > Sharon Osborn > DNAX > ScheringPlough BioPharma > Palo Alto, CA > > > Date: Tue, 10 May 2005 10:37:23 -0400 > From: "Favara, Cynthia (NIH/NIAID)" > Subject: [Histonet] slide labeller > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain > > I am interested in an automated slide labeler. I would like the capacity > to > do multiple slides with the same information as quiet as possible and of > course I have limited space. Any suggestions would be appreciated. > > > > I also have tops to the bottle for a Ventana special stainer. I am happy > to > send to anyone interested. > > > > c > > > > Cynthia Favara > NIAID/NIH/RML/LPVD > 903 South 4th Street > Hamilton, MT 59840 > 406-363-9317 > > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. > If you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited > -- Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 3 > Date: Tue, 10 May 2005 11:25:43 -0600 > From: Gayle Callis > Subject: Re: [Histonet] Asbestos studies > To: mtitford@aol.com, Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > If I remember correctly, an iron stain will identify asbestos fibers > nicely > in a section. Histonet had discussion on asbestos fibers some time ago, > > may be worth a search in archives at www.histosearch.org > > At 09:52 AM 5/10/2005, you wrote: >> Fred Underwood asks about asbestos fibers in lung tissues. >> >> Asbestosis and mesothelioma studies are quite big business along the > Gulf >> Coast here with the local shipyards and men of the "Golden Generation" >> passing away. A lot of lawyers have made a lot of money and most > recently, >> a big lawsuit in Corpus Christi Texas accuses screening companies of >> misdiagnosis. >> However, onto things histological: asbestos fibers can be easily seen > in >> an H&E section as brown, cylindrical, and beaded. In our laboratory we >> digest lung tissue in bleach and then filter the remains through a >> millipore. Once dried, it can be placed on a slide, the filter > dissolved >> in chloroform and mounted with a coverslip. The fibers are then > counted. >> The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >> asbestos-associated diseases" 2ed edition Spinger. 2004 >> >> Mike Titford >> USA Pathology >> Mobile AL USA >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 4 > Date: Tue, 10 May 2005 11:28:28 -0600 > From: Gayle Callis > Subject: [Histonet] Control for Mast cells > To: "Anna Inman" , > Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Skin contains mast cells in connective tissues and have seen them in > connective tissues surrounding bones. > > At 10:15 AM 5/10/2005, you wrote: >> --> >> >> I am having difficulty getting a control for Mast cells. Any > suggestions? >> >> >> >> Anna Inman B.S., HT (ASCP) >> >> SMH Pathology >> >> Anna.Inman@stmarygj.org >> >> >> >> >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle > Callis >> Sent: Tuesday, May 10, 2005 8:51 AM >> To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >> Subject: Re: [Histonet] Giemsa Stain for Mast Cells >> >> >> >> Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is >> >> Churukian Shenk method for mast cells. You don't have to worry about > so >> >> many other colors from Giemsa. If you want these, I will be happy to >> >> attach privately. >> >> >> >> At 07:45 AM 5/10/2005, you wrote: >> >>> Netters, >> >>> >> >>> >> >>> >> >>> I am looking for a Giemsa stain for Mast Cells that does not have a >> >>> pinkish hue like the Gaffney Giemsa. We are looking to have the Mast >> >>> Cells pop out at us on low power. Any suggestions/protocols? >> >>> >> >>> >> >>> >> >>> Thanks, >> >>> >> >>> >> >>> >> >>> Tom >> >>> >> >>> >> >>> >> >>> Tom Galati >> >>> >> >>> Laboratory Director >> >>> >> >>> HSRL, Inc.- A GLP Compliant Contract Laboratory >> >>> >> >>> 137 South Main Street >> >>> >> >>> Woodstock, Virginia 22664 >> >>> >> >>> (540)459-8211 >> >>> >> >>> Fax: (540)459-8217 >> >>> >> >>> tomgalati@hsrl.org >> >>> >> >>> www.hsrl.org >> >>> >> >>> >> >>> >> >>> _______________________________________________ >> >>> Histonet mailing list >> >>> Histonet@lists.utsouthwestern.edu >> >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> Gayle Callis >> >> MT,HT,HTL(ASCP) >> >> Research Histopathology Supervisor >> >> Veterinary Molecular Biology >> >> Montana State University - Bozeman >> >> PO Box 173610 >> >> Bozeman MT 59717-3610 >> >> 406 994-6367 (lab with voice mail) >> >> 406 994-4303 (FAX) >> >> >> >> >> >> >> >> _______________________________________________ >> >> Histonet mailing list >> >> Histonet@lists.utsouthwestern.edu >> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> This electronic message and all contents contain information from St. >> Mary's Hospital which may be attorney-client privileged, confidential > or >> otherwise protected from disclosure. The information is intended to be > >> for the addressee only. If you are not the addressee, any disclosure, >> copy, distribution or use of the contents of this message is >> prohibited. If you have received this electronic message in error, > please >> notify the sender immediately and destroy the original message and all >> copies. Thank you > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 5 > Date: Tue, 10 May 2005 13:30:54 -0400 > From: "Jeffrey Wu" > Subject: [Histonet] Picrosirius red stain > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format="flowed" > > > Hello all, > > I am trying to quantitate the amount of fibrosis in mouse > cardiac > tissue. I based my method on Dr Kiernan's excellent tutorial > on > picrosirius red staining (posted in 2000). Briefly, slides > are > deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); > 75% > ethanol (1x, 3 min); and running distilled water (10-15 min). > Next, > they are stained in saturated picrosirius red for 60 minutes > and > washed in acetic acid (2x, 3 min). They are dehydrated in 75% > ethanol > (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 > min). > Finally, the slides are mounted with Permount. > > For image processing, I am using circularly polarized light. > Under > the microscope, the collagen stains from pink to dark red > (almost > black), depending on its content; however, the background > tissue > appears green/blue. It is problematic because I quantify > the > collagen using ImagePro Plus software, in which I select > certain > colors. Especially with the pink and some dark blues, there > is > overlap between the collagen and tissue, causing overestimation > of > collagen. (Sorry for the long explanation.) I guess what I > am > seeking is the flaw in my procedure, causing the other tissue not > to > stain yellow. > > On a side note, I am using the correct picrosirius red stain, and > the > solution is saturated with crystals at the bottom. I > have > tried modifying washing times without any change. I used a short > (2 > min) 0.2% phosphomolybdic acid wash, which is supposed to > reduce > background, with no success as well. > > Once again, sorry for the long question. Any help with my > problem > would be greatly appreciated. Please do not hesitate to email me > with > any questions or anything that I have not made clear. Thank you > in > advance. > > J Wu > > > ------------------------------ > > Message: 6 > Date: Tue, 10 May 2005 13:52:27 -0400 > From: "Jim Staruk" > Subject: RE: [Histonet] Control for Mast cells > To: "'Anna Inman'" , > > Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> > Content-Type: text/plain; charset=us-ascii > > Anyone in diagnostic veterinary services gets mast cell tumors all of > the > time. I'll be glad to share some with someone in need (and I won't even > charge you). > > Jim > > ______________________ > Jim Staruk > Mass Histology Service > www.masshistology.com > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle > Callis > Sent: Tuesday, May 10, 2005 1:28 PM > To: Anna Inman; Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Control for Mast cells > > Skin contains mast cells in connective tissues and have seen them in > connective tissues surrounding bones. > > At 10:15 AM 5/10/2005, you wrote: >> --> >> >> I am having difficulty getting a control for Mast cells. Any > suggestions? >> >> >> >> Anna Inman B.S., HT (ASCP) >> >> SMH Pathology >> >> Anna.Inman@stmarygj.org > > > > > > ------------------------------ > > Message: 7 > Date: Tue, 10 May 2005 13:58:36 -0400 > From: John Kiernan > Subject: Re: [Histonet] mast cell in guinea pig > To: Histonet > Message-ID: <4280F64C.CD1F5085@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > > > John Kiernan wrote: >> >> Probably you didn't see any mast cells in the >> sections of guinea-pig tissues because there >> were none to be seen. >> >> Mouse and rat mast cells are unusual in that their >> granules are preserved and rendered insoluble by >> aqueous fixatives such as buffered formaldehyde. >> In most other species aqueous fixatives dissolve >> out the mast cell granules. Alcoholic fixatives (eg >> Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre >> etc) will preserve and insolubilize mast cell granules >> of any species. >> >> For more on this, see "The Mast Cells" by Hans Selye >> (1965) Chapter 3. Also the "Notes & Queries section in >> the current issue of Biotechnic & Histochemistry (Vol 80 >> No 1, p.43-45). The latter is available at the publisher's >> web site: >> > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c12 > 4b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1, > 1;homemain,1,1; >> >> The metachromasia is due to heparin, the major >> macromolecular anion of the granules. The staining >> properties of heparin are similar to those of >> cartilage matrix. Both materials carry a lot >> of sulphate-ester groups. Unfortunately heparin >> (except that of rats and mice) is water-soluble. >> I do not know if any tryptase or chymase immunoreactivity >> remains in mast cells whose granules have been >> extracted by an aqueous fixative. >> -- >> ------------------------------- >> John A. Kiernan >> Department of Anatomy and Cell Biology >> The University of Western Ontario >> London, Canada N6A 5C1 >> kiernan[AT]uwo.ca >> http://publish.uwo.ca/~jkiernan/ >> http://instruct.uwo.ca/anatomy/530/index.htm >> _______________________________ >> Elizabeth Chlipala wrote: >>> >>> Hello All >>> >>> I'm looking for a stain that will identify mast cells in guinea > pigs. I >>> have run both giemsa (modified dif quick) and 0.4% Toluidine Blue. > I >>> normally get very good staining of mast cells with the toluidine > blue. >>> I normally run this stain on mouse and rat tissue. This is the > first >>> time I have tried this stain in guinea pigs. Upon review of the > slides >>> I could not located one mast cell in 11 lung sections. I know they > have >>> to be there. Does the metachromatic nature of the t. blue stain > have to >>> do with the amount of histamine present in the cells? I have read > in >>> the literature that guinea pigs and humans have less histamine > present >>> in their mast cells than rats, hamsters and mice. Is anyone aware > of a >>> modification that will stain mast cells in guinea pig? Any help > would >>> be appreciated. I would prefer to stick with a histochemical > method. >>> I'm aware of Immunohistochemical staining for mast cell tryptase, > but in >>> researching this I could not find any references for guinea pig > tissue. >>> >>> Thanks in advance. >>> >>> Liz >>> >>> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >>> Manager >>> Premier Laboratory, LLC >>> P.O. Box 18592 >>> Boulder, Colorado 80308 >>> Office: (303) 735-5001 >>> Fax: (303) 735-3540 >>> liz@premierlab.com >>> www.premierlab.com >>> >>> Ship to Address: >>> Premier Laboratory >>> University of Colorado >>> MCDB, Room A3B40 >>> Boulder, Colorado 80309 >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed May 11 12:02:24 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: Heat inactivating serum Re: [Histonet] New rabbit antibody. In-Reply-To: <004001c55640$ef5f67f0$f623c084@DanaM> References: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> <6.0.0.22.1.20050511093412.01b4b0f8@gemini.msu.montana.edu> <004001c55640$ef5f67f0$f623c084@DanaM> Message-ID: <6.0.0.22.1.20050511102303.01b90c90@gemini.msu.montana.edu> To heat inactivate complement found in pooled serums. Complement has been known to cause nonspecific background staining. You can read more about this in free Immunochemical Staining Methods Manual, 3rd Edition, DAKO. They have this on their website or you can access a hard copy. At 09:48 AM 5/11/2005, you wrote: >i am curious as this has come up with a flow cytometry protocol as well. >why do you heat inactivate serum for a staining procedure? >thanks a lot, >dana Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From kbroomal <@t> NEMOURS.ORG Wed May 11 12:08:16 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Re: floating petri dish snap freezing question Message-ID: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49E@wlmmsx01.nemours.org> Unfortunately I don't see anything there to read. Perhaps you need a subscription to see that section? -----Original Message----- From: Greg Dobbin [mailto:dobbin@upei.ca] Sent: Wednesday, May 11, 2005 1:48 PM To: gcallis@montana.edu; kbroomal@NEMOURS.ORG Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: floating petri dish snap freezing question Dear Kristen/ Gayle, To see my detailed reply to a similar question on cryotomy, check the following reference: Biotechnic & Histochemistry 2003, 78(5): 279. The "Notes and Queries" section. Cheers! Greg Date sent: Wed, 11 May 2005 10:17:30 -0600 To: "Kristen Broomall" , Histonet@lists.utsouthwestern.edu From: Gayle Callis Copies to: Subject: [Histonet] Re: floating petri dish snap freezing question From tpmorken <@t> labvision.com Wed May 11 12:16:34 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] RE: RVU units Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CCEF@usca0082k08.labvision.apogent.com> Barry, This is the same problem that CAP ran into with it's old workload recording system. In trying to compare the same procedure between labs it became apparent that there is no standard way to handle any specimen or diagnostic procedure. The CAP kept adding all kinds of items for workload units in an attempt to add flexibility and accuracy. In the end it became a ridiculous exercise in which we were counting every time a person put on or took off gloves, wrote on a work-request list, or even walked to another lab to pick up a specimen. In the end the whole thing was abandoned as unworkable. While some kind of standard measure would be desireable, it just doesn't seem to be practical. Of course an institution has to meet certain bureaucratic goals, so this sort of thing will keep people busy for years. Maybe we just count it as part of the 30% of our medical costs that is devoted to paperwork. Tim Morken Free webhosting for US State Histotechnology Societies: http://www.labvisioncorp.com/demowebsite/index.cfm -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rittman, Barry R Sent: Wednesday, May 11, 2005 10:02 AM To: histonet Subject: RE: [Histonet] RE: RVU units Patti You have raised an interesting question that I feel should be discussed on Histonet. There seems to be a national tendency to use RVUs to compare different procedures and provide some type of standardization. The same process is being examined in order to compare teaching loads. On the surface it would appear to be desirable. However there are some drawbacks that I have seen in discussions of this topic for teaching. If x number of units are for a lecture given for the first time this does not take into account the skill of the teacher, the availability of resources or the different fields of teaching, biochemistry, histology etc. I would suggest that the same applies to your question. While it is nice to be able to compare the RVU of a specific technique that you are carrying out with that used by other laboratories in the field it may not be a fair comparison. The actual cost to you depends on several factors such as, skill of the individual, efficiency of technique and equipment (which may depend on newness of machine etc), availability of equipment and resources etc., etc. This is a complex problem that I feel would be in laboratories interest to handle at a local level. Just my opinion. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Wednesday, May 11, 2005 11:42 AM To: histonet Subject: Re: [Histonet] RE: RVU units I would like to see the answer posted on the histonet, too. Patti Loykasek PhenoPath Laboratories Seattle, WA > I need to set up two tests and need to assign RVU units to two CPT > codes in order to get the tests I need in our billing system. I > understand this may be a touchy subject but if anyone is willing to > share how many RVU units they assigned to the CPT codes for 88189 Flow > cytometry interpretation and 88358 DNA ploidy Digital image analysis I > would appreciate it greatly. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Wednesday, May 11, 2005 11:10 AM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 18, Issue 14 > > Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > > To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > > You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > > When replying, please edit your Subject line so it is more specific > than "Re: Contents of Histonet digest..." > > > Today's Topics: > > 1. Mast cell stain (Osborn, Sharon) > 2. Slide labelers (Osborn, Sharon) > 3. Re: Asbestos studies (Gayle Callis) > 4. Control for Mast cells (Gayle Callis) > 5. Picrosirius red stain (Jeffrey Wu) > 6. RE: Control for Mast cells (Jim Staruk) > 7. Re: mast cell in guinea pig (John Kiernan) > 8. RE: Asbestos studies (Shirley Powell) > 9. Final notice: Missouri Society for Histotechnology Spring > Symposium (Johnson, Teri) > 10. Master's of Histology (Farley, Sunni R) > 11. job (cynthia haynes) > 12. Re: Slide Labeller (MajorFocus@aol.com) > 13. Re: Mast cell stain (Laurie Reilly) > 14. Re: job (Patti Loykasek) > 15. Mast Cell/Giemsa clarification (HSRL) > 16. RE: Master's of Histology (Kemlo Rogerson) > 17. (no subject) (eswary) > 18. EDTA (Bernadette Weston) > 19. macrophages in mouse FFPE > (wasielewski.reinhard.von@mh-hannover.de) > 20. Gayle - floating petri dish snap freezing question > (Kristen Broomall) > 21. RE: macrophages in mouse FFPE (Flynn, Evelyn) > 22. New rabbit antibody... but what do I use as negative > (isotype) control? (- -) > 23. RE: job (Margaryan, Naira) > 24. Unsubscribe (Flores, Teresa) > 25. Unsubscribe (Flores, Teresa) > 26. Help!!! (vsailes@nd.edu) > 27. job listing (LINDA MARGRAF) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Tue, 10 May 2005 13:14:25 -0400 > From: "Osborn, Sharon" > Subject: [Histonet] Mast cell stain > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> > Content-Type: text/plain > > Tom, > > Luna's Toluidine Blue Method for Mast Cells is a superior demonstration > of > mast cells using a pink background. These literally pop up under low > power. The method is located on pages 311-312 of Luna's > Histopathologic Methods > and > color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a > copy of the procedure, private email me and I will send to you. > > sharon osborn > DNAX ScheringPlough BioPharma > Palo Alto, CA > > Subject: [Histonet] Giemsa Stain for Mast Cells > Netters, > I am looking for a Giemsa stain for Mast Cells that does not have a > pinkish hue like the Gaffney Giemsa. We are looking to have the Mast > Cells pop > out > at us on low power. Any suggestions/protocols? > Thanks, > Tom > Tom Galati > Laboratory Director > HSRL, Inc.- A GLP Compliant Contract Laboratory > 137 South Main Street > Woodstock, Virginia 22664 > (540)459-8211 > Fax: (540)459-8217 > tomgalati@hsrl.org > www.hsrl.org > > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. > If you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited > -- Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 2 > Date: Tue, 10 May 2005 13:21:48 -0400 > From: "Osborn, Sharon" > Subject: [Histonet] Slide labelers > To: "'histonet@lists.utsouthwestern.edu'" > > Message-ID: > <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> > Content-Type: text/plain > > Cynthia, > Contact your Leica representative and ask for a demonstration of their > slide printers. I think you will be pleasantly surprised as to the > amount of information and quietness. I have used several different > slide > labelers including the etch-a-sketch types and ink printer types; this > one > is the best of all I have used. Its footprint is a little larger than > the > others and well worth it. Sakura and Leica partnered on developing > these; > their main difference is in the software that is provided. Sakura uses > off > the shelf software while Leica specifically built the software on theirs > and > it is customizable for stand alone or for integration with MIS of an > institution. And, the company service will also make a difference. > So, I encourage you to demo several different types of slide labelers > and find the one that best suits your needs. Sharon Osborn > DNAX > ScheringPlough BioPharma > Palo Alto, CA > > > Date: Tue, 10 May 2005 10:37:23 -0400 > From: "Favara, Cynthia (NIH/NIAID)" > Subject: [Histonet] slide labeller > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain > > I am interested in an automated slide labeler. I would like the capacity > to > do multiple slides with the same information as quiet as possible and of > course I have limited space. Any suggestions would be appreciated. > > > > I also have tops to the bottle for a Ventana special stainer. I am happy > to > send to anyone interested. > > > > c > > > > Cynthia Favara > NIAID/NIH/RML/LPVD > 903 South 4th Street > Hamilton, MT 59840 > 406-363-9317 > > > > ********************************************************************* > This message and any attachments are solely for the intended recipient. > If you are not the intended recipient, disclosure, copying, use or > distribution of the information included in this message is prohibited > -- Please immediately and permanently delete. > > > > > ------------------------------ > > Message: 3 > Date: Tue, 10 May 2005 11:25:43 -0600 > From: Gayle Callis > Subject: Re: [Histonet] Asbestos studies > To: mtitford@aol.com, Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > If I remember correctly, an iron stain will identify asbestos fibers > nicely in a section. Histonet had discussion on asbestos fibers some > time ago, > > may be worth a search in archives at www.histosearch.org > > At 09:52 AM 5/10/2005, you wrote: >> Fred Underwood asks about asbestos fibers in lung tissues. >> >> Asbestosis and mesothelioma studies are quite big business along the > Gulf >> Coast here with the local shipyards and men of the "Golden Generation" >> passing away. A lot of lawyers have made a lot of money and most > recently, >> a big lawsuit in Corpus Christi Texas accuses screening companies of >> misdiagnosis. However, onto things histological: asbestos fibers can >> be easily seen > in >> an H&E section as brown, cylindrical, and beaded. In our laboratory we >> digest lung tissue in bleach and then filter the remains through a >> millipore. Once dried, it can be placed on a slide, the filter > dissolved >> in chloroform and mounted with a coverslip. The fibers are then > counted. >> The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >> asbestos-associated diseases" 2ed edition Spinger. 2004 >> >> Mike Titford >> USA Pathology >> Mobile AL USA _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 4 > Date: Tue, 10 May 2005 11:28:28 -0600 > From: Gayle Callis > Subject: [Histonet] Control for Mast cells > To: "Anna Inman" , > Histonet@lists.utsouthwestern.edu > Message-ID: > <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Skin contains mast cells in connective tissues and have seen them in > connective tissues surrounding bones. > > At 10:15 AM 5/10/2005, you wrote: >> --> >> >> I am having difficulty getting a control for Mast cells. Any > suggestions? >> >> >> >> Anna Inman B.S., HT (ASCP) >> >> SMH Pathology >> >> Anna.Inman@stmarygj.org >> >> >> >> >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle > Callis >> Sent: Tuesday, May 10, 2005 8:51 AM >> To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >> Subject: Re: [Histonet] Giemsa Stain for Mast Cells >> >> >> >> Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is >> >> Churukian Shenk method for mast cells. You don't have to worry about > so >> >> many other colors from Giemsa. If you want these, I will be happy to >> >> attach privately. >> >> >> >> At 07:45 AM 5/10/2005, you wrote: >> >>> Netters, >> >>> >> >>> >> >>> >> >>> I am looking for a Giemsa stain for Mast Cells that does not have a >> >>> pinkish hue like the Gaffney Giemsa. We are looking to have the Mast >> >>> Cells pop out at us on low power. Any suggestions/protocols? >> >>> >> >>> >> >>> >> >>> Thanks, >> >>> >> >>> >> >>> >> >>> Tom >> >>> >> >>> >> >>> >> >>> Tom Galati >> >>> >> >>> Laboratory Director >> >>> >> >>> HSRL, Inc.- A GLP Compliant Contract Laboratory >> >>> >> >>> 137 South Main Street >> >>> >> >>> Woodstock, Virginia 22664 >> >>> >> >>> (540)459-8211 >> >>> >> >>> Fax: (540)459-8217 >> >>> >> >>> tomgalati@hsrl.org >> >>> >> >>> www.hsrl.org >> >>> >> >>> >> >>> >> >>> _______________________________________________ >> >>> Histonet mailing list >> >>> Histonet@lists.utsouthwestern.edu >> >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> Gayle Callis >> >> MT,HT,HTL(ASCP) >> >> Research Histopathology Supervisor >> >> Veterinary Molecular Biology >> >> Montana State University - Bozeman >> >> PO Box 173610 >> >> Bozeman MT 59717-3610 >> >> 406 994-6367 (lab with voice mail) >> >> 406 994-4303 (FAX) >> >> >> >> >> >> >> >> _______________________________________________ >> >> Histonet mailing list >> >> Histonet@lists.utsouthwestern.edu >> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> This electronic message and all contents contain information from St. >> Mary's Hospital which may be attorney-client privileged, confidential > or >> otherwise protected from disclosure. The information is intended to be > >> for the addressee only. If you are not the addressee, any disclosure, >> copy, distribution or use of the contents of this message is >> prohibited. If you have received this electronic message in error, > please >> notify the sender immediately and destroy the original message and all >> copies. Thank you > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > > > ------------------------------ > > Message: 5 > Date: Tue, 10 May 2005 13:30:54 -0400 > From: "Jeffrey Wu" > Subject: [Histonet] Picrosirius red stain > To: histonet@lists.utsouthwestern.edu > Message-ID: > Content-Type: text/plain; format="flowed" > > > Hello all, > > I am trying to quantitate the amount of fibrosis in mouse > cardiac > tissue. I based my method on Dr Kiernan's excellent tutorial > on > picrosirius red staining (posted in 2000). Briefly, slides > are > deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); > 75% ethanol (1x, 3 min); and running distilled water (10-15 min). > Next, > they are stained in saturated picrosirius red for 60 minutes > and > washed in acetic acid (2x, 3 min). They are dehydrated in 75% > ethanol > (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 > min). > Finally, the slides are mounted with Permount. > > For image processing, I am using circularly polarized light. Under > the microscope, the collagen stains from pink to dark red > (almost > black), depending on its content; however, the background > tissue > appears green/blue. It is problematic because I quantify > the > collagen using ImagePro Plus software, in which I select > certain > colors. Especially with the pink and some dark blues, there > is > overlap between the collagen and tissue, causing overestimation > of > collagen. (Sorry for the long explanation.) I guess what I > am > seeking is the flaw in my procedure, causing the other tissue not > to > stain yellow. > > On a side note, I am using the correct picrosirius red stain, and > the > solution is saturated with crystals at the bottom. I > have > tried modifying washing times without any change. I used a short > (2 > min) 0.2% phosphomolybdic acid wash, which is supposed to > reduce background, with no success as well. > > Once again, sorry for the long question. Any help with my > problem would be greatly appreciated. Please do not hesitate to > email me with > any questions or anything that I have not made clear. Thank you > in > advance. > > J Wu > > > ------------------------------ > > Message: 6 > Date: Tue, 10 May 2005 13:52:27 -0400 > From: "Jim Staruk" > Subject: RE: [Histonet] Control for Mast cells > To: "'Anna Inman'" , > > Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> > Content-Type: text/plain; charset=us-ascii > > Anyone in diagnostic veterinary services gets mast cell tumors all of > the time. I'll be glad to share some with someone in need (and I > won't even > charge you). > > Jim > > ______________________ > Jim Staruk > Mass Histology Service > www.masshistology.com > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle > Callis > Sent: Tuesday, May 10, 2005 1:28 PM > To: Anna Inman; Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Control for Mast cells > > Skin contains mast cells in connective tissues and have seen them in > connective tissues surrounding bones. > > At 10:15 AM 5/10/2005, you wrote: >> --> >> >> I am having difficulty getting a control for Mast cells. Any > suggestions? >> >> >> >> Anna Inman B.S., HT (ASCP) >> >> SMH Pathology >> >> Anna.Inman@stmarygj.org > > > > > > ------------------------------ > > Message: 7 > Date: Tue, 10 May 2005 13:58:36 -0400 > From: John Kiernan > Subject: Re: [Histonet] mast cell in guinea pig > To: Histonet > Message-ID: <4280F64C.CD1F5085@uwo.ca> > Content-Type: text/plain; charset=us-ascii > > > > John Kiernan wrote: >> >> Probably you didn't see any mast cells in the >> sections of guinea-pig tissues because there >> were none to be seen. >> >> Mouse and rat mast cells are unusual in that their >> granules are preserved and rendered insoluble by >> aqueous fixatives such as buffered formaldehyde. >> In most other species aqueous fixatives dissolve >> out the mast cell granules. Alcoholic fixatives (eg >> Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre >> etc) will preserve and insolubilize mast cell granules >> of any species. >> >> For more on this, see "The Mast Cells" by Hans Selye >> (1965) Chapter 3. Also the "Notes & Queries section in >> the current issue of Biotechnic & Histochemistry (Vol 80 >> No 1, p.43-45). The latter is available at the publisher's web site: >> > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c12 > 4b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1, > 1;homemain,1,1; >> >> The metachromasia is due to heparin, the major macromolecular anion >> of the granules. The staining properties of heparin are similar to >> those of cartilage matrix. Both materials carry a lot >> of sulphate-ester groups. Unfortunately heparin >> (except that of rats and mice) is water-soluble. >> I do not know if any tryptase or chymase immunoreactivity >> remains in mast cells whose granules have been >> extracted by an aqueous fixative. >> -- >> ------------------------------- >> John A. Kiernan >> Department of Anatomy and Cell Biology >> The University of Western Ontario >> London, Canada N6A 5C1 >> kiernan[AT]uwo.ca >> http://publish.uwo.ca/~jkiernan/ >> http://instruct.uwo.ca/anatomy/530/index.htm >> _______________________________ >> Elizabeth Chlipala wrote: >>> >>> Hello All >>> >>> I'm looking for a stain that will identify mast cells in guinea > pigs. I >>> have run both giemsa (modified dif quick) and 0.4% Toluidine Blue. > I >>> normally get very good staining of mast cells with the toluidine > blue. >>> I normally run this stain on mouse and rat tissue. This is the > first >>> time I have tried this stain in guinea pigs. Upon review of the > slides >>> I could not located one mast cell in 11 lung sections. I know they > have >>> to be there. Does the metachromatic nature of the t. blue stain > have to >>> do with the amount of histamine present in the cells? I have read > in >>> the literature that guinea pigs and humans have less histamine > present >>> in their mast cells than rats, hamsters and mice. Is anyone aware > of a >>> modification that will stain mast cells in guinea pig? Any help > would >>> be appreciated. I would prefer to stick with a histochemical > method. >>> I'm aware of Immunohistochemical staining for mast cell tryptase, > but in >>> researching this I could not find any references for guinea pig > tissue. >>> >>> Thanks in advance. >>> >>> Liz >>> >>> Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >>> Manager >>> Premier Laboratory, LLC >>> P.O. Box 18592 >>> Boulder, Colorado 80308 >>> Office: (303) 735-5001 >>> Fax: (303) 735-3540 >>> liz@premierlab.com >>> www.premierlab.com >>> >>> Ship to Address: >>> Premier Laboratory >>> University of Colorado >>> MCDB, Room A3B40 >>> Boulder, Colorado 80309 >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed May 11 12:18:29 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] CCR7 antibody attn: Tim Coskran Message-ID: <6.0.0.22.1.20050511111518.01b715a8@gemini.msu.montana.edu> Tim, Not sure if it works on mouse, but there is a new rabbit monoclonal antibody. Interestingly, this is one company I did not dump into spam killing files. Go to "Epitomics" , who knows it might be worth a try!!! Plus there are more antibodies than this one. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed May 11 12:33:32 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] molds for RE: floating petri dish snap freezing question In-Reply-To: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49D@wlmmsx01.nemours. org> References: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49D@wlmmsx01.nemours.org> Message-ID: <6.0.0.22.1.20050511111915.01b27440@gemini.msu.montana.edu> Kristen, I think other plastic molds work just as well with this particular freezing setup. We snap freeze mouse heads (huge!) using Peel away molds (bigger and deeper) with this method, no problems. We like Tissue Tek plastic molds only because the plastic is thinner, less stiff. With the Liq N2 canoeing petri dish, you can use glass coverslip, put OCT on coverslip, embed tissue and let it freeze, to remove tiny block, warm back of glass slightly!!!! Peggy Wenk taught this one, clever lady! Plastic coverslips are available, and with canoeing petri dish, very easy snap freezing. Good thing you experienced the interface problem before embarking on whole study - I chopped blocks and ruined some precious tissues, usually when someone else did the freezing and not aware of this problem. Rules became, if you snap freeze, you cut your own blocks!!! Gayle At 10:46 AM 5/11/2005, you wrote: >That does help. Thanks! > >I was afraid that if I put the tissue too close to the bottom of the mold, >it would be affected somehow or other. I don't think our plastic molds are >Tissue Tek (you mentioned other brands being too thick), but an H&E section >from one of the samples I froze yesterday seems to be okay. Hoping they >continue to be okay... > >I like this technique. It's very easy. > >"you do not want an interface of two OCT layers, they will snap part during >sectioning at >times." Yeah, tried that when I was experimenting. Not good! Haha. They >popped apart as soon as I got it out of the mold. Good thing I tried that >practicing before putting real tissue in there. > >Thanks again! You're just a wealth a info! > >Kristen From gcallis <@t> montana.edu Wed May 11 12:37:05 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:04 2005 Subject: Biotechnic and Histochemistry access RE: [Histonet] Re: floating petri dish snap freezing question In-Reply-To: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49E@wlmmsx01.nemours. org> References: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49E@wlmmsx01.nemours.org> Message-ID: <6.0.0.22.1.20050511113610.01b26d50@gemini.msu.montana.edu> Same problem here. At 11:08 AM 5/11/2005, you wrote: >Unfortunately I don't see anything there to read. Perhaps you need a >subscription to see that section? > >-----Original Message----- >From: Greg Dobbin [mailto:dobbin@upei.ca] >Sent: Wednesday, May 11, 2005 1:48 PM >To: gcallis@montana.edu; kbroomal@NEMOURS.ORG >Cc: Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Re: floating petri dish snap freezing question > > >Dear Kristen/ Gayle, >To see my detailed reply to a similar question on cryotomy, check >the following reference: >Biotechnic & Histochemistry 2003, 78(5): 279. >The "Notes and Queries" section. >Cheers! >Greg Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From TJJ <@t> Stowers-Institute.org Wed May 11 12:50:25 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] New rabbit antibody... but what do I use as negative control Message-ID: I agree with Gayle, bought the ChromePure from Jackson for mouse, rabbit, rat, and goat. They come at a whopping 11+ mg/ml, so we diluted much of it out to 1 mg/ml and aliquotted that in our freezer to make our other dilutions from there. We did keep some at the concentrated 11+ mg/ml just in case you get that odd antibody that has you use some ungodly concentration for it to work. Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From pex0220 <@t> yahoo.com.cn Wed May 11 12:58:23 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] background!! Message-ID: <20050511175824.587.qmail@web15504.mail.cnb.yahoo.com> Hello, all. I am doing double immunoflourescence in bone sections, but I find that there is so much background and there are so many spots, I have used the normal serum matched to the hosts of secondary antibodies for blocking ( 5% donkey serum in PBS), spinined secondary antibodies over before incubating secondary antibodies and added washing times, but so much background still appears, I think that it isnot autofluorescence due to no background in negative sections, I am really sad because I can not find an effective method to reduce background. Any suggestions will be helpful! I am looking forward for your help! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From cgfields <@t> lexhealth.org Wed May 11 13:03:26 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] HPV antibody Message-ID: Our Immuno tech is currently buying the antibody for H Pylori from CelMarque. If has worked well until they reconfigured it. She said no one else makes the antibody. If anyone has information on this we would appreciate the help. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From amoklebu <@t> seattlecca.org Wed May 11 13:03:08 2005 From: amoklebu <@t> seattlecca.org (Moklebust, Amanda C) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Luna's toluidine blue; also Ventana Benchmark XT m orphology Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948013CF45C@wala01.seattlecca.org> Regarding the morphology changes in your patient tissue. You may want to examine how the slides are dried. I have worked in labs where controls were precut and dried in a 37 degree oven overnight. When we received an IHC order, we placed the patient tissue on the lower portion of a control slide. Then the slides were dried again. I am not currently using a Benchmark for IHC staining, but I thought those instruments were going to include protocols for drying and deparaffinizing slides. If your control is already dried and your patient tissue still has water around it when it is loaded on the instrument, it could be one avenue for more troubleshooting. Good Luck, Amanda Moklebust Seattle Cancer Care Alliance -----Original Message----- From: Malam Jacqueline [mailto:Jacqueline.Malam@rli.mbht.nhs.uk] Sent: Wednesday, May 11, 2005 8:57 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Luna's toluidine blue; also Ventana Benchmark XT morphology I would appreciate it if I could also be sent details of Luna's toluidine blue stain for mast cells, and any others that are considered superior so I can compare with our current method. Thanks! Also - do any of you users of Ventana's Benchmark or Benchmark XT autoimmunostainer have a problem with poor morphology after heat retrieval - usually affecting collagen / fibro-fatty tissues, and also the occasional section detachment? We trialled the Benchmark which was fine but the Benchmark XT which we purchased a year ago produces this problem. It does not affect our positive control tissue (or very little) and I have been through every possibility within the department and with Ventana but to no avail. All ideas / clues welcome! Jacqui Malam Royal Lancaster Infirmary DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From pex0220 <@t> yahoo.com.cn Wed May 11 13:06:58 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] ALP (alkaline phosphatase) expression! Message-ID: <20050511180659.43103.qmail@web15506.mail.cnb.yahoo.com> Hello, all, alkaline phosphatase ( ALP) is a marker gene of osteoblast, it is mainly expressed in cytoplasm in vitro. Recently I do immunofluorescence in bone sections, but I donot find positive expression in osteoblasts, only it is found in hypertrophic cartilage and part bone marrow, so I am not sure that where ALP is located in bone sections. If you have some experience in it, can you do me a favor? Thank you! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From Rcartun <@t> harthosp.org Wed May 11 13:20:51 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] HPV antibody Message-ID: We use DakoCytomation's H. pylori polyclonal antibody (B0471) with excellent results. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Carole Fields 05/11/05 02:03PM >>> Our Immuno tech is currently buying the antibody for H Pylori from CelMarque. If has worked well until they reconfigured it. She said no one else makes the antibody. If anyone has information on this we would appreciate the help. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From DGNajarian <@t> comcast.net Wed May 11 13:23:28 2005 From: DGNajarian <@t> comcast.net (Dennis Najarian) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] HPV antibody References: Message-ID: <000901c55656$8dc6a830$6501a8c0@chemicon.com> http://search.chemicon.com/exec/?q=h+pylori ----- Original Message ----- From: "Carole Fields" To: Sent: Wednesday, May 11, 2005 2:03 PM Subject: [Histonet] HPV antibody > Our Immuno tech is currently buying the antibody for H Pylori from > CelMarque. If has worked well until they reconfigured it. She said no > one > else makes the antibody. If anyone has information on this we would > appreciate the help. > > Carole Fields, HT,ASCP > Pathology Supervisor > Lexington Medical Center > 2720 Sunset Blvd. > W. Columbia, SC 29169 > > > > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed May 11 13:38:18 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Masson's trichrome References: <6.1.1.1.1.20050504131027.019835c0@pop3.uqar.qc.ca> Message-ID: <4282511A.EB80B428@uwo.ca> Dear Julien Lambrey de Souza, I have done a Masson variant closely similar to the one you refer to, and am an admirer (and lucky owner) of Gabe's big 1976 book. I suspect that the cause of your trouble is the fixation. Were your specimens fixed in a solution that had formaldehyde as the only chemically active ingredient? Gabe (like Mallory, Heidenhain and Masson) fixed specimens in mixtures appropriate to trichrome staining methods. Such mixtures contain acetic acid and either picric acid, mercuric chloride or potassium dichromate. Examples are Bouin, Heidenhain's SUSA and Zenker. Sections of formaldehyde-fixed tissue do not give the crisp colour distinctions that make trichrome preparations so beautiful and informative. In clinical laboratories, where specimens are almost always fixed in neutral (usually phosphate-buffered) formaldehyde, the hydrated sections can be exposed to a trichrome-friendly fixative compound before doing a method like Masson's. This trick usually allows the dyes to distribute themselves correctly into cytoplasm and collagen. The customary pre-treatment is immersion of rehydrated slides in Bouin's fixative either overnight at room temperature or for an hour or two in an oven at about 60C. My (unpublished and probably not original) observations indicate that saturated aqueous picric acid is just as effective as Bouin (thus avoiding fumes of formaldehyde and acetic acid). If your specimens were not fixed in formaldehyde, provide more information and keep on asking. There are (I thinnk) a few thousands of Histonetters. Someone else may have encountered and solved your problem. John Kiernan Anatomy, UWO, London, Canada __________________________________________ Julien LambreyDeSouza wrote: > > Hello, > > I am trying out Masson's trichrome for the first time. I used the Goldner > modification from Gabe 1976, with Fuschine-Ponceau, orang-G phosphomolybdic > and Light green. We are staining 7 micron sections of entire fish larvae > heads. When following the protocol (5 minutes in each color followed by a > 1% acetic water rinse) I get a general green staining of all tissues. The > only way to differentiate cartilage from the rest in our section is by > recognizing cellular topography, same as with Hematox-eosine staining, > therefore loosing the whole point in using Masson's trichrome. I seem to be > doing something wrong. > > Would anybody familiar with the technique be inclined to share their > expertise or protocol? I would be happy to send a pic of our staining > result by email to have input on the possible problem. > > Thanks, > > Julien. > Julien Lambrey de Souza > Assistant de recherche, Biologiste M.Sc. > Biologie ?volutive. > Universit? du Qu?bec ? Rimouski > D?partement de Biologie, Chimie et Sciences sant? > 300, all?e des Ursulines > Rimouski (Qu?bec) Canada G5L 3A1 > T?l.: (418) 723-1986 ext. 1714 > Fax.: (418) 724-1849 > Courriel: julien_lambreydesouza@uqar.qc.ca ------------------------------------------------------ From GDawson <@t> dynacaremilwaukee.com Wed May 11 13:37:44 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] HPV antibody Message-ID: Same here. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Wednesday, May 11, 2005 12:21 PM To: cgfields@lexhealth.org; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] HPV antibody We use DakoCytomation's H. pylori polyclonal antibody (B0471) with excellent results. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> Carole Fields 05/11/05 02:03PM >>> Our Immuno tech is currently buying the antibody for H Pylori from CelMarque. If has worked well until they reconfigured it. She said no one else makes the antibody. If anyone has information on this we would appreciate the help. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed May 11 13:40:07 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:04 2005 Subject: Sodium citrate. Was Re: [Histonet] (no subject) References: <200505111707.AA254607564@carif.com.my> Message-ID: <42825187.7B778204@uwo.ca> "No subject" Grrrrr! Trisodium citrate alone would make an unbuffered alkaline solution. Some antigens need an alkaline retrieval solution, and there are a few that require an acidic solution. pH 6 is satisfactory for most, and is less likely to remove the sections from the slides than a solution of more extreme (especially alkaline) pH. For a short article about how antigen retrieval might work, look at the spring newsletter of Region IX of the NSH: http://www.nshregionix.org/springn05ewsletter.pdf John Kiernan Anatomy, UWO, London, Canada _____________________________ eswary wrote: > > hi, > > does anyone know the advantage of using 0.01M sodium citrate over 0.01M > citric acid buffer, pH 6, for antigen retrieval on paraffin embedded > sections? also, is the sodium constituent important? would it matter if > i used trisodium citrate instead of monosodium? > > thanks. > > et > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BrealK <@t> Alexian.net Wed May 11 13:50:10 2005 From: BrealK <@t> Alexian.net (Kari Breal) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Job opening in Chicago Suburbs Message-ID: Alexian Brothers Medical Center in Elk Grove Village, IL has a full time histologist position open. Great benefits, competitive salary and wonderful place to work. Please direct any questions to Kari Breal @ 847-437-5500 ext 5155. From Laurie.Pereira <@t> sdcounty.ca.gov Wed May 11 14:25:30 2005 From: Laurie.Pereira <@t> sdcounty.ca.gov (Pereira, Laurie ) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] fontana masson Message-ID: Hey everyone, Let me start by saying this is my first time doing a Fontana Masson stain. The pathologist was looking for a melanoma but decided later that it might be a venereal disease in a dog. We make up our own solutions in the laboratory and our ammonium hydroxide is old. It took at least 20ml to get the solution to change initially! With that to say, my control and tissue sample have this non-specific brown pigment in the background. It is not just in the cytoplasm of the cells. It even occurs in the emtpy space of a hair follicle and exists in the background where tissue doesn't exist. My control is a skin control and the melanin stained beautifully black. What is this pigment? I hope someone can help! Thanks in advance! Laurie L. Pereira, RVT SDCADDL (County Veterinarian) 5555 Overland Avenue Suite 4103 San Diego, CA. 92123 Phone: 858-694-3384 Fax: 858-571-4268 From ftulenko06 <@t> jcu.edu Wed May 11 14:26:17 2005 From: ftulenko06 <@t> jcu.edu (ftulenko06@jcu.edu) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] reconstructing 3D models from serial sections Message-ID: Hello histonetters, First of all, thank you for your past advice. I am currently attempting to make 3-D models from serially sectioned specimens. I was wondering if anyone can suggest good, user-friendly software for doing this. I have been trying to use winsurf, but the program crashes often. Thank you for your time in responding, Frank From KPercival <@t> wyeth.com Wed May 11 14:38:32 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] hpv antibody Message-ID: Be careful, HPV is not the same as H.Pylori. Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com From cgfields <@t> lexhealth.org Wed May 11 14:41:41 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Thank you all for the great response Message-ID: I forwarded all the responses to our immuno tech. Thank you all for the great response...I'm sure it will help. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 (803) 936-8214 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From dcrippen <@t> buckinstitute.org Wed May 11 14:41:48 2005 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] reconstructing 3D models from serial sections Message-ID: Dear Frank, We routinely do this for 3-D rendering and measurements. We use Bitplane, Imaris Suite with Autoaligner. You can check out their website http://www.bitplane.com/ The software is fairly expensive, but I think you can get a personal version for less. Maintenance is also a bit costly, but the technicians are often very helpful. Otherwise, I think (though have not personally tried) that ImageJ from NIH (which is free) has similar capabilities. Good luck, Danielle Crippen Morphology and Imaging Core Manager Buck Institute for Age Research 8001 Redwood Blvd. Novato, CA 94945 415-209-2046 dcrippen@buckinstitute.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of ftulenko06@jcu.edu Sent: Wednesday, May 11, 2005 12:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] reconstructing 3D models from serial sections Hello histonetters, First of all, thank you for your past advice. I am currently attempting to make 3-D models from serially sectioned specimens. I was wondering if anyone can suggest good, user-friendly software for doing this. I have been trying to use winsurf, but the program crashes often. Thank you for your time in responding, Frank _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Patty.Lott <@t> ORTHO.UAB.EDU Wed May 11 15:41:21 2005 From: Patty.Lott <@t> ORTHO.UAB.EDU (Patty Lott) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] unsubscribe Message-ID: <85F6C7A1330E794DB8540AFD001CC77E05329071@rosco.ortho.uab.edu> Patty Lott, Laboratory Supervisor Orthopaedic Research Laboratory Center for Metabolic Bone Disease Laboratory University of Alabama at Birmingham LHRB B37 0007 1919 7th Ave. South Birmingham, AL 35294 (205) 934-2007 From RCHIOVETTI <@t> aol.com Wed May 11 15:53:47 2005 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] background!! Message-ID: In a message dated 5/11/2005 10:59:30 AM US Mountain Standard Time, pex0220@yahoo.com.cn writes: > I think that it isnot autofluorescence due to no background in negative > sections, I am really sad because I can not find an effective method to reduce > background. Any suggestions will be helpful! I am looking forward for your > help! > Guofeng, I don't know whether this will help or not, but it is worth a try. I used to use this technique to solve background problems for immunoelectron microscopy: After both the primary and secondary antibody steps, rinse a few times in normal PBS, then rinse a few more times in PBS with a very high concentration of salt. I used 5 Normal PBS, meaning the saline contained 5 times the amount of salt it should have. After you rinse with the concentrated salt solution, be sure to wash a few times in regular PBS, then with PBS containing the blocker. You have to return the slide to a normal concentration of saline before proceeding to the next steps. Normal saline is about 150 mM, or 0.9% NaCl by weight. Try making up a saline solution that contains about ** 4.5% ** NaCl by weight (750 mM). This worked very well (when nothing else would) at the EM level, so there's a chance it will work at the light level as well. This tip comes from a physical biochemistry lab where I was a postdoc. We did a lot of column chromatography, and we used high salt washes to clean up our columns. The salt probably alters the conformations of the antibodies and probes so much that only the specifically interacting ones are able to hang on in the concentrated salt. Good luck. Let us know if this solves the problem. Bob Robert (Bob) Chiovetti, Ph.D. Independent Consultant for The Science, Technology and Industrial Sectors 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA From conlonb <@t> comcast.net Wed May 11 16:37:57 2005 From: conlonb <@t> comcast.net (Brian D. Conlon) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] prostate needle biopsies Message-ID: <000c01c55671$b4784040$6401a8c0@gateway> hi everyone, our lab has a question about prostate needle biopsies. the pa's submit two cores per block and we embed them with a tamper so they are on the same level. we do three levels on each block. is there a protocol as to how many microns should be between each level? sometimes the tissue is so thin that there is nothing left in the block for recuts or ihc. we would like to have each tech cut with the same method so that there will be greater consistency. thanks so much everybody. paula From jlflow2 <@t> uky.edu Fri May 6 14:58:48 2005 From: jlflow2 <@t> uky.edu (Jennifer L Flowers) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Sakura Sapphatome SS-45 Message-ID: <1115409528.433df9fcjlflow2@uky.edu> Hello, I am trying to locate a dealer for a Sakura Sapphatome SS-45 knife. Sakura Finetech in the US does not know anything about this microtome knife. Are these knives still available? The old ones I have found in our lab work well but are getting dull. Any help is greatly appreciated! Thanks, Jen Flowers Department of Plant Pathology University of Kentucky jlflow2@uky.edu From funderwood <@t> mcohio.org Mon May 9 13:50:43 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] asbestos fibers Message-ID: Hi All, Does someone have a technic for demonstrating asbesots fibers in lung. Possibly by isolating them through digestion of the tissue. Thanks in advance. Fred Underwood HT(ASCP) Montgomery County Coroner Dayton, OH From cjswartz <@t> msn.com Mon May 9 19:30:42 2005 From: cjswartz <@t> msn.com (Cheryl Swartzentruver) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Integrated stainer and coverslipper Message-ID: We are currently looking for a coverslipper, and are considering both tape and glass. Because our stainer is also near the end of its useful life, we are looking at both Leica and Sakura integrated systems. Leica's glass coverslipper can be integrated with the autostainer or multistainer, and sakura will have a tape coverslipper and stainer available next month. They will also have a glass coverslipper available next year, which will also be able to be integated with the stainer. Has anyone on the list used the Leica combo? Cheryl Swartzentruver HT(ASCP) From gliuygao <@t> hotmail.com Wed May 11 19:10:29 2005 From: gliuygao <@t> hotmail.com (yan gao) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Mono clone phospho-MEK antibody Message-ID: I am looking for phospho-MEK mono clone antibody for stain raf pathway. I do know cell signaling technology sales it. Where else? Thanks. Yan Gao From jessgrocki <@t> yahoo.com Wed May 11 20:21:50 2005 From: jessgrocki <@t> yahoo.com (Jessica Piche) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] Disposing of Blocks Message-ID: <20050512012150.40502.qmail@web81703.mail.yahoo.com> Hi Everyone, How are we supposed to get rid of patient blocks once the required time to keep them ends? We are throwing them out in the regular trash and are wondering if regulations might have changed. Thanks, Jessica Piche-Grocki, HT(ASCP) From arvind <@t> nbrc.res.in Wed May 11 23:42:02 2005 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] freezing fetus brain tisue for storing Message-ID: can any one please suggest method to freeze the fetal brain tissue so that it can be used later for cryosectioning and IHC , brain samples are prenatal ranging from 3 months to full term thanks in advance Arvind Singh Pundir NBRC,Haryana , India From Kemlo.Rogerson <@t> elht.nhs.uk Thu May 12 01:45:07 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] asbestos fibers Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2AA@bhrv-nt-11.bhrv.nwest.nhs.uk> Burn the tissue? Asbestos doesn't, burn that is. -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: 09 May 2005 19:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] asbestos fibers Hi All, Does someone have a technic for demonstrating asbesots fibers in lung. Possibly by isolating them through digestion of the tissue. Thanks in advance. Fred Underwood HT(ASCP) Montgomery County Coroner Dayton, OH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tanni.Ahmed <@t> intervet.com Thu May 12 03:07:41 2005 From: Tanni.Ahmed <@t> intervet.com (Ahmed, T (Tanni)) Date: Fri Sep 16 15:25:04 2005 Subject: [Histonet] RE: MSc Histopathology Message-ID: <09E7875E806DFB4BBECBADBF966BDBB6AAFD55@mksn79.d50.intra> Dear Sunni and histonetters, Re: postgraduate histology training. I am intending to start a part time Histopathology postgraduate Msc course with Imperial College uni (imperial.ac.uk) this coming October. I am currently working for a veterinary co'(who are sponsoring and funding the course), where I have been setting up and running the histolab. I had previously worked as a trainee in a private healthcare company, also in histopath - (human histology) for one year after completing my degree in Biochemistry, two years ago. I decided the NHS training didn't suit what I wanted to do...it is a grading system here in the UK, where you are trained in various areas such as immunology, heamatology etc. However, you require a Biomedical Science degree to start off with - the Institute of BMS would not accept my degree, unless I did a top-up post-grad diploma and worked in an NHS lab for 2 years...so I took plan B, the backdoor.... Initially I was unsure whether to go ahead with a postgrad - in terms of becoming specialised in that particular field, however it made sense as my degree only had one histology module. I think a taught course like this, would be ideal, providing a thorough foundation in the fundamental mechanisms underpinning pathology, both practically and theoretically. The majority of students will predominantly be from the NHS, with human histology being the core area of teaching. However, I believe the fundamentals of histology is similar for both human/animal histopath and the course is also designed to accommodate applicants in the research field. The course entry requirements are an upper second class degree or above here, however they often take students who have lower qualifications if we feel that they are suitable. The course is always run on a day release basis - therefore you can get best of both worlds, studying and working fulltime. The next two years are going to be hectic, but I hope all worth it in the end as I will definetely get a lot out of it. The course has been and is currently very successful being run alongside a Clinical Cytology course here and they are always looking to improve the course and keep it relevant and up-to-date for techniques etc. All students have to complete a laboratory-based project of their choice and are expected to have a local supervisor and deal with Ethics etc although they will help wherever we can. Im considering doing a avian related project...this may change in 5 months though! Hope this helps.... Tanni Tanni S Ahmed Scientific Officer - Histopathology, R&D Intervet UK Ltd. Walton Manor, Walton, Milton Keynes, Buckinghamshire, MK7 7AJ, UK. Tel. +44(0)1908 685552/685543 Fax +44(0)1908 685614 E-mail: tanni.ahmed@intervet.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: 11 May 2005 16:14 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 18, Issue 14 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Mast cell stain (Osborn, Sharon) 2. Slide labelers (Osborn, Sharon) 3. Re: Asbestos studies (Gayle Callis) 4. Control for Mast cells (Gayle Callis) 5. Picrosirius red stain (Jeffrey Wu) 6. RE: Control for Mast cells (Jim Staruk) 7. Re: mast cell in guinea pig (John Kiernan) 8. RE: Asbestos studies (Shirley Powell) 9. Final notice: Missouri Society for Histotechnology Spring Symposium (Johnson, Teri) 10. Master's of Histology (Farley, Sunni R) 11. job (cynthia haynes) 12. Re: Slide Labeller (MajorFocus@aol.com) 13. Re: Mast cell stain (Laurie Reilly) 14. Re: job (Patti Loykasek) 15. Mast Cell/Giemsa clarification (HSRL) 16. RE: Master's of Histology (Kemlo Rogerson) 17. (no subject) (eswary) 18. EDTA (Bernadette Weston) 19. macrophages in mouse FFPE (wasielewski.reinhard.von@mh-hannover.de) 20. Gayle - floating petri dish snap freezing question (Kristen Broomall) 21. RE: macrophages in mouse FFPE (Flynn, Evelyn) 22. New rabbit antibody... but what do I use as negative (isotype) control? (- -) 23. RE: job (Margaryan, Naira) 24. Unsubscribe (Flores, Teresa) 25. Unsubscribe (Flores, Teresa) 26. Help!!! (vsailes@nd.edu) 27. job listing (LINDA MARGRAF) ---------------------------------------------------------------------- Message: 1 Date: Tue, 10 May 2005 13:14:25 -0400 From: "Osborn, Sharon" Subject: [Histonet] Mast cell stain To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> Content-Type: text/plain Tom, Luna's Toluidine Blue Method for Mast Cells is a superior demonstration of mast cells using a pink background. These literally pop up under low power. The method is located on pages 311-312 of Luna's Histopathologic Methods and color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a copy of the procedure, private email me and I will send to you. sharon osborn DNAX ScheringPlough BioPharma Palo Alto, CA Subject: [Histonet] Giemsa Stain for Mast Cells Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 2 Date: Tue, 10 May 2005 13:21:48 -0400 From: "Osborn, Sharon" Subject: [Histonet] Slide labelers To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> Content-Type: text/plain Cynthia, Contact your Leica representative and ask for a demonstration of their slide printers. I think you will be pleasantly surprised as to the amount of information and quietness. I have used several different slide labelers including the etch-a-sketch types and ink printer types; this one is the best of all I have used. Its footprint is a little larger than the others and well worth it. Sakura and Leica partnered on developing these; their main difference is in the software that is provided. Sakura uses off the shelf software while Leica specifically built the software on theirs and it is customizable for stand alone or for integration with MIS of an institution. And, the company service will also make a difference. So, I encourage you to demo several different types of slide labelers and find the one that best suits your needs. Sharon Osborn DNAX ScheringPlough BioPharma Palo Alto, CA Date: Tue, 10 May 2005 10:37:23 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: [Histonet] slide labeller To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I am interested in an automated slide labeler. I would like the capacity to do multiple slides with the same information as quiet as possible and of course I have limited space. Any suggestions would be appreciated. I also have tops to the bottle for a Ventana special stainer. I am happy to send to anyone interested. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 3 Date: Tue, 10 May 2005 11:25:43 -0600 From: Gayle Callis Subject: Re: [Histonet] Asbestos studies To: mtitford@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the >Gulf >Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most recently, >a big lawsuit in Corpus Christi Texas accuses screening companies of >misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen in >an H&E section as brown, cylindrical, and beaded. In our laboratory we >digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter dissolved >in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 4 Date: Tue, 10 May 2005 11:28:28 -0600 From: Gayle Callis Subject: [Histonet] Control for Mast cells To: "Anna Inman" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any > suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org > > > > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis >Sent: Tuesday, May 10, 2005 8:51 AM >To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Giemsa Stain for Mast Cells > > > >Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is > >Churukian Shenk method for mast cells. You don't have to worry about >so > >many other colors from Giemsa. If you want these, I will be happy to > >attach privately. > > > >At 07:45 AM 5/10/2005, you wrote: > > >Netters, > > > > > > > > > > > >I am looking for a Giemsa stain for Mast Cells that does not have a > > >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast > > >Cells pop out at us on low power. Any suggestions/protocols? > > > > > > > > > > > >Thanks, > > > > > > > > > > > >Tom > > > > > > > > > > > >Tom Galati > > > > > >Laboratory Director > > > > > >HSRL, Inc.- A GLP Compliant Contract Laboratory > > > > > >137 South Main Street > > > > > >Woodstock, Virginia 22664 > > > > > >(540)459-8211 > > > > > >Fax: (540)459-8217 > > > > > >tomgalati@hsrl.org > > > > > >www.hsrl.org > > > > > > > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Gayle Callis > >MT,HT,HTL(ASCP) > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 (lab with voice mail) > >406 994-4303 (FAX) > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >This electronic message and all contents contain information from St. >Mary's Hospital which may be attorney-client privileged, confidential or >otherwise protected from disclosure. The information is intended to be >for the addressee only. If you are not the addressee, any disclosure, >copy, distribution or use of the contents of this message is >prohibited. If you have received this electronic message in error, please >notify the sender immediately and destroy the original message and all >copies. Thank you Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Tue, 10 May 2005 13:30:54 -0400 From: "Jeffrey Wu" Subject: [Histonet] Picrosirius red stain To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" Hello all, I am trying to quantitate the amount of fibrosis in mouse cardiac tissue. I based my method on Dr Kiernan's excellent tutorial on picrosirius red staining (posted in 2000). Briefly, slides are deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); 75% ethanol (1x, 3 min); and running distilled water (10-15 min). Next, they are stained in saturated picrosirius red for 60 minutes and washed in acetic acid (2x, 3 min). They are dehydrated in 75% ethanol (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 min). Finally, the slides are mounted with Permount. For image processing, I am using circularly polarized light. Under the microscope, the collagen stains from pink to dark red (almost black), depending on its content; however, the background tissue appears green/blue. It is problematic because I quantify the collagen using ImagePro Plus software, in which I select certain colors. Especially with the pink and some dark blues, there is overlap between the collagen and tissue, causing overestimation of collagen. (Sorry for the long explanation.) I guess what I am seeking is the flaw in my procedure, causing the other tissue not to stain yellow. On a side note, I am using the correct picrosirius red stain, and the solution is saturated with crystals at the bottom. I have tried modifying washing times without any change. I used a short (2 min) 0.2% phosphomolybdic acid wash, which is supposed to reduce background, with no success as well. Once again, sorry for the long question. Any help with my problem would be greatly appreciated. Please do not hesitate to email me with any questions or anything that I have not made clear. Thank you in advance. J Wu ------------------------------ Message: 6 Date: Tue, 10 May 2005 13:52:27 -0400 From: "Jim Staruk" Subject: RE: [Histonet] Control for Mast cells To: "'Anna Inman'" , Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> Content-Type: text/plain; charset=us-ascii Anyone in diagnostic veterinary services gets mast cell tumors all of the time. I'll be glad to share some with someone in need (and I won't even charge you). Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 1:28 PM To: Anna Inman; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Control for Mast cells Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any > suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org ------------------------------ Message: 7 Date: Tue, 10 May 2005 13:58:36 -0400 From: John Kiernan Subject: Re: [Histonet] mast cell in guinea pig To: Histonet Message-ID: <4280F64C.CD1F5085@uwo.ca> Content-Type: text/plain; charset=us-ascii John Kiernan wrote: > > Probably you didn't see any mast cells in the > sections of guinea-pig tissues because there > were none to be seen. > > Mouse and rat mast cells are unusual in that their > granules are preserved and rendered insoluble by > aqueous fixatives such as buffered formaldehyde. > In most other species aqueous fixatives dissolve > out the mast cell granules. Alcoholic fixatives (eg > Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre > etc) will preserve and insolubilize mast cell granules > of any species. > > For more on this, see "The Mast Cells" by Hans Selye > (1965) Chapter 3. Also the "Notes & Queries section in > the current issue of Biotechnic & Histochemistry (Vol 80 > No 1, p.43-45). The latter is available at the publisher's web site: > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c12 4b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1, 1;homemain,1,1; > > The metachromasia is due to heparin, the major > macromolecular anion of the granules. The staining > properties of heparin are similar to those of > cartilage matrix. Both materials carry a lot > of sulphate-ester groups. Unfortunately heparin > (except that of rats and mice) is water-soluble. > I do not know if any tryptase or chymase immunoreactivity remains in > mast cells whose granules have been extracted by an aqueous fixative. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Elizabeth Chlipala wrote: > > > > Hello All > > > > I'm looking for a stain that will identify mast cells in guinea > > pigs. I have run both giemsa (modified dif quick) and 0.4% > > Toluidine Blue. I normally get very good staining of mast cells > > with the toluidine blue. I normally run this stain on mouse and rat > > tissue. This is the first time I have tried this stain in guinea > > pigs. Upon review of the slides I could not located one mast cell > > in 11 lung sections. I know they have to be there. Does the > > metachromatic nature of the t. blue stain have to do with the amount > > of histamine present in the cells? I have read in the literature > > that guinea pigs and humans have less histamine present in their > > mast cells than rats, hamsters and mice. Is anyone aware of a > > modification that will stain mast cells in guinea pig? Any help > > would be appreciated. I would prefer to stick with a histochemical > > method. I'm aware of Immunohistochemical staining for mast cell > > tryptase, but in researching this I could not find any references > > for guinea pig tissue. > > > > Thanks in advance. > > > > Liz > > > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > > Manager > > Premier Laboratory, LLC > > P.O. Box 18592 > > Boulder, Colorado 80308 > > Office: (303) 735-5001 > > Fax: (303) 735-3540 > > liz@premierlab.com > > www.premierlab.com > > > > Ship to Address: > > Premier Laboratory > > University of Colorado > > MCDB, Room A3B40 > > Boulder, Colorado 80309 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ ------------------------------ Message: 8 Date: Tue, 10 May 2005 14:15:25 -0400 From: Shirley Powell Subject: RE: [Histonet] Asbestos studies To: 'Gayle Callis' , mtitford@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <01LO3GS2JNLI8WX3KK@Macon2.Mercer.edu> Content-Type: text/plain; charset=us-ascii The histonet archives can be found at www.histosearch.com. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 12:26 PM To: mtitford@aol.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Asbestos studies If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the >Gulf Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most >recently, a big lawsuit in Corpus Christi Texas accuses screening >companies of misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen >in an H&E section as brown, cylindrical, and beaded. In our laboratory >we digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter >dissolved in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 10 May 2005 14:06:06 -0500 From: "Johnson, Teri" Subject: [Histonet] Final notice: Missouri Society for Histotechnology Spring Symposium To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" The Missouri Society for Histotechnology Spring Symposium When: May 20-21, 2005 Where: The Lodge of the Four Seasons, Lake Ozark, Missouri (800) 843-5253 www.4seasonsresort.com Note: Rooms for 'The Lodge' may be fully booked so here are 2 alternative places to stay: The Country Club (800) 964-6698 (approximately 1.5 miles away) The Resort at Port Arrowhead (800) 532-3575 (approximately 3 miles away) For additional meeting information please contact: Sharon Walsh Phone: (636) 305-9650 email: userwalsh@aol.com or Rosetta Barkley Phone: (913) 588-2737 email: rbarkley2@kumc.edu We invite you to participate in a unique learning experience. The program is outstanding this year and should have something for everyone. In addition there will be social activities and vendor displays. Please Note: All workshops are CEU approved. Friday Schedule: 8:30am "Ergonomics in the Workplace", Jan Minshew, Leica Microsystems 9:15am "Case Studies in Forensic Pathology", Dr. Michael Graham, St. Louis University 10:45am "Histochemistry of Special Stains", Jerry Fredenburgh 1:30pm-4:30pm Workshops #1, #2, or #3 (Your choice) 6:00-8:00pm - Cruise on the "Lake of the Ozarks" Come enjoy the lake and visit with exhibitor's and friends. Saturday Schedule: 8:15am "Creutzfeldt-Jakob Disease: Tissue Handling and Testing in the United Kingdom", Konnie Zietner, Nebraska Medical Center 9:15am "Histological Techniques in Hematopathology", Dr. Sharad Mather 10:45am "Applications for Microwave Decalcification Procedures", Skip Brown 1:30pm-4:30pm Workshops #4, #5, or #6 (Your choice) Registration Fees: Members - $50, Non-Members - $60 Additional Banquet Ticket - $25 2005 WORKSHOP DESCRIPTIONS Workshop#1 Theory of Routine & Microwave Fixation & Processing - Skip Brown & Jerry Fredenburgh The process of tissue fixation and processing will be illustrated; first from a molecular level to demonstrate what is actually happening at a micro level in the tissue; then from a macro level to show the gross effects on the tissue. Accelerated procedures will be introduced as a way to speed up the process without detrimental effects to the tissue. Workshop #2 (Sponsored by Ventana Medical Systems) A Tour Guide to Immunohistochemistry: How to get there and what to do when you arrive. - Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Immunohistochemistry is defined as, "A method of detecting the presence of specific proteins in cells or tissues." If only it was that easy. A better definition may be, "A method of standardizing each and every step in and out of your control working toward a complex chemical construction of molecules to identify specific proteins in a variety of tissues in a variety of stages in order to answer specific questions." This class will not only identify the building blocks of immunohistochemistry, from rabbit monoclonal antibodies to polymer detection kits, but we will also review how each step before the staining can affect those building blocks. This class will also address some basic dilution techniques for concentrated antibodies, the use of positive and negative controls, where to start trouble shooting your new antibody. Finally, we will discuss the utility of IHC in the clinical and research setting and its relevance to drug discovery and disease treatment. Workshop #3 Immunocytochemical and Immunohistochemical assays. A Pathologist's perspective, Lourdes R. Ylagan MD, Washington University. This workshop will show a brief overview of both immunohistochemical and immunocytochemical techniques. It will also show examples of how double immunostaining techniques can be helpful in both histologic and cytologic samples. It will also show the ways in which immunochemistry is used in: (1) the elucidation of the site of origin of a poorly differentiated neoplasm in the setting of a metastasis, (2) detection of antigens present in the surface of tumor cells amenable to antibody-mediated chemotherapeutic agents, (3) detection of tumor markers known to be potential poor prognostic indicators in tumors. Workshop #4 (Sponsored by Leica Microsystems) Quality and Skill in Microtomy Technique - Jan Minshew, Leica MicroSystems This session will present an understanding of the physical dynamics of microtomy, and discuss the effects of external variables such as knife angle, room temperature, inadequate instrumentation, etc. Workshop #5 (Sponsored by Ventana Medical Systems) Special Stains: the Once and Future King? Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Special Stains are an art of using dyes to stain specific tissues and/or cellular structures that were once thought of as a major advancement in the histology lab. Now we have much more complex chemicals to stain proteins and genes within those cells and tissues, yet we still find a vast usage of special stains in the histology lab. In fact, some of the specials stains have become so routine that they are oftentimes considered as routine as the H&E in certain circumstances. Why do we still utilize special stains when we have far less hazardous chemicals that can pinpoint much more specific things? This class will address that question. We will discuss where special stains has been, why they have been around for so long, and where we see them going. This class will break down the most frequently used special stains; discuss their bio-chemical reactions and their utility. We will then compare those special stains to their relevant immunohistochemistry and in-situ hybridization counterparts to illustrate their utility and futility . Workshop #6 >From Bench Tech to Management, Konnie Zietner, Nebraska Medical Center Basic principles and skills you will need when transitioning from technician to management. ------------------------------ Message: 10 Date: Tue, 10 May 2005 12:34:23 -0700 From: "Farley, Sunni R" Subject: [Histonet] Master's of Histology To: histonet@lists.utsouthwestern.edu Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948323349@wala01.seattlecca.org> Content-Type: text/plain; charset="ISO-8859-1" I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. ------------------------------ Message: 11 Date: Tue, 10 May 2005 14:54:16 -0700 (PDT) From: cynthia haynes Subject: [Histonet] job To: Histonet@lists.utsouthwestern.edu Message-ID: <20050510215416.51974.qmail@web41710.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi! We are looking for people who might be interested in a pool position in the Chicago land area. Please contact me Cynthia Haynes at 1-773-484-4133. Thanks in Advance Cynthia Haynes H.T. __________________________________ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. http://mobile.yahoo.com/learn/mail ------------------------------ Message: 12 Date: Tue, 10 May 2005 18:56:59 EDT From: MajorFocus@aol.com Subject: [Histonet] Re: Slide Labeller To: cfavara@niaid.nih.gov Cc: histonet@lists.utsouthwestern.edu Message-ID: <1f4.978eaa2.2fb2963b@aol.com> Content-Type: text/plain; charset="US-ASCII" I have used the Thermo Microwriter as well which is quite nice. If you are interested in an inexpensive software program for generating slide labels, check out our website for the Histo-Labels program. It will also interface with most slide and cassette printers/etchers as well. _www.majorfocus.com_ (http://www.majorfocus.com) Greg L. Good, HT(ASCP) Major Focus (800) 888-1152 ------------------------------ Message: 13 Date: Wed, 11 May 2005 09:36:52 +1000 From: Laurie Reilly Subject: Re: [Histonet] Mast cell stain To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5.1.0.14.0.20050511092655.019a6ec0@mail.jcu.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi Histonetters, We routinely stain all dog skin lesions with Toluidine Blue and it highlights the Mast Cells. They stain metachromatically and show up with red-purple granules in the cytoplasm. These biopsies are all fixed in 10% Neutral Buffered Formalin. The method we use is from Gretchen Humason, "Animal tissue Techniques", 3rd edition. It uses a solution of 0.2% Tol Blue in 60% Ethanol for 2 minutes. Regards, Laurie. At 01:14 PM 05/10/05 -0400, Osborn, Sharon wrote: >Tom, > >Luna's Toluidine Blue Method for Mast Cells is a superior demonstration >of mast cells using a pink background. These literally pop up under >low power. The method is located on pages 311-312 of Luna's >Histopathologic Methods and color Atlas of special Stains and Tissue >Artifacts; 1992. If you wish a copy of the procedure, private email me >and I will send to you. > >sharon osborn >DNAX ScheringPlough BioPharma >Palo Alto, CA > >Subject: [Histonet] Giemsa Stain for Mast Cells >Netters, >I am looking for a Giemsa stain for Mast Cells that does not have a >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast >Cells pop out at us on low power. Any suggestions/protocols? Thanks, >Tom >Tom Galati >Laboratory Director >HSRL, Inc.- A GLP Compliant Contract Laboratory >137 South Main Street >Woodstock, Virginia 22664 >(540)459-8211 >Fax: (540)459-8217 >tomgalati@hsrl.org >www.hsrl.org > > > >********************************************************************* >This message and any attachments are solely for the intended recipient. >If >you are not the intended recipient, disclosure, copying, use or >distribution of the information included in this message is prohibited -- >Please immediately and permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. ------------------------------ Message: 14 Date: Tue, 10 May 2005 16:41:50 -0700 From: Patti Loykasek Subject: Re: [Histonet] job To: histonet Message-ID: Content-Type: text/plain; charset="US-ASCII" You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Tue, 10 May 2005 20:32:13 -0400 From: "HSRL" Subject: [Histonet] Mast Cell/Giemsa clarification To: Message-ID: <000601c555c0$e5a31840$0200a8c0@HSRLMAIN> Content-Type: text/plain; charset="US-ASCII" Netters, I agree with all of you about the Tol Blue stain being a good mast cell stain. However, we are specifically looking for a Giemsa stain for mast cell granules. We already do the Tol Blue, and the Pinacynol Erythrosinate stains for mast cells. Thanks again, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ------------------------------ Message: 16 Date: Wed, 11 May 2005 07:42:03 +0100 From: Kemlo Rogerson Subject: RE: [Histonet] Master's of Histology To: "'Farley, Sunni R'" , histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2A7@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain In the UK you can obtain a Master of Science Degree by pursuing a research project. In fact you could always take a Master's or Doctorate if you could find a Uni, a Supervisor and something of interest to research. I feel that postgraduate degrees have become easier to obtain, but then we old people always say that. In the UK NVQ's and other vocational qualifications are becoming more popular as it is really a judgement call, and I'm going to get into trouble, but maybe postgraduate degrees could be thought of as an overkill for many positions in the Laboratory. Do we really have that much to offer MSc's and PHD's? Other than funding them to get the qualification; I fess up to having an MSc, but...... -----Original Message----- From: Farley, Sunni R [mailto:sfarley@seattlecca.org] Sent: 10 May 2005 20:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Master's of Histology I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Wed, 11 May 2005 17:07:14 +0800 From: "eswary" Subject: [Histonet] (no subject) To: Message-ID: <200505111707.AA254607564@carif.com.my> Content-Type: text/plain; charset=us-ascii hi, does anyone know the advantage of using 0.01M sodium citrate over 0.01M citric acid buffer, pH 6, for antigen retrieval on paraffin embedded sections? also, is the sodium constituent important? would it matter if i used trisodium citrate instead of monosodium? thanks. et ------------------------------ Message: 18 Date: Wed, 11 May 2005 08:31:25 -0400 From: "Bernadette Weston" Subject: [Histonet] EDTA To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed We can no longer supply EDTA for our bone marrow procedures, can some one recommend a substitution for aspirate collection? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton, OH ------------------------------ Message: 19 Date: Wed, 11 May 2005 14:40:33 +0200 From: wasielewski.reinhard.von@mh-hannover.de Subject: [Histonet] macrophages in mouse FFPE To: histonet@lists.utsouthwestern.edu Message-ID: <42821961.31188.FCDE1BE2@localhost> Content-Type: text/plain; charset=US-ASCII Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski ------------------------------ Message: 20 Date: Wed, 11 May 2005 08:46:42 -0400 From: "Kristen Broomall" Subject: [Histonet] Gayle - floating petri dish snap freezing question To: Histonet@lists.utsouthwestern.edu Message-ID: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours.org> Content-Type: text/plain; charset=iso-8859-1 Gayle, I tried out your snap freezing method with the floating petri dish in the liquid nitrogen yesterday for some itsy bitsy GI biopsies. I really like it! I have a question though. When using this method, do you place your sample in the bottom of your mold, then OCT on top, or put a dab (or more) of OCT, then the tissue, ... or does it really matter? Thanks, Kristen Broomall, HT (ASCP) ------------------------------ Message: 21 Date: Wed, 11 May 2005 09:21:33 -0400 From: "Flynn, Evelyn" Subject: RE: [Histonet] macrophages in mouse FFPE To: wasielewski.reinhard.von@mh-hannover.de, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Dear Reinhard, I have had good results with rat anti-mouse Mac-3 antibody from PharMingen (Cat. # 550292). Regards, Evelyn Flynn, M.A. Children's Hospital, Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of wasielewski.reinhard.von@mh-hannover.de Sent: Wed 5/11/2005 8:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] macrophages in mouse FFPE Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Wed, 11 May 2005 07:08:31 -0700 (PDT) From: - - Subject: [Histonet] New rabbit antibody... but what do I use as negative (isotype) control? To: histonet@lists.utsouthwestern.edu Message-ID: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi everyone, I have a new antibody that I was given. It is a rabbit polyclonal anti-mouse antibody. Usually I use a specific isotype control, but I do not have info on how this antibody was made other than the epitope(s) that it should adhere to... but I want a negative control and was wondering if I can just use regular rabbit serum (diluted) as a negative seeing that it should contain the isotype IgG I am using??? I think I am a bit confused here.... I need assistance. Can I use rabbit serum as a negative control? What dilution do I use if that is the case? Anything else I should know before I proceed? Thank you all!! --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! ------------------------------ Message: 23 Date: Wed, 11 May 2005 09:26:25 -0500 From: "Margaryan, Naira" Subject: RE: [Histonet] job To: "Patti Loykasek" , "histonet" Message-ID: <63B8B599DE283148B92E83C78B32C15DB9164E@cmhexbe2.childrensmemorial.org> Content-Type: text/plain What do you mean Pool position? Thanks, Naira -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Tuesday, May 10, 2005 6:42 PM To: histonet Subject: Re: [Histonet] job You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. ------------------------------ Message: 24 Date: Wed, 11 May 2005 09:28:22 -0500 From: "Flores, Teresa" Subject: [Histonet] Unsubscribe To: histonet@lists.utsouthwestern.edu Cc: histonet-request@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, "'histonet@pathology.swmed.edu'" Message-ID: Content-Type: text/plain Please unsubscribe. ------------------------------ Message: 25 Date: Wed, 11 May 2005 09:28:22 -0500 From: "Flores, Teresa" Subject: [Histonet] Unsubscribe To: histonet@lists.utsouthwestern.edu Cc: histonet-request@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, "'histonet@pathology.swmed.edu'" Message-ID: Content-Type: text/plain Please unsubscribe. ------------------------------ Message: 26 Date: Wed, 11 May 2005 09:44:37 -0500 From: vsailes@nd.edu Subject: [Histonet] Help!!! To: histonet@lists.utsouthwestern.edu Message-ID: <1115822677.42821a55c5ced@webmail.nd.edu> Content-Type: text/plain; charset=ISO-8859-1 Hello Histonetters, I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues. I'm new to working out conditons for antibody protocols and could use some help.I'm testing the antibody at different dilutions and I appear to have positive staining but I'm also seeing small areas in the negative that shows positive staining. Liver is my control and I use steamer (HIER) for 30 minutes, my steps include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, Secondary antibody(Biotin), Streptavidin, Chromogen, Counterstain My concern is that I'm using buffers that contain BSA and I'm wondering if that is causing staining to appear or should I be using avidin/biotin block along with a peroxide block. Any suggestion would be helpful. Thank you for your help in advance. Valerie ------------------------------ Message: 27 Date: Wed, 11 May 2005 10:05:33 -0500 From: "LINDA MARGRAF" Subject: [Histonet] job listing To: Message-ID: Content-Type: text/plain; charset=US-ASCII Lead and/or Bench Histology Technician Scott & White is seeking a Lead and/or Bench Histology Technician. Responsible for histology laboratory operations including testing, training of support personnel, QA, personnel utilization and regulatory compliance. Selected candidate will also participate in routine histology laboratory functions. Requirements: An Associate's degree; HT(ASCP); 3 to 5 years experience. Just an hour north of Austin, Scott & White offers highly competitive salaries, comprehensive benefits and advancement opportunities. Candidates should send resumes to: ghollie@swmail.sw.org. Phone: 800.527.JOBS. Fax: 254.724.5591. Apply in person or mail resume to: Human Resources Dept., 2401 S. 31st St., Temple, TX 77508. www.sw.org An equal opportunity employer/tobacco-free environment. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 14 **************************************** -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- From Tanni.Ahmed <@t> intervet.com Thu May 12 03:11:41 2005 From: Tanni.Ahmed <@t> intervet.com (Ahmed, T (Tanni)) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] RE: MSc Histopathology Message-ID: <09E7875E806DFB4BBECBADBF966BDBB6AAFD5B@mksn79.d50.intra> Dear Sunni and histonetters, Re: postgraduate histology training. I am intending to start a part time Histopathology postgraduate Msc course with Imperial College uni (imperial.ac.uk) this coming October. I am currently working for a veterinary co'(who are sponsoring and funding the course), where I have been setting up and running the histolab. I had previously worked as a trainee in a private healthcare company, also in histopath - (human histology) for one year after completing my degree in Biochemistry, two years ago. I decided the NHS training didn't suit what I wanted to do...it is a grading system here in the UK, where you are trained in various areas such as immunology, heamatology etc. However, you require a Biomedical Science degree to start off with - the Institute of BMS would not accept my degree, unless I did a top-up post-grad diploma and worked in an NHS lab for 2 years...so I took plan B, the backdoor.... Initially I was unsure whether to go ahead with a postgrad - in terms of becoming specialised in that particular field, however it made sense as my degree only had one histology module. I think a taught course like this, would be ideal, providing a thorough foundation in the fundamental mechanisms underpinning pathology, both practically and theoretically. The majority of students will predominantly be from the NHS, with human histology being the core area of teaching. However, I believe the fundamentals of histology is similar for both human/animal histopath and the course is also designed to accommodate applicants in the research field. The course entry requirements are an upper second class degree or above here, however they often take students who have lower qualifications if we feel that they are suitable. The course is always run on a day release basis - therefore you can get best of both worlds, studying and working fulltime. The next two years are going to be hectic, but I hope all worth it in the end as I will definetely get a lot out of it. The course has been and is currently very successful being run alongside a Clinical Cytology course here and they are always looking to improve the course and keep it relevant and up-to-date for techniques etc. All students have to complete a laboratory-based project of their choice and are expected to have a local supervisor and deal with Ethics etc although they will help wherever we can. Im considering doing a avian related project...this may change in 5 months though! Hope this helps.... Tanni Tanni S Ahmed Scientific Officer - Histopathology, R&D Intervet UK Ltd. Walton Manor, Walton, Milton Keynes, Buckinghamshire, MK7 7AJ, UK. Tel. +44(0)1908 685552/685543 Fax +44(0)1908 685614 E-mail: tanni.ahmed@intervet.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: 11 May 2005 16:14 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 18, Issue 14 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Mast cell stain (Osborn, Sharon) 2. Slide labelers (Osborn, Sharon) 3. Re: Asbestos studies (Gayle Callis) 4. Control for Mast cells (Gayle Callis) 5. Picrosirius red stain (Jeffrey Wu) 6. RE: Control for Mast cells (Jim Staruk) 7. Re: mast cell in guinea pig (John Kiernan) 8. RE: Asbestos studies (Shirley Powell) 9. Final notice: Missouri Society for Histotechnology Spring Symposium (Johnson, Teri) 10. Master's of Histology (Farley, Sunni R) 11. job (cynthia haynes) 12. Re: Slide Labeller (MajorFocus@aol.com) 13. Re: Mast cell stain (Laurie Reilly) 14. Re: job (Patti Loykasek) 15. Mast Cell/Giemsa clarification (HSRL) 16. RE: Master's of Histology (Kemlo Rogerson) 17. (no subject) (eswary) 18. EDTA (Bernadette Weston) 19. macrophages in mouse FFPE (wasielewski.reinhard.von@mh-hannover.de) 20. Gayle - floating petri dish snap freezing question (Kristen Broomall) 21. RE: macrophages in mouse FFPE (Flynn, Evelyn) 22. New rabbit antibody... but what do I use as negative (isotype) control? (- -) 23. RE: job (Margaryan, Naira) 24. Unsubscribe (Flores, Teresa) 25. Unsubscribe (Flores, Teresa) 26. Help!!! (vsailes@nd.edu) 27. job listing (LINDA MARGRAF) ---------------------------------------------------------------------- Message: 1 Date: Tue, 10 May 2005 13:14:25 -0400 From: "Osborn, Sharon" Subject: [Histonet] Mast cell stain To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397F8E@PALMSG30.us.schp.com> Content-Type: text/plain Tom, Luna's Toluidine Blue Method for Mast Cells is a superior demonstration of mast cells using a pink background. These literally pop up under low power. The method is located on pages 311-312 of Luna's Histopathologic Methods and color Atlas of special Stains and Tissue Artifacts; 1992. If you wish a copy of the procedure, private email me and I will send to you. sharon osborn DNAX ScheringPlough BioPharma Palo Alto, CA Subject: [Histonet] Giemsa Stain for Mast Cells Netters, I am looking for a Giemsa stain for Mast Cells that does not have a pinkish hue like the Gaffney Giemsa. We are looking to have the Mast Cells pop out at us on low power. Any suggestions/protocols? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 2 Date: Tue, 10 May 2005 13:21:48 -0400 From: "Osborn, Sharon" Subject: [Histonet] Slide labelers To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <29B25753F6B1D51196110002A589D44402397F8F@PALMSG30.us.schp.com> Content-Type: text/plain Cynthia, Contact your Leica representative and ask for a demonstration of their slide printers. I think you will be pleasantly surprised as to the amount of information and quietness. I have used several different slide labelers including the etch-a-sketch types and ink printer types; this one is the best of all I have used. Its footprint is a little larger than the others and well worth it. Sakura and Leica partnered on developing these; their main difference is in the software that is provided. Sakura uses off the shelf software while Leica specifically built the software on theirs and it is customizable for stand alone or for integration with MIS of an institution. And, the company service will also make a difference. So, I encourage you to demo several different types of slide labelers and find the one that best suits your needs. Sharon Osborn DNAX ScheringPlough BioPharma Palo Alto, CA Date: Tue, 10 May 2005 10:37:23 -0400 From: "Favara, Cynthia (NIH/NIAID)" Subject: [Histonet] slide labeller To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I am interested in an automated slide labeler. I would like the capacity to do multiple slides with the same information as quiet as possible and of course I have limited space. Any suggestions would be appreciated. I also have tops to the bottle for a Ventana special stainer. I am happy to send to anyone interested. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. ------------------------------ Message: 3 Date: Tue, 10 May 2005 11:25:43 -0600 From: Gayle Callis Subject: Re: [Histonet] Asbestos studies To: mtitford@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050510112352.01b41688@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the >Gulf >Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most recently, >a big lawsuit in Corpus Christi Texas accuses screening companies of >misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen in >an H&E section as brown, cylindrical, and beaded. In our laboratory we >digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter dissolved >in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 4 Date: Tue, 10 May 2005 11:28:28 -0600 From: Gayle Callis Subject: [Histonet] Control for Mast cells To: "Anna Inman" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20050510112558.01b62930@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any > suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org > > > > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis >Sent: Tuesday, May 10, 2005 8:51 AM >To: histosci@shentel.net; Histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Giemsa Stain for Mast Cells > > > >Carsons pH 4.3 toluidine blue stain is excellent for Mast cells as is > >Churukian Shenk method for mast cells. You don't have to worry about >so > >many other colors from Giemsa. If you want these, I will be happy to > >attach privately. > > > >At 07:45 AM 5/10/2005, you wrote: > > >Netters, > > > > > > > > > > > >I am looking for a Giemsa stain for Mast Cells that does not have a > > >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast > > >Cells pop out at us on low power. Any suggestions/protocols? > > > > > > > > > > > >Thanks, > > > > > > > > > > > >Tom > > > > > > > > > > > >Tom Galati > > > > > >Laboratory Director > > > > > >HSRL, Inc.- A GLP Compliant Contract Laboratory > > > > > >137 South Main Street > > > > > >Woodstock, Virginia 22664 > > > > > >(540)459-8211 > > > > > >Fax: (540)459-8217 > > > > > >tomgalati@hsrl.org > > > > > >www.hsrl.org > > > > > > > > > > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Gayle Callis > >MT,HT,HTL(ASCP) > >Research Histopathology Supervisor > >Veterinary Molecular Biology > >Montana State University - Bozeman > >PO Box 173610 > >Bozeman MT 59717-3610 > >406 994-6367 (lab with voice mail) > >406 994-4303 (FAX) > > > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >This electronic message and all contents contain information from St. >Mary's Hospital which may be attorney-client privileged, confidential or >otherwise protected from disclosure. The information is intended to be >for the addressee only. If you are not the addressee, any disclosure, >copy, distribution or use of the contents of this message is >prohibited. If you have received this electronic message in error, please >notify the sender immediately and destroy the original message and all >copies. Thank you Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 5 Date: Tue, 10 May 2005 13:30:54 -0400 From: "Jeffrey Wu" Subject: [Histonet] Picrosirius red stain To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" Hello all, I am trying to quantitate the amount of fibrosis in mouse cardiac tissue. I based my method on Dr Kiernan's excellent tutorial on picrosirius red staining (posted in 2000). Briefly, slides are deparaffinated in xylene (2x, 6 min); 100% ethanol (2x, 3 min); 75% ethanol (1x, 3 min); and running distilled water (10-15 min). Next, they are stained in saturated picrosirius red for 60 minutes and washed in acetic acid (2x, 3 min). They are dehydrated in 75% ethanol (1x, 3 min); 100% ethanol (2x, 3 min); and xylene (1x, 3 min). Finally, the slides are mounted with Permount. For image processing, I am using circularly polarized light. Under the microscope, the collagen stains from pink to dark red (almost black), depending on its content; however, the background tissue appears green/blue. It is problematic because I quantify the collagen using ImagePro Plus software, in which I select certain colors. Especially with the pink and some dark blues, there is overlap between the collagen and tissue, causing overestimation of collagen. (Sorry for the long explanation.) I guess what I am seeking is the flaw in my procedure, causing the other tissue not to stain yellow. On a side note, I am using the correct picrosirius red stain, and the solution is saturated with crystals at the bottom. I have tried modifying washing times without any change. I used a short (2 min) 0.2% phosphomolybdic acid wash, which is supposed to reduce background, with no success as well. Once again, sorry for the long question. Any help with my problem would be greatly appreciated. Please do not hesitate to email me with any questions or anything that I have not made clear. Thank you in advance. J Wu ------------------------------ Message: 6 Date: Tue, 10 May 2005 13:52:27 -0400 From: "Jim Staruk" Subject: RE: [Histonet] Control for Mast cells To: "'Anna Inman'" , Message-ID: <0IGA002BACE3E8G4@vms048.mailsrvcs.net> Content-Type: text/plain; charset=us-ascii Anyone in diagnostic veterinary services gets mast cell tumors all of the time. I'll be glad to share some with someone in need (and I won't even charge you). Jim ______________________ Jim Staruk Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 1:28 PM To: Anna Inman; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Control for Mast cells Skin contains mast cells in connective tissues and have seen them in connective tissues surrounding bones. At 10:15 AM 5/10/2005, you wrote: >--> > > I am having difficulty getting a control for Mast cells. Any > suggestions? > > > >Anna Inman B.S., HT (ASCP) > >SMH Pathology > >Anna.Inman@stmarygj.org ------------------------------ Message: 7 Date: Tue, 10 May 2005 13:58:36 -0400 From: John Kiernan Subject: Re: [Histonet] mast cell in guinea pig To: Histonet Message-ID: <4280F64C.CD1F5085@uwo.ca> Content-Type: text/plain; charset=us-ascii John Kiernan wrote: > > Probably you didn't see any mast cells in the > sections of guinea-pig tissues because there > were none to be seen. > > Mouse and rat mast cells are unusual in that their > granules are preserved and rendered insoluble by > aqueous fixatives such as buffered formaldehyde. > In most other species aqueous fixatives dissolve > out the mast cell granules. Alcoholic fixatives (eg > Carnoy, or the "alcoholic Bouin" fluids: Dubosq, Gendre > etc) will preserve and insolubilize mast cell granules > of any species. > > For more on this, see "The Mast Cells" by Hans Selye > (1965) Chapter 3. Also the "Notes & Queries section in > the current issue of Biotechnic & Histochemistry (Vol 80 > No 1, p.43-45). The latter is available at the publisher's web site: > http://journalsonline.tandf.co.uk/app/home/journal.asp?wasp=5a0cd1389c12 4b68b94584a9e888d3f3&referrer=parent&backto=searchpublicationsresults,1, 1;homemain,1,1; > > The metachromasia is due to heparin, the major > macromolecular anion of the granules. The staining > properties of heparin are similar to those of > cartilage matrix. Both materials carry a lot > of sulphate-ester groups. Unfortunately heparin > (except that of rats and mice) is water-soluble. > I do not know if any tryptase or chymase immunoreactivity remains in > mast cells whose granules have been extracted by an aqueous fixative. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Elizabeth Chlipala wrote: > > > > Hello All > > > > I'm looking for a stain that will identify mast cells in guinea > > pigs. I have run both giemsa (modified dif quick) and 0.4% > > Toluidine Blue. I normally get very good staining of mast cells > > with the toluidine blue. I normally run this stain on mouse and rat > > tissue. This is the first time I have tried this stain in guinea > > pigs. Upon review of the slides I could not located one mast cell > > in 11 lung sections. I know they have to be there. Does the > > metachromatic nature of the t. blue stain have to do with the amount > > of histamine present in the cells? I have read in the literature > > that guinea pigs and humans have less histamine present in their > > mast cells than rats, hamsters and mice. Is anyone aware of a > > modification that will stain mast cells in guinea pig? Any help > > would be appreciated. I would prefer to stick with a histochemical > > method. I'm aware of Immunohistochemical staining for mast cell > > tryptase, but in researching this I could not find any references > > for guinea pig tissue. > > > > Thanks in advance. > > > > Liz > > > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > > Manager > > Premier Laboratory, LLC > > P.O. Box 18592 > > Boulder, Colorado 80308 > > Office: (303) 735-5001 > > Fax: (303) 735-3540 > > liz@premierlab.com > > www.premierlab.com > > > > Ship to Address: > > Premier Laboratory > > University of Colorado > > MCDB, Room A3B40 > > Boulder, Colorado 80309 > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ ------------------------------ Message: 8 Date: Tue, 10 May 2005 14:15:25 -0400 From: Shirley Powell Subject: RE: [Histonet] Asbestos studies To: 'Gayle Callis' , mtitford@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <01LO3GS2JNLI8WX3KK@Macon2.Mercer.edu> Content-Type: text/plain; charset=us-ascii The histonet archives can be found at www.histosearch.com. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Tuesday, May 10, 2005 12:26 PM To: mtitford@aol.com; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Asbestos studies If I remember correctly, an iron stain will identify asbestos fibers nicely in a section. Histonet had discussion on asbestos fibers some time ago, may be worth a search in archives at www.histosearch.org At 09:52 AM 5/10/2005, you wrote: >Fred Underwood asks about asbestos fibers in lung tissues. > >Asbestosis and mesothelioma studies are quite big business along the >Gulf Coast here with the local shipyards and men of the "Golden Generation" >passing away. A lot of lawyers have made a lot of money and most >recently, a big lawsuit in Corpus Christi Texas accuses screening >companies of misdiagnosis. >However, onto things histological: asbestos fibers can be easily seen >in an H&E section as brown, cylindrical, and beaded. In our laboratory >we digest lung tissue in bleach and then filter the remains through a >millipore. Once dried, it can be placed on a slide, the filter >dissolved in chloroform and mounted with a coverslip. The fibers are then counted. >The reference is Roggli VL, Oury, TD and Sporn TA " Pathology of >asbestos-associated diseases" 2ed edition Spinger. 2004 > >Mike Titford >USA Pathology >Mobile AL USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Tue, 10 May 2005 14:06:06 -0500 From: "Johnson, Teri" Subject: [Histonet] Final notice: Missouri Society for Histotechnology Spring Symposium To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" The Missouri Society for Histotechnology Spring Symposium When: May 20-21, 2005 Where: The Lodge of the Four Seasons, Lake Ozark, Missouri (800) 843-5253 www.4seasonsresort.com Note: Rooms for 'The Lodge' may be fully booked so here are 2 alternative places to stay: The Country Club (800) 964-6698 (approximately 1.5 miles away) The Resort at Port Arrowhead (800) 532-3575 (approximately 3 miles away) For additional meeting information please contact: Sharon Walsh Phone: (636) 305-9650 email: userwalsh@aol.com or Rosetta Barkley Phone: (913) 588-2737 email: rbarkley2@kumc.edu We invite you to participate in a unique learning experience. The program is outstanding this year and should have something for everyone. In addition there will be social activities and vendor displays. Please Note: All workshops are CEU approved. Friday Schedule: 8:30am "Ergonomics in the Workplace", Jan Minshew, Leica Microsystems 9:15am "Case Studies in Forensic Pathology", Dr. Michael Graham, St. Louis University 10:45am "Histochemistry of Special Stains", Jerry Fredenburgh 1:30pm-4:30pm Workshops #1, #2, or #3 (Your choice) 6:00-8:00pm - Cruise on the "Lake of the Ozarks" Come enjoy the lake and visit with exhibitor's and friends. Saturday Schedule: 8:15am "Creutzfeldt-Jakob Disease: Tissue Handling and Testing in the United Kingdom", Konnie Zietner, Nebraska Medical Center 9:15am "Histological Techniques in Hematopathology", Dr. Sharad Mather 10:45am "Applications for Microwave Decalcification Procedures", Skip Brown 1:30pm-4:30pm Workshops #4, #5, or #6 (Your choice) Registration Fees: Members - $50, Non-Members - $60 Additional Banquet Ticket - $25 2005 WORKSHOP DESCRIPTIONS Workshop#1 Theory of Routine & Microwave Fixation & Processing - Skip Brown & Jerry Fredenburgh The process of tissue fixation and processing will be illustrated; first from a molecular level to demonstrate what is actually happening at a micro level in the tissue; then from a macro level to show the gross effects on the tissue. Accelerated procedures will be introduced as a way to speed up the process without detrimental effects to the tissue. Workshop #2 (Sponsored by Ventana Medical Systems) A Tour Guide to Immunohistochemistry: How to get there and what to do when you arrive. - Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Immunohistochemistry is defined as, "A method of detecting the presence of specific proteins in cells or tissues." If only it was that easy. A better definition may be, "A method of standardizing each and every step in and out of your control working toward a complex chemical construction of molecules to identify specific proteins in a variety of tissues in a variety of stages in order to answer specific questions." This class will not only identify the building blocks of immunohistochemistry, from rabbit monoclonal antibodies to polymer detection kits, but we will also review how each step before the staining can affect those building blocks. This class will also address some basic dilution techniques for concentrated antibodies, the use of positive and negative controls, where to start trouble shooting your new antibody. Finally, we will discuss the utility of IHC in the clinical and research setting and its relevance to drug discovery and disease treatment. Workshop #3 Immunocytochemical and Immunohistochemical assays. A Pathologist's perspective, Lourdes R. Ylagan MD, Washington University. This workshop will show a brief overview of both immunohistochemical and immunocytochemical techniques. It will also show examples of how double immunostaining techniques can be helpful in both histologic and cytologic samples. It will also show the ways in which immunochemistry is used in: (1) the elucidation of the site of origin of a poorly differentiated neoplasm in the setting of a metastasis, (2) detection of antigens present in the surface of tumor cells amenable to antibody-mediated chemotherapeutic agents, (3) detection of tumor markers known to be potential poor prognostic indicators in tumors. Workshop #4 (Sponsored by Leica Microsystems) Quality and Skill in Microtomy Technique - Jan Minshew, Leica MicroSystems This session will present an understanding of the physical dynamics of microtomy, and discuss the effects of external variables such as knife angle, room temperature, inadequate instrumentation, etc. Workshop #5 (Sponsored by Ventana Medical Systems) Special Stains: the Once and Future King? Chris Moore, BS World Wide Project Manager, Special Stains, Ventana Medical Systems, Inc. Special Stains are an art of using dyes to stain specific tissues and/or cellular structures that were once thought of as a major advancement in the histology lab. Now we have much more complex chemicals to stain proteins and genes within those cells and tissues, yet we still find a vast usage of special stains in the histology lab. In fact, some of the specials stains have become so routine that they are oftentimes considered as routine as the H&E in certain circumstances. Why do we still utilize special stains when we have far less hazardous chemicals that can pinpoint much more specific things? This class will address that question. We will discuss where special stains has been, why they have been around for so long, and where we see them going. This class will break down the most frequently used special stains; discuss their bio-chemical reactions and their utility. We will then compare those special stains to their relevant immunohistochemistry and in-situ hybridization counterparts to illustrate their utility and futility . Workshop #6 >From Bench Tech to Management, Konnie Zietner, Nebraska Medical Center Basic principles and skills you will need when transitioning from technician to management. ------------------------------ Message: 10 Date: Tue, 10 May 2005 12:34:23 -0700 From: "Farley, Sunni R" Subject: [Histonet] Master's of Histology To: histonet@lists.utsouthwestern.edu Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948323349@wala01.seattlecca.org> Content-Type: text/plain; charset="ISO-8859-1" I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. ------------------------------ Message: 11 Date: Tue, 10 May 2005 14:54:16 -0700 (PDT) From: cynthia haynes Subject: [Histonet] job To: Histonet@lists.utsouthwestern.edu Message-ID: <20050510215416.51974.qmail@web41710.mail.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi! We are looking for people who might be interested in a pool position in the Chicago land area. Please contact me Cynthia Haynes at 1-773-484-4133. Thanks in Advance Cynthia Haynes H.T. __________________________________ Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. http://mobile.yahoo.com/learn/mail ------------------------------ Message: 12 Date: Tue, 10 May 2005 18:56:59 EDT From: MajorFocus@aol.com Subject: [Histonet] Re: Slide Labeller To: cfavara@niaid.nih.gov Cc: histonet@lists.utsouthwestern.edu Message-ID: <1f4.978eaa2.2fb2963b@aol.com> Content-Type: text/plain; charset="US-ASCII" I have used the Thermo Microwriter as well which is quite nice. If you are interested in an inexpensive software program for generating slide labels, check out our website for the Histo-Labels program. It will also interface with most slide and cassette printers/etchers as well. _www.majorfocus.com_ (http://www.majorfocus.com) Greg L. Good, HT(ASCP) Major Focus (800) 888-1152 ------------------------------ Message: 13 Date: Wed, 11 May 2005 09:36:52 +1000 From: Laurie Reilly Subject: Re: [Histonet] Mast cell stain To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <5.1.0.14.0.20050511092655.019a6ec0@mail.jcu.edu.au> Content-Type: text/plain; charset="us-ascii"; format=flowed Hi Histonetters, We routinely stain all dog skin lesions with Toluidine Blue and it highlights the Mast Cells. They stain metachromatically and show up with red-purple granules in the cytoplasm. These biopsies are all fixed in 10% Neutral Buffered Formalin. The method we use is from Gretchen Humason, "Animal tissue Techniques", 3rd edition. It uses a solution of 0.2% Tol Blue in 60% Ethanol for 2 minutes. Regards, Laurie. At 01:14 PM 05/10/05 -0400, Osborn, Sharon wrote: >Tom, > >Luna's Toluidine Blue Method for Mast Cells is a superior demonstration >of mast cells using a pink background. These literally pop up under >low power. The method is located on pages 311-312 of Luna's >Histopathologic Methods and color Atlas of special Stains and Tissue >Artifacts; 1992. If you wish a copy of the procedure, private email me >and I will send to you. > >sharon osborn >DNAX ScheringPlough BioPharma >Palo Alto, CA > >Subject: [Histonet] Giemsa Stain for Mast Cells >Netters, >I am looking for a Giemsa stain for Mast Cells that does not have a >pinkish hue like the Gaffney Giemsa. We are looking to have the Mast >Cells pop out at us on low power. Any suggestions/protocols? Thanks, >Tom >Tom Galati >Laboratory Director >HSRL, Inc.- A GLP Compliant Contract Laboratory >137 South Main Street >Woodstock, Virginia 22664 >(540)459-8211 >Fax: (540)459-8217 >tomgalati@hsrl.org >www.hsrl.org > > > >********************************************************************* >This message and any attachments are solely for the intended recipient. >If >you are not the intended recipient, disclosure, copying, use or >distribution of the information included in this message is prohibited -- >Please immediately and permanently delete. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. ------------------------------ Message: 14 Date: Tue, 10 May 2005 16:41:50 -0700 From: Patti Loykasek Subject: Re: [Histonet] job To: histonet Message-ID: Content-Type: text/plain; charset="US-ASCII" You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 15 Date: Tue, 10 May 2005 20:32:13 -0400 From: "HSRL" Subject: [Histonet] Mast Cell/Giemsa clarification To: Message-ID: <000601c555c0$e5a31840$0200a8c0@HSRLMAIN> Content-Type: text/plain; charset="US-ASCII" Netters, I agree with all of you about the Tol Blue stain being a good mast cell stain. However, we are specifically looking for a Giemsa stain for mast cell granules. We already do the Tol Blue, and the Pinacynol Erythrosinate stains for mast cells. Thanks again, Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 137 South Main Street Woodstock, Virginia 22664 (540)459-8211 Fax: (540)459-8217 tomgalati@hsrl.org www.hsrl.org ------------------------------ Message: 16 Date: Wed, 11 May 2005 07:42:03 +0100 From: Kemlo Rogerson Subject: RE: [Histonet] Master's of Histology To: "'Farley, Sunni R'" , histonet@lists.utsouthwestern.edu Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2A7@bhrv-nt-11.bhrv.nwest.nhs.uk> Content-Type: text/plain In the UK you can obtain a Master of Science Degree by pursuing a research project. In fact you could always take a Master's or Doctorate if you could find a Uni, a Supervisor and something of interest to research. I feel that postgraduate degrees have become easier to obtain, but then we old people always say that. In the UK NVQ's and other vocational qualifications are becoming more popular as it is really a judgement call, and I'm going to get into trouble, but maybe postgraduate degrees could be thought of as an overkill for many positions in the Laboratory. Do we really have that much to offer MSc's and PHD's? Other than funding them to get the qualification; I fess up to having an MSc, but...... -----Original Message----- From: Farley, Sunni R [mailto:sfarley@seattlecca.org] Sent: 10 May 2005 20:34 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Master's of Histology I have recently learned that you can earn a Master's of Histology/Cell Biology - does anybody have any information and/or recommendations on earning this degree? Thank you ahead for your time! Sunni Sunni R. Farley Histotechnician Clinical Pathology Mailstop G1-201 Seattle Cancer Care Alliance 825 Eastlake Ave East Seattle, WA 98109 (206) 288-1312 This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 17 Date: Wed, 11 May 2005 17:07:14 +0800 From: "eswary" Subject: [Histonet] (no subject) To: Message-ID: <200505111707.AA254607564@carif.com.my> Content-Type: text/plain; charset=us-ascii hi, does anyone know the advantage of using 0.01M sodium citrate over 0.01M citric acid buffer, pH 6, for antigen retrieval on paraffin embedded sections? also, is the sodium constituent important? would it matter if i used trisodium citrate instead of monosodium? thanks. et ------------------------------ Message: 18 Date: Wed, 11 May 2005 08:31:25 -0400 From: "Bernadette Weston" Subject: [Histonet] EDTA To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed We can no longer supply EDTA for our bone marrow procedures, can some one recommend a substitution for aspirate collection? Bernadette Weston HT Histology Supervisor Barberton Citizens Hospital Barberton, OH ------------------------------ Message: 19 Date: Wed, 11 May 2005 14:40:33 +0200 From: wasielewski.reinhard.von@mh-hannover.de Subject: [Histonet] macrophages in mouse FFPE To: histonet@lists.utsouthwestern.edu Message-ID: <42821961.31188.FCDE1BE2@localhost> Content-Type: text/plain; charset=US-ASCII Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski ------------------------------ Message: 20 Date: Wed, 11 May 2005 08:46:42 -0400 From: "Kristen Broomall" Subject: [Histonet] Gayle - floating petri dish snap freezing question To: Histonet@lists.utsouthwestern.edu Message-ID: <2B9D24FC863EC841B161CBA3BA4D5B3C0181D49A@wlmmsx01.nemours.org> Content-Type: text/plain; charset=iso-8859-1 Gayle, I tried out your snap freezing method with the floating petri dish in the liquid nitrogen yesterday for some itsy bitsy GI biopsies. I really like it! I have a question though. When using this method, do you place your sample in the bottom of your mold, then OCT on top, or put a dab (or more) of OCT, then the tissue, ... or does it really matter? Thanks, Kristen Broomall, HT (ASCP) ------------------------------ Message: 21 Date: Wed, 11 May 2005 09:21:33 -0400 From: "Flynn, Evelyn" Subject: RE: [Histonet] macrophages in mouse FFPE To: wasielewski.reinhard.von@mh-hannover.de, histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Dear Reinhard, I have had good results with rat anti-mouse Mac-3 antibody from PharMingen (Cat. # 550292). Regards, Evelyn Flynn, M.A. Children's Hospital, Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of wasielewski.reinhard.von@mh-hannover.de Sent: Wed 5/11/2005 8:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] macrophages in mouse FFPE Hi Histonetters, I am looking for a suitable antibody working in FFPE mouse tissue to detect macrophages (reproducibly and reliable). Many thanks in advance Reinhard PD Dr. med. Reinhard von Wasielewski _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 22 Date: Wed, 11 May 2005 07:08:31 -0700 (PDT) From: - - Subject: [Histonet] New rabbit antibody... but what do I use as negative (isotype) control? To: histonet@lists.utsouthwestern.edu Message-ID: <20050511140831.96586.qmail@web31706.mail.mud.yahoo.com> Content-Type: text/plain; charset=us-ascii Hi everyone, I have a new antibody that I was given. It is a rabbit polyclonal anti-mouse antibody. Usually I use a specific isotype control, but I do not have info on how this antibody was made other than the epitope(s) that it should adhere to... but I want a negative control and was wondering if I can just use regular rabbit serum (diluted) as a negative seeing that it should contain the isotype IgG I am using??? I think I am a bit confused here.... I need assistance. Can I use rabbit serum as a negative control? What dilution do I use if that is the case? Anything else I should know before I proceed? Thank you all!! --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! ------------------------------ Message: 23 Date: Wed, 11 May 2005 09:26:25 -0500 From: "Margaryan, Naira" Subject: RE: [Histonet] job To: "Patti Loykasek" , "histonet" Message-ID: <63B8B599DE283148B92E83C78B32C15DB9164E@cmhexbe2.childrensmemorial.org> Content-Type: text/plain What do you mean Pool position? Thanks, Naira -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Tuesday, May 10, 2005 6:42 PM To: histonet Subject: Re: [Histonet] job You can count me in if you mean moving to a warmer climate, more sunshine, and definite pool/sun time! Patti Loykasek Seattle, WA > Hi! > > We are looking for people who might be interested in a > pool position in the Chicago land area. > Please contact me Cynthia Haynes at 1-773-484-4133. > > Thanks in Advance > > Cynthia Haynes H.T. > > > > __________________________________ > Yahoo! Mail Mobile > Take Yahoo! Mail with you! Check email on your mobile phone. > http://mobile.yahoo.com/learn/mail > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. ------------------------------ Message: 24 Date: Wed, 11 May 2005 09:28:22 -0500 From: "Flores, Teresa" Subject: [Histonet] Unsubscribe To: histonet@lists.utsouthwestern.edu Cc: histonet-request@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, "'histonet@pathology.swmed.edu'" Message-ID: Content-Type: text/plain Please unsubscribe. ------------------------------ Message: 25 Date: Wed, 11 May 2005 09:28:22 -0500 From: "Flores, Teresa" Subject: [Histonet] Unsubscribe To: histonet@lists.utsouthwestern.edu Cc: histonet-request@lists.utsouthwestern.edu, histonet-bounces@lists.utsouthwestern.edu, "'histonet@pathology.swmed.edu'" Message-ID: Content-Type: text/plain Please unsubscribe. ------------------------------ Message: 26 Date: Wed, 11 May 2005 09:44:37 -0500 From: vsailes@nd.edu Subject: [Histonet] Help!!! To: histonet@lists.utsouthwestern.edu Message-ID: <1115822677.42821a55c5ced@webmail.nd.edu> Content-Type: text/plain; charset=ISO-8859-1 Hello Histonetters, I'm working on an IHC protocol for Albumin -- staining FFPE mouse tissues. I'm new to working out conditons for antibody protocols and could use some help.I'm testing the antibody at different dilutions and I appear to have positive staining but I'm also seeing small areas in the negative that shows positive staining. Liver is my control and I use steamer (HIER) for 30 minutes, my steps include a 30 min quench in 1.6% hydrogen peroxide, NS block, Antibody, Secondary antibody(Biotin), Streptavidin, Chromogen, Counterstain My concern is that I'm using buffers that contain BSA and I'm wondering if that is causing staining to appear or should I be using avidin/biotin block along with a peroxide block. Any suggestion would be helpful. Thank you for your help in advance. Valerie ------------------------------ Message: 27 Date: Wed, 11 May 2005 10:05:33 -0500 From: "LINDA MARGRAF" Subject: [Histonet] job listing To: Message-ID: Content-Type: text/plain; charset=US-ASCII Lead and/or Bench Histology Technician Scott & White is seeking a Lead and/or Bench Histology Technician. Responsible for histology laboratory operations including testing, training of support personnel, QA, personnel utilization and regulatory compliance. Selected candidate will also participate in routine histology laboratory functions. Requirements: An Associate's degree; HT(ASCP); 3 to 5 years experience. Just an hour north of Austin, Scott & White offers highly competitive salaries, comprehensive benefits and advancement opportunities. Candidates should send resumes to: ghollie@swmail.sw.org. Phone: 800.527.JOBS. Fax: 254.724.5591. Apply in person or mail resume to: Human Resources Dept., 2401 S. 31st St., Temple, TX 77508. www.sw.org An equal opportunity employer/tobacco-free environment. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 14 **************************************** -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- From p.j.bergin <@t> qmul.ac.uk Thu May 12 03:24:29 2005 From: p.j.bergin <@t> qmul.ac.uk (p.j.bergin@qmul.ac.uk) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Journal of histotechnology paper search Message-ID: <1115886269.428312bdca5b5@webapps.qmul.ac.uk> Hi everyone, I was hoping that someone out there could give me some advice on how to search for an article from the journal of histotechnology. I have had some staining done and want to reference the original paper for the method- apparently its 'Leung K and Gibbon KJ, A rapid staining method for Helicobacter pylori in gastric biopsies, Journal of Histotechnology, June 1996, v 19, No.2 pp 131'. Unfortunately the paper doesn't show up on medline- and I can't seem to find an archive for J Histotech that goes back that far so I can check the reference is correct. The website is great for the journal, but its archive only goes back to 1999 (so far) and they don't have contents for past issues. Thanks for any help, Phil From lpwenk <@t> sbcglobal.net Thu May 12 05:23:13 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Journal of histotechnology paper search In-Reply-To: <1115886269.428312bdca5b5@webapps.qmul.ac.uk> Message-ID: Contact the National Society for Histotechnology. They publish the journal. 301-262-6221 histo@nsh.org Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of p.j.bergin@qmul.ac.uk Sent: Thursday, May 12, 2005 4:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Journal of histotechnology paper search Hi everyone, I was hoping that someone out there could give me some advice on how to search for an article from the journal of histotechnology. I have had some staining done and want to reference the original paper for the method- apparently its 'Leung K and Gibbon KJ, A rapid staining method for Helicobacter pylori in gastric biopsies, Journal of Histotechnology, June 1996, v 19, No.2 pp 131'. Unfortunately the paper doesn't show up on medline- and I can't seem to find an archive for J Histotech that goes back that far so I can check the reference is correct. The website is great for the journal, but its archive only goes back to 1999 (so far) and they don't have contents for past issues. Thanks for any help, Phil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KPercival <@t> wyeth.com Thu May 12 06:41:41 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] prostate needle biopsies Message-ID: Brian, There is no standard protocol for how many microns there should be between levels because, as you say, each block is different. What we did in our lab, when I was working in a clinical pathology lab, was to save the tissue between levels and use that for IHC requests. On rare occaisions, we would decolorize an H&E level and use that for IHC or a special stain. We started putting all three levels on the same slide after a time in order to reduce costs. We did not embed with a tamper because the tissue cores were so delicate. We also soaked our blocks on a cold plate with a little detergent (Joy dishwasing soap) in water. The blocks cut great, no wrinkles and no recut requests for additional H&E's due to poor quality. All of our techs did this and we had consistent and high-quality results. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> "Brian D. Conlon" 5/11/2005 5:37:57 PM >>> hi everyone, our lab has a question about prostate needle biopsies. the pa's submit two cores per block and we embed them with a tamper so they are on the same level. we do three levels on each block. is there a protocol as to how many microns should be between each level? sometimes the tissue is so thin that there is nothing left in the block for recuts or ihc. we would like to have each tech cut with the same method so that there will be greater consistency. thanks so much everybody. paula _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KPercival <@t> wyeth.com Thu May 12 06:44:41 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Disposing of Blocks Message-ID: Jessica, Check with your state and local countyj/community rules, too. We (in Massachusetts) had to comply with local regualtions and "red-bag" or biohazard our blocks for disposal. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> Jessica Piche 5/11/2005 9:21:50 PM >>> Hi Everyone, How are we supposed to get rid of patient blocks once the required time to keep them ends? We are throwing them out in the regular trash and are wondering if regulations might have changed. Thanks, Jessica Piche-Grocki, HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shawnster73 <@t> aol.com Thu May 12 06:55:28 2005 From: shawnster73 <@t> aol.com (shawnster73@aol.com) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Boston Message-ID: <8C725009FF7C80E-EE8-2DD84@FWM-R41.sysops.aol.com> My husband is taking a position in the Boston area and I am interested if anyone knows of some Histotech job openings in that area and what the salary range is for that part of the country. From GauchV <@t> mail.amc.edu Thu May 12 07:45:06 2005 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Disposing of Blocks Message-ID: Jessica, We dispose of our blocks and slides once it is past the necessary hold period, in double red bags inside of hard fiber boxes and they are picked up by your EHS department for disposal. Vicki Gauch AMCH Albany, NY >>> Jessica Piche 5/11/2005 9:21:50 PM >>> Hi Everyone, How are we supposed to get rid of patient blocks once the required time to keep them ends? We are throwing them out in the regular trash and are wondering if regulations might have changed. Thanks, Jessica Piche-Grocki, HT(ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu May 12 07:58:56 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Journal of histotechnology paper search In-Reply-To: <200505121027.j4CARkpe081439@pro12.abac.com> Message-ID: <200505121259.j4CCx9Y1070876@pro12.abac.com> JOH is applying for indexing in Medline which we do not yet have. Hopefully soon we will be indexed. We are also planning to expand the JOH website but that too will be in the future. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Thursday, May 12, 2005 3:23 AM To: p.j.bergin@qmul.ac.uk; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Journal of histotechnology paper search Contact the National Society for Histotechnology. They publish the journal. 301-262-6221 histo@nsh.org Peggy Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of p.j.bergin@qmul.ac.uk Sent: Thursday, May 12, 2005 4:24 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Journal of histotechnology paper search Hi everyone, I was hoping that someone out there could give me some advice on how to search for an article from the journal of histotechnology. I have had some staining done and want to reference the original paper for the method- apparently its 'Leung K and Gibbon KJ, A rapid staining method for Helicobacter pylori in gastric biopsies, Journal of Histotechnology, June 1996, v 19, No.2 pp 131'. Unfortunately the paper doesn't show up on medline- and I can't seem to find an archive for J Histotech that goes back that far so I can check the reference is correct. The website is great for the journal, but its archive only goes back to 1999 (so far) and they don't have contents for past issues. Thanks for any help, Phil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgfields <@t> lexhealth.org Thu May 12 08:25:17 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Histo. cutting requirements Message-ID: Awhile back on the HistoNet there was information about average cutting requirements for Histo techs...... time cutting one block, etc. Does anyone remember where that information is located? THX in advance. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From JWEEMS <@t> sjha.org Thu May 12 08:38:10 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Touch Prep CPT Code Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45CC4@sjhaexc02.sjha.org> Do you guys charge for touch preps? If so, what CPT code? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From Diane.Gladney <@t> se.amedd.army.mil Thu May 12 08:52:07 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Touch Prep CPT Code Message-ID: <4D55B2E997EFAE4DA6081DDE100B83025049DA@amedmlsermc133.amed.ds.army.mil> I would really like this information, also. Thanks, Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Thursday, May 12, 2005 9:38 AM To: Histonet Subject: [Histonet] Touch Prep CPT Code Do you guys charge for touch preps? If so, what CPT code? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From la.sebree <@t> hosp.wisc.edu Thu May 12 09:22:07 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] AA Amyloid Message-ID: One of our pathologists wants us to bring on AA Amyloid antibody for investigating cases of amyloidosis. What are people using with good results? We have Ventana automated immunostainers. Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From gu.lang <@t> gmx.at Thu May 12 10:06:49 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] giemsa stain, green-blue acid mucin Message-ID: Hallo, Last time we stained a gastric biopsy with Giemsa. We found cells with green-blue acid mucin. I have never seen this before. It looks like the mucins in the movat-stain. Can anyone tell me the theoretical background about this? We do the Giemsa on the Ventana-stainer, with following differentiation in A.d. with a few drops glacial acetic acid. Thank you Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cgfields <@t> lexhealth.org Thu May 12 10:16:23 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:05 2005 Subject: FW: [Histonet] Histo. cutting requirements Message-ID: Thank you for the information on productivity. The information was in the Journal of Histotechnology, December 2004. It is not on line yet. Good article. Thanks again Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 -----Original Message----- From: Nita Searcy [mailto:NSEARCY@swmail.sw.org] Sent: Thursday, May 12, 2005 9:58 AM To: Carole Fields Subject: Re: [Histonet] Histo. cutting requirements NSH did a workload study and published in journal >>> Carole Fields 05/12/05 8:25 AM >>> Awhile back on the HistoNet there was information about average cutting requirements for Histo techs...... time cutting one block, etc. Does anyone remember where that information is located? THX in advance. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From gcallis <@t> montana.edu Thu May 12 10:24:51 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Journal of histotechnology paper search In-Reply-To: <1115886269.428312bdca5b5@webapps.qmul.ac.uk> References: <1115886269.428312bdca5b5@webapps.qmul.ac.uk> Message-ID: <6.0.0.22.1.20050512091333.01b5f528@gemini.msu.montana.edu> Phil, If you send me your postal mailing address, I will make a copy and send it to you. I have all JOH issues since publication started in 1977. I could try and scan it, then attach, but my scanner is at home, journals at work. I am more than happy to help. I was just in the Sakura Finetek website, http://www.sakuraus.com/ and while searching Histo Logic archives, I ran across additional articles on Helicobacter staining. I found these under staining and quality assurance topics. JOH is still building their archives - only begun in the last few years. At 02:24 AM 5/12/2005, you wrote: >Hi everyone, > >I was hoping that someone out there could give me some advice on how to search >for an article from the journal of histotechnology. I have had some staining >done and want to reference the original paper for the method- apparently its >'Leung K and Gibbon KJ, A rapid staining method for Helicobacter pylori in >gastric biopsies, Journal of Histotechnology, June 1996, v 19, No.2 pp 131'. >Unfortunately the paper doesn't show up on medline- and I can't seem to >find an >archive for J Histotech that goes back that far so I can check the >reference is >correct. > >The website is great for the journal, but its archive only goes back to >1999 (so >far) and they don't have contents for past issues. > >Thanks for any help, > >Phil > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From algranth <@t> u.arizona.edu Thu May 12 11:10:04 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde Message-ID: <4.3.2.7.2.20050512090428.024f4918@algranth.inbox.email.arizona.edu> One of the labs here is closing and they have a case of formaldehyde, 37.5%, that they are trying to give away. They have had it in their lab since 1993. The bottles have not been opened. Is it still good to use? Andi ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From algranth <@t> u.arizona.edu Thu May 12 11:50:53 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde In-Reply-To: <6.0.0.22.1.20050512102407.01b0e680@gemini.msu.montana.edu> References: <4.3.2.7.2.20050512090428.024f4918@algranth.inbox.email.ari zona.edu> <4.3.2.7.2.20050512090428.024f4918@algranth.inbox.email.arizona.edu> Message-ID: <4.3.2.7.2.20050512094909.00cc8008@algranth.inbox.email.arizona.edu> Thanks for the quick answers - you pretty much echoed what I thought - call our waste disposal guys and get rid of it. Believe it or not, there is no exp. date on the bottles. Andi At 10:25 AM 5/12/2005 -0600, you wrote: >Andi, > >Since 1993, I would give it to chemical safety for disposal!!! Don't >take a chance on your projects. Maybe it is good but after 12 years, I >would dump it. > >Gayle > >At 10:10 AM 5/12/2005, you wrote: >>One of the labs here is closing and they have a case of formaldehyde, >>37.5%, that they are trying to give away. They have had it in their lab >>since 1993. The bottles have not been opened. Is it still good to use? >>Andi >>..................................................................... >>: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : >>: Sr. Research Specialist University of Arizona : >>: (office: AHSC 4212) P.O. Box 245044 : >>: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : >>: (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : >>:...................................................................: >> http://www.cba.arizona.edu/histology-lab.html >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From jkiernan <@t> uwo.ca Thu May 12 11:56:46 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Journal of histotechnology paper search References: <1115886269.428312bdca5b5@webapps.qmul.ac.uk> Message-ID: <42838ACE.B85F48EC@uwo.ca> A search with www.Scopus.com brought up: ----- Journal of Histotechnology Volume 19, Issue 2, June 1996, Pages 131-132 Rapid staining method for Helicobacter pylori in gastric biopsies Leung, J.K.ab, Gibbon, K.J.a., Vartanian, R.K.a a Anatomic Pathology, Vancouver Hosp. and Hlth. Sci. Ctr., Vancouver, BC, Canada b Vancouver Hosp. and Hlth. Sci. Ctr., Anatomic Pathology, 855 W. 12th Street, Vancouver, BC V5Z 1M9, Canada Abstract Helicobucter pytori is a gram negative bacterium that has been shown to be the causative organism in some gastric and duodenal ulcers. It is the most common cause of antral gastritis and has been linked to the development of distal gastric carcinoma and low grade gastric lymphoma. Identification in a gastric biopsy and prompt treatment with antibiotics, can lead to a speedy recovery for the patient. Unfortunately this organism stains poorly with standard hematoxylin and eosin techniques and special staining methods can be lengthy. The usual demonstration methods are Steiner and Romanowski type stains. Toluidine blue has been used as a rapid method for the demonstration of these organisms, but the contrast is generally very poor. We have developed a counterstain for toluidine blue that shows deep blue organisms on a yellow background. The combined procedure takes about 30 min. Author Keywords alcian yellow; gastric biopsy; Helicobacter pylori; toluidine blue Matched Index Keywords: helicobacter pylori See the Extended format page for all index keywords in this document ------ I couldn't find the full text on line. Anyone who has been a member of the NSH since June 1996 should have the journal. Scopus is an excellent literature searching service, even better than Web of Science, but your library has to subscribe to it. Your library at Queen Mary College subscribes to Web of Science (which, unlike Medline, covers all journals). They may well be testing Scopus, as our library is. It's very new. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ p.j.bergin@qmul.ac.uk wrote: > > Hi everyone, > > I was hoping that someone out there could give me some advice on how to search > for an article from the journal of histotechnology. I have had some staining > done and want to reference the original paper for the method- apparently its > 'Leung K and Gibbon KJ, A rapid staining method for Helicobacter pylori in > gastric biopsies, Journal of Histotechnology, June 1996, v 19, No.2 pp 131'. > Unfortunately the paper doesn't show up on medline- and I can't seem to find an > archive for J Histotech that goes back that far so I can check the reference is > correct. > > The website is great for the journal, but its archive only goes back to 1999 (so > far) and they don't have contents for past issues. > > Thanks for any help, > > Phil > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plucas <@t> biopath.org Thu May 12 12:28:30 2005 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: <003101c55718$046051a0$7c01a8c0@biopath.org> If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group From jqb7 <@t> cdc.gov Thu May 12 12:37:23 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: We leave ours on continuously. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Paula Lucas Sent: Thu 5/12/2005 1:28 PM To: Histonet (E-mail) Cc: Subject: [Histonet] Oven for Histology If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Thu May 12 12:38:35 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: We keep our Desert Chamber on all the time. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paula Lucas Sent: Thursday, May 12, 2005 12:29 PM To: Histonet (E-mail) Subject: [Histonet] Oven for Histology If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu May 12 12:41:59 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45CD4@sjhaexc02.sjha.org> We keep ours on all the time. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Paula Lucas Sent: Thursday, May 12, 2005 1:29 PM To: Histonet (E-mail) Subject: [Histonet] Oven for Histology If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From Jackie.O'Connor <@t> abbott.com Thu May 12 12:42:25 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: I'm curious as to whether or not you have a paraffin accumulation on the heating element - if it's in the bottom of the oven - you probably do. I have an oven that's older than me - I leave it on all the time (it's been on for 3 years now) with no adverse affects. I put my slide racks on paper towels to absorb the melting paraffin, and throw them away when I remove the slides. I could get a new oven - but if it ain't broke . . . . . . Jackie O' "Paula Lucas" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/12/2005 12:28 PM To: "Histonet \(E-mail\)" cc: Subject: [Histonet] Oven for Histology If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Thu May 12 12:48:29 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde References: <4.3.2.7.2.20050512090428.024f4918@algranth.inbox.email.arizona.edu> Message-ID: <428396ED.9DEF8B2B@uwo.ca> Two things could have happened to unopened formalin in 12 years: 1. Polymerization (to paraformaldehyde). This is evident as a white precipitate. It slightly reduces the concentration in the liquid, but that does not matter for fixation. Polymerization is accelerated by low room temperature, and it is claimed that the process can be reversed by autoclaving(paper in Stain Technol about 40 years ago). 2. Cannizzaro's reaction, in which 2 molecules of formaldehyde react together, producing one molecule each of methanol and formic acid. This happens in all formaldehyde solutions and causes lowering of the pH. This doesn't matter if you make a neutral buffered fixative solution. Small amounts of methanol and formate ions are not going to change the fixative properties. Bottom line: OK to use, but be sure to check the pH of the working fixative solution and adjust if necessary. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Andrea Grantham wrote: > > One of the labs here is closing and they have a case of formaldehyde, > 37.5%, that they are trying to give away. They have had it in their lab > since 1993. The bottles have not been opened. Is it still good to use? > Andi > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tpmorken <@t> labvision.com Thu May 12 13:08:04 2005 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328CD08@usca0082k08.labvision.apogent.com> John, you said " Small amounts of methanol and formate ions are not going to change the fixative properties." But after 12 years will it really be a "small amount?" How do we know what percentage of the solution will have been converted? Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Thursday, May 12, 2005 10:48 AM To: Andrea Grantham Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quick question about formaldehyde Two things could have happened to unopened formalin in 12 years: 1. Polymerization (to paraformaldehyde). This is evident as a white precipitate. It slightly reduces the concentration in the liquid, but that does not matter for fixation. Polymerization is accelerated by low room temperature, and it is claimed that the process can be reversed by autoclaving(paper in Stain Technol about 40 years ago). 2. Cannizzaro's reaction, in which 2 molecules of formaldehyde react together, producing one molecule each of methanol and formic acid. This happens in all formaldehyde solutions and causes lowering of the pH. This doesn't matter if you make a neutral buffered fixative solution. Small amounts of methanol and formate ions are not going to change the fixative properties. Bottom line: OK to use, but be sure to check the pH of the working fixative solution and adjust if necessary. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Andrea Grantham wrote: > > One of the labs here is closing and they have a case of formaldehyde, > 37.5%, that they are trying to give away. They have had it in their > lab since 1993. The bottles have not been opened. Is it still good to > use? Andi > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Thu May 12 13:12:14 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45CD8@sjhaexc02.sjha.org> Also, John said "formalin" - was the solution formalin or 37% formaldehyde - without buffers? Would that make a difference? Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, Tim - Labvision Sent: Thursday, May 12, 2005 2:08 PM To: 'John Kiernan'; Andrea Grantham Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] quick question about formaldehyde John, you said " Small amounts of methanol and formate ions are not going to change the fixative properties." But after 12 years will it really be a "small amount?" How do we know what percentage of the solution will have been converted? Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Thursday, May 12, 2005 10:48 AM To: Andrea Grantham Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quick question about formaldehyde Two things could have happened to unopened formalin in 12 years: 1. Polymerization (to paraformaldehyde). This is evident as a white precipitate. It slightly reduces the concentration in the liquid, but that does not matter for fixation. Polymerization is accelerated by low room temperature, and it is claimed that the process can be reversed by autoclaving(paper in Stain Technol about 40 years ago). 2. Cannizzaro's reaction, in which 2 molecules of formaldehyde react together, producing one molecule each of methanol and formic acid. This happens in all formaldehyde solutions and causes lowering of the pH. This doesn't matter if you make a neutral buffered fixative solution. Small amounts of methanol and formate ions are not going to change the fixative properties. Bottom line: OK to use, but be sure to check the pH of the working fixative solution and adjust if necessary. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Andrea Grantham wrote: > > One of the labs here is closing and they have a case of formaldehyde, > 37.5%, that they are trying to give away. They have had it in their > lab since 1993. The bottles have not been opened. Is it still good to > use? Andi > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From funderwood <@t> mcohio.org Thu May 12 13:17:45 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: I have mine on a timer. The slides dry overnight, the oven swithces off before I arrive in the morning. I've had no problems with the operation of the oven. Fred >>> "Paula Lucas" 05/12/05 01:28PM >>> If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From t-sherman <@t> comcast.net Thu May 12 13:34:58 2005 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Re: Histonet Digest, Vol 18, Issue 18 Message-ID: Hello Andrea, I think I'll offer a counterpoint to dumping the chemical. Test it to see if it's still good. If you do not have access to a lab (or curious chemistry classroom) nearby that can perform the test, there are kits available to do such analysis. For example, I did a quick Google search and found this: http://omnicontrols.com/lists/Hanna_Chem_color_titratTestKit.html#FORMALDEHYDE%20TEST%20KIT Granted, their test only has a range up to 10% and not 37%... but I didn't search for very long. I don't know this company from Adam but many companies offer such titration kits. If the solution is still tightly sealed and there is no precipitate in the bottle, there is not much reason to send it to some hazardous waste site; afterall, it's still just a noxious chemical with its properties intact. If the organic content or buffer is unable to escape from its container, and the container is inert (glass would be) then the solution should maintain its viability. Also, you might even consider using any unused titration tests on your "good" stuff just to confirm... sort of a quality control confirmation that a) you are doing the test properly and b) the good stuff isn't just foul-smelling water. :) Fundamentally, I dislike wasting such chemicals particularly when the typically overlooked expense of handling and disposal is incorporated. Certainly I'd confirm its quality before using it on critical tissue, but I'd still test it before dumping such a generous amount. If you still opt to dump it, maybe you could call some research laboratory or other university (not Arizona) that could put it to use first? Of course, I'm ignoring any legalities and expense of such a transfer so this may be a moot point. At any rate, just plunking down another $0.02. Todd Sherman HistoSoft Corporation -- >>>>>>>> www.histosoft.com <<<<<<<<< <<<<<<<< Biology In A New Form (c) >>>>>>> On Thu, 12 May 2005 12:00:15 -0500, wrote: > > Today's Topics: > > 15. quick question about formaldehyde (Andrea Grantham) > > ---------------------------------------------------------------------- > > Message: 15 > Date: Thu, 12 May 2005 09:10:04 -0700 > From: Andrea Grantham > Subject: [Histonet] quick question about formaldehyde > To: histonet@lists.utsouthwestern.edu > Message-ID: > <4.3.2.7.2.20050512090428.024f4918@algranth.inbox.email.arizona.edu> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > One of the labs here is closing and they have a case of formaldehyde, > 37.5%, that they are trying to give away. They have had it in their lab > since 1993. The bottles have not been opened. Is it still good to use? > Andi > ..................................................................... > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > : Sr. Research Specialist University of Arizona : > : (office: AHSC 4212) P.O. Box 245044 : > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > :...................................................................: > http://www.cba.arizona.edu/histology-lab.html > > ---------------------------------------------------------------------- From ander093 <@t> tc.umn.edu Thu May 12 13:45:20 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:25:05 2005 Subject: Fwd: Re: [Histonet] Oven for Histology Message-ID: <6.1.2.0.0.20050512134502.028b9890@ander093.email.umn.edu> > > >Ovens are left on--never turned off except to clean. > > >At 12:28 PM 5/12/05, you wrote: >>If someone could offer an opinion or if any one has experienced this type of >>situation, I would appreciate the feedback: >> >>We have an oven that is about 4 to 5 years old, which we use to dry our >>histology slides. The heating element is going out already and my concern >>is that it's relatively new for having this problem. >> >>We turn the oven off before we leave for the day and then turn it back on >>the following morning. >> >>I'm wondering if turning on/off would cause problems or it really doesn't >>matter. >> >>Do you keep your ovens on overnight or off? >> >>Thanks in advance. It's greatly appreciated. >> >>Paula >>Bio-Path Medical Group >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From plucas <@t> biopath.org Thu May 12 14:13:55 2005 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven Responses Message-ID: <005c01c55726$be77e770$7c01a8c0@biopath.org> Thanks to all who responded. This is a great resource for help/feedback. Paula From TillRenee <@t> uams.edu Thu May 12 14:20:41 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Ki-67 Message-ID: What is a good Ki-67 for rat or mouse tissues? I am mostly concerned with the rat. Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 From froyer <@t> bitstream.net Thu May 12 14:37:34 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology In-Reply-To: <003101c55718$046051a0$7c01a8c0@biopath.org> References: <003101c55718$046051a0$7c01a8c0@biopath.org> Message-ID: <4283B07E.2090706@bitstream.net> You are correct. The heating element on an oven that is 4-5 years old should not be "going out" ...if it has not been artificially corroded. Solvents, oxidizers, paraffin, etc. that has been spilled or dripped onto the heating element will most certainly harm it. Also, you did not mention what temperature you are routinely using. From what I see on the HistNet, most folks run anywhere from 50 degree C to 70 degree C. The higher the temp, the shorter the life of the element... but even at 70 degrees, an oven heating element should last for many years. This depends on the "oven". What is the temperature range, as specified in your operator's manual for your oven?. Most "ovens" are Room Temp. to 200 to 300 degrees C. Check your "oven" to make sure that you are not actually using an "Incubator" Standard Incubators have a temperature range of Room temp to 65-70 degrees. If in fact you have an "incubator" and are running at 65 degrees, you are almost at the max of the temperature range set for that heating element. It will therefore burn out much quicker. Oven heating elements (200-300 degrees) are made of sterner stuff than Incubator elements. Another thing to check is if it is, in fact, the heating element that is going out. It could be an electrical problem elsewhere in the unit.... the thermostat as example. In my years of experience, I have not seen a major shelf- life difference between those who leave their ovens 'ON' 24/7 and those who turn them off at the end of each day. Just a few tips in the wind... ~ Ford Ford M. Royer, MT(ASCP) Midwest Science & Biocenter Minneapolis, MN 800-745-4869 Paula Lucas wrote: >If someone could offer an opinion or if any one has experienced this type of >situation, I would appreciate the feedback: > >We have an oven that is about 4 to 5 years old, which we use to dry our >histology slides. The heating element is going out already and my concern >is that it's relatively new for having this problem. > >We turn the oven off before we leave for the day and then turn it back on >the following morning. > >I'm wondering if turning on/off would cause problems or it really doesn't >matter. > >Do you keep your ovens on overnight or off? > >Thanks in advance. It's greatly appreciated. > >Paula >Bio-Path Medical Group > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From anh2006 <@t> med.cornell.edu Thu May 12 14:55:34 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Ki-67 - for rat tissue In-Reply-To: References: Message-ID: MIB-5 is the Ki67 antibody of choice for rat tissue. DAKO sells it. At 2:20 PM -0500 5/12/05, Till, Renee wrote: >What is a good Ki-67 for rat or mouse tissues? I am mostly concerned >with the rat. > -- From Charlene.Henry <@t> STJUDE.ORG Thu May 12 15:11:56 2005 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] TDT over PAP Stain Message-ID: <5CB39BCA5724F349BCB748675C6CA1A202C75710@SJMEMXMB02.stjude.sjcrh.local> Has anyone ever ran a TdT stain on top of a PAP stained cytospin? We have a visiting international outreach pathologist that wants to be able to run a TdT on the same cytospin that has been stained with a PAP stain. Any suggestions? Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 From JMacDonald <@t> mtsac.edu Thu May 12 15:26:03 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] asbestos fibers In-Reply-To: <1030B679AD69D6119C3F00080210DD9D05A3F2AA@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: I believe asbestos will stain with a Perl's Prussian Blue reaction. Kemlo Rogerson Sent by: histonet-bounces@lists.utsouthwestern.edu 05/11/2005 11:45 PM To 'Fred Underwood' , histonet@lists.utsouthwestern.edu cc Subject RE: [Histonet] asbestos fibers Burn the tissue? Asbestos doesn't, burn that is. -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: 09 May 2005 19:51 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] asbestos fibers Hi All, Does someone have a technic for demonstrating asbesots fibers in lung. Possibly by isolating them through digestion of the tissue. Thanks in advance. Fred Underwood HT(ASCP) Montgomery County Coroner Dayton, OH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Thu May 12 15:42:40 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] TDT over PAP Stain Message-ID: We do it for other markers; why not try it (that is, of course, you do Tdt IHC on a routine basis). Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Henry, Charlene" 05/12/05 04:11PM >>> Has anyone ever ran a TdT stain on top of a PAP stained cytospin? We have a visiting international outreach pathologist that wants to be able to run a TdT on the same cytospin that has been stained with a PAP stain. Any suggestions? Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From ktournear <@t> cox.net Thu May 12 17:55:41 2005 From: ktournear <@t> cox.net (ktournear@cox.net) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Job Opportunity-Tucson, AZ Message-ID: <20050512225540.SPGG550.fed1rmmtao12.cox.net@smtp.west.cox.net> Specialists in Dermatology, located in sunny Tucson, Arizona is seeking to fill a full time Histology/Mohs Technican position. Specialists in Dermatology offers a wide range of skin care. In addition, we offer skin care products such as a line of make-up, as well as laser procedures, chemical procedures, and cosmetic procedures. We offer a competitive salary, medical benefits, 401K, and profit sharing. Relocation assistance offered with commitment. In addition to this, we have a new facility with an excellent team of employees and 5 wonderful doctors. Minimum qualifications: minimum 2 years histology experience in a private or clinical lab setting, HT/HTL certification, experience with Mohs procedure preferred, but not necessary, will train. Job description: Grossing skin specimens, tissue processing, maintenance of all histology equipment, embedding, cutting, staining (H&E and special stains), process microscopic slides for pathological evaluation. Assist Mohs surgeon, cut frozen sections related to Mohs procedure. Visit our web site at: www.clearskindoctor.com If interested, please fax or email resume to: Susan Britt-Roby, C.P.C e-mail: manager@clearskindoctor.com fax#: 520-319-1101 From Adesupod <@t> aol.com Thu May 12 19:06:18 2005 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] LOOKING FOR NEW JOB Message-ID: Netters, I am looking for a new job opportunity. My pathologist passed away few months ago, and the hospital management have now decided to close down the pathology dept. My name is Adesupo Adesuyi. I have BS in Medical Technology and I am HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years experience as an histotech. I presently work in Texas, but I am willing to relocate. My contact infos are; House #: 830-768-0859 Cell #: 830-422-9038 E/Mail: adesupod@aol.com Adesupo Adesuyi Pathology Dept, Val Verde Reg.Med.Center, Del Rio, Tx 78840. From djohnson14 <@t> hotmail.com Thu May 12 19:16:50 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] LOOKING FOR NEW JOB In-Reply-To: Message-ID: Depending on what you want to make, you could get a job anywhere. It seems everyone is hiring Unfortunately, it doesnt seem that salarys are rising despite the lack of histotechs. Sad and scary situation if you ask me. I am not much for unions but now i see why they are valuable. Just my opinion. >From: Adesupod@aol.com >To: histonet@lists.utsouthwestern.edu >CC: Adesupod@aol.com >Subject: [Histonet] LOOKING FOR NEW JOB >Date: Thu, 12 May 2005 20:06:18 EDT > > > Netters, > I am looking for a new job opportunity. My pathologist passed >away few months ago, and the hospital management have now decided to close >down >the pathology dept. > My name is Adesupo Adesuyi. I have BS in Medical Technology and I am >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years experience >as an >histotech. > I presently work in Texas, but I am willing to relocate. > My contact infos are; > > House #: 830-768-0859 > > Cell #: 830-422-9038 > > E/Mail: adesupod@aol.com > > Adesupo Adesuyi > Pathology Dept, > Val Verde Reg.Med.Center, > Del Rio, Tx 78840. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ulphani <@t> yahoo.com Thu May 12 21:22:53 2005 From: ulphani <@t> yahoo.com (Joseph Ulphani) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] (no subject) Message-ID: <20050513022253.19733.qmail@web30514.mail.mud.yahoo.com> Anyone knows the method for staining a whole tissue ( e.g. kidney or heart) for adrenergic nerves on the surface of the tissue? --------------------------------- Do you Yahoo!? Yahoo! Mail - Find what you need with new enhanced search. Learn more. From Kim.Osullivan <@t> med.monash.edu.au Fri May 13 00:34:43 2005 From: Kim.Osullivan <@t> med.monash.edu.au (Kim O'Sullivan) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Immunohistowax supplier Message-ID: <130.194.114.57.1115962219.00414@my.monash.edu.au> Hi all, Can anyone tell me if they know of an australian supplier of immunohistowax, and immunohistofix- apparently T cell staining is successful using this method with the exception of GK1.4(anti CD4). For us to import it from the UK would cost the same amount as the product. Also any feed back from people who have used it would be useful, particulary if they have used a rotary microtome to cut the tissue with instead of a sliding microtome (don't want to buy a sliding microtome if you can cut it with a rotary one). Thanks Kim From bruyntjes <@t> voeding.tno.nl Fri May 13 01:56:29 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Ki-67 Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DB6@ntexch1.voeding.tno.nl> Renee I use an antibody from Labvision/Neomarkers on rat tissues. It is a rabbit monoclonal and it works great. HIER in citrate pH 6.0 for 15 minutes. Joost Bruijntjes TNO Zeist Holland -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Till, Renee Sent: donderdag 12 mei 2005 21:21 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Ki-67 What is a good Ki-67 for rat or mouse tissues? I am mostly concerned with the rat. Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From HornHV <@t> archildrens.org Fri May 13 08:34:49 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Oven for Histology Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDD3D@EMAIL.archildrens.org> We have 2 ovens that are on continuously. I've worked here 15 years and they were both here when I started this job. In fact, one is so old, I think it came over on the Ark. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital - Changing Children's Lives Phone - 501.364.4240 Fax - 501.364.3912 Visit us on the web at www. archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Thursday, May 12, 2005 1:18 PM To: plucas@biopath.org; histonet@pathology.swmed.edu Subject: Re: [Histonet] Oven for Histology I have mine on a timer. The slides dry overnight, the oven swithces off before I arrive in the morning. I've had no problems with the operation of the oven. Fred >>> "Paula Lucas" 05/12/05 01:28PM >>> If someone could offer an opinion or if any one has experienced this type of situation, I would appreciate the feedback: We have an oven that is about 4 to 5 years old, which we use to dry our histology slides. The heating element is going out already and my concern is that it's relatively new for having this problem. We turn the oven off before we leave for the day and then turn it back on the following morning. I'm wondering if turning on/off would cause problems or it really doesn't matter. Do you keep your ovens on overnight or off? Thanks in advance. It's greatly appreciated. Paula Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From GDawson <@t> dynacaremilwaukee.com Fri May 13 09:06:49 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] low salary & unions Message-ID: Dave, My lab has unionized histotechs and I'd say we are in the lower half of salary ranges for our area. As far as the union here goes, it seems they are good at collecting dues and at keeping garbage employees employed. Not only do unions allow bad employees to keep their jobs, they make it difficult to reward exceptional employees for going above and beyond the call of duty (since all employees are given union negotiated raises/benefits/etc..). This might be a grass is always greener thing so don't be so sure that a union will help the histotech's position, that is not always the case. My Opinion, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Dave Johnson [mailto:djohnson14@hotmail.com] Sent: Thursday, May 12, 2005 6:17 PM To: Adesupod@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] LOOKING FOR NEW JOB Depending on what you want to make, you could get a job anywhere. It seems everyone is hiring Unfortunately, it doesnt seem that salarys are rising despite the lack of histotechs. Sad and scary situation if you ask me. I am not much for unions but now i see why they are valuable. Just my opinion. >From: Adesupod@aol.com >To: histonet@lists.utsouthwestern.edu >CC: Adesupod@aol.com >Subject: [Histonet] LOOKING FOR NEW JOB >Date: Thu, 12 May 2005 20:06:18 EDT > > > Netters, > I am looking for a new job opportunity. My pathologist passed >away few months ago, and the hospital management have now decided to close >down >the pathology dept. > My name is Adesupo Adesuyi. I have BS in Medical Technology and I am >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years experience >as an >histotech. > I presently work in Texas, but I am willing to relocate. > My contact infos are; > > House #: 830-768-0859 > > Cell #: 830-422-9038 > > E/Mail: adesupod@aol.com > > Adesupo Adesuyi > Pathology Dept, > Val Verde Reg.Med.Center, > Del Rio, Tx 78840. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pex0220 <@t> yahoo.com.cn Fri May 13 09:15:00 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] immunofluorescence background!! Message-ID: <20050513141501.87066.qmail@web15501.mail.cnb.yahoo.com> At first, I will thank Dr. Gayle and Dr. Bob for their good suggestions. But I find that there are so many background and so many spots due to the problems of primary antibodies possibly, because there is seldom backgound in my negative sections.Recently I check some data about reducing background, but many data only show the common causes of background from secondary antibodies alone, so I have no ideas about primary antibodies. In addition, I am sure my primary antibodies are good, they come from polysciences and acris. others have used them to get good results. Next time I will try a method from Dr.Bob. But I am not sure if it is OK. Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From 1dpeterson <@t> meriter.com Fri May 13 10:15:59 2005 From: 1dpeterson <@t> meriter.com (Peterson, Dan) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Slide drying oven Message-ID: <328CBAE62F31C642B422970E879DFADC01A68C4E@pcwex01> Paula Have your Bio-med Dept. check the fan on the oven. My guess is that it's clogged with dust. The same thing used to happen to ours. Daniel R Peterson HT(ASCP) Histopathology Section Head General Medical Laboratories (608)-267-6557 1dpeterson@meriter.com CONFIDENTIALITY NOTICE: This message (including any attachments) is intended for the sole use of the individual and entity to whom it is addressed. This message may contain information that is confidential and is protected by law. If you are not the intended recipient, you are hereby notified that any disclosure, copying or distribution of this message is strictly prohibited. If you received this message in error, please immediately notify the sender by reply email and then delete the message. Thank you. From ian.montgomery <@t> bio.gla.ac.uk Fri May 13 10:28:29 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Fri Sep 16 15:25:05 2005 Subject: Fwd: [Histonet] low salary & unions Message-ID: <6.2.1.2.2.20050513162104.0301deb0@udcf.gla.ac.uk> Glen, Your attitude is typical of the anti-union stance. Unions are bad, employers are good and without them employees would have a better life. RUBBISH, without properly organised Trades Unions it would be a swift return to Victorian values of poverty wages and misery. Why are you mentioning bad employees, do we not have bad employers who collect fat cat wages for minimal effort. Of course we do or are you really blinkered to your surroundings. Ian. >Dave, > >My lab has unionized histotechs and I'd say we are in the lower half of >salary ranges for our area. As far as the union here goes, it seems they >are good at collecting dues and at keeping garbage employees employed. Not >only do unions allow bad employees to keep their jobs, they make it >difficult to reward exceptional employees for going above and beyond the >call of duty (since all employees are given union negotiated >raises/benefits/etc..). > >This might be a grass is always greener thing so don't be so sure that a >union will help the histotech's position, that is not always the case. > >My Opinion, > >Glen Dawson BS, HT & QIHC (ASCP) >IHC Manager >Milwaukee, WI > >-----Original Message----- >From: Dave Johnson [mailto:djohnson14@hotmail.com] >Sent: Thursday, May 12, 2005 6:17 PM >To: Adesupod@aol.com >Cc: histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] LOOKING FOR NEW JOB > > >Depending on what you want to make, you could get a job anywhere. It seems >everyone is hiring >Unfortunately, it doesnt seem that salarys are rising despite the lack of >histotechs. > >Sad and scary situation if you ask me. I am not much for unions but now i >see why they are valuable. > >Just my opinion. > > >From: Adesupod@aol.com > >To: histonet@lists.utsouthwestern.edu > >CC: Adesupod@aol.com > >Subject: [Histonet] LOOKING FOR NEW JOB > >Date: Thu, 12 May 2005 20:06:18 EDT > > > > > > Netters, > > I am looking for a new job opportunity. My pathologist passed > >away few months ago, and the hospital management have now decided to close > >down > >the pathology dept. > > My name is Adesupo Adesuyi. I have BS in Medical Technology and I am > >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years experience > > >as an > >histotech. > > I presently work in Texas, but I am willing to relocate. > > My contact infos are; > > > > House #: 830-768-0859 > > > > Cell #: 830-422-9038 > > > > E/Mail: adesupod@aol.com > > > > Adesupo Adesuyi > > Pathology Dept, > > Val Verde Reg.Med.Center, > > Del Rio, Tx 78840. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From jkiernan <@t> uwo.ca Fri May 13 10:30:38 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde References: <83AACDB0810528418AA106F9AE9B7F7EA45CD8@sjhaexc02.sjha.org> Message-ID: <4284C81E.B6D2A050@uwo.ca> Dear Joyce, Formalin = 37% formaldehyde; so no, it doesn't make a difference. Formalin is not buffered; it does contain about 10% methanol, which is put in to retard polymerization. When diluted to make a 4% formaldehyde fixative, the methanol concentration is 1%. Buffering of the dilute solution offsets pH changes due to the Cannizzaro reaction. It also inhibits the formation of blood-derived "formalin pigment" which forms after fixation in an acidic formaldehyde solution. Tim Morken is correct in saying we don't know the extent of chemical change in 12 year-old formalin. The fact that there's no expiry date sugggests that it's not much. For what it's worth, I've used formalin that's more than 5 years old and fixation has been OK. John Kiernan london, Canada ---------------------------------------------- "Weems, Joyce" wrote: > > Also, John said "formalin" - was the solution formalin or 37% formaldehyde - without buffers? Would that make a difference? > > Joyce > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, > Tim - Labvision > Sent: Thursday, May 12, 2005 2:08 PM > To: 'John Kiernan'; Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] quick question about formaldehyde > > John, you said " Small amounts of methanol and formate ions are not going > to change the fixative properties." > > But after 12 years will it really be a "small amount?" How do we know what > percentage of the solution will have been converted? > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan > Sent: Thursday, May 12, 2005 10:48 AM > To: Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick question about formaldehyde > > Two things could have happened to unopened > formalin in 12 years: > 1. Polymerization (to paraformaldehyde). This is > evident as a white precipitate. It slightly > reduces the concentration in the liquid, > but that does not matter for fixation. > Polymerization is accelerated by low room > temperature, and it is claimed that the > process can be reversed by autoclaving(paper > in Stain Technol about 40 years ago). > 2. Cannizzaro's reaction, in which 2 molecules > of formaldehyde react together, producing > one molecule each of methanol and formic > acid. This happens in all formaldehyde solutions > and causes lowering of the pH. This doesn't matter > if you make a neutral buffered fixative solution. > Small amounts of methanol and formate ions are not > going to change the fixative properties. > Bottom line: OK to use, but be sure to check the > pH of the working fixative solution and adjust if > necessary. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Andrea Grantham wrote: > > > > One of the labs here is closing and they have a case of formaldehyde, > > 37.5%, that they are trying to give away. They have had it in their > > lab since 1993. The bottles have not been opened. Is it still good to > > use? Andi > > ..................................................................... > > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > > : Sr. Research Specialist University of Arizona : > > : (office: AHSC 4212) P.O. Box 245044 : > > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > > :...................................................................: > > http://www.cba.arizona.edu/histology-lab.html > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. > Thank you. Saint Joseph -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ From froyer <@t> bitstream.net Fri May 13 10:57:25 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] low salary & unions In-Reply-To: References: Message-ID: <4284CE65.60303@bitstream.net> Do be careful of what you wish for... or if not careful, at least extremely knowledgeable. Always remember that ANY contract that you enter into is a two-way street. Many workers (laboratory or otherwise) do not always understand this. If you become unionized, the union will sit down with management and negotiate the terms of the contract (you personally, are not invited to these sessions). If they have sold themselves to the workers that they will see that they get you higher wages, just remember that if management concedes to that demand, they are going to want something from the union in return . It is a give and take process. You may end up with a slightly higher hourly wage, but lose your leisurely two-a-day coffee breaks and/or your dental benefits. For certain, you would/could lose the "open door" policy with management that you now enjoy and are accustom to. I am not anti-union by any stretch of the imagination. Do not get me wrong. Just... 'Caveat Emptor' ~ Ford Ford M. Royer, B.Sc. M.T.(ASCP) Midwest Science & Biocenter Minneapolis, MN Dawson, Glen wrote: >Dave, > >My lab has unionized histotechs and I'd say we are in the lower half of >salary ranges for our area. As far as the union here goes, it seems they >are good at collecting dues and at keeping garbage employees employed. Not >only do unions allow bad employees to keep their jobs, they make it >difficult to reward exceptional employees for going above and beyond the >call of duty (since all employees are given union negotiated >raises/benefits/etc..). > >This might be a grass is always greener thing so don't be so sure that a >union will help the histotech's position, that is not always the case. > >My Opinion, > >Glen Dawson BS, HT & QIHC (ASCP) >IHC Manager >Milwaukee, WI > >-----Original Message----- >From: Dave Johnson [mailto:djohnson14@hotmail.com] >Sent: Thursday, May 12, 2005 6:17 PM >To: Adesupod@aol.com >Cc: histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] LOOKING FOR NEW JOB > > >Depending on what you want to make, you could get a job anywhere. It seems >everyone is hiring >Unfortunately, it doesnt seem that salarys are rising despite the lack of >histotechs. > >Sad and scary situation if you ask me. I am not much for unions but now i >see why they are valuable. > >Just my opinion. > > > >>From: Adesupod@aol.com >>To: histonet@lists.utsouthwestern.edu >>CC: Adesupod@aol.com >>Subject: [Histonet] LOOKING FOR NEW JOB >>Date: Thu, 12 May 2005 20:06:18 EDT >> >> >> Netters, >> I am looking for a new job opportunity. My pathologist passed >>away few months ago, and the hospital management have now decided to close >>down >>the pathology dept. >> My name is Adesupo Adesuyi. I have BS in Medical Technology and I am >>HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years experience >> >> > > > >>as an >>histotech. >> I presently work in Texas, but I am willing to relocate. >> My contact infos are; >> >> House #: 830-768-0859 >> >> Cell #: 830-422-9038 >> >> E/Mail: adesupod@aol.com >> >> Adesupo Adesuyi >> Pathology Dept, >> Val Verde Reg.Med.Center, >> Del Rio, Tx 78840. >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From jdmd77 <@t> hotmail.com Fri May 13 10:49:12 2005 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Fri Sep 16 15:25:05 2005 Subject: Fwd: [Histonet] low salary & unions In-Reply-To: <6.2.1.2.2.20050513162104.0301deb0@udcf.gla.ac.uk> Message-ID: As a pathologist, I've heard from many histotechnicians and histotechnologists that in addition to the pay scale - underappreciation from pathologists (and/or administration) adds insult to injury. If you had a comfortable working environment (not crowded or "production line" mentality) working with pathologists and administrators that realized the inherent reliance upon histotechnologists for each case... What is the pay scale that you would allow you to feel fairly compensated? Generously compensated? How much paid vacation and PTO time do you desire? What benefits are essential for you? Yes, I'm really asking and I'd love to hear how to provide a happy, well compensated work environment. Julia Dahl, M.D. Mosaic Gastrointestinal Research Consortium Memphis, Tennessee >From: Ian Montgomery >To: histonet@lists.utsouthwestern.edu >Subject: Fwd: [Histonet] low salary & unions >Date: Fri, 13 May 2005 16:28:29 +0100 >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc9-f17.hotmail.com >with Microsoft SMTPSVC(6.0.3790.211); Fri, 13 May 2005 08:29:12 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1DWc5v-0000Iq-Tj; Fri, 13 May >2005 10:29:01 -0500 >Received: from [199.165.152.166] (helo=swlx166.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DWc5Y-0000IK-D1for >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:43 -0500 >Received: from [130.209.16.18] (helo=lenzie.cent.gla.ac.uk)by >swlx166.swmed.edu with esmtp (Exim 4.44) id 1DWc5X-0000I5-FGfor >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:32 -0500 >Received: from monty.bio.gla.ac.uk (montydesk.ibls.gla.ac.uk >[130.209.46.83])by lenzie.cent.gla.ac.uk (8.11.7/8.11.2) with ESMTP id >j4DFSUB19208for ;Fri, 13 May 2005 >16:28:30 +0100 (BST) >X-Message-Info: gUeNUVfFqHBxE7JvKVytr2DPrM5qbybavSJerH9UTDQ= >X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 >X-Scan-Signature: d57e7743657f0a8bf60f191863530969 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 4239ed49a0af0eb50142010c0dad1a43 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: * >X-Spam-Status: No, hits=1.1 required=5.5 tests=MAILTO_TO_SPAM_ADDR >autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 13 May 2005 15:29:12.0313 (UTC) >FILETIME=[83A15690:01C557D0] > >Glen, > Your attitude is typical of the anti-union stance. Unions are bad, >employers are good and without them employees would have a better life. >RUBBISH, without properly organised Trades Unions it would be a swift >return to Victorian values of poverty wages and misery. Why are you >mentioning bad employees, do we not have bad employers who collect fat cat >wages for minimal effort. Of course we do or are you really blinkered to >your surroundings. >Ian. > >>Dave, >> >>My lab has unionized histotechs and I'd say we are in the lower half of >>salary ranges for our area. As far as the union here goes, it seems they >>are good at collecting dues and at keeping garbage employees employed. >>Not >>only do unions allow bad employees to keep their jobs, they make it >>difficult to reward exceptional employees for going above and beyond the >>call of duty (since all employees are given union negotiated >>raises/benefits/etc..). >> >>This might be a grass is always greener thing so don't be so sure that a >>union will help the histotech's position, that is not always the case. >> >>My Opinion, >> >>Glen Dawson BS, HT & QIHC (ASCP) >>IHC Manager >>Milwaukee, WI >> >>-----Original Message----- >>From: Dave Johnson [mailto:djohnson14@hotmail.com] >>Sent: Thursday, May 12, 2005 6:17 PM >>To: Adesupod@aol.com >>Cc: histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] LOOKING FOR NEW JOB >> >> >>Depending on what you want to make, you could get a job anywhere. It >>seems >>everyone is hiring >>Unfortunately, it doesnt seem that salarys are rising despite the lack of >>histotechs. >> >>Sad and scary situation if you ask me. I am not much for unions but now i >>see why they are valuable. >> >>Just my opinion. >> >> >From: Adesupod@aol.com >> >To: histonet@lists.utsouthwestern.edu >> >CC: Adesupod@aol.com >> >Subject: [Histonet] LOOKING FOR NEW JOB >> >Date: Thu, 12 May 2005 20:06:18 EDT >> > >> > >> > Netters, >> > I am looking for a new job opportunity. My pathologist >>passed >> >away few months ago, and the hospital management have now decided to >>close >> >down >> >the pathology dept. >> > My name is Adesupo Adesuyi. I have BS in Medical Technology and I >>am >> >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years >>experience >> >> >as an >> >histotech. >> > I presently work in Texas, but I am willing to relocate. >> > My contact infos are; >> > >> > House #: 830-768-0859 >> > >> > Cell #: 830-422-9038 >> > >> > E/Mail: adesupod@aol.com >> > >> > Adesupo Adesuyi >> > Pathology Dept, >> > Val Verde Reg.Med.Center, >> > Del Rio, Tx 78840. >> >_______________________________________________ >> >Histonet mailing list >> >Histonet@lists.utsouthwestern.edu >> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07623 975451 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri May 13 11:35:30 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45CF4@sjhaexc02.sjha.org> For all my years I have believed that formaldehyde was the 37% concentrate without buffers, and that formalin was 10% buffered... learn something new every day! Thanks John! -----Original Message----- From: John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, May 13, 2005 11:31 AM To: Weems, Joyce Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quick question about formaldehyde Dear Joyce, Formalin = 37% formaldehyde; so no, it doesn't make a difference. Formalin is not buffered; it does contain about 10% methanol, which is put in to retard polymerization. When diluted to make a 4% formaldehyde fixative, the methanol concentration is 1%. Buffering of the dilute solution offsets pH changes due to the Cannizzaro reaction. It also inhibits the formation of blood-derived "formalin pigment" which forms after fixation in an acidic formaldehyde solution. Tim Morken is correct in saying we don't know the extent of chemical change in 12 year-old formalin. The fact that there's no expiry date sugggests that it's not much. For what it's worth, I've used formalin that's more than 5 years old and fixation has been OK. John Kiernan london, Canada ---------------------------------------------- "Weems, Joyce" wrote: > > Also, John said "formalin" - was the solution formalin or 37% formaldehyde - without buffers? Would that make a difference? > > Joyce > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, > Tim - Labvision > Sent: Thursday, May 12, 2005 2:08 PM > To: 'John Kiernan'; Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] quick question about formaldehyde > > John, you said " Small amounts of methanol and formate ions are not going > to change the fixative properties." > > But after 12 years will it really be a "small amount?" How do we know what > percentage of the solution will have been converted? > > Tim Morken > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan > Sent: Thursday, May 12, 2005 10:48 AM > To: Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick question about formaldehyde > > Two things could have happened to unopened > formalin in 12 years: > 1. Polymerization (to paraformaldehyde). This is > evident as a white precipitate. It slightly > reduces the concentration in the liquid, > but that does not matter for fixation. > Polymerization is accelerated by low room > temperature, and it is claimed that the > process can be reversed by autoclaving(paper > in Stain Technol about 40 years ago). > 2. Cannizzaro's reaction, in which 2 molecules > of formaldehyde react together, producing > one molecule each of methanol and formic > acid. This happens in all formaldehyde solutions > and causes lowering of the pH. This doesn't matter > if you make a neutral buffered fixative solution. > Small amounts of methanol and formate ions are not > going to change the fixative properties. > Bottom line: OK to use, but be sure to check the > pH of the working fixative solution and adjust if > necessary. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Andrea Grantham wrote: > > > > One of the labs here is closing and they have a case of formaldehyde, > > 37.5%, that they are trying to give away. They have had it in their > > lab since 1993. The bottles have not been opened. Is it still good to > > use? Andi > > ..................................................................... > > : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > > : Sr. Research Specialist University of Arizona : > > : (office: AHSC 4212) P.O. Box 245044 : > > : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > > : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : > > :...................................................................: > > http://www.cba.arizona.edu/histology-lab.html > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. > Thank you. Saint Joseph -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From ttruscot <@t> vetmed.wsu.edu Fri May 13 12:36:44 2005 From: ttruscot <@t> vetmed.wsu.edu (Truscott, Tom) Date: Fri Sep 16 15:25:05 2005 Subject: Fwd: [Histonet] low salary & unions Message-ID: <95111571D1C6B5498C64F68DD0769BB506BEFF@cvm36.vetmed.wsu.edu> Hi Dr. Dahl, I think that the Federal GS 9 pay scale is fair at the lower end and generous at the higher end for my job in research. I am an HT with a BS degree in Animal Science. My job entails collecting live animal biopsies and collecting tissues at necropsy, then all trimming, processing, microtomy and immunostaining and some slide screening. I operate the lab myself except for some excellent student help. A good health insurance is necessary. 2 to 3 weeks vacation is good. Flex time is helpful. When I worked in a private hospital histology lab, the stress was higher and the pay poorer. A pat on the back for a job well done was only from the best pathologist. As a pathologist starts relying less on his or her ability to diagnose a case from an H&E and more on the ability of a technician to perform a myriad of diagnostic stains, then the tech should start earning a greater percentage of the income that the pathologist makes. If a union can't get it's members above ave. pay and benefits, then it shouldn't charge them dues. Thanks for your concern. Tom Truscott -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julia Dahl Sent: Friday, May 13, 2005 7:49 AM To: ian.montgomery@bio.gla.ac.uk; histonet@lists.utsouthwestern.edu Subject: RE: Fwd: [Histonet] low salary & unions As a pathologist, I've heard from many histotechnicians and histotechnologists that in addition to the pay scale - underappreciation from pathologists (and/or administration) adds insult to injury. If you had a comfortable working environment (not crowded or "production line" mentality) working with pathologists and administrators that realized the inherent reliance upon histotechnologists for each case... What is the pay scale that you would allow you to feel fairly compensated? Generously compensated? How much paid vacation and PTO time do you desire? What benefits are essential for you? Yes, I'm really asking and I'd love to hear how to provide a happy, well compensated work environment. Julia Dahl, M.D. Mosaic Gastrointestinal Research Consortium Memphis, Tennessee >From: Ian Montgomery >To: histonet@lists.utsouthwestern.edu >Subject: Fwd: [Histonet] low salary & unions >Date: Fri, 13 May 2005 16:28:29 +0100 >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc9-f17.hotmail.com >with Microsoft SMTPSVC(6.0.3790.211); Fri, 13 May 2005 08:29:12 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1DWc5v-0000Iq-Tj; Fri, 13 May >2005 10:29:01 -0500 >Received: from [199.165.152.166] (helo=swlx166.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DWc5Y-0000IK-D1for >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:43 -0500 >Received: from [130.209.16.18] (helo=lenzie.cent.gla.ac.uk)by >swlx166.swmed.edu with esmtp (Exim 4.44) id 1DWc5X-0000I5-FGfor >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:32 -0500 >Received: from monty.bio.gla.ac.uk (montydesk.ibls.gla.ac.uk >[130.209.46.83])by lenzie.cent.gla.ac.uk (8.11.7/8.11.2) with ESMTP id >j4DFSUB19208for ;Fri, 13 May 2005 >16:28:30 +0100 (BST) >X-Message-Info: gUeNUVfFqHBxE7JvKVytr2DPrM5qbybavSJerH9UTDQ= >X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 >X-Scan-Signature: d57e7743657f0a8bf60f191863530969 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 4239ed49a0af0eb50142010c0dad1a43 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: * >X-Spam-Status: No, hits=1.1 required=5.5 tests=MAILTO_TO_SPAM_ADDR >autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 13 May 2005 15:29:12.0313 (UTC) >FILETIME=[83A15690:01C557D0] > >Glen, > Your attitude is typical of the anti-union stance. Unions are bad, >employers are good and without them employees would have a better life. >RUBBISH, without properly organised Trades Unions it would be a swift >return to Victorian values of poverty wages and misery. Why are you >mentioning bad employees, do we not have bad employers who collect fat cat >wages for minimal effort. Of course we do or are you really blinkered to >your surroundings. >Ian. > >>Dave, >> >>My lab has unionized histotechs and I'd say we are in the lower half of >>salary ranges for our area. As far as the union here goes, it seems they >>are good at collecting dues and at keeping garbage employees employed. >>Not >>only do unions allow bad employees to keep their jobs, they make it >>difficult to reward exceptional employees for going above and beyond the >>call of duty (since all employees are given union negotiated >>raises/benefits/etc..). >> >>This might be a grass is always greener thing so don't be so sure that a >>union will help the histotech's position, that is not always the case. >> >>My Opinion, >> >>Glen Dawson BS, HT & QIHC (ASCP) >>IHC Manager >>Milwaukee, WI >> >>-----Original Message----- >>From: Dave Johnson [mailto:djohnson14@hotmail.com] >>Sent: Thursday, May 12, 2005 6:17 PM >>To: Adesupod@aol.com >>Cc: histonet@lists.utsouthwestern.edu >>Subject: RE: [Histonet] LOOKING FOR NEW JOB >> >> >>Depending on what you want to make, you could get a job anywhere. It >>seems >>everyone is hiring >>Unfortunately, it doesnt seem that salarys are rising despite the lack of >>histotechs. >> >>Sad and scary situation if you ask me. I am not much for unions but now i >>see why they are valuable. >> >>Just my opinion. >> >> >From: Adesupod@aol.com >> >To: histonet@lists.utsouthwestern.edu >> >CC: Adesupod@aol.com >> >Subject: [Histonet] LOOKING FOR NEW JOB >> >Date: Thu, 12 May 2005 20:06:18 EDT >> > >> > >> > Netters, >> > I am looking for a new job opportunity. My pathologist >>passed >> >away few months ago, and the hospital management have now decided to >>close >> >down >> >the pathology dept. >> > My name is Adesupo Adesuyi. I have BS in Medical Technology and I >>am >> >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years >>experience >> >> >as an >> >histotech. >> > I presently work in Texas, but I am willing to relocate. >> > My contact infos are; >> > >> > House #: 830-768-0859 >> > >> > Cell #: 830-422-9038 >> > >> > E/Mail: adesupod@aol.com >> > >> > Adesupo Adesuyi >> > Pathology Dept, >> > Val Verde Reg.Med.Center, >> > Del Rio, Tx 78840. >> >_______________________________________________ >> >Histonet mailing list >> >Histonet@lists.utsouthwestern.edu >> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >Dr. Ian Montgomery, >Histotechnology, >Graham Kerr Building, >Institute of Biomedical & Life Sciences, >University of Glasgow, >Glasgow, >G12 8QQ. >Tel: 0141 339 8855 >Office: 4652 >Lab: 6644. >Pager: 07623 975451 >e-mail: ian.montgomery@bio.gla.ac.uk > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ander093 <@t> tc.umn.edu Fri May 13 12:43:53 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde In-Reply-To: <6.1.2.0.0.20050513124205.028b0260@ander093.email.umn.edu> References: <83AACDB0810528418AA106F9AE9B7F7EA45CF4@sjhaexc02.sjha.org> <6.1.2.0.0.20050513124205.028b0260@ander093.email.umn.edu> Message-ID: <6.1.2.0.0.20050513124342.028b0110@ander093.email.umn.edu> At 12:43 PM 5/13/05, LuAnn Anderson wrote: >Hi Joyce, >That is exactly what we were taught. formaldhyde=37% and formalin=10% >LuAnn > > > >>For all my years I have believed that formaldehyde was the 37% >>concentrate without buffers, and that formalin was 10% buffered... learn >>something new every day! Thanks John! -----Original Message----- From: >>John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, May 13, 2005 11:31 >>AM To: Weems, Joyce Cc: histonet@lists.utsouthwestern.edu Subject: Re: >>[Histonet] quick question about formaldehyde Dear Joyce, Formalin = 37% >>formaldehyde; so no, it doesn't make a difference. Formalin is not >>buffered; it does contain about 10% methanol, which is put in to retard >>polymerization. When diluted to make a 4% formaldehyde fixative, the >>methanol concentration is 1%. Buffering of the dilute solution offsets pH >>changes due to the Cannizzaro reaction. It also inhibits the formation of >>blood-derived "formalin pigment" which forms after fixation in an acidic >>formaldehyde solution. Tim Morken is correct in saying we don't know the >>extent of chemical change in 12 year-old formalin. The fact that there's >>no expiry date sugggests that it's not much. For what it's worth, I've >>used formalin that's more than 5 years old and fixation has been >>OK. John Kiernan london, >>Canada ---------------------------------------------- "Weems, Joyce" >>wrote: > > Also, John said "formalin" - was the solution formalin or 37% >>formaldehyde - without buffers? Would that make a difference? > > >>Joyce > > -----Original Message----- > From: >>histonet-bounces@lists.utsouthwestern.edu > >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, > >>Tim - Labvision > Sent: Thursday, May 12, 2005 2:08 PM > To: 'John >>Kiernan'; Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > >>Subject: RE: [Histonet] quick question about formaldehyde > > John, you >>said " Small amounts of methanol and formate ions are not going > to >>change the fixative properties." > > But after 12 years will it really >>be a "small amount?" How do we know what > percentage of the solution >>will have been converted? > > Tim Morken > > -----Original Message----- > >>From: histonet-bounces@lists.utsouthwestern.edu > >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John >>Kiernan > Sent: Thursday, May 12, 2005 10:48 AM > To: Andrea Grantham > >>Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick >>question about formaldehyde > > Two things could have happened to >>unopened > formalin in 12 years: > 1. Polymerization (to >>paraformaldehyde). This is > evident as a white precipitate. It >>slightly > reduces the concentration in the liquid, > but that does >>not matter for fixation. > Polymerization is accelerated by low >>room > temperature, and it is claimed that the > process can be >>reversed by autoclaving(paper > in Stain Technol about 40 years >>ago). > 2. Cannizzaro's reaction, in which 2 molecules > of >>formaldehyde react together, producing > one molecule each of methanol >>and formic > acid. This happens in all formaldehyde solutions > and >>causes lowering of the pH. This doesn't matter > if you make a neutral >>buffered fixative solution. > Small amounts of methanol and formate >>ions are not > going to change the fixative properties. > Bottom line: >>OK to use, but be sure to check the > pH of the working fixative solution >>and adjust if > necessary. > -- > ------------------------------- > John >>A. Kiernan > Department of Anatomy and Cell Biology > The University of >>Western Ontario > London, Canada N6A >>5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > >>http://instruct.uwo.ca/anatomy/530/index.htm > >>_______________________________ > Andrea Grantham wrote: > > > > One of >>the labs here is closing and they have a case of formaldehyde, > > 37.5%, >>that they are trying to give away. They have had it in their > > lab >>since 1993. The bottles have not been opened. Is it still good to > > >>use? Andi > > >>..................................................................... > > >>: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > > >>: Sr. Research Specialist University of Arizona : > > >>: (office: AHSC 4212) P.O. Box 245044 : > > >>: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > > >>: >>(FAX: 520-626-2097) (email: algranth@u.arizona.edu) >>: > > >>:...................................................................: > > >> http://www.cba.arizona.edu/histology-lab.html > > > > >>_______________________________________________ > > Histonet mailing >>list > > Histonet@lists.utsouthwestern.edu > > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >>_______________________________________________ > Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >>_______________________________________________ > Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >>Confidentiality Notice ** The information contained in this message may >>be privileged and is confidential information intended for the use of the >>addressee listed above. If you are neither the intended recipient nor the >>employee or agent responsible for delivering this message to the intended >>recipient, you are hereby notified that any disclosure, copying, >>distribution or the taking of any action in reliance on the contents of >>this information is strictly prohibited. If you have received this >>communication in error, please notify us immediately by replying to the >>message and deleting it from your computer. > Thank you. Saint Joseph -- >>------------------------------- John A. Kiernan Department of Anatomy and >>Cell Biology The University of Western Ontario London, Canada N6A >>5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ >>http://instruct.uwo.ca/anatomy/530/index.htm >>_______________________________ Confidentiality Notice ** The information >>contained in this message may be privileged and is confidential >>information intended for the use of the addressee listed above. If you >>are neither the intended recipient nor the employee or agent responsible >>for delivering this message to the intended recipient, you are hereby >>notified that any disclosure, copying, distribution or the taking of any >>action in reliance on the contents of this information is strictly >>prohibited. If you have received this communication in error, please >>notify us immediately by replying to the message and deleting it from >>your computer. Thank you. Saint Josephs Health System, >>Inc._______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Debbiejsiena <@t> aol.com Fri May 13 12:40:11 2005 From: Debbiejsiena <@t> aol.com (Debbiejsiena@aol.com) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Histology Lab Supervisor Wanted - Dallas, TX Message-ID: <13e.132d423b.2fb6407b@aol.com> Hi All Tissue Techniques Pathology Labs in Dallas, Texas is looking for a supervisor. Private lab good working conditions, good pay, flexable schedule when needed. If interested please apply as follows: Debbie Siena HT (ASCP) QIHC Cell: 520-360-3013 Office: 972-241-6277 Fax: 972-241-4747 E:mail: tissuetech@juno.com From Jackie.O'Connor <@t> abbott.com Fri May 13 12:54:40 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde Message-ID: Formaldehyde is a gas. Full strength formalin is 37-40% formaldehyde gas in water. 10% Formalin is in effect 3.7 - 4% formaldehyde. 10% Neutral Buffered Formalin is just that - 10% formalin buffered to a neutral pH, usually with sodium phosphates. Jacqueline M. O'Connor HT(ASCP) Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com LuAnn Anderson Sent by: histonet-bounces@lists.utsouthwestern.edu 05/13/2005 12:43 PM To: histonet@lists.utsouthwestern.edu cc: Subject: RE: [Histonet] quick question about formaldehyde At 12:43 PM 5/13/05, LuAnn Anderson wrote: >Hi Joyce, >That is exactly what we were taught. formaldhyde=37% and formalin=10% >LuAnn > > > >>For all my years I have believed that formaldehyde was the 37% >>concentrate without buffers, and that formalin was 10% buffered... learn >>something new every day! Thanks John! -----Original Message----- From: >>John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, May 13, 2005 11:31 >>AM To: Weems, Joyce Cc: histonet@lists.utsouthwestern.edu Subject: Re: >>[Histonet] quick question about formaldehyde Dear Joyce, Formalin = 37% >>formaldehyde; so no, it doesn't make a difference. Formalin is not >>buffered; it does contain about 10% methanol, which is put in to retard >>polymerization. When diluted to make a 4% formaldehyde fixative, the >>methanol concentration is 1%. Buffering of the dilute solution offsets pH >>changes due to the Cannizzaro reaction. It also inhibits the formation of >>blood-derived "formalin pigment" which forms after fixation in an acidic >>formaldehyde solution. Tim Morken is correct in saying we don't know the >>extent of chemical change in 12 year-old formalin. The fact that there's >>no expiry date sugggests that it's not much. For what it's worth, I've >>used formalin that's more than 5 years old and fixation has been >>OK. John Kiernan london, >>Canada ---------------------------------------------- "Weems, Joyce" >>wrote: > > Also, John said "formalin" - was the solution formalin or 37% >>formaldehyde - without buffers? Would that make a difference? > > >>Joyce > > -----Original Message----- > From: >>histonet-bounces@lists.utsouthwestern.edu > >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, > >>Tim - Labvision > Sent: Thursday, May 12, 2005 2:08 PM > To: 'John >>Kiernan'; Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > >>Subject: RE: [Histonet] quick question about formaldehyde > > John, you >>said " Small amounts of methanol and formate ions are not going > to >>change the fixative properties." > > But after 12 years will it really >>be a "small amount?" How do we know what > percentage of the solution >>will have been converted? > > Tim Morken > > -----Original Message----- > >>From: histonet-bounces@lists.utsouthwestern.edu > >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John >>Kiernan > Sent: Thursday, May 12, 2005 10:48 AM > To: Andrea Grantham > >>Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick >>question about formaldehyde > > Two things could have happened to >>unopened > formalin in 12 years: > 1. Polymerization (to >>paraformaldehyde). This is > evident as a white precipitate. It >>slightly > reduces the concentration in the liquid, > but that does >>not matter for fixation. > Polymerization is accelerated by low >>room > temperature, and it is claimed that the > process can be >>reversed by autoclaving(paper > in Stain Technol about 40 years >>ago). > 2. Cannizzaro's reaction, in which 2 molecules > of >>formaldehyde react together, producing > one molecule each of methanol >>and formic > acid. This happens in all formaldehyde solutions > and >>causes lowering of the pH. This doesn't matter > if you make a neutral >>buffered fixative solution. > Small amounts of methanol and formate >>ions are not > going to change the fixative properties. > Bottom line: >>OK to use, but be sure to check the > pH of the working fixative solution >>and adjust if > necessary. > -- > ------------------------------- > John >>A. Kiernan > Department of Anatomy and Cell Biology > The University of >>Western Ontario > London, Canada N6A >>5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > >>http://instruct.uwo.ca/anatomy/530/index.htm > >>_______________________________ > Andrea Grantham wrote: > > > > One of >>the labs here is closing and they have a case of formaldehyde, > > 37.5%, >>that they are trying to give away. They have had it in their > > lab >>since 1993. The bottles have not been opened. Is it still good to > > >>use? Andi > > >>..................................................................... > > >>: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > > >>: Sr. Research Specialist University of Arizona : > > >>: (office: AHSC 4212) P.O. Box 245044 : > > >>: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > > >>: >>(FAX: 520-626-2097) (email: algranth@u.arizona.edu) >>: > > >>:...................................................................: > > >> http://www.cba.arizona.edu/histology-lab.html > > > > >>_______________________________________________ > > Histonet mailing >>list > > Histonet@lists.utsouthwestern.edu > > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >>_______________________________________________ > Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >>_______________________________________________ > Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >>Confidentiality Notice ** The information contained in this message may >>be privileged and is confidential information intended for the use of the >>addressee listed above. If you are neither the intended recipient nor the >>employee or agent responsible for delivering this message to the intended >>recipient, you are hereby notified that any disclosure, copying, >>distribution or the taking of any action in reliance on the contents of >>this information is strictly prohibited. If you have received this >>communication in error, please notify us immediately by replying to the >>message and deleting it from your computer. > Thank you. Saint Joseph -- >>------------------------------- John A. Kiernan Department of Anatomy and >>Cell Biology The University of Western Ontario London, Canada N6A >>5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ >>http://instruct.uwo.ca/anatomy/530/index.htm >>_______________________________ Confidentiality Notice ** The information >>contained in this message may be privileged and is confidential >>information intended for the use of the addressee listed above. If you >>are neither the intended recipient nor the employee or agent responsible >>for delivering this message to the intended recipient, you are hereby >>notified that any disclosure, copying, distribution or the taking of any >>action in reliance on the contents of this information is strictly >>prohibited. If you have received this communication in error, please >>notify us immediately by replying to the message and deleting it from >>your computer. Thank you. Saint Josephs Health System, >>Inc._______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ander093 <@t> tc.umn.edu Fri May 13 13:13:08 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] quick question about formaldehyde In-Reply-To: References: Message-ID: <6.1.2.0.0.20050513131040.04dc9db0@ander093.email.umn.edu> Yes, formaldehyde is a gas. Maybe back in the "old" days it was termed that way for the pupose of distinguishing between the two. I was taught that way in school and the pathologists always used those terms to specify differnences between formalin (10%) and formaldehyde (37-40%). At 12:54 PM 5/13/05, Jackie M O'Connor wrote: >Formaldehyde is a gas. Full strength formalin is 37-40% formaldehyde gas >in water. 10% Formalin is in effect 3.7 - 4% formaldehyde. >10% Neutral Buffered Formalin is just that - 10% formalin buffered to a >neutral pH, usually with sodium phosphates. > >Jacqueline M. O'Connor HT(ASCP) >Assistant Scientist >GPRD Cancer Research >Abbott Laboratories, Abbott Park, IL >Jackie.OConnor@abbott.com > > >LuAnn Anderson >Sent by: histonet-bounces@lists.utsouthwestern.edu > >05/13/2005 12:43 PM > > To: histonet@lists.utsouthwestern.edu > cc: > Subject: RE: [Histonet] quick question about formaldehyde > > >At 12:43 PM 5/13/05, LuAnn Anderson wrote: > >Hi Joyce, > >That is exactly what we were taught. formaldhyde=37% and formalin=10% > >LuAnn > > > > > > > >>For all my years I have believed that formaldehyde was the 37% > >>concentrate without buffers, and that formalin was 10% buffered... learn > >>something new every day! Thanks John! -----Original Message----- From: > >>John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, May 13, 2005 11:31 > >>AM To: Weems, Joyce Cc: histonet@lists.utsouthwestern.edu Subject: Re: > >>[Histonet] quick question about formaldehyde Dear Joyce, Formalin = 37% > >>formaldehyde; so no, it doesn't make a difference. Formalin is not > >>buffered; it does contain about 10% methanol, which is put in to retard > >>polymerization. When diluted to make a 4% formaldehyde fixative, the > >>methanol concentration is 1%. Buffering of the dilute solution offsets pH > >>changes due to the Cannizzaro reaction. It also inhibits the formation of > >>blood-derived "formalin pigment" which forms after fixation in an acidic > >>formaldehyde solution. Tim Morken is correct in saying we don't know the > >>extent of chemical change in 12 year-old formalin. The fact that there's > >>no expiry date sugggests that it's not much. For what it's worth, I've > >>used formalin that's more than 5 years old and fixation has been > >>OK. John Kiernan london, > >>Canada ---------------------------------------------- "Weems, Joyce" > >>wrote: > > Also, John said "formalin" - was the solution formalin or 37% > >>formaldehyde - without buffers? Would that make a difference? > > > >>Joyce > > -----Original Message----- > From: > >>histonet-bounces@lists.utsouthwestern.edu > > >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Morken, > > >>Tim - Labvision > Sent: Thursday, May 12, 2005 2:08 PM > To: 'John > >>Kiernan'; Andrea Grantham > Cc: histonet@lists.utsouthwestern.edu > > >>Subject: RE: [Histonet] quick question about formaldehyde > > John, you > >>said " Small amounts of methanol and formate ions are not going > to > >>change the fixative properties." > > But after 12 years will it really > >>be a "small amount?" How do we know what > percentage of the solution > >>will have been converted? > > Tim Morken > > -----Original Message----- > > >>From: histonet-bounces@lists.utsouthwestern.edu > > >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John > >>Kiernan > Sent: Thursday, May 12, 2005 10:48 AM > To: Andrea Grantham > > >>Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick > >>question about formaldehyde > > Two things could have happened to > >>unopened > formalin in 12 years: > 1. Polymerization (to > >>paraformaldehyde). This is > evident as a white precipitate. It > >>slightly > reduces the concentration in the liquid, > but that does > >>not matter for fixation. > Polymerization is accelerated by low > >>room > temperature, and it is claimed that the > process can be > >>reversed by autoclaving(paper > in Stain Technol about 40 years > >>ago). > 2. Cannizzaro's reaction, in which 2 molecules > of > >>formaldehyde react together, producing > one molecule each of methanol > >>and formic > acid. This happens in all formaldehyde solutions > and > >>causes lowering of the pH. This doesn't matter > if you make a neutral > >>buffered fixative solution. > Small amounts of methanol and formate > >>ions are not > going to change the fixative properties. > Bottom line: > >>OK to use, but be sure to check the > pH of the working fixative solution > >>and adjust if > necessary. > -- > ------------------------------- > John > >>A. Kiernan > Department of Anatomy and Cell Biology > The University of > >>Western Ontario > London, Canada N6A > >>5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > > >>http://instruct.uwo.ca/anatomy/530/index.htm > > >>_______________________________ > Andrea Grantham wrote: > > > > One of > >>the labs here is closing and they have a case of formaldehyde, > > 37.5%, > >>that they are trying to give away. They have had it in their > > lab > >>since 1993. The bottles have not been opened. Is it still good to > > > >>use? Andi > > > >>..................................................................... > > > >>: Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : > > > >>: Sr. Research Specialist University of Arizona : > > > >>: (office: AHSC 4212) P.O. Box 245044 : > > > >>: (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : > > > >>: > >>(FAX: 520-626-2097) (email: algranth@u.arizona.edu) > >>: > > > >>:...................................................................: > > > >> http://www.cba.arizona.edu/histology-lab.html > > > > > >>_______________________________________________ > > Histonet mailing > >>list > > Histonet@lists.utsouthwestern.edu > > > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >>_______________________________________________ > Histonet mailing list > > >>Histonet@lists.utsouthwestern.edu > > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >>_______________________________________________ > Histonet mailing list > > >>Histonet@lists.utsouthwestern.edu > > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >>Confidentiality Notice ** The information contained in this message may > >>be privileged and is confidential information intended for the use of the > >>addressee listed above. If you are neither the intended recipient nor the > >>employee or agent responsible for delivering this message to the intended > >>recipient, you are hereby notified that any disclosure, copying, > >>distribution or the taking of any action in reliance on the contents of > >>this information is strictly prohibited. If you have received this > >>communication in error, please notify us immediately by replying to the > >>message and deleting it from your computer. > Thank you. Saint Joseph -- > >>------------------------------- John A. Kiernan Department of Anatomy and > >>Cell Biology The University of Western Ontario London, Canada N6A > >>5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ > >>http://instruct.uwo.ca/anatomy/530/index.htm > >>_______________________________ Confidentiality Notice ** The information > >>contained in this message may be privileged and is confidential > >>information intended for the use of the addressee listed above. If you > >>are neither the intended recipient nor the employee or agent responsible > >>for delivering this message to the intended recipient, you are hereby > >>notified that any disclosure, copying, distribution or the taking of any > >>action in reliance on the contents of this information is strictly > >>prohibited. If you have received this communication in error, please > >>notify us immediately by replying to the message and deleting it from > >>your computer. Thank you. Saint Josephs Health System, > >>Inc._______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jdmd77 <@t> hotmail.com Fri May 13 13:21:11 2005 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Thank you RE: Salary and Benefits Questions In-Reply-To: <95111571D1C6B5498C64F68DD0769BB506BEFF@cvm36.vetmed.wsu.edu> Message-ID: Thank you to all who have posted! I agree completely that pathologists and PAs should have a course in Histology. Where I trained, I was fortunate enough to do just this - and it has made a lot of difference in how I gross, how I teach grossing and how I talk with my clinical colleagues about quality and turnaround time. Again, my thanks for your taking the time to share your perspective - I will certainly incorporate your influence in planning for recruitment and retention of your valuable talents! Julia Dahl, M.D. Mosaic Gastrointestinal Research Consortium >From: "Truscott, Tom" >To: "Julia Dahl" >,, >Subject: RE: Fwd: [Histonet] low salary & unions >Date: Fri, 13 May 2005 10:36:44 -0700 >MIME-Version: 1.0 >Received: from cvm36.vetmed.wsu.edu ([134.121.128.6]) by mc1-f8.hotmail.com >with Microsoft SMTPSVC(6.0.3790.211); Fri, 13 May 2005 10:37:09 -0700 >X-Message-Info: JGTYoYF78jGEhCL50HbvXKZEc+z0eBZREmA8KVl1tnQ= >Content-class: urn:content-classes:message >X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 >X-MS-Has-Attach: X-MS-TNEF-Correlator: Thread-Topic: Fwd: [Histonet] low >salary & unions >Thread-Index: AcVX01gnisokgYADSKaqKJEyI9GvQAACqPEQ >Return-Path: ttruscot@vetmed.wsu.edu >X-OriginalArrivalTime: 13 May 2005 17:37:09.0347 (UTC) >FILETIME=[637FC330:01C557E2] > >Hi Dr. Dahl, I think that the Federal GS 9 pay scale is fair at the >lower end and generous at the higher end for my job in research. I am an >HT with a BS degree in Animal Science. My job entails collecting live >animal biopsies and collecting tissues at necropsy, then all trimming, >processing, microtomy and immunostaining and some slide screening. I >operate the lab myself except for some excellent student help. A good >health insurance is necessary. 2 to 3 weeks vacation is good. Flex time >is helpful. When I worked in a private hospital histology lab, the >stress was higher and the pay poorer. A pat on the back for a job well >done was only from the best pathologist. As a pathologist starts relying >less on his or her ability to diagnose a case from an H&E and more on >the ability of a technician to perform a myriad of diagnostic stains, >then the tech should start earning a greater percentage of the income >that the pathologist makes. If a union can't get it's members above ave. >pay and benefits, then it shouldn't charge them dues. Thanks for your >concern. Tom Truscott > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julia >Dahl >Sent: Friday, May 13, 2005 7:49 AM >To: ian.montgomery@bio.gla.ac.uk; histonet@lists.utsouthwestern.edu >Subject: RE: Fwd: [Histonet] low salary & unions > >As a pathologist, I've heard from many histotechnicians and >histotechnologists that in addition to the pay scale - underappreciation > >from pathologists (and/or administration) adds insult to injury. > >If you had a comfortable working environment (not crowded or "production > >line" mentality) working with pathologists and administrators that >realized >the inherent reliance upon histotechnologists for each case... > >What is the pay scale that you would allow you to feel fairly >compensated? >Generously compensated? > >How much paid vacation and PTO time do you desire? > >What benefits are essential for you? > > >Yes, I'm really asking and I'd love to hear how to provide a happy, well > >compensated work environment. > >Julia Dahl, M.D. >Mosaic Gastrointestinal Research Consortium >Memphis, Tennessee > > > >From: Ian Montgomery > >To: histonet@lists.utsouthwestern.edu > >Subject: Fwd: [Histonet] low salary & unions > >Date: Fri, 13 May 2005 16:28:29 +0100 > >MIME-Version: 1.0 > >Received: from swlx162.swmed.edu ([199.165.152.162]) by >mc9-f17.hotmail.com > >with Microsoft SMTPSVC(6.0.3790.211); Fri, 13 May 2005 08:29:12 -0700 > >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by > >swlx162.swmed.edu with esmtp (Exim 4.34)id 1DWc5v-0000Iq-Tj; Fri, 13 >May > >2005 10:29:01 -0500 > >Received: from [199.165.152.166] (helo=swlx166.swmed.edu)by > >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DWc5Y-0000IK-D1for > >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:43 -0500 > >Received: from [130.209.16.18] (helo=lenzie.cent.gla.ac.uk)by > >swlx166.swmed.edu with esmtp (Exim 4.44) id 1DWc5X-0000I5-FGfor > >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:32 -0500 > >Received: from monty.bio.gla.ac.uk (montydesk.ibls.gla.ac.uk > >[130.209.46.83])by lenzie.cent.gla.ac.uk (8.11.7/8.11.2) with ESMTP id > >j4DFSUB19208for ;Fri, 13 May 2005 > >16:28:30 +0100 (BST) > >X-Message-Info: gUeNUVfFqHBxE7JvKVytr2DPrM5qbybavSJerH9UTDQ= > >X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 > >X-Scan-Signature: d57e7743657f0a8bf60f191863530969 > >X-BeenThere: histonet@lists.utsouthwestern.edu > >X-Mailman-Version: 2.1.5 > >Precedence: list > >List-Id: For the exchange of information pertaining to histotechnology > >andrelated fields > >List-Unsubscribe: > >, > > > >List-Archive: > >List-Post: > >List-Help: > > >List-Subscribe: > >,tonet-request@lists.utsouthwestern.edu?subject=subscribe> > >Errors-To: histonet-bounces@lists.utsouthwestern.edu > >X-Scan-Signature: 4239ed49a0af0eb50142010c0dad1a43 > >X-SA-Exim-Connect-IP: 127.0.0.1 > >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu > >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on >swlx162.swmed.edu > >X-Spam-Level: * > >X-Spam-Status: No, hits=1.1 required=5.5 tests=MAILTO_TO_SPAM_ADDR > >autolearn=no version=2.64 > >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) > >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) > >Return-Path: histonet-bounces@lists.utsouthwestern.edu > >X-OriginalArrivalTime: 13 May 2005 15:29:12.0313 (UTC) > >FILETIME=[83A15690:01C557D0] > > > >Glen, > > Your attitude is typical of the anti-union stance. Unions are >bad, > >employers are good and without them employees would have a better life. > > >RUBBISH, without properly organised Trades Unions it would be a swift > >return to Victorian values of poverty wages and misery. Why are you > >mentioning bad employees, do we not have bad employers who collect fat >cat > >wages for minimal effort. Of course we do or are you really blinkered >to > >your surroundings. > >Ian. > > > >>Dave, > >> > >>My lab has unionized histotechs and I'd say we are in the lower half >of > >>salary ranges for our area. As far as the union here goes, it seems >they > >>are good at collecting dues and at keeping garbage employees employed. > > >>Not > >>only do unions allow bad employees to keep their jobs, they make it > >>difficult to reward exceptional employees for going above and beyond >the > >>call of duty (since all employees are given union negotiated > >>raises/benefits/etc..). > >> > >>This might be a grass is always greener thing so don't be so sure that >a > >>union will help the histotech's position, that is not always the case. > >> > >>My Opinion, > >> > >>Glen Dawson BS, HT & QIHC (ASCP) > >>IHC Manager > >>Milwaukee, WI > >> > >>-----Original Message----- > >>From: Dave Johnson [mailto:djohnson14@hotmail.com] > >>Sent: Thursday, May 12, 2005 6:17 PM > >>To: Adesupod@aol.com > >>Cc: histonet@lists.utsouthwestern.edu > >>Subject: RE: [Histonet] LOOKING FOR NEW JOB > >> > >> > >>Depending on what you want to make, you could get a job anywhere. It > >>seems > >>everyone is hiring > >>Unfortunately, it doesnt seem that salarys are rising despite the lack >of > >>histotechs. > >> > >>Sad and scary situation if you ask me. I am not much for unions but >now i > >>see why they are valuable. > >> > >>Just my opinion. > >> > >> >From: Adesupod@aol.com > >> >To: histonet@lists.utsouthwestern.edu > >> >CC: Adesupod@aol.com > >> >Subject: [Histonet] LOOKING FOR NEW JOB > >> >Date: Thu, 12 May 2005 20:06:18 EDT > >> > > >> > > >> > Netters, > >> > I am looking for a new job opportunity. My pathologist > > >>passed > >> >away few months ago, and the hospital management have now decided to > > >>close > >> >down > >> >the pathology dept. > >> > My name is Adesupo Adesuyi. I have BS in Medical Technology and >I > >>am > >> >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years > >>experience > >> > >> >as an > >> >histotech. > >> > I presently work in Texas, but I am willing to relocate. > >> > My contact infos are; > >> > > >> > House #: 830-768-0859 > >> > > >> > Cell #: 830-422-9038 > >> > > >> > E/Mail: adesupod@aol.com > >> > > >> > Adesupo Adesuyi > >> > Pathology Dept, > >> > Val Verde Reg.Med.Center, > >> > Del Rio, Tx 78840. > >> >_______________________________________________ > >> >Histonet mailing list > >> >Histonet@lists.utsouthwestern.edu > >> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >> > >> > >> > >>_______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >> > >>_______________________________________________ > >>Histonet mailing list > >>Histonet@lists.utsouthwestern.edu > >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >Dr. Ian Montgomery, > >Histotechnology, > >Graham Kerr Building, > >Institute of Biomedical & Life Sciences, > >University of Glasgow, > >Glasgow, > >G12 8QQ. > >Tel: 0141 339 8855 > >Office: 4652 > >Lab: 6644. > >Pager: 07623 975451 > >e-mail: ian.montgomery@bio.gla.ac.uk > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ander093 <@t> tc.umn.edu Fri May 13 13:41:40 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Thank you RE: Salary and Benefits Questions In-Reply-To: References: <95111571D1C6B5498C64F68DD0769BB506BEFF@cvm36.vetmed.wsu.edu> Message-ID: <6.1.2.0.0.20050513134125.04e05d90@ander093.email.umn.edu> AMEN!!! Thank you Dr. Dahl!!! At 01:21 PM 5/13/05, Julia Dahl wrote: >Thank you to all who have posted! > >I agree completely that pathologists and PAs should have a course in >Histology. Where I trained, I was fortunate enough to do just this - and >it has made a lot of difference in how I gross, how I teach grossing and >how I talk with my clinical colleagues about quality and turnaround time. > >Again, my thanks for your taking the time to share your perspective - I >will certainly incorporate your influence in planning for recruitment and >retention of your valuable talents! > >Julia Dahl, M.D. >Mosaic Gastrointestinal Research Consortium > >>From: "Truscott, Tom" >>To: "Julia Dahl" >>,, >>Subject: RE: Fwd: [Histonet] low salary & unions >>Date: Fri, 13 May 2005 10:36:44 -0700 >>MIME-Version: 1.0 >>Received: from cvm36.vetmed.wsu.edu ([134.121.128.6]) by >>mc1-f8.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Fri, 13 May 2005 >>10:37:09 -0700 >>X-Message-Info: JGTYoYF78jGEhCL50HbvXKZEc+z0eBZREmA8KVl1tnQ= >>Content-class: urn:content-classes:message >>X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 >>X-MS-Has-Attach: X-MS-TNEF-Correlator: Thread-Topic: Fwd: [Histonet] low >>salary & unions >>Thread-Index: AcVX01gnisokgYADSKaqKJEyI9GvQAACqPEQ >>Return-Path: ttruscot@vetmed.wsu.edu >>X-OriginalArrivalTime: 13 May 2005 17:37:09.0347 (UTC) >>FILETIME=[637FC330:01C557E2] >> >>Hi Dr. Dahl, I think that the Federal GS 9 pay scale is fair at the >>lower end and generous at the higher end for my job in research. I am an >>HT with a BS degree in Animal Science. My job entails collecting live >>animal biopsies and collecting tissues at necropsy, then all trimming, >>processing, microtomy and immunostaining and some slide screening. I >>operate the lab myself except for some excellent student help. A good >>health insurance is necessary. 2 to 3 weeks vacation is good. Flex time >>is helpful. When I worked in a private hospital histology lab, the >>stress was higher and the pay poorer. A pat on the back for a job well >>done was only from the best pathologist. As a pathologist starts relying >>less on his or her ability to diagnose a case from an H&E and more on >>the ability of a technician to perform a myriad of diagnostic stains, >>then the tech should start earning a greater percentage of the income >>that the pathologist makes. If a union can't get it's members above ave. >>pay and benefits, then it shouldn't charge them dues. Thanks for your >>concern. Tom Truscott >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Julia >>Dahl >>Sent: Friday, May 13, 2005 7:49 AM >>To: ian.montgomery@bio.gla.ac.uk; histonet@lists.utsouthwestern.edu >>Subject: RE: Fwd: [Histonet] low salary & unions >> >>As a pathologist, I've heard from many histotechnicians and >>histotechnologists that in addition to the pay scale - underappreciation >> >>from pathologists (and/or administration) adds insult to injury. >> >>If you had a comfortable working environment (not crowded or "production >> >>line" mentality) working with pathologists and administrators that >>realized >>the inherent reliance upon histotechnologists for each case... >> >>What is the pay scale that you would allow you to feel fairly >>compensated? >>Generously compensated? >> >>How much paid vacation and PTO time do you desire? >> >>What benefits are essential for you? >> >> >>Yes, I'm really asking and I'd love to hear how to provide a happy, well >> >>compensated work environment. >> >>Julia Dahl, M.D. >>Mosaic Gastrointestinal Research Consortium >>Memphis, Tennessee >> >> >> >From: Ian Montgomery >> >To: histonet@lists.utsouthwestern.edu >> >Subject: Fwd: [Histonet] low salary & unions >> >Date: Fri, 13 May 2005 16:28:29 +0100 >> >MIME-Version: 1.0 >> >Received: from swlx162.swmed.edu ([199.165.152.162]) by >>mc9-f17.hotmail.com >> >with Microsoft SMTPSVC(6.0.3790.211); Fri, 13 May 2005 08:29:12 -0700 >> >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >> >swlx162.swmed.edu with esmtp (Exim 4.34)id 1DWc5v-0000Iq-Tj; Fri, 13 >>May >> >2005 10:29:01 -0500 >> >Received: from [199.165.152.166] (helo=swlx166.swmed.edu)by >> >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DWc5Y-0000IK-D1for >> >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:43 -0500 >> >Received: from [130.209.16.18] (helo=lenzie.cent.gla.ac.uk)by >> >swlx166.swmed.edu with esmtp (Exim 4.44) id 1DWc5X-0000I5-FGfor >> >histonet@lists.utsouthwestern.edu; Fri, 13 May 2005 10:28:32 -0500 >> >Received: from monty.bio.gla.ac.uk (montydesk.ibls.gla.ac.uk >> >[130.209.46.83])by lenzie.cent.gla.ac.uk (8.11.7/8.11.2) with ESMTP id >> >j4DFSUB19208for ;Fri, 13 May 2005 >> >16:28:30 +0100 (BST) >> >X-Message-Info: gUeNUVfFqHBxE7JvKVytr2DPrM5qbybavSJerH9UTDQ= >> >X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 >> >X-Scan-Signature: d57e7743657f0a8bf60f191863530969 >> >X-BeenThere: histonet@lists.utsouthwestern.edu >> >X-Mailman-Version: 2.1.5 >> >Precedence: list >> >List-Id: For the exchange of information pertaining to histotechnology >> >andrelated fields >> >List-Unsubscribe: >> >, >> > >> >List-Archive: >> >List-Post: >> >List-Help: >> >> >List-Subscribe: >> >, >> tonet-request@lists.utsouthwestern.edu?subject=subscribe> >> >Errors-To: histonet-bounces@lists.utsouthwestern.edu >> >X-Scan-Signature: 4239ed49a0af0eb50142010c0dad1a43 >> >X-SA-Exim-Connect-IP: 127.0.0.1 >> >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >> >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on >>swlx162.swmed.edu >> >X-Spam-Level: * >> >X-Spam-Status: No, hits=1.1 required=5.5 tests=MAILTO_TO_SPAM_ADDR >> >autolearn=no version=2.64 >> >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >> >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >> >Return-Path: histonet-bounces@lists.utsouthwestern.edu >> >X-OriginalArrivalTime: 13 May 2005 15:29:12.0313 (UTC) >> >FILETIME=[83A15690:01C557D0] >> > >> >Glen, >> > Your attitude is typical of the anti-union stance. Unions are >>bad, >> >employers are good and without them employees would have a better life. >> >> >RUBBISH, without properly organised Trades Unions it would be a swift >> >return to Victorian values of poverty wages and misery. Why are you >> >mentioning bad employees, do we not have bad employers who collect fat >>cat >> >wages for minimal effort. Of course we do or are you really blinkered >>to >> >your surroundings. >> >Ian. >> > >> >>Dave, >> >> >> >>My lab has unionized histotechs and I'd say we are in the lower half >>of >> >>salary ranges for our area. As far as the union here goes, it seems >>they >> >>are good at collecting dues and at keeping garbage employees employed. >> >> >>Not >> >>only do unions allow bad employees to keep their jobs, they make it >> >>difficult to reward exceptional employees for going above and beyond >>the >> >>call of duty (since all employees are given union negotiated >> >>raises/benefits/etc..). >> >> >> >>This might be a grass is always greener thing so don't be so sure that >>a >> >>union will help the histotech's position, that is not always the case. >> >> >> >>My Opinion, >> >> >> >>Glen Dawson BS, HT & QIHC (ASCP) >> >>IHC Manager >> >>Milwaukee, WI >> >> >> >>-----Original Message----- >> >>From: Dave Johnson [mailto:djohnson14@hotmail.com] >> >>Sent: Thursday, May 12, 2005 6:17 PM >> >>To: Adesupod@aol.com >> >>Cc: histonet@lists.utsouthwestern.edu >> >>Subject: RE: [Histonet] LOOKING FOR NEW JOB >> >> >> >> >> >>Depending on what you want to make, you could get a job anywhere. It >> >>seems >> >>everyone is hiring >> >>Unfortunately, it doesnt seem that salarys are rising despite the lack >>of >> >>histotechs. >> >> >> >>Sad and scary situation if you ask me. I am not much for unions but >>now i >> >>see why they are valuable. >> >> >> >>Just my opinion. >> >> >> >> >From: Adesupod@aol.com >> >> >To: histonet@lists.utsouthwestern.edu >> >> >CC: Adesupod@aol.com >> >> >Subject: [Histonet] LOOKING FOR NEW JOB >> >> >Date: Thu, 12 May 2005 20:06:18 EDT >> >> > >> >> > >> >> > Netters, >> >> > I am looking for a new job opportunity. My pathologist >> >> >>passed >> >> >away few months ago, and the hospital management have now decided to >> >> >>close >> >> >down >> >> >the pathology dept. >> >> > My name is Adesupo Adesuyi. I have BS in Medical Technology and >>I >> >>am >> >> >HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years >> >>experience >> >> >> >> >as an >> >> >histotech. >> >> > I presently work in Texas, but I am willing to relocate. >> >> > My contact infos are; >> >> > >> >> > House #: 830-768-0859 >> >> > >> >> > Cell #: 830-422-9038 >> >> > >> >> > E/Mail: adesupod@aol.com >> >> > >> >> > Adesupo Adesuyi >> >> > Pathology Dept, >> >> > Val Verde Reg.Med.Center, >> >> > Del Rio, Tx 78840. >> >> >_______________________________________________ >> >> >Histonet mailing list >> >> >Histonet@lists.utsouthwestern.edu >> >> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> >> >> >> >>_______________________________________________ >> >>Histonet mailing list >> >>Histonet@lists.utsouthwestern.edu >> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >> >>Histonet mailing list >> >>Histonet@lists.utsouthwestern.edu >> >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > >> >Dr. Ian Montgomery, >> >Histotechnology, >> >Graham Kerr Building, >> >Institute of Biomedical & Life Sciences, >> >University of Glasgow, >> >Glasgow, >> >G12 8QQ. >> >Tel: 0141 339 8855 >> >Office: 4652 >> >Lab: 6644. >> >Pager: 07623 975451 >> >e-mail: ian.montgomery@bio.gla.ac.uk >> > >> > >> >_______________________________________________ >> >Histonet mailing list >> >Histonet@lists.utsouthwestern.edu >> >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lsb <@t> jax.org Fri May 13 14:12:09 2005 From: lsb <@t> jax.org (Lesley S. Bechtold) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Green Mountain Logic Message-ID: <6.2.1.2.2.20050513151102.02c5bc48@aretha.jax.org> Hi, Does anyone out there have any experience with a customized LIMS known as LABPas? It's made by Green Mountain Logic. They're in Vermont. TIA! Lesley Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 From marktarango <@t> earthlink.net Fri May 6 20:23:41 2005 From: marktarango <@t> earthlink.net (Mark Tarango) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Re: Histonet Digest, Vol 18, Issue 20 References: <200505131000.1dwDwuI03NZFmR0@bunting.mail.pas.earthlink.net> Message-ID: <001b01c552a3$68296f60$62b6a6ac@TARANGO> Ian, No way, unions make you work somewhere forever to get a raise....and they start you out on the bottom. They have silly rules. Employers can aren't all good, but the bottom line is if you're needed, you can do tons better negociating on your own. Because histologists are in demand, we have the upper hand in negociations. There is no way I'd be making as much I do being for 25 years old and not being in the field for 30+ years if I were working somewhere where unions were the rule. Mark >Date: Fri, 13 May 2005 16:28:29 +0100 >From: Ian Montgomery >Subject: Fwd: [Histonet] low salary & unions >To: histonet@lists.utsouthwestern.edu >Message-ID: <6.2.1.2.2.20050513162104.0301deb0@udcf.gla.ac.uk> >Content-Type: text/plain; charset="us-ascii"; format=flowed > >Glen, > Your attitude is typical of the anti-union stance. Unions are bad, >employers are good and without them employees would have a better life. >RUBBISH, without properly organised Trades Unions it would be a swift >return to Victorian values of poverty wages and misery. Why are you >mentioning bad employees, do we not have bad employers who collect fat cat >wages for minimal effort. Of course we do or are you really blinkered to >your surroundings. >Ian. From PatPatterson <@t> mhd.com Fri May 13 15:59:00 2005 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Histology Position Message-ID: <293C7C19EFF7D611AE1A0002A53F81140FA63532@omega.mhd.com> Histology Technician: Named one of the "Best Places to work 2005" by the Dallas Business Journal. Our dedicated team is very outspoken in their appreciation of the excellent clinical environment we foster and the ongoing commitment Methodist has made to patient satisfaction. Our dual focus on out patients and our employees is working - 2002 Methodist Health System earned the Gold Seal of Approval form the Joint Commission on Accreditation of Healthcare Organizations and more than 28 percent of our new hires come from employee referral! If excellent patient care is one of your priorities for career satisfaction, then you belong at Methodist. Join our Histology team at Methodist Dallas Medical Center. We're seeking a full-time Histology Technician on the day shift in our Anatomic Pathology Section. Job responsibilities include processing, embedding, sectioning, routine, special and Immunofluoresence staining, assisting in gross room, frozen sectioning and computer entry into the MediTech lab information system. Experience with microwave technology would be an added plus. Requires HT/HTL ASCP registry or eligible. Minimum 2-year experience desired. Pat Patterson Lab Manager Phone 214-947-3538 Fax: 214-947-3524 patpatterson@mhd.com or contact Methodist Dallas Medical Center Phone 214-947-6510 Fax 214-947-6529 E-mail lorenadavila@mhd.com *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From jkiernan <@t> uwo.ca Fri May 13 23:49:37 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] methasol fast blue References: <000601c55491$d6e7e540$ed54bec1@ugent.be> Message-ID: <42858361.21E98F63@uwo.ca> Dear Manuelle, You didn't give details of your reference. Is it Maxwell, A (1963) The alcian dyes applied to gastric mucosa. Stain Technol 38: 286-287? This paper showed that gastric mucus (a neutral carbohydrate macromolecule, normally alcian-negative) could be oxidized with periodic acid and then treated with bisulphite to generate aldehyde bisulphite compounds. These are strong acids. As such, they can be stained with alcian dyes even at low pH. This principle is exploited to stain gastric mucus yellow (with alcian yellow) as a contrasting background to Helicobacter organisms subsequently stained at a lower pH with a blue cationic dye. Look up: http://stainsfile.info/StainsFile/stain/micro/heliaytb.htm and check out its main reference (Leung & Gibbons 1996 J. Histotechnol. 19: 131ff) I cannot find "methasol fast blue" as the name of a dye. Lillie (in the 9th edition of Conn's Biological Stains, 9th edn, 1977) had an entry for "methazol fast blue 2GI" (CI 74360, Solvent blue 24). He briefly argued that the name probably applied to a solvent dye equivalent to luxol fast blue MBS. A dye with a "luxol" name will never be used to stain mucus. The "luxol" dyes are used to stain myelin. Don't look for "links on the web where I can buy this dye". Check out the original published account and review it in the light of more recent work. You are at a major European university. Use its libraty to find the information. A name like "methasol" or "methazol" almost certainly indicates a solvent dye (used to stain lipids). John Kiernan London, Canada __________________________ Manuelle De Bock wrote: > > I am trying to stain collected gastric tissue cells with Maxwell's staining procedure (1963) > but we don't have methasol fast blue in our lab. I also can't find any links on the web where I can buy this dye. can someone help me with this? Is it the same dye as Luxol fast blue? > > Kind regards > Manuelle > > Manuelle De Bock, dierenarts (DVM) > Laboratory of Veterinary Pathology > Department of Pathology, Bacteriology and Avian Diseases > Faculty of Veterinary Medicine > Ghent University > Salisburylaan 133 > B9820 Merelbeke - Belgium > Tel 0032(0)9 264 7745 > Fax 0032(0)9 264 7789 From conlonb <@t> comcast.net Sat May 14 15:43:05 2005 From: conlonb <@t> comcast.net (Brian D. Conlon) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] (no subject) Message-ID: <004301c558c5$8b3783c0$6401a8c0@gateway> From sladd <@t> hsc.usf.edu Mon May 16 06:40:05 2005 From: sladd <@t> hsc.usf.edu (Ladd, Sharron) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Need IHC done...willing to pay Message-ID: <841D767DCDE87C49A02DA6DFC0BABABF67471F@COMEXCHANGE.hscnet.hsc.usf.edu> Dear Histonetters, I received the following plea for help: "I have a grant going out for July 1 and I am under the wire for some last preliminary data including slides. I will need some routine brain histology including H & E, GFAP, CD3, CD4, CD8, and CD 56 on several specimens between June 15 and June 25." Is there anyone out there who can do this? If so, please send me your contact information and price list (if available). Thanks so much!! Sharron Tampa, FL From sjchtascp <@t> yahoo.com Mon May 16 07:33:27 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Wisconsin Position Message-ID: <20050516123327.10416.qmail@web90208.mail.scd.yahoo.com> Newcomer Supply, Madison WI 800-383-7799 --------------------------------- Yahoo! Mail Mobile Take Yahoo! Mail with you! Check email on your mobile phone. From Hedley.Glencross <@t> CMMC.nhs.uk Mon May 16 08:01:51 2005 From: Hedley.Glencross <@t> CMMC.nhs.uk (Glencross Hedley (RW3) CM&MC Manchester) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] RE: formalin Message-ID: Hi everyone It has always been my understanding that "formalin" like "Hoover" & "Sellotape" (and countless other words) is just a trade name. This describes a 37% solution of formaldehyde gas in water, and we should be talking about 3.7% (4%) formaldehyde and not 10% formalin. Regards Hedley Glencross Manchester Cytology Centre UK For all my years I have believed that formaldehyde was the 37% concentrate without buffers, and that formalin was 10% buffered... learn something new every day! Thanks John! -----Original Message----- From: John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, May 13, 2005 11:31 AM To: Weems, Joyce Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quick question about formaldehyde Dear Joyce, Formalin = 37% formaldehyde; so no, it doesn't make a difference. Formalin is not buffered; it does contain about 10% methanol, which is put in to retard polymerization. When diluted to make a 4% formaldehyde fixative, the methanol concentration is 1%. Buffering of the dilute solution offsets pH changes due to the Cannizzaro reaction. It also inhibits the formation of blood-derived "formalin pigment" which forms after fixation in an acidic formaldehyde solution. Tim Morken is correct in saying we don't know the extent of chemical change in 12 year-old formalin. The fact that there's no expiry date sugggests that it's not much. For what it's worth, I've used formalin that's more than 5 years old and fixation has been OK. John Kiernan london, Canada From aep10 <@t> cornell.edu Mon May 16 08:32:07 2005 From: aep10 <@t> cornell.edu (Anna Elisse Beaudin) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] in situ with anti-dig HRP? Message-ID: <1906.128.253.96.40.1116250327.squirrel@128.253.96.40> Hello, I am currently using Dig-labeled sense and as probes on fresh frozen mouse brain cryosections. I have been using anti-dig AP, but I find it takes a long time to develop the signal with NBT/BCIP (this is a rare mRNA transcript), and also find that the sections get much darker over time (someone else brought this up on histonet recently). I am considering using Dako's anti-dig HRP, but I was wondering if (and where in the protocol) I then needed to include a peroxidase blocking step. Also, is it necessary to do a TSA amplification with anti-dig HRP, or can I still get clean signal with just a DAB incubation? I would greatly appreciate anyone's advice on this matter! Thanks! Anna Beaudin Division of Nutritional Sciences Cornell University Ithaca, NY From pmarcum <@t> vet.upenn.edu Mon May 16 08:41:08 2005 From: pmarcum <@t> vet.upenn.edu (pmarcum@vet.upenn.edu) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] RE: formalin In-Reply-To: References: Message-ID: <1116250868.4288a2f40d60c@imp.vet.upenn.edu> Hi Hedley and All, Over the years we have adopted in the US and a great deal of literature the use of formalin to cover most formaldehyde solutions. Yes, it is wrong however, it is sometimes difficult to change. What is interesting to me is paraformaldehyde powder solutions are generally called 4% as it is made (4gm to 100mL of 60C water). It is difficult to get an understanding that anything starting as paraformaldehyde powder will end up as a formaldehyde solution. I tend to think of formaldehyde as 37 to 40% gas in water and formalin as what is usually called 10% Neutral Buffered Formalin, as it is 10mL of 37% to 40% formaldehdye in 90mL of buffer or water. I know it is really approximately 3.7% however it is not clear to a lot of people that this what we are using. In speaking with people as a technical person these terms were always sticky and required an explanation so we could talk about the same concentration no matter what the starting material. Even the histology textbooks do not explain this well. Also John said on Friday that formaldehye contained 10% methanol. I have done a good deal of referencing on this subject for a seminar and it can range from 5% to 15% methanol depending on the manufacturer. It is the same with pre-made 10% NBF. The methanol precentage will depend on when the methanol is added and if it is only in the 37% formaldehyde or added to the final NBF solution at the time of manufacture. Reading the MSDS to determine exactly what amonut of methanol is contained in a solution is critcal for some processes and should be carefully monitored when vendor changes are made and problems arise. Pam Marcum Quoting "Glencross Hedley (RW3) CM&MC Manchester" : > Hi everyone > > It has always been my understanding that "formalin" like "Hoover" & > "Sellotape" (and countless other words) is just a trade name. This > describes a 37% solution of formaldehyde gas in water, and we should be > talking about 3.7% (4%) formaldehyde and not 10% formalin. > > Regards > > Hedley Glencross > > Manchester Cytology Centre UK > > For all my years I have believed that formaldehyde was the 37% > concentrate without buffers, and that formalin was 10% buffered... learn > something new every day! > > Thanks John! > > -----Original Message----- > From: John A. Kiernan [mailto:jkiernan@uwo.ca] > Sent: Friday, May 13, 2005 11:31 AM > To: Weems, Joyce > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick question about formaldehyde > > > Dear Joyce, > > Formalin = 37% formaldehyde; so no, it doesn't > make a difference. Formalin is not buffered; it > does contain about 10% methanol, which is put > in to retard polymerization. When diluted to > make a 4% formaldehyde fixative, the methanol > concentration is 1%. Buffering of the dilute > solution offsets pH changes due to the > Cannizzaro reaction. It also inhibits the > formation of blood-derived "formalin pigment" > which forms after fixation in an acidic > formaldehyde solution. > > Tim Morken is correct in saying we don't know > the extent of chemical change in 12 year-old > formalin. The fact that there's no expiry date > sugggests that it's not much. For what it's > worth, I've used formalin that's more than 5 > years old and fixation has been OK. > > John Kiernan > london, Canada > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Kemlo.Rogerson <@t> elht.nhs.uk Mon May 16 08:36:15 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] RE: formalin Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2C3@bhrv-nt-11.bhrv.nwest.nhs.uk> "Formaldehyde, as 4% buffered formaldehyde (10% buffered formalin), is the most widely employed universal fixative particularly for routine paraffin embedded sections. It is a gas with a very pungent odour, soluble in water to a maximum extent of 40% by weight and is sold as such under the name of formaldehyde (40%) or formalin (a colourless liquid). Formaldehyde is also obtainable in a stable solid form composed of high molecular weight polymers known as paraformaldehyde. Heated paraformaldehyde generates pure gaseous formaldehyde which, when dissolved in water, reverts mostly to the monomeric form. Aqueous formaldehyde exists principally in the form of its monohydrate, methylene glycol, CH2(OH)2, and as low molecular weight polymeric hydrates or polyoxymethylene glycols. It has been suggested that the hydrated form, methylene glycol, is the reactive component of formaldehyde but this has been disputed2" Fixation and fixatives Anthony S-Y Leong So as it is a gas it must be held in solution by atmospheric pressure? If that atmospheric is raised or lowered then lesser or greater amount of gas can be held in solution; the same with increasing or decreasing temperatures. So is formalin 40% formaldehyde gas up Everest or is it much less? Atmospheric pressure is reduced so the Saturated Vapour Pressure of the solution may exceed barometric pressure and the gas is driven off until the pressures equilibrate; plus it is very cold. Do you have to make a stronger dilution up Everest to end up with a 4% buffered formaldehyde? Are there any Histo Labs up there? Who cares? Kemlo Rogerson Cellular Pathology Manager East Lancashire Hospitals NHS Trust DD. 01254-294162 Mobile 0774-9754194 -----Original Message----- From: Glencross Hedley (RW3) CM&MC Manchester [mailto:Hedley.Glencross@CMMC.nhs.uk] Sent: 16 May 2005 14:02 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: formalin Hi everyone It has always been my understanding that "formalin" like "Hoover" & "Sellotape" (and countless other words) is just a trade name. This describes a 37% solution of formaldehyde gas in water, and we should be talking about 3.7% (4%) formaldehyde and not 10% formalin. Regards Hedley Glencross Manchester Cytology Centre UK For all my years I have believed that formaldehyde was the 37% concentrate without buffers, and that formalin was 10% buffered... learn something new every day! Thanks John! -----Original Message----- From: John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: Friday, May 13, 2005 11:31 AM To: Weems, Joyce Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] quick question about formaldehyde Dear Joyce, Formalin = 37% formaldehyde; so no, it doesn't make a difference. Formalin is not buffered; it does contain about 10% methanol, which is put in to retard polymerization. When diluted to make a 4% formaldehyde fixative, the methanol concentration is 1%. Buffering of the dilute solution offsets pH changes due to the Cannizzaro reaction. It also inhibits the formation of blood-derived "formalin pigment" which forms after fixation in an acidic formaldehyde solution. Tim Morken is correct in saying we don't know the extent of chemical change in 12 year-old formalin. The fact that there's no expiry date sugggests that it's not much. For what it's worth, I've used formalin that's more than 5 years old and fixation has been OK. John Kiernan london, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pzeitlow <@t> bbpllab.com Mon May 16 10:16:28 2005 From: pzeitlow <@t> bbpllab.com (Pat Zeitlow) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] CA19-9 on XT Message-ID: <813FB33DA405334F947F8BFC6EBD0B2A0BEC00@bbplsrv1.bbpl> Does anyone have CA19-9 protocol that works well on Benchmark XT? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. From relia1 <@t> earthlink.net Mon May 16 11:37:13 2005 From: relia1 <@t> earthlink.net (Pam Barker) Date: Fri Sep 16 15:25:05 2005 Subject: [Histonet] Relia's Job Opportunity Update 5/16/05 Message-ID: Hi Histonetters! My name is Pam Barker and I am a recruiter that has specialized in permanent placement in the histology profession for the past 2 ? years. All of the positions I work with are fulltime 40 hour per week positions with top hospitals, labs and doctors offices. My clients offer excellent compensation including competitive salaries, great benefits, relocation assistance and in some cases sign on bonuses. My services are FREE of charge to you. All of my fees are paid by my clients, the facilities that I represent. I represent companies nationwide that are in need of histology supervisors, histotechnologists and histo technicians. Here is a list of my most exciting current openings: 1. Histology Supervisor ? Central Florida (Florida Supervisor?s license required) 2. Histo Tech ? Minnesota 3. Histo Tech ? South Florida (current Florida license required) 4. Histo Tech ? Southeastern Florida (current Florida license required) 5. Histo Tech ? Illinois 6. Histo Tech ? Michigan 7. Histo Tech - Massachusetts If you are interested in any of these positions please call me at 866-607-3542 or e-mail me at relia1@earthlink.net If you would like you can e-mail me your resume and a number where I can reach you at a time that is convenient for you. If you are interested in looking into new job opportunities in other areas that are not mentioned above please contact me as well. I work with facilities nationwide and I will keep your resume confidential. I will only represent you to jobs you tell me you are interested in looking into. Remember It never hurts to keep an eye open even if you are happy in your present job. Also if you know anyone else that might be interested I would really appreciate it if you would pass my information along to them as well. Thank you, Pam ? 866-607-3542 (866-60RELIA) Thank You! Pam Barker President RELIA Specialists in Allied Healthcare Recruiting 5703 Red Bug Lake Road #330 Winter Springs, FL 32708-4969 Phone: (407)657-2027 Cell: (407)353-5070 FAX: (407)678-2788 E-mail: relia1@earthlink.net From juan.gutierrez <@t> christushealth.org Mon May 16 12:04:07 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] CA19-9 on XT Message-ID: We use DAKO's primary on a regular Benchmark at 1:50 for 32 min no pretreatment. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Zeitlow Sent: Monday, May 16, 2005 10:16 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CA19-9 on XT Does anyone have CA19-9 protocol that works well on Benchmark XT? Pat Z Department of Molecular Pathology Boyce and Bynum Pathology Labs, P.C. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From innomedical <@t> charter.net Mon May 16 12:14:46 2005 From: innomedical <@t> charter.net (Innovative Medical Recruiting) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Histology Supervisor - New York City Message-ID: <010f01c55a3a$c347e900$63e6b944@cx657389a> We are conducting a search for a qualified Histology Supervisor for a very large, state-of-the-art, commercial laboratory in New York City. My client is a multi-billion dollar biotechnology company, servicing patients in over 80 countries, and dedicated to making a major positive impact on the lives of people with serious diseases. For over 20 years my client's products and services have focused on the treatment of rare inherited disorders, kidney disease, orthopedics, transplant and immune disease, cancer, and diagnostic testing. Their commitment today continues with a substantial research and development program focused on genetic diseases, immune system disorders, heart disease, and cancer. A brief description of the job is listed below: JOB SUMMARY Supervises and is responsible for all histology procedures and operation of the Histology Laboratory. Trains and coordinates staff to ensure the highest quality work as well as an excellent turn-around-time. TECHNICAL RESPONSIBILITIES Routine Duties: 1. Oversees and/or performs all procedures under the scope of the Histology laboratory. 2. Oversees the staining of H & E's, embedding, slide set up and positive control production, cutting and all "Special Stains". 3. Sets up R & D assays and reviews protocol changes or new reagent tests to be added to the daily caseload. 4. Trains new employees, administers competency examinations and documents all continuing education. Reagent Preparation: 1. Oversees the maintenance of stock solutions and reagents preparations. 2. Oversees the proper labeling and dating of prepared reagents and solutions. 3. Tests and validates all new reagents. Quality Control: 1. Evaluates and documents quality and quantity of work daily. 2. Ensures documentation of daily performance of reagents and equipment. 3. Revises and updates Standard Operating Procedures Manual for the Histology Laboratory. 4. Checks that the QC record keeping of equipment is accurately recorded and reviews documents on a monthly basis. 5. Selects, tests and coordinates efforts to maintain the positive control tissue bank for the Histotechnicians. 6. Ensures that all new reagents are validated before use as clinical reagents. 7. Performs validations for reagents, procedures and/or equipment. 8. Reviews and spot-checks slides for staining and compliance with procedures. Trouble Shooting: Is able to perform trouble-shooting procedures for the Histology lab and to instruct others on same. ADMINISTRATIVE RESPONSIBILITIES: 1. Coordinates and trains, either directly or through lead/senior personnel in the Histology Laboratory and insures proper coverage using FTE's and part time personnel. 2. Record-keeping: a. Maintains numeric data on lab test output. b. Maintains up-to-date files for all staff according to QA standards. 3. Inventory Control: Ensures a consistent supply of reagents and consumables by submitting supply requisitions in a timely manner. 4. Quality Assurance: a. Follows all Quality Assurance procedures as mandated by laboratory licensing agencies. For example: 1. Generates incident reports as needed. 2. Responsible for development and implementation of all corrective actions. 3. Expected to attend all monthly QA meetings and report lab incidents, QI statistics, and trends. 4. Reports to staff the findings of monthly QA meetings. 5. Communicates with staff through use of communication log and/or lab meetings. 5. Answers phone inquiries and takes messages. ENVIRONMENT/SAFETY: 1. Responsible for the knowledge and practice of safety SOP's in the laboratory. 2. Maintains a neat and orderly work area. 3. Controls exposure to hazardous chemicals and other safety hazards such as: a. Carcinogens b. Strong and obnoxious fumes c.Sharp and dangerous blades and needles d. Biohazard exposure to fresh/frozen specimens PERSONAL WORK CONTACTS: 1. Reports to and takes direction from the Director of Technical Services. 2. Takes direction from the professional and medical staff. 3. Trains and supervises technical staff assigned to the Histology laboratory. 4. Interfaces with other laboratory and operational departments for specimen processing and other cooperative efforts. 5. Maintains a pleasant, cooperative and respectful relationship with co-workers and all staff. JOB QUALIFICATIONS: 1. Bachelor's degree in the biological sciences is required. 2. Five years of progressive histology and/or pathology laboratory experience. 3. Good working knowledge of microanatomy and familiarity with the staining characteristics of the antibodies currently in use. 4. ASCP certification is preferred. 5. Good communication skills 6. Thrives on the opportunity to excel. Salary/Benefits: Very competitive base salary ($85K - 95K range), along with some of the best healthcare, retirement, and 401(k) benefits in the industry. All qualified and interested candidates should attach a resume as a Word document to dale@innomedical.com for more information. Dale Busbee, Owner Innovative Medical Recruiting, LLC (985) 641-8817 www.innomedical.com From azdudley <@t> hotmail.com Mon May 16 12:19:48 2005 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Charging for decal In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7EA458FB@sjhaexc02.sjha.org> Message-ID: joyce, we bill decal per specimen. anita dudley providence hosp mobile ala >From: "Weems, Joyce" >To: "Histonet" >Subject: [Histonet] Charging for decal >Date: Fri, 8 Apr 2005 10:57:02 -0400 > >Do you folks bill decal per block or per specimen? > >Thanks >Joyce > >Joyce Weems >Pathology Manager >Saint Joseph's Hospital >5665 Peachtree Dunwoody Rd NE >Atlanta, GA 30342 >404-851-7376 - Phone >404-851-7831 - Fax > > > >Confidentiality Notice ** The information contained in this message may be >privileged and is confidential information intended for the use of the >addressee listed above. If you are neither the intended recipient nor the >employee or agent responsible for delivering this message to the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or the taking of any action in reliance on the contents of >this information is strictly prohibited. If you have received this >communication in error, please notify us immediately by replying to the >message and deleting it from your computer. >Thank you. Saint Josephs Health System, Inc. > ><< disclaimer.txt >> >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From VBlanc <@t> asterand.com Mon May 16 13:05:28 2005 From: VBlanc <@t> asterand.com (Vici Blanc) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: Need IHC done...willing to pay VENDOR RESPONSE Message-ID: <56E63440ABF1FC4897947D5BA4CFA44F04F9304E@ATL1VEXC005.usdom004.tco.tc> Dear Sharron, Asterand provides fee for service IHC staining. I realize I am in breach of Histonet protocol by responding to all with a commercial email-please forgive the transgression. I didn't see a direct email address. You may reach me as outlined below. Hope we can help you, Vici -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ladd, Sharron Sent: Monday, May 16, 2005 7:40 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Need IHC done...willing to pay Dear Histonetters, I received the following plea for help: "I have a grant going out for July 1 and I am under the wire for some last preliminary data including slides. I will need some routine brain histology including H & E, GFAP, CD3, CD4, CD8, and CD 56 on several specimens between June 15 and June 25." Is there anyone out there who can do this? If so, please send me your contact information and price list (if available). Thanks so much!! Sharron Tampa, FL Victoria M. Blanc, Ph.D. Director of Experimental Pathology Services Asterand, Inc 440 Burroughs TechOne, suite 501 Detroit, MI 48202 Phone: 313-263-0960, ext. 240 Fax: 313-263-0961 www.asterand.com This e-mail message, together with any attachments, contains information belonging to Asterand, Inc. that may be confidential, proprietary, copyrighted and/or legally protected or privileged, and is intended for the sole use of the individual or entity named on this message. If you are not the intended recipient, and have received this e-mail message in error, please immediately return this message to its original sender and permanently delete it from your electronic and paper records. Thank You. From jkiernan <@t> uwo.ca Mon May 16 13:10:37 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: formalin [Longish] References: <1030B679AD69D6119C3F00080210DD9D05A3F2C3@bhrv-nt-11.bhrv.nwest.nhs.uk> Message-ID: <4288E21D.2A936674@uwo.ca> Kemlo, I think your atmospheric pressure ideas would be true only if our atmosphere consisted largely of gaseous formaldehyde, which fortunately is not the case. (Formaldehyde molecules are said to be present in interstellar space; perhaps that's why there isn't any life out there among the fixed stars.) The gas dissolves in water because it's very soluble and also because, as you point out, there is a chemical reaction that forms methylene hydrate. In 37% formaldehyde (neat formalin) 37% of the weight is derived from formaldehyde gas and 63% from water. 40% is the wt/vol expression (There are 40 g of formaldehyde in 100 ml of the 37% wt/wt solution.) Despite the smell, not much formaldehyde escapes into the air. When formalin evaporates, most of the formaldehyde ends up as the solid polymer paraformaldehyde. Evaporation might occur more quickly on top of Mt Everest than at sea level, especially on a sunny day with a brisk wind; I'm sure the YETIS (Yogic Engineering and Technical Institute for Sherpas) up there know this, and fix their specimens in capped jars and vials. It is possible to distil aqueous solutions of formaldehyde, and Walker's book ("Formaldehyde", 3rd ed 1964) gives tabulated data for the composition of distillates obtained under different conditions. Regarding the quotation, "It has been suggested that the hydrated form, methylene glycol, is the reactive component ..." this was certainly once believed (see for example, Walker's book and the first 3 editions of Pearse's Histochemistry). The more recent view is that the methylene glycol constitutes a reserve, always in equilibrium with a very small concentration of dissolved formaldehyde molecules and water. When a formaldehyde molecule is pulled out of the solution (by combining with part of a protein molecule, for example), it is immediately replaced from the reservoir of methylene glycol. I do not know enough chemistry to fully understand the experimental evidence, but it was published in chemical journals, presumably with knowledgeable chemists doing the peer-reviewing. For a recent review of fixation, see D. Hopwood's chapter in Bancroft & Gamble's "Theory and Practice of histological Techniques" (2002). The fixation chapter in the 4th edition of Pearse (Vol 1, 1980) is older but seems still to be up to date. The short review "Formaldehyde fixation" by Fox et al (1985) J Histochem Cytochem 33:845-853 is worth reading because it covers the earliest applications as well as the current understanding of th chemistry and its practical implications. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Kemlo Rogerson wrote: > > "Formaldehyde, as 4% buffered formaldehyde (10% buffered formalin), is the > most widely employed universal fixative particularly for routine paraffin > embedded sections. It is a gas with a very pungent odour, soluble in water > to a maximum extent of 40% by weight and is sold as such under the name of > formaldehyde (40%) or formalin (a colourless liquid). Formaldehyde is also > obtainable in a stable solid form composed of high molecular weight polymers > known as paraformaldehyde. Heated paraformaldehyde generates pure gaseous > formaldehyde which, when dissolved in water, reverts mostly to the monomeric > form. Aqueous formaldehyde exists principally in the form of its > monohydrate, methylene glycol, CH2(OH)2, and as low molecular weight > polymeric hydrates or polyoxymethylene glycols. It has been suggested that > the hydrated form, methylene glycol, is the reactive component of > formaldehyde but this has been disputed2" > > Fixation and fixatives > Anthony S-Y Leong > > So as it is a gas it must be held in solution by atmospheric pressure? If > that atmospheric is raised or lowered then lesser or greater amount of gas > can be held in solution; the same with increasing or decreasing > temperatures. So is formalin 40% formaldehyde gas up Everest or is it much > less? Atmospheric pressure is reduced so the Saturated Vapour Pressure of > the solution may exceed barometric pressure and the gas is driven off until > the pressures equilibrate; plus it is very cold. Do you have to make a > stronger dilution up Everest to end up with a 4% buffered formaldehyde? Are > there any Histo Labs up there? Who cares? > > Kemlo Rogerson > Cellular Pathology Manager > East Lancashire Hospitals NHS Trust > DD. 01254-294162 > Mobile 0774-9754194 > > > -----Original Message----- > From: Glencross Hedley (RW3) CM&MC Manchester > [mailto:Hedley.Glencross@CMMC.nhs.uk] > Sent: 16 May 2005 14:02 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: formalin > > Hi everyone > > It has always been my understanding that "formalin" like "Hoover" & > "Sellotape" (and countless other words) is just a trade name. This > describes a 37% solution of formaldehyde gas in water, and we should be > talking about 3.7% (4%) formaldehyde and not 10% formalin. > > Regards > > Hedley Glencross > > Manchester Cytology Centre UK > > For all my years I have believed that formaldehyde was the 37% > concentrate without buffers, and that formalin was 10% buffered... learn > something new every day! > > Thanks John! > > -----Original Message----- > From: John A. Kiernan [mailto:jkiernan@uwo.ca] > Sent: Friday, May 13, 2005 11:31 AM > To: Weems, Joyce > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick question about formaldehyde > > Dear Joyce, > > Formalin = 37% formaldehyde; so no, it doesn't > make a difference. Formalin is not buffered; it > does contain about 10% methanol, which is put > in to retard polymerization. When diluted to > make a 4% formaldehyde fixative, the methanol > concentration is 1%. Buffering of the dilute > solution offsets pH changes due to the > Cannizzaro reaction. It also inhibits the > formation of blood-derived "formalin pigment" > which forms after fixation in an acidic > formaldehyde solution. > > Tim Morken is correct in saying we don't know > the extent of chemical change in 12 year-old > formalin. The fact that there's no expiry date > sugggests that it's not much. For what it's > worth, I've used formalin that's more than 5 > years old and fixation has been OK. > > John Kiernan > london, Canada From Adesupod <@t> aol.com Mon May 16 13:56:42 2005 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] LOOKING FOR A NEW JOB Message-ID: <126.5d2be61c.2fba46ea@aol.com> ?????? ???? Netters, ???????????? I am looking for a new job opportunity. My pathologist passed away few months ago, and the hospital management have now decided to close down the pathology dept. ?? My name is Adesupo Adesuyi. I? have BS in Medical Technology and I am? HT(ASCP) and HTL(ASCP)certified histotech. I have about 15-years experience as an histotech. ??? I presently work in Texas, but I am willing to relocate. ???? My contact infos are; ????? House #:??? 830-768-0859 ????? Cell #:??????? 830-422-9038 ? ????? E/Mail:?????? adesupod@aol.com ??????? Adesupo Adesuyi ?????? Pathology Dept, ????? Val Verde Reg.Med.Center, ????? Del Rio, Tx 78840. From ernestinemiddleton <@t> yahoo.ca Mon May 16 21:10:19 2005 From: ernestinemiddleton <@t> yahoo.ca (Ernestine Middleton) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] positions opening Message-ID: <20050517021019.62095.qmail@web51503.mail.yahoo.com> Hi; Montefiore Medical Center, Bronx, NY has 3 jobs opening in Surgical Pathology. Two positions will be designated for the grossing lab. and one is for the histology lab. These are Technologist positions. HT/HTL is required for Histology. They will accept MT for the grossing lab. Fax resume to: Ernestine Middleton 718-547-1920 e-mail: ernestinemiddleton@yahoo.ca --------------------------------- Post your free ad now! Yahoo! Canada Personals From leahcox27 <@t> yahoo.com Mon May 16 23:20:49 2005 From: leahcox27 <@t> yahoo.com (Leah Cox) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Wanted: Position in Arizona Message-ID: <20050517042049.20825.qmail@web50201.mail.yahoo.com> Hello Histonet, I have been working in Seattle as a Histologist for a little over two years. I am not certified yet and would like to enroll in the Phoenix College Histology Program in Arizona. Does anyone know of any employers near Phoenix that will hire a non-certified tech? I will also work as a Lab Assistant. Thanks! Leah Cox __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 17 02:20:43 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Kemlo Rogerson) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: formalin [Longish] Message-ID: <1030B679AD69D6119C3F00080210DD9D05A3F2CC@bhrv-nt-11.bhrv.nwest.nhs.uk> WOW! Give up, you're the man!!!! -----Original Message----- From: John A. Kiernan [mailto:jkiernan@uwo.ca] Sent: 16 May 2005 19:11 To: Kemlo Rogerson Cc: Glencross Hedley (RW3) Central Manchester & Manchester Children's; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: formalin [Longish] Kemlo, I think your atmospheric pressure ideas would be true only if our atmosphere consisted largely of gaseous formaldehyde, which fortunately is not the case. (Formaldehyde molecules are said to be present in interstellar space; perhaps that's why there isn't any life out there among the fixed stars.) The gas dissolves in water because it's very soluble and also because, as you point out, there is a chemical reaction that forms methylene hydrate. In 37% formaldehyde (neat formalin) 37% of the weight is derived from formaldehyde gas and 63% from water. 40% is the wt/vol expression (There are 40 g of formaldehyde in 100 ml of the 37% wt/wt solution.) Despite the smell, not much formaldehyde escapes into the air. When formalin evaporates, most of the formaldehyde ends up as the solid polymer paraformaldehyde. Evaporation might occur more quickly on top of Mt Everest than at sea level, especially on a sunny day with a brisk wind; I'm sure the YETIS (Yogic Engineering and Technical Institute for Sherpas) up there know this, and fix their specimens in capped jars and vials. It is possible to distil aqueous solutions of formaldehyde, and Walker's book ("Formaldehyde", 3rd ed 1964) gives tabulated data for the composition of distillates obtained under different conditions. Regarding the quotation, "It has been suggested that the hydrated form, methylene glycol, is the reactive component ..." this was certainly once believed (see for example, Walker's book and the first 3 editions of Pearse's Histochemistry). The more recent view is that the methylene glycol constitutes a reserve, always in equilibrium with a very small concentration of dissolved formaldehyde molecules and water. When a formaldehyde molecule is pulled out of the solution (by combining with part of a protein molecule, for example), it is immediately replaced from the reservoir of methylene glycol. I do not know enough chemistry to fully understand the experimental evidence, but it was published in chemical journals, presumably with knowledgeable chemists doing the peer-reviewing. For a recent review of fixation, see D. Hopwood's chapter in Bancroft & Gamble's "Theory and Practice of histological Techniques" (2002). The fixation chapter in the 4th edition of Pearse (Vol 1, 1980) is older but seems still to be up to date. The short review "Formaldehyde fixation" by Fox et al (1985) J Histochem Cytochem 33:845-853 is worth reading because it covers the earliest applications as well as the current understanding of th chemistry and its practical implications. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Kemlo Rogerson wrote: > > "Formaldehyde, as 4% buffered formaldehyde (10% buffered formalin), is the > most widely employed universal fixative particularly for routine paraffin > embedded sections. It is a gas with a very pungent odour, soluble in water > to a maximum extent of 40% by weight and is sold as such under the name of > formaldehyde (40%) or formalin (a colourless liquid). Formaldehyde is also > obtainable in a stable solid form composed of high molecular weight polymers > known as paraformaldehyde. Heated paraformaldehyde generates pure gaseous > formaldehyde which, when dissolved in water, reverts mostly to the monomeric > form. Aqueous formaldehyde exists principally in the form of its > monohydrate, methylene glycol, CH2(OH)2, and as low molecular weight > polymeric hydrates or polyoxymethylene glycols. It has been suggested that > the hydrated form, methylene glycol, is the reactive component of > formaldehyde but this has been disputed2" > > Fixation and fixatives > Anthony S-Y Leong > > So as it is a gas it must be held in solution by atmospheric pressure? If > that atmospheric is raised or lowered then lesser or greater amount of gas > can be held in solution; the same with increasing or decreasing > temperatures. So is formalin 40% formaldehyde gas up Everest or is it much > less? Atmospheric pressure is reduced so the Saturated Vapour Pressure of > the solution may exceed barometric pressure and the gas is driven off until > the pressures equilibrate; plus it is very cold. Do you have to make a > stronger dilution up Everest to end up with a 4% buffered formaldehyde? Are > there any Histo Labs up there? Who cares? > > Kemlo Rogerson > Cellular Pathology Manager > East Lancashire Hospitals NHS Trust > DD. 01254-294162 > Mobile 0774-9754194 > > > -----Original Message----- > From: Glencross Hedley (RW3) CM&MC Manchester > [mailto:Hedley.Glencross@CMMC.nhs.uk] > Sent: 16 May 2005 14:02 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: formalin > > Hi everyone > > It has always been my understanding that "formalin" like "Hoover" & > "Sellotape" (and countless other words) is just a trade name. This > describes a 37% solution of formaldehyde gas in water, and we should be > talking about 3.7% (4%) formaldehyde and not 10% formalin. > > Regards > > Hedley Glencross > > Manchester Cytology Centre UK > > For all my years I have believed that formaldehyde was the 37% > concentrate without buffers, and that formalin was 10% buffered... learn > something new every day! > > Thanks John! > > -----Original Message----- > From: John A. Kiernan [mailto:jkiernan@uwo.ca] > Sent: Friday, May 13, 2005 11:31 AM > To: Weems, Joyce > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] quick question about formaldehyde > > Dear Joyce, > > Formalin = 37% formaldehyde; so no, it doesn't > make a difference. Formalin is not buffered; it > does contain about 10% methanol, which is put > in to retard polymerization. When diluted to > make a 4% formaldehyde fixative, the methanol > concentration is 1%. Buffering of the dilute > solution offsets pH changes due to the > Cannizzaro reaction. It also inhibits the > formation of blood-derived "formalin pigment" > which forms after fixation in an acidic > formaldehyde solution. > > Tim Morken is correct in saying we don't know > the extent of chemical change in 12 year-old > formalin. The fact that there's no expiry date > sugggests that it's not much. For what it's > worth, I've used formalin that's more than 5 > years old and fixation has been OK. > > John Kiernan > london, Canada From jnocito <@t> satx.rr.com Tue May 17 06:26:15 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Indirect Immunofluorescence Message-ID: <01c701c55ad3$3d0eb320$2423f318@yourxhtr8hvc4p> Hello Histo land, My pathologists are interested in starting indirect IF. I thought this was going to be an easy task, however, they tell me that the method they want is using whole serum. (Well just hush my mouth and take away my QIHC). Any ideas? The want to pemphoid? panels and Pemphigus IgA? Heeeeeeeeeeeeeeeellllllllllllllllllllllllllllllllllllpppppppp. As always, thank you for allowing me to tap into the great resource in the world and for your help. As a follow up, I want to thank everyone for their help with the RPMI answers. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX From TillRenee <@t> uams.edu Tue May 17 08:10:26 2005 From: TillRenee <@t> uams.edu (Till, Renee) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] pten Message-ID: Is anyone doing PTEN staining? We have been using a Santa Cruz antibody for a while, and it works, though there has always seemed to be background. We had been counting everything as nuclear, but now they are saying pten can be nuclear or cytoplasmic and we will have to go back and see if it can be differentiated. Has anyone else noticed staining in both areas and had any luck counting them separately? Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 (501) 364-2774 From dsnider <@t> shrinenet.org Tue May 17 10:17:50 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Keeping blocks/slides Message-ID: Hell netters, I would like to know if there is a rule or regulation in any of the governing bodies, concerning the length of time a facility must retain the blocks /slides. I know each facility seems to have different rules for this, but I don't know if there is an actual rule anywhere. Thanks, Deanna Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From BlazekL <@t> childrensdayton.org Tue May 17 10:27:25 2005 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Keeping blocks/slides Message-ID: Deanna,This is the CAP guideline for retention of records, blocks and slides. Hope this helps.Linda ANP.12500 Phase II N/A YES NO Are surgical pathology records and materials retained for an appropriate period? NOTE: Minimum requirements for surgical pathology, providing these are not less stringent than state and federal regulations, are: 1. Accession log records 2 years2. Wet tissue (stock bottle) 2 weeks after final report3. Paraffin blocks 10 years4. Glass slides and reports 10 years The retention period should be extended, when appropriate, to provide documentation for adequate quality control and medical care. There must be a documented policy for protecting and preserving the integrity and retrieval of surgical pathology materials and records. COMMENTARY: N/A REFERENCE: College of American Pathologists. http://www.cap.org/apps/docs/laboratory_accreditation/RetentionGuidelines010405.pdf. Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> "Snider, Deanna" 05/17/2005 11:17 AM >>> Hell netters, I would like to know if there is a rule or regulation in any of the governing bodies, concerning the length of time a facility must retain the blocks /slides. I know each facility seems to have different rules for this, but I don't know if there is an actual rule anywhere. Thanks, Deanna Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From peterntreed <@t> hotmail.com Tue May 17 10:31:10 2005 From: peterntreed <@t> hotmail.com (Peter Reed) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Oocyte (frog) embedding and sectioning Message-ID: I'm fishing for any insights into embedding and sectioning Xenopus oocytes. I have two main problems: Oocytes don't always stay in the block, as if they are not perfused with paraffin; Sections shatter on the blade or on water bath before mounting to slide. Have attempted several strategies: pretreatment with proteinase K to remove membrane, fixing in MEMFA 1h followed by PBS rinses and PBS storage, fixing in MEMFA overnight then proceeding directly to dehydration and embedding, using Paraplast and PolyFin, changing blade angle between 2.5 and 10 degrees. Thanks, Peter Reed Stem Cell Institute University of Minnesota peterntreed@hotmail.com _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From petepath <@t> yahoo.com Tue May 17 11:19:24 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Oocyte (frog) embedding and sectioning Message-ID: <20050517161924.32811.qmail@web30408.mail.mud.yahoo.com> Have you tried frozen sections? Would FS preps satisfy your needs? From azdudley <@t> hotmail.com Tue May 17 11:26:04 2005 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Keeping blocks/slides In-Reply-To: Message-ID: deanna, the cap requires blocks be kept for 10 yrs. and slides for 10 yrs. anita dudley providence hosp mobile al >From: "Snider, Deanna" >To: "'histonet@lists.utsouthwestern.edu'" > >Subject: [Histonet] Keeping blocks/slides >Date: Tue, 17 May 2005 11:17:50 -0400 > >Hell netters, >I would like to know if there is a rule or regulation in any of the >governing bodies, concerning the length of time a facility must retain the >blocks /slides. I know each facility seems to have different rules for >this, but I don't know if there is an actual rule anywhere. >Thanks, >Deanna > >Shriners Hospital for Children >Research Dept. >Cincinnatti, Oh >513-872-6388 > >CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may >contain confidential and privileged information for the use of the >designated recipients. If you are not the intended recipient, (or >authorized to receive for the recipient) you are hereby notified that you >have received this communication in error and that any review, disclosure, >dissemination, distribution or copying of it or its contents is prohibited. >If you have received this communication in error, please destroy all copies >of this communication and any attachments and contact the sender by reply >e-mail or telephone (813) 281-0300. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Tue May 17 12:59:03 2005 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Keeping blocks/slides In-Reply-To: Message-ID: There are federal guidelines, state guidelines and CAP guidelines. It will depend on your accreditation and what state you are in. "Snider, Deanna" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/17/2005 08:17 AM To "'histonet@lists.utsouthwestern.edu'" cc Subject [Histonet] Keeping blocks/slides Hell netters, I would like to know if there is a rule or regulation in any of the governing bodies, concerning the length of time a facility must retain the blocks /slides. I know each facility seems to have different rules for this, but I don't know if there is an actual rule anywhere. Thanks, Deanna Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LuckG <@t> empirehealth.org Tue May 17 13:45:06 2005 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Locate Mary Bryhan Message-ID: Good Morning, I'm trying to locate Mary Bryhan. Mary are you out there? Do any of you know how I might get in touch with Mary? Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org From mcauliff <@t> umdnj.edu Tue May 17 13:53:30 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] (no subject) In-Reply-To: <20050513022253.19733.qmail@web30514.mail.mud.yahoo.com> References: <20050513022253.19733.qmail@web30514.mail.mud.yahoo.com> Message-ID: <428A3DAA.6020900@umdnj.edu> Maybe. I don't think you will find much on the surface of the kidney but the surface of the heart should have plenty. Go to the medical library and get any of several massive tomes on "The Heart". Early on there will be a chapter on anatomy which will include innervation. There will be references to original papers so you can find out who did the work and what method they used. The multi volume "Handbook of Physiology" will also have copious information on the innervation of the heart and references to the methods used. You could try using an antibody to tyrosine hydroxylase to show adrenergic nerves. You could look up a 1975 paper by Herdman and Taylor in Stain Technology, vol 50, page 37. Many years ago a colleague of mine used that method, with considerable modification, to show re-innervation of wounds in mouse skin whole mounts. Leuco-methylene blue is another possible try. Geoff Joseph Ulphani wrote: >Anyone knows the method for staining a whole tissue ( e.g. kidney or heart) for adrenergic nerves on the surface of the tissue? > > > >--------------------------------- >Do you Yahoo!? > Yahoo! Mail - Find what you need with new enhanced search. Learn more. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From pathrm35 <@t> adelphia.net Tue May 17 14:08:55 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] CAP inspection Message-ID: <17262211.1116356936005.JavaMail.root@web1.mail.adelphia.net> Fellow Techs, We recently had a CAP inspection (and passed!) but were cited for GEN 20380 " no graphical tools to communicate quality findings". We thought this was N/A for a histology lab, but apparently not. Does anyone have any ideas on what graphs are needed? Also, we had an MT assist the MD during the inspection. The MT was the one who cited us.Wouldn't it be more appropriate to have an HTor HTL assist with the inspection of a histology lab? Any input is greatly appreciated. Ron Martin From portera203 <@t> yahoo.com Tue May 17 15:29:54 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Oocyte (frog) embedding and sectioning In-Reply-To: 6667 Message-ID: <20050517202954.67641.qmail@web40903.mail.yahoo.com> Peter - we have done goldfish oocytes and they sound very similar to what you are working with. We make sure that they are in a cluster / wrapped in lens paper or biopsy paper / placed between two biopsy sponges. We then process with a routine automated processor overnight program that makes kind of a little flat slab of eggs. Embed using Surgipath paraffin infiltration/embedding system. We trim the blocks as normal. We then place them on cold trays kind of letting the condensation from the warm blocks going onto the cold tray moisten them up slightly. Cut them on rotary microtomes at 4 or 5 microns using low profile dura edge blades with standard angle for dispo blades. They will cut somewhat well at that point. Make sure they don't over soak if you are soaking - that will cause them to fall out of the blocks. They will shred and look like they are going to be horrible but actually turn out pretty good. Put them on adhesive slides, air dry them until they are white, place in a 56-60 oven to dry overnight and routine stain with H&E. Hope this helps out - I know they are difficult and I imagine that the frogs are even smaller than the fish!! Good Luck, Peter Reed wrote:I'm fishing for any insights into embedding and sectioning Xenopus oocytes. I have two main problems: Oocytes don't always stay in the block, as if they are not perfused with paraffin; Sections shatter on the blade or on water bath before mounting to slide. Have attempted several strategies: pretreatment with proteinase K to remove membrane, fixing in MEMFA 1h followed by PBS rinses and PBS storage, fixing in MEMFA overnight then proceeding directly to dehydration and embedding, using Paraplast and PolyFin, changing blade angle between 2.5 and 10 degrees. Thanks, Peter Reed Stem Cell Institute University of Minnesota peterntreed@hotmail.com _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP), QIHC Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu --------------------------------- Do you Yahoo!? Read only the mail you want - Yahoo! Mail SpamGuard. From portera203 <@t> yahoo.com Tue May 17 15:31:27 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Locate Mary Bryhan In-Reply-To: 6667 Message-ID: <20050517203127.7939.qmail@web40907.mail.yahoo.com> Greg - Mary is usually on the listserver. I am sure you will be able to reach her. She is at a hospital in Petoskey, Mi. If she sees your posting she will answer. I used to work with Mary at Ingham Medical Center. "Luck, Greg D." wrote:Good Morning, I'm trying to locate Mary Bryhan. Mary are you out there? Do any of you know how I might get in touch with Mary? Thanks, Greg Greg Luck, BS, HT(ASCP) Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 Phone 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP), QIHC Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu --------------------------------- Discover Yahoo! Use Yahoo! to plan a weekend, have fun online & more. Check it out! From mmcgraw <@t> seattlecca.org Tue May 17 15:36:16 2005 From: mmcgraw <@t> seattlecca.org (McGraw, Medea J) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Looking for reliable methods for H. Pylori using a silver stain t echnique Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B948033A89DF@wala01.seattlecca.org> I am looking for a reliable method(s) used in detecting H. Pylori using any silver technique. I would appreciate your input in the best reliable, reproducible methods. Thanks. Medea J. McGraw , B.S. HT (ASCP) SCCA Pathology Department Desk: 206-288-1358 Main Lab: 206-288-1355 Email: mmcgraw@seattlecca.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From escott8 <@t> houston.rr.com Tue May 17 19:30:07 2005 From: escott8 <@t> houston.rr.com (Edward Scott) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Creating a schedule for employees Message-ID: <001901c55b40$be8021b0$0d817446@thescotts> I have been given a position for another histotech. My medical director wants this person to come in at 4:00 AM. This will increase my staff to 3 full time techs. I will make the 4th tech.(I am not in the schedule all the time) My problem is trying to come up with a schedule that will work with the third person. We process about 8,000 cases per year. She wants at least 1 tray of biopsies out by 8:30 am. I would appreciate if anyone could suggst some possible schedules that they are currently using. I know that person will be responsible for coming in early to embed the tissue and start cutting biopsies. I feel that coming in at 5:00 am would be better. Any help in this matter would be greatly appreciated. Alliosn Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 713-566-5287 From doscwk <@t> nus.edu.sg Tue May 17 22:28:34 2005 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Double fluorescent stains for apoptosis Message-ID: Hi, We=92re doing a study on the effect of compression and degeneration of = rabbit intervertebral discs, and we're using the In Situ Cell Death = Detection Kit, TMR red (Roche) for the detection of apoptosis by = fluorescent microscopy. As our lab is new to this technique, we would = be grateful for any advice on the correct apoptosis stain/kits to use. =20 Our problem in using this kit is that we get a large number of positive = stained cells (all red). As we would like to find out the percentage of = apoptotic cells, we need a protocol which can give us a double stain = which according to one of the papers is bright green (tunnel positive) = and light red (unaffected cells). =20 Any advice and protocols would be greatly appreciated. Thanks in advance Julee Chan Orthopaedic Surgery Histology Lab National University of Singapore=20 --=20 No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.308 / Virus Database: 266.11.12 - Release Date: 5/17/2005 =20 From jkiernan <@t> uwo.ca Tue May 17 23:54:09 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Double fluorescent stains for apoptosis References: Message-ID: <428ACA71.6CE8BF2B@uwo.ca> Dear Chan Wai Kam, Your enquiry clearly indicates that you don't know how your bought kit works. The TUNEL (not tunnel! It's an acronym for Terminal Uridine Nick-End Labelling) family of methods has many shortcomings, all thoroughly documented in peer-reviewed papers that are cited in textbooks. TUNEL, intelligently used, has its place, alongside other techniques that indicate modes of cell death. Your "we need a protocol" plea is a way of saying "Tell me exactly what to do" instead of going to the library and spending half a week immersing yourself in the published literature of cell death and the methods used to recognize how it happens. Are you a clinical resident required to do a research project in a few weeks? This is the impression that I get from your email. The only sensible reply is "Go to the library and tell your boss to go there too". -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Chan Wai Kam wrote: > > Hi, > > We?re doing a study on the effect of compression and degeneration of rabbit intervertebral discs, and we're using the In Situ Cell Death Detection Kit, TMR red (Roche) for the detection of apoptosis by fluorescent microscopy. As our lab is new to this technique, we would be grateful for any advice on the correct apoptosis stain/kits to use. > > Our problem in using this kit is that we get a large number of positive stained cells (all red). As we would like to find out the percentage of apoptotic cells, we need a protocol which can give us a double stain which according to one of the papers is bright green (tunnel positive) and light red (unaffected cells). > > Any advice and protocols would be greatly appreciated. > > Thanks in advance > Julee Chan > Orthopaedic Surgery > Histology Lab > National University of Singapore > From arvind <@t> nbrc.res.in Wed May 18 07:50:48 2005 From: arvind <@t> nbrc.res.in (Arvind Pundir) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] job opportunity Message-ID: R/Sir, i have been working at National Brain Research Centre in INDIA as histo tech for the past 2 yrs can i also apply for the same with regards, Arvind Singh Pundir, NBRC, Manesar, Gurgaon, INDIA-122050 From JNocito <@t> Pathreflab.com Wed May 18 08:08:22 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] CAP inspection In-Reply-To: <17262211.1116356936005.JavaMail.root@web1.mail.adelphia.net> Message-ID: Ron, we were cited for the same thing. Cytology had pictures and graphs, but histo didn't. I just made up something. we perform our on logging in, so I take a sampling of the number of patients logged in to the number of misspelled patients, threw it in an Excel spreadsheet and created a graph (color of course so these inspectors {and I use the term loosely}). Maybe you can track the turn around time of cases, how many special stains or immunos are ordered against how many total cases. I don't like it either, but there is no regulation stating that a histology tech has to inspect a histology lab. One of the many issues I have with CAP. As usual, the opinions of the author in no way reflects the opinions of his employers, their lawyers, grandmothers, distant cousins, etc., etc., etc., Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of pathrm35@adelphia.net Sent: Tuesday, May 17, 2005 2:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CAP inspection Fellow Techs, We recently had a CAP inspection (and passed!) but were cited for GEN 20380 " no graphical tools to communicate quality findings". We thought this was N/A for a histology lab, but apparently not. Does anyone have any ideas on what graphs are needed? Also, we had an MT assist the MD during the inspection. The MT was the one who cited us.Wouldn't it be more appropriate to have an HTor HTL assist with the inspection of a histology lab? Any input is greatly appreciated. Ron Martin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mario <@t> bu.edu Wed May 18 08:15:31 2005 From: mario <@t> bu.edu (Mario Muscedere) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Methacarn Solution Message-ID: Hi All, I've heard a lot about the plusses of using methacarn for IHC, from this board and others. However I tried twice to make up a solution of methacarn (60% absolute MeOH / 30% chloroform / 10% glacial acetic acid), but the result is a milky solution filled with small flecks of what appear to be white precipitate. I'm assuming this isn't normal. Has anyone had any experience with this sort of problem and know what I'm doing wrong? Mario Muscedere??? ? ? Biology Department Boston University 5 Cummington Street Boston, MA, 02215? Office: 409 BRB Office #: 617-353-6977 From kelly.mcqueeney <@t> bms.com Wed May 18 08:20:32 2005 From: kelly.mcqueeney <@t> bms.com (Kelly D Mcqueeney) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Double fluorescent stains for apoptosis In-Reply-To: <428ACA71.6CE8BF2B@uwo.ca> References: <428ACA71.6CE8BF2B@uwo.ca> Message-ID: <428B4120.3060006@bms.com> The reason people subscribe to histonet is to reduce time spent "going to the library and spending half a week immersing yourself in the published literature of cell death and the methods used to recognize how it happens." This is achieved by asking your peers and "experts" for help. You should copy and send this response to everyone asking advice when they can simply take a week and look it up. I know I don't have that kind of time, and it is clear that Chan Wai Kam does not either. Chan Wai Kam: I would help you, my friend, but I do not have experience with the Roche kit. Kelly McQueeney Research Scientist Bristol-Myers Squibb John Kiernan wrote: >Dear Chan Wai Kam, > >Your enquiry clearly indicates that you don't know how >your bought kit works. The TUNEL (not tunnel! It's an >acronym for Terminal Uridine Nick-End Labelling) family >of methods has many shortcomings, all thoroughly >documented in peer-reviewed papers that are cited in >textbooks. TUNEL, intelligently used, has its place, >alongside other techniques that indicate modes of >cell death. > >Your "we need a protocol" plea is a way of saying >"Tell me exactly what to do" instead of going to the >library and spending half a week immersing yourself >in the published literature of cell death and the >methods used to recognize how it happens. > >Are you a clinical resident required to do a research >project in a few weeks? This is the impression that >I get from your email. The only sensible reply is >"Go to the library and tell your boss to go there >too". > > From TJJ <@t> Stowers-Institute.org Wed May 18 08:56:44 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Bringing pH up Message-ID: I've been handed a request to make 1% toluidine blue "buffered to pH 11.6 with sodium carbonate." I always thought sodium carbonate was a neutral pH. Is it possible to bring the pH of this dye solution up with sodium carbonate, and if so, what %, N, or M solution would you recommend using to do this? Interestingly in this procedure, a "1% ammonia" solution is used as a rinse. I tend to think the dye solution is buffered with sodium carbonate and the pH is brought up to 11.6 with ammonium hydroxide, although I have no reference other than the quoted text for clarification. Thanks for any advice you can offer! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Kristopher.Kalleberg <@t> unilever.com Wed May 18 09:01:26 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] distilled water Message-ID: I am looking to get a distilled water sytem in my lab, but the question arose if I need 1Mohm or if 18Mohm is required. I perform routine routine histology experiments, including IHC and immunofluorescence on skin. Does anyone know if the 1 Mohm system would suffice or if I will need to get the 18Mohm system. Any help would be greatly appreciated. Thank you. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381-5765 From bsylinda <@t> aol.com Wed May 18 09:47:23 2005 From: bsylinda <@t> aol.com (bsylinda@aol.com) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] (no subject) Message-ID: <8C729CFA2ADCFF4-848-41DA8@MBLK-M30.sysops.aol.com> Hello Histoland, Does anyone know of a lab that performs Insitu-Hybrid stain to rule out Von Hippel Lindau. Any help will be greatly appreciated. Thanks, Sylinda Battle HT (ASCP) Longview, Tx 903-232-3722 903-232-3839 fax From jkiernan <@t> uwo.ca Wed May 18 09:56:18 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Bringing pH up References: <0IGO008TQUWAJA70@groucho.mail.uwo.ca> Message-ID: <428B5792.A3D1EFA8@uwo.ca> Sodium carbonate is an alkali. (The Merck index gives 11.6 as the pH of an aqueous solution of unspecified strength.) A 1.0M solution of ammonia has pH 11.6; this is equivalent to concentrated ammonium hydroxide diluted to about 16 volumes with water. The pH will be more stable with sodium carbonate because it isn't losing ammonia (gas) or absorbing carbon dioxide from the air. Neither compound constitutes a buffer. Buffers based on carbonate-bicarbonate combinations are effective in the range pH 9.2-11.0, and ammonia/ammonium chloride mixtures are for pH 8.3-10.8 A simple buffer that's effective in the range of pH 11-12 is the Na2HPO4-NaOH system. For pH 11.6 take 50 ml 0.05M Na2HPO4 and add 13.5 ml of 0.1M NaOH (Perrin & Dempsey 1974 Buffers for pH and Metal Ion Control. London: Chapman & Hall Science Paperbacks, p.151) An alkaline solution of toluidine blue does not need to be buffered. Above pH 9 it will stain everything. The higher pH may help the dye penetrate into epoxy-embedded semi-thin sections. Penetration can also be enhanced by exposing the section (before staining) to sodium ethoxide, which is made by dissolving NaOH pellets in 100% ethanol. This "etches" (partly depolymerizes) the resin. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "Johnson, Teri" wrote: > > I've been handed a request to make 1% toluidine blue "buffered to pH > 11.6 with sodium carbonate." I always thought sodium carbonate was a > neutral pH. Is it possible to bring the pH of this dye solution up with > sodium carbonate, and if so, what %, N, or M solution would you > recommend using to do this? > > Interestingly in this procedure, a "1% ammonia" solution is used as a > rinse. I tend to think the dye solution is buffered with sodium > carbonate and the pH is brought up to 11.6 with ammonium hydroxide, > although I have no reference other than the quoted text for > clarification. Thanks for any advice you can offer! > > > Teri Johnson > Managing Director Histology Facility > Stowers Institute for Medical Research > 1000 E. 50th St. > Kansas City, Missouri 64110 > tjj@stowers-institute.org > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mstahl <@t> bvha.org Wed May 18 11:01:30 2005 From: mstahl <@t> bvha.org (Stahl, Michael) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Block disposal Message-ID: <4C878E714B21EB4F8EB159777B8822EE287E8F@bvfyms01.net.bvha.org> Help! Our off site storage facility is overflowing with old paraffin blocks. We know the 10 year limit according to CAP. Our dilemma is how to dispose of them. Correct me us if were wrong, but we have a bio hazard company, and were thinking red back the blocks, and place in the provided bio hazard containers. Any thoughts? Thanks Michael P.Stahl HT(ASCP) Blanchard Valley Regional Health Center Findlay, Ohio 45840 mstahl@bvha.org From Frederick.Fifield <@t> sunhealth.org Wed May 18 12:16:32 2005 From: Frederick.Fifield <@t> sunhealth.org (Frederick.Fifield@sunhealth.org) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: Creating a schedule for employees In-Reply-To: <20050518165558.0FE9CA006F@mx.z108.zixworks.com> Message-ID: Hi Alliosn We have a person that comes in at 4am. This person embeds our bone marrows and biopsies and then starts cutting them. At 5am our second tech arrives and begins to do the bulk of the embedding. Our third tech arrives at 6am and starts cutting. Most mornings we have plenty of work for our pathologists who begin arriving at 7:30 am. Our last tech arrives at 7am and fills in where needed depending on the nature of the workload that day. Most days we have all our slides out by 10-10:30. We do around 17,000 cases/year. Fred Fifield HT (ASCP) Pathology Section Manager Sun Health - Boswell Memorial Hospital (623) 876-5338 I have been given a position for another histotech. My medical director wants this person to come in at 4:00 AM. This will increase my staff to 3 full time techs. I will make the 4th tech.(I am not in the schedule all the time) My problem is trying to come up with a schedule that will work with the third person. We process about 8,000 cases per year. She wants at least 1 tray of biopsies out by 8:30 am. I would appreciate if anyone could suggst some possible schedules that they are currently using. I know that person will be responsible for coming in early to embed the tissue and start cutting biopsies. I feel that coming in at 5:00 am would be better. Any help in this matter would be greatly appreciated. Alliosn Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas 713-566-5287 ******************************************************************************* The information contained in this transmission may be legally privileged and/or confidential information. Any dissemination, distribution or copying of this transmission by anyone other than the intended recipient is strictly prohibited. If you receive this in error, please inform the sender immediately and remove any record of this message. ******************************************************************************* For more information about Sun Health, visit us at: www.sunhealth.org From mprice26 <@t> juno.com Wed May 18 15:11:06 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: software programs for Breast Panels Message-ID: <20050518.131148.7119.63254@webmail26.nyc.untd.com> Hi histonetters, I am interested in finding out all the different software programs for helping to interpret ER,PR,Her/2-Neu, p53 and Ki-67. Are there any other than Chromovision? How many labs are just interpreting using the manual or just eyeballing it method for interpretation? Thank you. Marsha Price From dw18 <@t> uchicago.edu Wed May 18 16:11:19 2005 From: dw18 <@t> uchicago.edu (David A. Wright) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: crud in Methacarn Message-ID: Hi folks Mario, my guess is you've read that Methacarn should be made up freshly and so you're doing it on a small scale in a plastic (polystyrene) tube [e.g. a clear (not translucent) graduated 15ml conical tube] AND you're putting the chloroform in first. Result: plastic dissolved in chloroform which then precipitates when diluted with the other components in which it is not soluble (...or maybe you are storing your chloroform in such plastic, with the same result, or pipeting it with a disposable, polystyrene pipet). Do you see any sign of crazing or etching of the tube/pipet afterwards? The solution could be as simple as adding the chloroform last, but try using polypropylene tubes instead, or better still, a small glass measuring cylinder and/or glass pipet. Cheers! -David =================================================== David A. Wright, PhD Section of Neurosurgery University of Chicago From mario <@t> bu.edu Wed May 18 17:01:13 2005 From: mario <@t> bu.edu (Mario Muscedere) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Methocarn Solution Message-ID: Hello All, I'm not sure if it was using plastic or old chloroform (I was using both) but I tried with fresh chemicals and a glass vial and it worked fine. Thanks for the helpful comments! Mario Muscedere??? ? ? Biology Department Boston University 5 Cummington Street Boston, MA, 02215? Office: 409 BRB Office #: 617-353-6977 From doscwk <@t> nus.edu.sg Wed May 18 20:50:29 2005 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Double fluorescent stains for apoptosis Message-ID: Hi Kelly, Thanks for your understanding. Frankly I did feel chastened and thought = I'd better go and do my homework more thoroughly. It's true that I had = hoped that some kind souls in histoland could offer me their advice on = the correct protocols or kits to use as the kits are so costly and I'm = wandering into unfamiliar territory. =20 I'm not a clinical but lab staff in research and my work has been mostly = on routine histology of decalcified tissues. I've no prior experience = in apoptosis and so find it a bit confusing with all the protocols I've = found on the web. The PI whose project I'm assisting isn't of much help = either as he was on a short attachment with our dept, finished the = experimental surgery part and left the bulk of the work for the lab = staff to complete. He's happily in some other country and would drop an = email every so often expecting results. So frankly I'm unsupervised and = trying to do the job as best as I can. =20 The Roche kit I used (In Situ Cell Death Detection Kit, TMR red for = fluorescent microscopy analysis) comes with a protocol but the results = do not seem accurate as there were too many cells stained positive. I = also realized too late that I should be going for something that shows = up the unaffected cells as well since we need to get a percentage. So = back to the web I went to do more searches but still not any wiser. And = that's when I thought about all those experts out there in histoland who = could perhaps point me in the right direction. =20 Thanks again, Wai Kam =20 -----Original Message----- From: kelly.mcqueeney@bms.com [mailto:kelly.mcqueeney@bms.com]=20 Sent: Wednesday, May 18, 2005 9:21 PM To: John Kiernan Cc: Chan Wai Kam; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Double fluorescent stains for apoptosis The reason people subscribe to histonet is to reduce time spent "going=20 to the library and spending half a week immersing yourself in the=20 published literature of cell death and the methods used to recognize how = it happens." This is achieved by asking your peers and "experts" for=20 help. You should copy and send this response to everyone asking advice=20 when they can simply take a week and look it up. I know I don't have=20 that kind of time, and it is clear that Chan Wai Kam does not either. Chan Wai Kam: I would help you, my friend, but I do not have experience = with the Roche kit. Kelly McQueeney Research Scientist Bristol-Myers Squibb John Kiernan wrote: >Dear Chan Wai Kam, > >Your enquiry clearly indicates that you don't know how >your bought kit works. The TUNEL (not tunnel! It's an >acronym for Terminal Uridine Nick-End Labelling) family=20 >of methods has many shortcomings, all thoroughly=20 >documented in peer-reviewed papers that are cited in >textbooks. TUNEL, intelligently used, has its place,=20 >alongside other techniques that indicate modes of >cell death.=20 > >Your "we need a protocol" plea is a way of saying=20 >"Tell me exactly what to do" instead of going to the >library and spending half a week immersing yourself >in the published literature of cell death and the >methods used to recognize how it happens.=20 > >Are you a clinical resident required to do a research >project in a few weeks? This is the impression that >I get from your email. The only sensible reply is >"Go to the library and tell your boss to go there=20 >too". > =20 > --=20 No virus found in this incoming message. Checked by AVG Anti-Virus. Version: 7.0.308 / Virus Database: 266.11.12 - Release Date: 5/17/2005 =20 --=20 No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.322 / Virus Database: 266.11.12 - Release Date: 5/17/2005 =20 From onep00 <@t> hotmail.com Wed May 18 21:15:06 2005 From: onep00 <@t> hotmail.com (Pam O'Neill) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] choosing a good cryostat Message-ID: Hello Histonetters- We have been approved for a new cryostat. Anyone have recommendations? We are a hospital lab and use our cryostat for frozen sections and kidney biopsies(at 4 microns). The boss is also concerned about decontamination because of new CAP requirements. Any suggestions are welcome. Pam O'Neill , Toledo, Ohio _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From jorge.tornero <@t> gmail.com Thu May 19 06:05:04 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Plastic sectioning basics Message-ID: <8c964a7905051904057f5237c2@mail.gmail.com> Hello, I am a newby in the histology business and working with anchovy gonads in the daily egg production method. We have to cut the gonads at 3 micron thick, so we use technovit 7100 resin for the blocks. We have a leica rm2145 microtome equipped with a tungsten-carbide knife. Until now, all my little knowledge about histotechnique has been adquired with the experience and with the old trial-and-error method, but I would like to ask you about severals aspects of the work. What's the correct angle for cutting this kind of plastic? How should be the maximum witdh of the block to be cutted? What's the "life expectance" of the edge of a tungsten-carbide knife when cutting resin? I think I've read something about lubricating the cut with water to ease the cut, but I have experienced that even when in some cases I have got some moisture over the knife, the contact of the slice with the moisture ruins it. But also I have experienced that a little breathing over the block just before the cut eases it and prevents from rolling. I would like to know about good books in spanish or in english about these particulars, but unhappily i do not have good access to libraries and this stuff, but anyway i'd appreciate comments about it. I have a lot more of questions to ask you, but i think it is enough now. Thank you very much and excuses for my bad english Jorge Tornero IEO - C?diz From kmerriam2003 <@t> yahoo.com Thu May 19 08:22:46 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Dendritic Cells in Intestine Message-ID: <20050519132247.94189.qmail@web50304.mail.yahoo.com> Hello everyone, Does anyone know of a special stain that will identify dendritic cells in the intestine? Kim Merriam Novartis Cambridge, MA __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From gcallis <@t> montana.edu Thu May 19 10:10:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: Dendritic cells intestine Message-ID: <6.0.0.22.1.20050519090824.01b5ab88@gemini.msu.montana.edu> Kim, Are you wanting to see murine dendritic cells? The best way is immunohistochemistry, and there are several monoclonal antibodies available. I am not sure they all work with FFPE, we only do frozen sections. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Thu May 19 10:32:27 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: GMA resin i.e. Technovits 7100 Message-ID: <6.0.0.22.1.20050519091105.01b62978@gemini.msu.montana.edu> You wrote: Hello, I am a newby in the histology business and working with anchovy gonads in the daily egg production method. We have to cut the gonads at 3 micron thick, so we use technovit 7100 resin for the blocks. We have a leica rm2145 microtome equipped with a tungsten-carbide knife. Until now, all my little knowledge about histotechnique has been adquired with the experience and with the old trial-and-error method, but I would like to ask you about severals aspects of the work. What's the correct angle for cutting this kind of plastic? How should be the maximum witdh of the block to be cutted? What's the "life expectance" of the edge of a tungsten-carbide knife when cutting resin? I think I've read something about lubricating the cut with water to ease the cut, but I have experienced that even when in some cases I have got some moisture over the knife, the contact of the slice with the moisture ruins it. But also I have experienced that a little breathing over the block just before the cut eases it and prevents from rolling. I would like to know about good books in spanish or in english about these particulars, but unhappily i do not have good access to libraries and this stuff, but anyway i'd appreciate comments about it. I have a lot more of questions to ask you, but i think it is enough now. Thank you very much and excuses for my bad english Jorge Tornero IEO - C?diz Technovits 7100 is glycol methacrylate, and it will not be necessary to wet the block face with anything. In general, this plastic is soft enough to be sectioned with a tungsten carbide blade, although Linda Jenkins has great success with disposable microtome blades. I am not sure what brand she uses. The plastic that requires wetting the block face for sectioning is methyl methacrylate (MMA) a much harder, more hydrophobic plastic than GMA. If you get the GMA block face too wet with water, it may make the sectioning more difficult, causing section to bunch up in a gooey mess. Some people have terrible problems with too much humidity when sectioning GMA, so just section without any wetting. Breathing on block face IS providing warm moist air. People section GMA with a tungsten carbide blade, but it must be very sharp, as with any blade. Have you reconditioned recently? If you are cutting very thick sections, it may contribute to rolling. We never cut GMA any thicker than 2 to 3um . It may be knife angle and sharpness of blade that contributes to rolling. I have heard of some people reversing a d profile tungsten carbide blade so you do not cut with largest angle facing you, rather the smaller angle on what is considered the back of the knife. There are also c profile tungsten carbide blades made, from DDK (Delaware Diamond Knives. We sometimes captured the section with forceps as it starts to come over blade, to insure it does not roll or do funky flyaway things. We rarely had what you experience, but we preferred glass triangular knives, freshly broken and extremely sharp for all our GMA sectioning, with a new knife for every block. From pruegg <@t> ihctech.net Thu May 19 11:15:46 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Removing dab Message-ID: <200505191615.j4JGFhwm025322@chip.viawest.net> I remember someone mentioning that DAB can be removed from a section when that is the only section one has to work with, how is that done again? Patsy From TJJ <@t> Stowers-Institute.org Thu May 19 12:10:49 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Bringing pH up Message-ID: John wrote: "Sodium carbonate is an alkali. (The Merck index gives 11.6 as the pH of an aqueous solution of unspecified strength.)" Interestingly, a 1M solution of this gave me a pH of 11.0. Increasing the concentration brought the pH down. 0.1M gave me a pH of 11.2, 0.05M held steady at pH 11.2. That was the extent of my testing as 11.2 was an acceptable number to the researcher. Thanks for your help, John! Teri From Melissa.Gonzalez <@t> cellgenesys.com Thu May 19 12:39:06 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Fluorescent Apoptosis Detection Message-ID: Hi Wai Kam My experience with the Roche kit has been pretty unsuccessful. Only because some lots work great and others don't. The reason you are seeing so many positive cells is that they manufacture the labeling enzyme pretty strong, and depending on the application/tissue, it can end up labeling all of your cells, instead of just some of them. Usually when I get a new kit, I dilute the TUNEL reaction mixture anywhere between 1:5-1:20 using their TUNEL dilution buffer. Use DNase as a positive control which each dilution. Regardless of the dilution, your positive should label ~100%. If it is less than 80% positive, you have diluted the reaction mixture too much. If you counterstain with DAPI, all your non-apoptic cells will have a blue nucleus. If you use the TMR kit, your apoptotic cells will have a red nucleus, and if you use the FITC kit, the positive cells will be green. Because of this hassle I now use an anti-Caspase 3 antibody which works great for detecting apoptosis. (from R&D systems) Good luck, Melissa Message: 5 Date: Thu, 19 May 2005 09:50:29 +0800 From: "Chan Wai Kam" Subject: RE: [Histonet] Double fluorescent stains for apoptosis To: Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="windows-1250" Hi Kelly, Thanks for your understanding. Frankly I did feel chastened and thought I'd better go and do my homework more thoroughly. It's true that I had hoped that some kind souls in histoland could offer me their advice on the correct protocols or kits to use as the kits are so costly and I'm wandering into unfamiliar territory. I'm not a clinical but lab staff in research and my work has been mostly on routine histology of decalcified tissues. I've no prior experience in apoptosis and so find it a bit confusing with all the protocols I've found on the web. The PI whose project I'm assisting isn't of much help either as he was on a short attachment with our dept, finished the experimental surgery part and left the bulk of the work for the lab staff to complete. He's happily in some other country and would drop an email every so often expecting results. So frankly I'm unsupervised and trying to do the job as best as I can. The Roche kit I used (In Situ Cell Death Detection Kit, TMR red for fluorescent microscopy analysis) comes with a protocol but the results do not seem accurate as there were too many cells stained positive. I also realized too late that I should be going for something that shows up the unaffected cells as well since we need to get a percentage. So back to the web I went to do more searches but still not any wiser. And that's when I thought about all those experts out there in histoland who could perhaps point me in the right direction. Thanks again, Wai Kam From jlinda <@t> ces.clemson.edu Thu May 19 13:01:37 2005 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: Plastic sectioning basics Message-ID: <5.2.1.1.2.20050519132045.05c70488@mailhost.ces.clemson.edu> Dear Jorge, You know, I never did care to eat anchovies and, apparently, they don't section well either! As Gayle said in her message, I do section GMA with disposable blades. I use a variety of high profile, stainless steel disposable blades...they all work well. I have a Leica 2155 and set the angle between 0 - 5. Is the 2145 a manual or motorized microtome? Hopefully, it is motorized as this makes sectioning GMA a lot easier. Our labs in South Carolina are very humid and we leave our GMA blocks under vacuum with Drierite (brandname dessicant) until sectioning each block. Unfortunately, there are no classical books devoted to plastic sectioning per se. You can get bits and pieces from some books. Try, "Handbook of Histology Methods for Bone and Cartilage" by An & Martin, Humana Press, 2003. Best wishes, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From BDUE <@t> PARTNERS.ORG Thu May 19 13:58:39 2005 From: BDUE <@t> PARTNERS.ORG (Due, Brice) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Removing dab Message-ID: <59C772E2D8EDF345AE1601F5B60B3CB5027A23@PHSXMB7.partners.org> Hello Patsy, because I was intrigued, I did the search for you at www.histosearch.com and from John Kiernan comes the following answer... Nothing will bemove the brown DAB product. You could rinse the sections in acid (e.g. about pH 2) to remove all bound antibodies, then immunostain again for the correct antigen, using a differently coloured detection system, such as AEC for peroxidase (red) or an alkaline phosphatase-based ABC or APAP kit (commonly purple or red). Nickel- or cobalt-enhanced DAB (blue-black) might be OK. John A. Kiernan Dept of Anatomy and Cell Biology University of Western Ontario LONDON, Canada. (kiernan@uwo.ca) DOH! -brice Brigham & Women's Hospital Boston -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Thursday, May 19, 2005 12:16 PM To: 'Histonet' Subject: [Histonet] Removing dab I remember someone mentioning that DAB can be removed from a section when that is the only section one has to work with, how is that done again? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Thu May 19 14:45:36 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Block disposal Message-ID: <5D2189E74151CC42BEC02906BA8996322B90B4@exchsrv01.barrynet.barry.edu> Why don't you give them to the Cooperative Human Tissue Network (3400 Spruce St., Philadelphia, PA 19104)? Academic research is starving for both normal and diseased human tissue. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stahl, Michael Sent: Wednesday, May 18, 2005 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Block disposal Help! Our off site storage facility is overflowing with old paraffin blocks. We know the 10 year limit according to CAP. Our dilemma is how to dispose of them. Correct me us if were wrong, but we have a bio hazard company, and were thinking red back the blocks, and place in the provided bio hazard containers. Any thoughts? Thanks Michael P.Stahl HT(ASCP) Blanchard Valley Regional Health Center Findlay, Ohio 45840 mstahl@bvha.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From brett_connolly <@t> merck.com Thu May 19 14:54:15 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Block disposal Message-ID: That's a great idea, Allen!...provided that appropriate patient consent forms were obtained. NDRI, National Disease Research Interchange is another option (www.ndri.com) Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith, Allen Sent: Thursday, May 19, 2005 3:46 PM To: Stahl, Michael Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Block disposal Why don't you give them to the Cooperative Human Tissue Network (3400 Spruce St., Philadelphia, PA 19104)? Academic research is starving for both normal and diseased human tissue. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stahl, Michael Sent: Wednesday, May 18, 2005 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Block disposal Help! Our off site storage facility is overflowing with old paraffin blocks. We know the 10 year limit according to CAP. Our dilemma is how to dispose of them. Correct me us if were wrong, but we have a bio hazard company, and were thinking red back the blocks, and place in the provided bio hazard containers. Any thoughts? Thanks Michael P.Stahl HT(ASCP) Blanchard Valley Regional Health Center Findlay, Ohio 45840 mstahl@bvha.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From KPercival <@t> wyeth.com Thu May 19 15:10:51 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Block disposal Message-ID: Allen, Michael, Better check with legal services. I think you have to get permission to give away those blocks even though they are ready for disposal. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> "Smith, Allen" 5/19/2005 3:45:36 PM >>> Why don't you give them to the Cooperative Human Tissue Network (3400 Spruce St., Philadelphia, PA 19104)? Academic research is starving for both normal and diseased human tissue. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stahl, Michael Sent: Wednesday, May 18, 2005 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Block disposal Help! Our off site storage facility is overflowing with old paraffin blocks. We know the 10 year limit according to CAP. Our dilemma is how to dispose of them. Correct me us if were wrong, but we have a bio hazard company, and were thinking red back the blocks, and place in the provided bio hazard containers. Any thoughts? Thanks Michael P.Stahl HT(ASCP) Blanchard Valley Regional Health Center Findlay, Ohio 45840 mstahl@bvha.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From swaram <@t> myrealbox.com Thu May 19 16:48:19 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Double fluorescent stains for apoptosis In-Reply-To: References: Message-ID: <428D09A3.2080904@myrealbox.com> Dear Chan, I wont try to evangelize or advise you on the benefits of using the library. But here is something you can do for TUNEL Roche Kit. I use one with fluorescein. But if you read the product inser that came with it, you will find something called TUNEL Dilution buffer. Use that to dilute your enzyme mixture in serial dilutions of 1:10 to 1:100 and so on. You will see that the background gradually fades away. Standardize it for your satisfaction. Also make sure that you have a good Block for the slides. Otherwise you will find the Fluorescein all over the place. http://www.roche-applied-science.com/pack-insert/1966006a.pdf Should work...! Best Wishes, Swaram And here is a previous reply which might be slightly more informative on the same subject Dear Toshi, TUNEL is not a waste of time, just don't use the Roche kit! I had the same problems with it: lots of normal looking nuclei staining positive. Now I am using the Trevigen TACS kit (in Canada sold through BIO/CAN)and it works great. You can buy individual components, no need to get the whole kit each time you run out of a component. I buy the TdT enzyme, biotinylated dNTP nucleotides, Co 2+ cations, Proteinase K, TdT Labeling buffer and TdT Stop buffer. I follow their protocol up to the Stop buffer step, then I vizualize the incorporated biotinylated nucleotides either with my usual IHC Streptavidin-peroxidase, Nova-Red incubations, or for fluorescence, with Streptavidin-Alexa from Molecular Probes. The staining is very clean with nuclei of apoptotic morphology staining positive. You can check my double immunofluorescence for TUNEL and cleaved caspase-3 in mouse thymus in our recent paper: Circulation 2005;111:1814-1821, Supplemental on-line Figure 5. TUNEL specifically stained nuclei in cells that had cytoplasm positive for cleaved caspase-3. Sarka Lhotak McMaster University, Hamilton, Ontario \Chan Wai Kam wrote: >Hi, > >We?re doing a study on the effect of compression and degeneration of rabbit intervertebral discs, and we're using the In Situ Cell Death Detection Kit, TMR red (Roche) for the detection of apoptosis by fluorescent microscopy. As our lab is new to this technique, we would be grateful for any advice on the correct apoptosis stain/kits to use. > >Our problem in using this kit is that we get a large number of positive stained cells (all red). As we would like to find out the percentage of apoptotic cells, we need a protocol which can give us a double stain which according to one of the papers is bright green (tunnel positive) and light red (unaffected cells). > >Any advice and protocols would be greatly appreciated. > >Thanks in advance >Julee Chan >Orthopaedic Surgery >Histology Lab >National University of Singapore > > > From swaram <@t> myrealbox.com Thu May 19 16:56:47 2005 From: swaram <@t> myrealbox.com (Swaram) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Double fluorescent stains for apoptosis In-Reply-To: References: Message-ID: <428D0B9F.1070700@myrealbox.com> Dear Chan, I wont try to evangelize or advise you on the benefits of using the library. But here is something you can do for TUNEL Roche Kit. I use one with fluorescein. But if you read the product inser that came with it, you will find something called TUNEL Dilution buffer. Use that to dilute your enzyme mixture in serial dilutions of 1:10 to 1:100 and so on. You will see that the background gradually fades away. Standardize it for your satisfaction. Also make sure that you have a good Block for the slides. Otherwise you will find the Fluorescein all over the place. http://www.roche-applied-science.com/pack-insert/1966006a.pdf Should work...! Best Wishes, Swaram And here is a previous reply which might be slightly more informative on the same subject Dear Toshi, TUNEL is not a waste of time, just don't use the Roche kit! I had the same problems with it: lots of normal looking nuclei staining positive. Now I am using the Trevigen TACS kit (in Canada sold through BIO/CAN)and it works great. You can buy individual components, no need to get the whole kit each time you run out of a component. I buy the TdT enzyme, biotinylated dNTP nucleotides, Co 2+ cations, Proteinase K, TdT Labeling buffer and TdT Stop buffer. I follow their protocol up to the Stop buffer step, then I vizualize the incorporated biotinylated nucleotides either with my usual IHC Streptavidin-peroxidase, Nova-Red incubations, or for fluorescence, with Streptavidin-Alexa from Molecular Probes. The staining is very clean with nuclei of apoptotic morphology staining positive. You can check my double immunofluorescence for TUNEL and cleaved caspase-3 in mouse thymus in our recent paper: Circulation 2005;111:1814-1821, Supplemental on-line Figure 5. TUNEL specifically stained nuclei in cells that had cytoplasm positive for cleaved caspase-3. Sarka Lhotak McMaster University, Hamilton, Ontario \Chan Wai Kam wrote: >Hi, > >We?re doing a study on the effect of compression and degeneration of rabbit intervertebral discs, and we're using the In Situ Cell Death Detection Kit, TMR red (Roche) for the detection of apoptosis by fluorescent microscopy. As our lab is new to this technique, we would be grateful for any advice on the correct apoptosis stain/kits to use. > >Our problem in using this kit is that we get a large number of positive stained cells (all red). As we would like to find out the percentage of apoptotic cells, we need a protocol which can give us a double stain which according to one of the papers is bright green (tunnel positive) and light red (unaffected cells). > >Any advice and protocols would be greatly appreciated. > >Thanks in advance >Julee Chan >Orthopaedic Surgery >Histology Lab >National University of Singapore > > > From marjoh3 <@t> telus.net Thu May 19 17:16:13 2005 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Swine Influenza by Immunohistochemistry Message-ID: <001a01c55cc0$5e5126a0$6401a8c0@VALUED20606295> Hi Histonetters, Our IHC Lab. would like to initiate the demonstration of Swine Influenza. If there are any Labs. currently staining for Swine Influenza, would you please include the detection system used and the source of primary antisera, including the dilution? Source of positive control blocks/slides would also be appreciated. Thankyou in advance. Marilyn Johnson Alberta Agriculture Agri-Food Systems Branch Edmonton, Alberta, Canada. From t-sherman <@t> comcast.net Thu May 19 22:07:55 2005 From: t-sherman <@t> comcast.net (Todd Sherman) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: Double fluorescent stains for apoptosis (Chan Wai Kam) Message-ID: Hello Julee, Since you are using a kit and someone selected it for TUNEL, maybe you could consult them first. Or, if that source is no longer available, try the vendor. They will have a technical staff to help you troubleshoot. I've done TUNEL staining but not with the kit you mention and not recently enough to provide dependable guidance. The vendor, or maybe manufacturer is more precise, earn their higher kit fees for such services so certainly query them for the subtleties of their product. Regards, Todd Todd Sherman President HistoSoft Corporation >>>>>>>> www.histosoft.com <<<<<<<< <<<<<<<< Biology In A New Form (c)>>>>>>>> On Wed, 18 May 2005 12:00:26 -0500, wrote: > > Today's Topics: > > 9. Double fluorescent stains for apoptosis (Chan Wai Kam) > 10. Re: Double fluorescent stains for apoptosis (John Kiernan) > > ---------------------------------------------------------------------- > Message: 9 Date: Wed, 18 May 2005 11:28:34 +0800 From: "Chan Wai Kam" Subject: [Histonet] Double fluorescent stains for apoptosis To: Message-ID: Content-Type: text/plain;charset="windows-1250" Hi, We?re doing a study on the effect of compression and degeneration of rabbit intervertebral discs, and we're using the In Situ Cell Death Detection Kit, TMR red (Roche) for the detection of apoptosis by fluorescent microscopy. As our lab is new to this technique, we would be grateful for any advice on the correct apoptosis stain/kits to use. Our problem in using this kit is that we get a large number of positive stained cells (all red). As we would like to find out the percentage of apoptotic cells, we need a protocol which can give us a double stain which according to one of the papers is bright green (tunnel positive) and light red (unaffected cells). Any advice and protocols would be greatly appreciated. Thanks in advance Julee Chan Orthopaedic Surgery Histology Lab National University of Singapore ------------------------------ Message: 10 Date: Wed, 18 May 2005 00:54:09 -0400 From: John Kiernan Subject: Re: [Histonet] Double fluorescent stains for apoptosis To: Chan Wai Kam Cc: histonet@lists.utsouthwestern.edu Message-ID: <428ACA71.6CE8BF2B@uwo.ca> Content-Type: text/plain; charset=iso-8859-1 Dear Chan Wai Kam, Your enquiry clearly indicates that you don't know how your bought kit works. The TUNEL (not tunnel! It's an acronym for Terminal Uridine Nick-End Labelling) family of methods has many shortcomings, all thoroughly documented in peer-reviewed papers that are cited in textbooks. TUNEL, intelligently used, has its place, alongside other techniques that indicate modes of cell death. Your "we need a protocol" plea is a way of saying "Tell me exactly what to do" instead of going to the library and spending half a week immersing yourself in the published literature of cell death and the methods used to recognize how it happens. Are you a clinical resident required to do a research project in a few weeks? This is the impression that I get from your email. The only sensible reply is "Go to the library and tell your boss to go there too". ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm ------------------------------ From barbara.bublava <@t> meduniwien.ac.at Fri May 20 00:56:54 2005 From: barbara.bublava <@t> meduniwien.ac.at (Barbara Bublava) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] PTAH - stability Message-ID: <001301c55d00$b9cb78b0$dd01a8c0@GERICHTS9XOZZ8> Dear Histonetters, I know lifetime is shorter, when chemically ripend: But how long will PTAH solution be stable if ripened with potassium permanganate. Days, Weeks, several Months - instead of years? Thanks for your help Barbara From JWEEMS <@t> sjha.org Fri May 20 06:30:02 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: Touch Prep CPT Code Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45D67@sjhaexc02.sjha.org> Are you all ignoring me? I will not go away! :>) Resending as I did not get a response. Happy weekend! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: Weems, Joyce Sent: Thursday, May 12, 2005 9:38 AM To: 'Histonet' Subject: Touch Prep CPT Code Do you guys charge for touch preps? If so, what CPT code? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. -------------- next part -------------- Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph?s Health System, Inc. From jerry.lyons <@t> ucd.ie Fri May 20 06:48:16 2005 From: jerry.lyons <@t> ucd.ie (Jeremiah Lyons) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] ELISA background Message-ID: <001b01c55d31$cfa0a6c0$c8882b89@DFFRCL0J> Hi all! I could do with some advice as regards eliminating background from my ELISA. I am detecting anti-Flk-1 antibody in mouse serum from mice vaccinated against this molecule. I know there is an immune response there but it is quite low, at best around 1:800. I am coating my plates with whole cell lysate which contains the flk-1 protein. I know the antigen is there as my positive control works well. I block for 2 hours using skimmilk 10%. I wash 5 times with a 90 second pause between each wash. My wash solution is PBS plus tween-20. I dilute my sera samples in skim milk 10% and leave at 4 degrees overnight. My secondary antibody is a polyclonal rabbit antimouse-HRP. My signal is positive but is quite weak, and it needs to be double the control absorbance to be conclusively positive. At the moment it is about 1.75 times the control, but the background on my control is quite high. Can anyone offer any suggestions? Thanking you in advance! From kmerriam2003 <@t> yahoo.com Fri May 20 06:48:51 2005 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Dendritic Cells in Intestine In-Reply-To: Message-ID: <20050520114851.40001.qmail@web50307.mail.yahoo.com> Julia, Gayle, I am not sure exactly what the researcher is looking for, she just asked me if I knew of a stain that would specifically identify these cells (FFPE mouse tissue). I do have a DVM-pathologist here that will be reading the slides. I had hoped there was some type of special stain that I could do (rather than going about antibody validation for just one slide). The researcher will need to check into this. It looks like we will probably need to do IHC on this one. Thanks for your input. Kim Julia Dahl wrote: Kim - There are two main populations of dendritic cells in the intestine - dendritic cells of macrophage lineage and dendritic cells of myofibroblast lineage. These populations serve some overlapping, but some distinct functions. Which types of cells/functions are you wanting to identify? That will specify the type of antibody that you will want to use. Do you have a pathologist on staff that understands the difference between these cells? If not, I can provide a more in depth consultation than the above. Julia Dahl, M.D. Mosaic Gastrointestinal Research Consortium Author: COLON in Histology for Pathologists, 3rd Edition; In press >From: Kim Merriam >To: Histonet >Subject: [Histonet] Dendritic Cells in Intestine >Date: Thu, 19 May 2005 06:22:46 -0700 (PDT) >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by mc2-f19.hotmail.com >with Microsoft SMTPSVC(6.0.3790.211); Thu, 19 May 2005 06:22:11 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1DYkzS-0005JK-DA; Thu, 19 May >2005 08:23:11 -0500 >Received: from [199.165.152.167] (helo=swlx167.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1DYkzA-0005JD-7Ofor >histonet@lists.utsouthwestern.edu; Thu, 19 May 2005 08:22:53 -0500 >Received: from [206.190.38.58] (helo=web50304.mail.yahoo.com)by >swlx167.swmed.edu with smtp (Exim 4.44) id 1DYkz9-000221-Shfor >histonet@lists.utsouthwestern.edu; Thu, 19 May 2005 08:22:48 -0500 >Received: (qmail 94191 invoked by uid 60001); 19 May 2005 13:22:47 -0000 >Received: from [205.181.102.120] by web50304.mail.yahoo.com via HTTP;Thu, >19 May 2005 06:22:46 PDT >X-Message-Info: x1iTvSDExI49dH/6Rnbrwl8LZ6pePfURD58c4AKmgCM= >Comment: DomainKeys? See http://antispam.yahoo.com/domainkeys >DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; s=s1024; >d=yahoo.com;b=JIAQnnQicQfnve4qSSqg9CBLZIlSs6ZTTreXgbVj24mRthlScuPG6wlU2zzfxlQ5Vkv1aSqnCzudPKCWUBelw8BoR8uTnlomtKIkWWxMGoZ0XBVMT+zSH2sH0XjEBJw0QmqpBN+kIn0EcVP5Sgcj0cA6yMHooIZG0zyCe9eUMKQ=; >X-Scan-Signature: 7bf0d7e947a05afa907d3090d45e1f8d >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 6e3e8a9b2a8472a697c9c9c003db8af6 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: *** >X-Spam-Status: No, hits=3.2 required=5.5 >tests=FROM_ENDS_IN_NUMS,RCVD_IN_DSBL,RCVD_IN_NJABL_PROXY autolearn=no >version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 19 May 2005 13:22:11.0345 (UTC) >FILETIME=[C3A86810:01C55C75] > >Hello everyone, > >Does anyone know of a special stain that will identify dendritic cells in >the intestine? > > > > >Kim Merriam >Novartis >Cambridge, MA >__________________________________________________ >Do You Yahoo!? >Tired of spam? Yahoo! Mail has the best spam protection around >http://mail.yahoo.com >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Kim Merriam Novartis Cambridge, MA --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From jorge.tornero <@t> gmail.com Fri May 20 07:33:34 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: Plastic sectioning basics In-Reply-To: <5.2.1.1.2.20050519132045.05c70488@mailhost.ces.clemson.edu> References: <5.2.1.1.2.20050519132045.05c70488@mailhost.ces.clemson.edu> Message-ID: <8c964a790505200533847ca8d@mail.gmail.com> Dear Linda, I have to thank you for your information. My microtome is not motorized, and I think i have a problem closer to yours, inCadiz we have a lot of humidity and I suspected a little about the fact that moisture could have influence in cutting, but also i thought than temperature could be another important fact, because it was easier for me to cut the block in winter than in summer. Also, i have thought that maybe the blocks need to "cure" a long time to be good to cut. Also I noticed that the harder the block, the easier to cut it. I tried also to try to freeze a block but in very high humidity conditions is not a good idea (condensation). I will try to dry the blocks more and see what happens. Thank you. 2005/5/19, Linda Jenkins : > Dear Jorge, > You know, I never did care to eat anchovies and, apparently, they > don't section well either! As Gayle said in her message, I do > section GMA with disposable blades. I use a variety of high profile, > stainless steel disposable blades...they all work well. I have a Leica > 2155 and set the angle between 0 - 5. Is the 2145 a manual or motorized > microtome? Hopefully, it is motorized as this makes sectioning GMA a lot > easier. Our labs in South Carolina are very humid and we leave our GMA > blocks under vacuum with Drierite (brandname dessicant) until sectioning > each block. > Unfortunately, there are no classical books devoted to plastic > sectioning per se. You can get bits and pieces from some books. Try, > "Handbook of Histology Methods for Bone and Cartilage" by An & Martin, > Humana Press, 2003. > Best wishes, > Linda > > > Linda Jenkins, HT > Clemson University > Dept. of Bioengineering > Clemson, SC 29634-0905 > 864.656.5553 > http://www.ces.clemson.edu/bio/research/histo.htm > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jorge.tornero <@t> gmail.com Fri May 20 07:51:46 2005 From: jorge.tornero <@t> gmail.com (Jorge Tornero) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Plastic sectioning basics In-Reply-To: <1116512467.428ca0d3ccba2@imp.vet.upenn.edu> References: <8c964a7905051904057f5237c2@mail.gmail.com> <1116512467.428ca0d3ccba2@imp.vet.upenn.edu> Message-ID: <8c964a7905052005512f0d3edd@mail.gmail.com> Dear Pamela and all, I thought that the "dance" (wich has happened to me a very few times) was caused by the turbulence inside the container of water, very small but enough to move such a small thing like a cut. I have other question concerning to the maximum width of the bolck to be cut. And also, wich molds do you use for your blocks. Actually we use molding cup trays from ems (ref. 70176-10 6x12x5mm) but we'll try to use other, the same kind but 13x19x5mm. I think it would be too wide and the cutting will we bad but I'll appreciate your opinions. Do someone have experience staining GMA with H&E? I have experienced some problems, and a big one is coming, because we are going to begin to use an autostainer XL from leica to do so. if someone has experience on it, please tell. Thank you very much. Jorge Tornero IEO - Cadiz Spain 2005/5/19, Pamela A. Marcum : > Hi Jorge, > > There really is no universal angle for plastics. The block size and type of > microtome and plastic are all different. I have always experimented until it > felt right and the sections were coming off well. As for water - the product > you are using is a gylcol methacrylate base and will react with water. If you > take a section off the knife with forceps and place it on a water bath or > container of water most of the time it will literally dance around the surface. > I have always picked GMA up from a water bath or container to allow this to > happen. If I placed on a drop of water on a slide it would go the edge and > just crumple up. You can add alcohol to the water bath or container at 50 ot > 60% and it will help stretch them as they dance. Plastics are very different > from paraffin and do not ribbon well as it is genreally one section at a time > with patience. > > The trick with water is used with methyl methacrylate MMA not GMA. If I can > assist your further or you have any questions please write me and I will > attempt to help you in any way I can. > > Best Regards, > From Rick.Couture <@t> vision-bio.com Fri May 20 07:54:04 2005 From: Rick.Couture <@t> vision-bio.com (Rick Couture) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] unsubscribe Message-ID: <000501c55d3b$007294d0$090318ac@visionusa.com.visionusa.com> Rick Couture Technical Service Manager Vision Biosystems 700 Longwater Drive Norwell Ma. 02061 781-616-1135 Rick.Couture@vision-bio.com From TMcNemar <@t> lmhealth.org Fri May 20 07:52:07 2005 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: Touch Prep CPT Code Message-ID: <6CD94D97ED7D924BA5C2B588FA9528213967C2@nt_exchange.lmhealth.org> We charge for touch preps and use code 88161. Tom Mc Nemar HT(ASCP) Histology Supervisor Licking Memorial Hospital Newark, Ohio 43055 -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Friday, May 20, 2005 7:30 AM To: Weems, Joyce; Histonet Subject: [Histonet] RE: Touch Prep CPT Code Are you all ignoring me? I will not go away! :>) Resending as I did not get a response. Happy weekend! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: Weems, Joyce Sent: Thursday, May 12, 2005 9:38 AM To: 'Histonet' Subject: Touch Prep CPT Code Do you guys charge for touch preps? If so, what CPT code? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From akbitting <@t> geisinger.edu Fri May 20 08:21:32 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] GRAC-TP gravity recycler Message-ID: Does anyone out there on Histonet use this recycler? What type of testimony can I get about it? Thanks. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From DOOLEEO <@t> shands.ufl.edu Fri May 20 09:27:11 2005 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Hansen stain Message-ID: Dear Histonetters, I am looking for some information on a Hansen stain for eosinophils. I have had several Doctors request it. Does any body have a working protcal for it or know a good substitute. Thanks From asmith <@t> mail.barry.edu Fri May 20 10:15:23 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] distilled water Message-ID: <5D2189E74151CC42BEC02906BA8996322B90B5@exchsrv01.barrynet.barry.edu> When I took physical chemistry, we considered 1 Mohm water pure enough for making solutions for conductometric titrations. 1 Mohm water is certainly pure enough for histochemistry. I suspect that the water I now use for histochemistry has a resistance somewhat less than 1 Mohm. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kristopher Kalleberg Sent: Wednesday, May 18, 2005 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] distilled water I am looking to get a distilled water sytem in my lab, but the question arose if I need 1Mohm or if 18Mohm is required. I perform routine routine histology experiments, including IHC and immunofluorescence on skin. Does anyone know if the 1 Mohm system would suffice or if I will need to get the 18Mohm system. Any help would be greatly appreciated. Thank you. Kris Kalleberg Histologist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 203 381-5765 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From Lynne.Bell <@t> hitchcock.org Fri May 20 10:14:53 2005 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: Touch Prep CPT Code Message-ID: Joyce, I will not ignore you.......Yes, we do charge for touch preps performed by the pathologist. We use the CPT code 88104 (cytopathology, fluids, washing or brushings, except cervical or vaginal; smears with interpretation). We stain them with our frozen section stain technique and then they are interpreted by the pathologist. Most often, we do touch preps along with a frozen section on lymph nodes). Lynne Lynne A. Bell, HT (ASCP) Laboratory Central Vermont Hospital P.O. Box 547 Barre, VT 05641 (802)433-6711 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Weems, Joyce Sent: Fri 5/20/2005 7:30 AM To: Weems, Joyce; Histonet Subject: [Histonet] RE: Touch Prep CPT Code Are you all ignoring me? I will not go away! :>) Resending as I did not get a response. Happy weekend! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: Weems, Joyce Sent: Thursday, May 12, 2005 9:38 AM To: 'Histonet' Subject: Touch Prep CPT Code Do you guys charge for touch preps? If so, what CPT code? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From sa.drew <@t> hosp.wisc.edu Fri May 20 10:20:06 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] (no subject) Message-ID: How do people deal with immuno requests on formic acid treated autopsy tissue from suspected CJD cases? Are there concerns with putting these slides through automated immunostainers? Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From liz <@t> premierlab.com Fri May 20 10:31:11 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] GRAC-TP gravity recycler In-Reply-To: Message-ID: <001401c55d50$f6945410$a7d48a80@AMY> Angela We use this recycler and have had been using it for about a year now. It works fine for us. We place our used 100% and 95% from the stainer, misc. special stains and tissue processor and get out recycled 95%. You can not put any alcohol that has xylene in it. So your deparaffinization alcohols can not go into the recycler. We are a small lab and we just replaced our first filter after a year. I only order 100% alcohol from a vendor and we rely on the recycler to provide us with 95% alcohol. We have a hydrometer that we use to test the recycled product and adjust accordingly. It works quickly. We like it or else I would not have ordered a second filter. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Friday, May 20, 2005 6:22 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GRAC-TP gravity recycler Does anyone out there on Histonet use this recycler? What type of testimony can I get about it? Thanks. Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Fri May 20 10:39:37 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Hansen stain Message-ID: It's actually Hansel's stain. We get ours from: Lide Laboratories, Inc. 401 4th Avenue S.W. New Prague, MN 56071 Ph./Fax: 952-758-9760 It's a quick stain and we just follow their procedure. Good luck. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elaine Dooley Sent: Friday, May 20, 2005 9:27 AM To: Histonet@pathology.swmed.edu Subject: [Histonet] Hansen stain Dear Histonetters, I am looking for some information on a Hansen stain for eosinophils. I have had several Doctors request it. Does any body have a working protcal for it or know a good substitute. Thanks _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ree3 <@t> leicester.ac.uk Fri May 20 10:53:39 2005 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Sudan black/myeloid Message-ID: A question was asked recently concerning Sudan black staining in myeloid leukaemias, were there any useful responses??,unable to find anything in histoarchive. Many thanks Richard Edwards MRC TOX UNIT LEICESTER..U.K... From sa.drew <@t> hosp.wisc.edu Fri May 20 11:16:34 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Forgot subject line before-Immunostains on suspected CJD Message-ID: How do people deal with immuno requests on formic acid treated autopsy tissue from suspected CJD cases? Are there concerns with putting these slides through automated immunostainers? Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From shive003 <@t> umn.edu Fri May 20 11:33:30 2005 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Porcine Parvovirus IHC Message-ID: <015001c55d59$a8abefc0$41065486@auxs.umn.edu> Has anyone had experience using anti-Porcine Parvovirus from VMRD for IHC purposes (successfully)? If so, could you tell me the approximate dilution range you use? That'll help me determine the quantity size of Ab I need to order. Thanks. Jan From cfavara <@t> niaid.nih.gov Fri May 20 11:43:14 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Forgot subject line before-Immunostains on suspect ed CJD Message-ID: Sally, If the formic acid was done according to protocol for prion deactivation then theoretically you should have no problem with using an automated stainer. There are probably a few things to keep in mind. How was the specimen cut? Were precautions taken at that time? In the current animal models the infectious agent needs to be ingested or inoculated IC to cause disease. The prion is bound to the tissue which is bound to the slide so my feeling is that things are pretty stable at this point. Alternatively you could stain by hand using disposable containers, segregate your waste and treat with Environ LpH which inactivated prions. If you do put it on an automated stainer you should collect your waste and treat to deactivate c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Drew Sally A. [mailto:sa.drew@hosp.wisc.edu] Sent: Friday, May 20, 2005 9:17 AM To: Histonet (Histonet) Subject: [Histonet] Forgot subject line before-Immunostains on suspected CJD How do people deal with immuno requests on formic acid treated autopsy tissue from suspected CJD cases? Are there concerns with putting these slides through automated immunostainers? Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri May 20 12:27:41 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Envrion LpH treatment of prion diseased tissues In-Reply-To: References: Message-ID: <6.0.0.22.1.20050520111220.01b50890@gemini.msu.montana.edu> Environ LpH (EPA reg No. 1043-118) via key word search into Steris website gave directions for use, sites handling deer, elk, beef and sheep tissues with following transmissable spongioform encephalopathies (TSEs). Chronic wasting disease (CWD) scrapie, bovine spongiform encephalopathy (BSE). Nothing was said about treating human tissues with CJD or CJD variant. However the following publication was worth reading on this new product as compared to sodium hydroxide or bleach treatments. Race RE, Raymond GJ. Inactivation of transmissible spongiform encephalopathy (Prion) agens by Environ LpH J Virology 78(4):2164-2165, 2004. Although the testing in this publication was done using hamster prion model, this agent is being used by hospitals, and other institutions for prion disease. Environ LpH was recommended but not other LpH products. It will be interesting to see if this product becomes the recommended means for dealing with prion diseases. Cynthia's advice is well taken AND I would be inclined to treat the fixative the tissue was in also. At 10:43 AM 5/20/2005, you wrote: >Sally, > >If the formic acid was done according to protocol for prion deactivation >then theoretically you should have no problem with using an automated >stainer. There are probably a few things to keep in mind. How was the >specimen cut? Were precautions taken at that time? In the current animal >models the infectious agent needs to be ingested or inoculated IC to cause >disease. The prion is bound to the tissue which is bound to the slide so my >feeling is that things are pretty stable at this point. Alternatively you >could stain by hand using disposable containers, segregate your waste and >treat with Environ LpH which inactivated prions. If you do put it on an >automated stainer you should collect your waste and treat to deactivate > >c > > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is confidential >and may contain sensitive information. It should not be used by anyone who >is not the original intended recipient. If you have received this e-mail in >error please inform the sender and delete it from your mailbox or any other >storage devices. National Institute of Allergy and Infectious Diseases shall >not accept liability for any statements made that are sender's own and not >expressly made on behalf of the NIAID by one of its representatives > > >-----Original Message----- >From: Drew Sally A. [mailto:sa.drew@hosp.wisc.edu] >Sent: Friday, May 20, 2005 9:17 AM >To: Histonet (Histonet) >Subject: [Histonet] Forgot subject line before-Immunostains on suspected CJD > >How do people deal with immuno requests on formic acid treated autopsy >tissue from >suspected CJD cases? Are there concerns with putting these slides >through >automated immunostainers? > > >Sally Ann Drew, MT(ASCP) >IHC/ISH Laboratory >University of Wisconsin Hosp. & Clinics >Madison, WI 53792 >(608)265-6596 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pex0220 <@t> yahoo.com.cn Fri May 20 12:29:15 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Immunofluorescence in bone sections! Message-ID: <20050520172915.73632.qmail@web15508.mail.cnb.yahoo.com> Hello, all. I still have no good results about double immunofluorescence in bone sections until now. I have modified my protocol according some suggestions, but my results are still bad, only cartilage staining is better. Today my professor shows me some embryo sections, she does double immunofluorescence in embryo sections, and many positive expressions in bone and bone marrow appear. But positive expression is very weak! In addition, I have used the protocol of my professor, but results are also not good. I do not know the reasons, maybe it is related with mouse age and decalcification, but I am not sure of the exact reasons. this is my protocol: 1. Not deparaffinization, only dry flat at 37degree to 40 degree for a week. 2. Xylol 3x ( every time for 5 min) 3. Gradient ethanol dehydration ( 99%,99%, 96%, 80%,70% and 50% for 3min) 4. Distilled water 1x ( every time for 5min) Incubate with 0.05% trypsin in 37 degree ( for 15 min) Wash with PBS Blocking with 5% normal serum matched to the host of secondary antibodies ( for 2h) Wash with PBS+0.05%Tween 20 3x (every time for 5min) Primary antibody for overnight in 4 degree Wash with PBS+0.05%Tween 20 3x (every time for 5min) Secondary antibody ( for 2h) Wash with PBS+0.05%Tween 20 3x (every time for 5min) Nuclear staining (DAPI) for 5min Coverslip I do not know what should I do. If possible, Can you show me your protocol? Thank you! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From petepath <@t> yahoo.com Fri May 20 12:56:49 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Touch Prep CPT Code Message-ID: <20050520175649.81255.qmail@web30408.mail.mud.yahoo.com> Joyce and Lynn, We use 88161 code for intraoperative touch prep. Lynn, it is the understanding of our billing and compliance advisors that you cannot bill for the intaoperative touch prep if you are all ready billing for the frozen section. If that is what you are doing I would check to see if there is a compliance issue. This is what we were told. If anyone knows different I am interested in hearing what your interpretaton is. Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From tompset2 <@t> msu.edu Fri May 20 13:32:57 2005 From: tompset2 <@t> msu.edu (Amber R Tompsett) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] American Optical 820 Microtome Manual Message-ID: Histonetters, I just inherited an American Optical 820 Microtome to perform my research with (being a graduate student, I don't have the funding to buy a new one). The microtome, however, didn't come with a manual. I searched the archives looking for the manual, but hit a dead end. Can anyone help? Thanks, Amber Tompsett Graduate Assistant Aquatic Toxicology Laboratory Department of Zoology Michigan State University East Lansing, MI 48825 From mprice26 <@t> juno.com Fri May 20 13:52:03 2005 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: PROBE OR ANTIBODY FOR CYSTIC FIBROSIS ON THIN PREP SMEARS Message-ID: <20050520.115215.11709.93997@webmail29.nyc.untd.com> Hi histonetters, We are looking into doing Cystic Fibrosis Screening on our thin prep smears. I am interested in hearing feedback from other labs as to how they are performing this IHC or In-Situ Hybridization? and What vendor are you buying your probes and/or antibodies from? Thank you. Marsha Price From portera203 <@t> yahoo.com Fri May 20 15:45:50 2005 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Forgot subject line before-Immunostains on suspected CJD In-Reply-To: 6667 Message-ID: <20050520204550.72050.qmail@web40901.mail.yahoo.com> We treat ours for 3 minutes in Formic Acid, rinse in running water for 10 minutes, place in buffer with surfactant - good to go. "Drew Sally A." wrote:How do people deal with immuno requests on formic acid treated autopsy tissue from suspected CJD cases? Are there concerns with putting these slides through automated immunostainers? Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP), QIHC Michigan State University - Department of Physiology Division of Human Pathology Investigative Histopathology Laboratory 2201 Biomedical Physical Sciences Room 2133 East Lansing, MI 48824-3320 Ph: 517-355-6475 ext 1480/1481 Fax: 517-432-1368 portera@msu.edu --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From histology.bc <@t> shaw.ca Sat May 21 09:59:35 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Asbestos fibres in tissues References: Message-ID: <428F4CD7.2050402@shaw.ca> Hi Fred, I know you gave already had several replies to your question, but I would like to add my 2 cents worth. Several years ago while I was still working in England, I was part of a major asbestososis-mesothelioma project. I was involved in examining tissues from several hundred autopsies for asbestos fibres and/or absbestos bodies. This project was initially sparked by the discovery of asbestos-associated diseases in elderly females none of whom had ever been employed in the industries traditionally associated with asbestos. To cut a long story short, these women, who were mainly housewives, had been exposed to powdered asbestos while assembling gas-masks during World War II. The masks were intended to be used in the event of "poisonous gas" attacks (which never occurred). The exposure to asbestos was quite brief, sometimes only a couple of weeks working on the assembly line. But, some 25-30 years later, they presented with classical asbestos-associated diseases. We did find patients with large numbers of asbestos fibres, but no asbestos bodies; patients with few fibres/bodies, but massive fibrosis/mesothelioma; and patients with large numbers of asbestos bodies, but little evidence of fibrosis, etc. We also found that specific types of asbestos were more likely to cause mesothelioma than other types. Asbestos is a silicate and consequently will polarize light, however, the fibres can be very fine and easily overlooked by the inexperienced. After period of time, the ends of the fibres are coated by an iron-containing protein material, producing the classical "dumbell" shape. Additional protein-iron material will produce a fully coated fibre. Coated fibres are no longer birefringent when viewed by polarized light. The pathologist in charge of the project, Stephen Phillips Jones, wrote several articles on these topics, and went on the become on of Europe's most respected asbestosis experts. As part of the project, we used a variety of procedures to find the most effective/efficient means of identifying the presence of asbestos fibres or bodies. The procedures we finally settled on require both fresh and fixed autopsy tissues. The procedure we used for fresh tissues is as follows: Slice through a large portion of lung tissue (we routinely used the left lower lobe) to expose the parenchyma. Squeeze the tissues very firmly to extract the tissue fluids, using a funnel to collect the "juices" into a large plastic centrifuge tube is most convenient. This is a messy, bloody, seemingly crude procedure, but it works very well. Add a lysing reagent to the "juices" to get rid of the red cells, I believe we used saponin, but any lysing agent will work. Centrifuge the tissue fluid, discard most of the supernatent and make wet preparations from the concentrate. No staining is required. Asbestos bodies, if present, are very conspicuous and easily identified. Asbestos fibres, much finer and easily overlooked, can be identified by the use of polarized light. The wet preps are not permanent and can be discarded after examination. The lung tissue must be unfixed. Autolysis and putrefaction do not have any effect on the asbestos content, although they make the procedure less pleasant. For fixed tissues, we used the following procedure. From each processed block of lung tissue, we cut a 30 micron section. This was dried as usual, dewaxed, coverslipped, and examined using brightfield and polarized light microscopy. Asbestos fibres or bodies were easily identified.The section does not require staining. As other Histonet members have commented, asbestos bodies are strongly Perls' Prussian blue positive due to the ferric iron present in the coating. However, the morphology of the asbestos body is unmistakable and does not really require confirmation by special stains. Some texts have recommended using microincineration to "cremate" the cellular tissue components and leave a residue of asbestos-containing ash that can be examined. We tried this method, but found it to be unreliable, time-consuming and generally a pain in the %$#. Hope these thoughts help, if you need any more suggestions, let me know. Sorry for the late response, but I was down in Florida for the past three weeks enjoying the beautiful weather, beaches, and scenery of the Gulf coast. Paul Bradbury Kamloops, BC, Canada Fred Underwood wrote: >Hi All, > > I asked this question earlier, but am not sure it got posted. I'm >looking for a technic to demonstrate asbestos fibers. Possbily by >digesting the tissue. Thanks in advance. > >Fred Underwood HT(ASCP) >Montgomery County Coroner >Dayton,OH > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From histojock <@t> hotmail.com Sat May 21 09:59:38 2005 From: histojock <@t> hotmail.com (Histo Jock) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Re: Immunostains on suspected CJD Message-ID: Just a note of caution here. If you decontaminate with "LpH" make absolutely certain that it's "Environ LpH" and not "LpH-SE", "LpH-AG" or any other similarly named product. There are several different products sold by Steris that contain the "LpH" name and all have different formulas. Only the stuff marked "Environ LpH" is effective against prions. Use caution when ordering to make certain you get the right stuff. See Race & Raymond, J Virol. 2004 February; 78(4): 2164–2165. HistoJock _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From jnocito <@t> satx.rr.com Sat May 21 12:02:23 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Hair Bulbs Message-ID: <024f01c55e26$dba02180$2423f318@yourxhtr8hvc4p> Howdy Histoland, I swear one day I'm gonna shoot me a pathologist. We are diving into the realm of hair pulls by dermatologists to look for fungus. Any ideas on how to handle these itsy bitsy specimens? Embedding tips? We were successful on the first specimen, but haven't even come close since then. I'm pulling my hair out. We are supposed to be a nice little lab that does routine histology. Don't the "reference" part fool ya. Ummmmmm, maybe some one should have told this pathologist that. As always, thanks in advance for your help and the routine disclaimer "The opinions of the author do not reflect the opinions of his employers, their lawyers, family members, distant cousins, yada yada yada. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX From Barry.R.Rittman <@t> uth.tmc.edu Sat May 21 18:21:03 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Hair Bulbs Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0F3539F@UTHEVS3.mail.uthouston.edu> Joe I assume that you are askig about embedding? Can't you treat the hair bulbs as whole specimens, attach them to a slide (agar perhaps) and then stain the entire hair bulb for fungus? Try this first and then shoot the pathologist. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Sat 5/21/2005 12:02 PM To: histonet Subject: [Histonet] Hair Bulbs Howdy Histoland, I swear one day I'm gonna shoot me a pathologist. We are diving into the realm of hair pulls by dermatologists to look for fungus. Any ideas on how to handle these itsy bitsy specimens? Embedding tips? We were successful on the first specimen, but haven't even come close since then. I'm pulling my hair out. We are supposed to be a nice little lab that does routine histology. Don't the "reference" part fool ya. Ummmmmm, maybe some one should have told this pathologist that. As always, thanks in advance for your help and the routine disclaimer "The opinions of the author do not reflect the opinions of his employers, their lawyers, family members, distant cousins, yada yada yada. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Sun May 22 09:54:58 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] Hair Bulbs In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F0F3539F@UTHEVS3.mail.uthouston.edu> Message-ID: Can I just shoot the pathologist first? Joe -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rittman, Barry R Sent: Saturday, May 21, 2005 6:21 PM To: Joe Nocito; histonet Subject: RE: [Histonet] Hair Bulbs Joe I assume that you are askig about embedding? Can't you treat the hair bulbs as whole specimens, attach them to a slide (agar perhaps) and then stain the entire hair bulb for fungus? Try this first and then shoot the pathologist. Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Sat 5/21/2005 12:02 PM To: histonet Subject: [Histonet] Hair Bulbs Howdy Histoland, I swear one day I'm gonna shoot me a pathologist. We are diving into the realm of hair pulls by dermatologists to look for fungus. Any ideas on how to handle these itsy bitsy specimens? Embedding tips? We were successful on the first specimen, but haven't even come close since then. I'm pulling my hair out. We are supposed to be a nice little lab that does routine histology. Don't the "reference" part fool ya. Ummmmmm, maybe some one should have told this pathologist that. As always, thanks in advance for your help and the routine disclaimer "The opinions of the author do not reflect the opinions of his employers, their lawyers, family members, distant cousins, yada yada yada. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sun May 22 12:24:16 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] looking for Moffit Cancer Center Investigator Message-ID: <200505221724.j4MHO91C020453@chip.viawest.net> Pardon me for using this forum for this but this is the only way I have to try and contact this person. An investigator from ?Moffit Cancer Center in ?Tampa, FL contacted me to do some IHC work on mouse brain tumors, he did not leave his contact information with me on the phone and has not emailed me with the details so I can discuss this with him. If you are still out there on histonet or if someone else knows who this is, please contact me at pruegg@ihctech.net I do need to buy rat anti-ms cd4, cd8, and cd56 for your work. Patsy Ruegg 720-859-4060 From AnthonyH <@t> chw.edu.au Sun May 22 19:23:28 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:06 2005 Subject: [Histonet] RE: Touch Prep CPT Code Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E25C@simba.kids> Joyce, I'm not ignoring you, we just don't have CPT Codes. Sorry Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Weems, Joyce [mailto:JWEEMS@sjha.org] Sent: Friday, 20 May 2005 9:30 PM To: Weems, Joyce; Histonet Subject: [Histonet] RE: Touch Prep CPT Code Are you all ignoring me? I will not go away! :>) Resending as I did not get a response. Happy weekend! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: Weems, Joyce Sent: Thursday, May 12, 2005 9:38 AM To: 'Histonet' Subject: Touch Prep CPT Code Do you guys charge for touch preps? If so, what CPT code? Thanks, Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From histology.bc <@t> shaw.ca Sun May 22 22:15:05 2005 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs References: Message-ID: <42914AB9.6010909@shaw.ca> Are they in season down in Texas? Or do you have open season on varmints? Joe Nocito wrote: >Can I just shoot the pathologist first? > >Joe > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rittman, >Barry R >Sent: Saturday, May 21, 2005 6:21 PM >To: Joe Nocito; histonet >Subject: RE: [Histonet] Hair Bulbs > > >Joe >I assume that you are askig about embedding? >Can't you treat the hair bulbs as whole specimens, attach them to a slide >(agar perhaps) and then stain the entire hair bulb for fungus? >Try this first and then shoot the pathologist. >Barry > >________________________________ > >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito >Sent: Sat 5/21/2005 12:02 PM >To: histonet >Subject: [Histonet] Hair Bulbs > > > >Howdy Histoland, > I swear one day I'm gonna shoot me a pathologist. We are diving into the >realm of hair pulls by dermatologists to look for fungus. Any ideas on how >to handle these itsy bitsy specimens? Embedding tips? We were successful on >the first specimen, but haven't even come close since then. I'm pulling my >hair out. > We are supposed to be a nice little lab that does routine histology. >Don't the "reference" part fool ya. Ummmmmm, maybe some one should have told >this pathologist that. > As always, thanks in advance for your help and the routine disclaimer > "The opinions of the author do not reflect the opinions of his >employers, their lawyers, family members, distant cousins, yada yada yada. > >Joe Nocito BS, HT(ASCP)QIHC >Histology Manager >Pathology Reference Lab >San Antonio, TX >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From bills <@t> icpmr.wsahs.nsw.gov.au Sun May 22 22:37:12 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs Message-ID: <000001c55f48$b5a99100$7d84080a@wsahs.nsw.gov.au> Joe, Our Microbiological/Fungal experts look at scrapings from hairs, skin, nails etc. It is almost impossible to orientate hairs to get cuts at full length. Many years ago I tried and nearly went insane, and these were in intact skin. I achieved about one in two hundred success and even then was only able to get about 5-6 serial sections from each hair. Bill Sinai Laboratory Manager Tissue Pathology, ICPMR Westmead NSW 2145 Australia Ph 02 9845 7774 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From Sjohnso616 <@t> aol.com Mon May 23 03:32:54 2005 From: Sjohnso616 <@t> aol.com (Sjohnso616@aol.com) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Position available In Southwest Fla Message-ID: Hi All, Sarasota Pathology in Southwest Florida is looking to hire a HT or HTL. Our successful candidate will have strong skills in embedding, cutting and special stains. We offer excellent salary and benefits as well as an abundance of SUN. Please forward your resume to: Nancy Day 2001 Webber St Sarasota Florida 34239-5737 Fax 941-362-8992 Phone: 941 362-8900 From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon May 23 03:41:39 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] H pylori stain Message-ID: We don't use a silver stain but a "Quick Diff" giemsa which is very good - I can give you this method if you want Jacqui Malam Royal Lancaster Infirmary DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Julie.Sanders <@t> med.va.gov Mon May 23 06:41:33 2005 From: Julie.Sanders <@t> med.va.gov (Julie.Sanders@med.va.gov) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Productivity Issues Message-ID: <457381D92B01BD44B21CF37CC02EBDFD02927636@vhacinexc2.v10.med.va.gov> Greetings Histonetters, I have a question about what are reasonable expectations for productivity. For example, if the majority of work done are biopsies, prostate needle bxs, livers, bone marrows and kidney biopsies is embedding 80 blocks an hour and cutting 40 per hour reasonable? Keep in mind that the bone marrows, livers, and kidneys require special stains/immunos and prostate bxs require 4 slides each. I read the article in NSH Journal about productivity and tried (and failed) to figure out what would be reasonable expectations from my techs for embedding and cutting. I wanted to include all the tasks required: labeling slides, facing blocks, cutting biopsies and cutting routine. I tried by adding all the numbers in the article (The Journal of Histotechnology,Vol. 27, No.4/ Dec. 2004) but obviously I'm not mathematically inclined. Any ideas/suggestions/answers would be greatly appreciated! Thanks, Julie Julie Sanders, BA,HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio From petepath <@t> yahoo.com Mon May 23 06:52:04 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs Message-ID: <20050523115204.17279.qmail@web30404.mail.mud.yahoo.com> Joe, If you can use frozen sections technique for this chore, I think you might find it easier to solve your embedding problem using my Precision Cryoembedding System. If your not familiar with it have a peek at the web site below. If you do decide to kill your pathologist, let me know , my residents are always looking for openings. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From Kemlo.Rogerson <@t> elht.nhs.uk Mon May 23 08:27:39 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Productivity Issues Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD188@elht-exch1.xelht.nhs.uk> Is that per person? 80 blocks per hour embedded, 40 cut, per person? If so that seems a bit high; but then Australian and American Cytologists can screen over 100 cervical smears per day. Maybe us Brits are whimps? -----Original Message----- From: Julie.Sanders@med.va.gov [mailto:Julie.Sanders@med.va.gov] Sent: 23 May 2005 12:42 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Productivity Issues Greetings Histonetters, I have a question about what are reasonable expectations for productivity. For example, if the majority of work done are biopsies, prostate needle bxs, livers, bone marrows and kidney biopsies is embedding 80 blocks an hour and cutting 40 per hour reasonable? Keep in mind that the bone marrows, livers, and kidneys require special stains/immunos and prostate bxs require 4 slides each. I read the article in NSH Journal about productivity and tried (and failed) to figure out what would be reasonable expectations from my techs for embedding and cutting. I wanted to include all the tasks required: labeling slides, facing blocks, cutting biopsies and cutting routine. I tried by adding all the numbers in the article (The Journal of Histotechnology,Vol. 27, No.4/ Dec. 2004) but obviously I'm not mathematically inclined. Any ideas/suggestions/answers would be greatly appreciated! Thanks, Julie Julie Sanders, BA,HT(ASCP) Supervisor, Anatomic Pathology VAMC, Cincinnati, Ohio _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bruyntjes <@t> voeding.tno.nl Mon May 23 08:37:35 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] GFAP - Rat Message-ID: <3B070848E7C2204F9DEB8BCFD767728001079DBE@ntexch1.voeding.tno.nl> Hi all Is anyone of you working with rat brain and GFAP? I have noticed that many GFAP antibodies do cross react with rat tissue, but maybe someone of you is working with rat tissue and does have a preference to a particular antibody. If so, can you share this knowledge with me? Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From kappeler <@t> patho.unibe.ch Mon May 23 09:26:21 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] GFAP - Rat References: <3B070848E7C2204F9DEB8BCFD767728001079DBE@ntexch1.voeding.tno.nl> Message-ID: <00de01c55fa3$7a446da0$27955c82@patho.unibe.ch> Hi Joost We have been using DakoCytomation's polyclonal rb-a-GFAP, Z 0334, with quite good success on rat brains. Dilution was 1:500-1:1000, pretreatment: HIER (citrate pH 6.0, pressure cooker). Hope this helps. Best regards Andi Kappeler, Ph.D. Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Bruijntjes, J.P." To: Sent: Monday, May 23, 2005 3:37 PM Subject: [Histonet] GFAP - Rat > Hi all > > > > Is anyone of you working with rat brain and GFAP? > > I have noticed that many GFAP antibodies do cross react with rat tissue, > but maybe someone of you is working with rat tissue and does have a > preference to a particular antibody. If so, can you share this knowledge > with me? > > > > Joost Bruijntjes > > TNO Quality of Life > > Zeist Holland > > > > > > > > > This e-mail and its contents are subject to the DISCLAIMER at > http://www.tno.nl/disclaimer/email.html > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From claudiamel2000 <@t> yahoo.com Mon May 23 10:05:04 2005 From: claudiamel2000 <@t> yahoo.com (claudia melidona) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Re: FISH Bechmark XT Message-ID: <20050523150504.13942.qmail@web60821.mail.yahoo.com> In need of some info if you please-- feedback from anyone using Bechmark XT for FISH .Hope everyone had a great week-end! Claudia --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From mcauliff <@t> umdnj.edu Mon May 23 10:33:14 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] GFAP - Rat In-Reply-To: <3B070848E7C2204F9DEB8BCFD767728001079DBE@ntexch1.voeding.tno.nl> References: <3B070848E7C2204F9DEB8BCFD767728001079DBE@ntexch1.voeding.tno.nl> Message-ID: <4291F7BA.2060808@umdnj.edu> I also have had excellent results with DAKO's rabbit anti-cow GFAP (Z0334). I use it with a Vector ABC Elite kit and DAB for the chromogen. For FFPE sections my primary AB dilutions are about 1:5000 when using nickel + cobalt to intensify the DAB, more concentrated without the nickel-cobalt. Geoff Bruijntjes, J.P. wrote: >Hi all > > > >Is anyone of you working with rat brain and GFAP? > >I have noticed that many GFAP antibodies do cross react with rat tissue, >but maybe someone of you is working with rat tissue and does have a >preference to a particular antibody. If so, can you share this knowledge >with me? > > > >Joost Bruijntjes > >TNO Quality of Life > >Zeist Holland > > > > > > > > >This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From andrae <@t> u.washington.edu Mon May 23 10:47:41 2005 From: andrae <@t> u.washington.edu (A. Erickson) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs In-Reply-To: <42914AB9.6010909@shaw.ca> References: <42914AB9.6010909@shaw.ca> Message-ID: you could sell tickets! andra erickson On Sun, 22 May 2005, Paul Bradbury wrote: > Are they in season down in Texas? Or do you have open season on varmints? > > Joe Nocito wrote: > >> Can I just shoot the pathologist first? >> >> Joe >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rittman, >> Barry R >> Sent: Saturday, May 21, 2005 6:21 PM >> To: Joe Nocito; histonet >> Subject: RE: [Histonet] Hair Bulbs >> >> >> Joe >> I assume that you are askig about embedding? >> Can't you treat the hair bulbs as whole specimens, attach them to a slide >> (agar perhaps) and then stain the entire hair bulb for fungus? >> Try this first and then shoot the pathologist. >> Barry >> >> ________________________________ >> >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito >> Sent: Sat 5/21/2005 12:02 PM >> To: histonet >> Subject: [Histonet] Hair Bulbs >> >> >> >> Howdy Histoland, >> I swear one day I'm gonna shoot me a pathologist. We are diving into the >> realm of hair pulls by dermatologists to look for fungus. Any ideas on how >> to handle these itsy bitsy specimens? Embedding tips? We were successful on >> the first specimen, but haven't even come close since then. I'm pulling my >> hair out. >> We are supposed to be a nice little lab that does routine histology. >> Don't the "reference" part fool ya. Ummmmmm, maybe some one should have >> told >> this pathologist that. >> As always, thanks in advance for your help and the routine disclaimer >> "The opinions of the author do not reflect the opinions of his >> employers, their lawyers, family members, distant cousins, yada yada yada. >> >> Joe Nocito BS, HT(ASCP)QIHC >> Histology Manager >> Pathology Reference Lab >> San Antonio, TX >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Allison_Scott <@t> hchd.tmc.edu Mon May 23 10:58:51 2005 From: Allison_Scott <@t> hchd.tmc.edu (Scott, Allison D) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Test Message-ID: CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. From Allison_Scott <@t> hchd.tmc.edu Mon May 23 10:59:59 2005 From: Allison_Scott <@t> hchd.tmc.edu (Scott, Allison D) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] FW: Who should be doing the hiring? Message-ID: -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:58 AM To: 'www.histonet@lists.utsouthwestern.edu' Subject: FW: Who should be doing the hiring? -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:56 AM To: 'www.histonet@lists.utsouthwestern.edu' Subject: FW: Who should be doing the hiring? -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:55 AM To: 'histonet@lists.utsouthwester.edu' Subject: Who should be doing the hiring? Hello to all in histoland. I have been finally given another position for a full time tech. My question is, who should ultimately make the decision on who gets the position. Should it be the supervisor, the medical director or the manager? I have been the supervisor here for the last 10 years. I made the decision on who we should hire for the last position that we had open. We now have a new medical director who feels that they should have the ultimate say in the decision. I feel that I should have the last say in the matter due to the fact that I will know what type of person would be best to work in the area, and besides I have to be the one that will supervise this person. The medical director and the manager will not be in the lab for the day to day operations. I'll take any feedback regarding this situation. Thanks Allison Scott CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. From kappeler <@t> patho.unibe.ch Mon May 23 11:26:55 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] GFAP - Rat References: <3B070848E7C2204F9DEB8BCFD767728004A69571@ntexch1.voeding.tno.nl> Message-ID: <012401c55fb4$4dd872f0$27955c82@patho.unibe.ch> Hi Joost We did compare 6 different pretreatmeant variants and had best results with HIER (citrate) in a pressure cooker, although we did see some faint staining without HIER. It may also depend on the way (time, fixative) your tissue samples have been fixed. If you have the possibility, run a comparison yourself to be sure you have the protocol that works best with your material. Best regards Andi ----- Original Message ----- From: "Bruijntjes, J.P." To: "Andi Kappeler" Sent: Monday, May 23, 2005 5:35 PM Subject: RE: [Histonet] GFAP - Rat > Hi Andy > > Thanks for your reply. You use HIER pretreatment (citrate). Is it really > necessary? Someone else recommended the same antibody and she did not > use a HIER technique. > > Joost Bruijntjes > > > -----Original Message----- > From: Andi Kappeler [mailto:kappeler@patho.unibe.ch] > Sent: maandag 23 mei 2005 16:26 > To: Bruijntjes, J.P.; Histonet > Subject: Re: [Histonet] GFAP - Rat > > Hi Joost > > We have been using DakoCytomation's polyclonal rb-a-GFAP, Z 0334, with > quite > good success on rat brains. Dilution was 1:500-1:1000, pretreatment: > HIER > (citrate pH 6.0, pressure cooker). Hope this helps. > > Best regards > Andi Kappeler, Ph.D. > Institute of Pathology, University of Bern, Switzerland > > ----- Original Message ----- > From: "Bruijntjes, J.P." > To: > Sent: Monday, May 23, 2005 3:37 PM > Subject: [Histonet] GFAP - Rat > > >> Hi all >> >> >> >> Is anyone of you working with rat brain and GFAP? >> >> I have noticed that many GFAP antibodies do cross react with rat > tissue, >> but maybe someone of you is working with rat tissue and does have a >> preference to a particular antibody. If so, can you share this > knowledge >> with me? >> >> >> >> Joost Bruijntjes >> >> TNO Quality of Life >> >> Zeist Holland >> >> >> >> >> >> >> >> >> This e-mail and its contents are subject to the DISCLAIMER at >> http://www.tno.nl/disclaimer/email.html >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > > This e-mail and its contents are subject to the DISCLAIMER at > http://www.tno.nl/disclaimer/email.html > From peoshel <@t> wisc.edu Mon May 23 11:53:16 2005 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs In-Reply-To: <42914AB9.6010909@shaw.ca> References: <42914AB9.6010909@shaw.ca> Message-ID: It's Texas, they have open season on *everything*. >Are they in season down in Texas? Or do you have open season on varmints? > >Joe Nocito wrote: > >>Can I just shoot the pathologist first? >> >>Joe >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rittman, >>Barry R >>Sent: Saturday, May 21, 2005 6:21 PM >>To: Joe Nocito; histonet >>Subject: RE: [Histonet] Hair Bulbs >> >> >>Joe >>I assume that you are askig about embedding? >>Can't you treat the hair bulbs as whole specimens, attach them to a slide >>(agar perhaps) and then stain the entire hair bulb for fungus? >>Try this first and then shoot the pathologist. >>Barry >> >>________________________________ >> >>From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito >>Sent: Sat 5/21/2005 12:02 PM >>To: histonet >>Subject: [Histonet] Hair Bulbs >> >> >> >>Howdy Histoland, >> I swear one day I'm gonna shoot me a pathologist. We are diving into the >>realm of hair pulls by dermatologists to look for fungus. Any ideas on how >>to handle these itsy bitsy specimens? Embedding tips? We were successful on >>the first specimen, but haven't even come close since then. I'm pulling my >>hair out. >> We are supposed to be a nice little lab that does routine histology. >>Don't the "reference" part fool ya. Ummmmmm, maybe some one should have told >>this pathologist that. >> As always, thanks in advance for your help and the routine disclaimer >> "The opinions of the author do not reflect the opinions of his >>employers, their lawyers, family members, distant cousins, yada yada yada. >> >>Joe Nocito BS, HT(ASCP)QIHC >>Histology Manager >>Pathology Reference Lab >>San Antonio, TX -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm From petepath <@t> yahoo.com Mon May 23 12:02:01 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Who should be doing the hiring? Message-ID: <20050523170201.48755.qmail@web30402.mail.mud.yahoo.com> Allison, Pathologist point of view. It is the pathologists job to keep you informed of their histology needs and any new programs, studies, antibodies, techniques, which will require new or additional staffing so that you can adjust your staffing in a timely manner. You should keep track of your changing workload so that you can predict need for additional staff in a timely manner. When it comes time to interview and hire you are best suited to recognize the skill or lack of it in a given candidate and how that person will fill the needs of your department. Checking references should be your job as your knowledge of histology will allow you to best understand the subliminal messages that often are the only negatives you are given. If the pathologist has the interest to call about references I would welcome his help. The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker. When it is time to make a decision I think your impressions and any impression the medical director has made should be discussed by both of you. If you have made a decision on a candidate that you are confident about, I would hope that your medical director would support you in this decision. If you have a medical director who does not care to learn about the candidates as much as he feels the need to control everything in his enviornment then this can lead to problems. Bottom line. You are responsible for the day to day workings of your lab, the working enviornment and quality of the work. You will have to live with the misery of bad hiring decision. You will be the one to feel the heat when mistakes are made or work is repeatedly late. You should be the one to decide who to hire. If you are relitively inexperienced then I would invite other input if it is a tough decision. I would foster a good working relationship with your medical director, keep him informed of your progress and when you have made your decision, discuss it with him and consider his input. If he has faith in you and he is doing his job as Director I hope he would respect your decision and allow you to do the job you were hired to do. If he feels the need to choose a different candidate which you have reservations about then I would discuss your reasoning in a collegeal manner. Explain the pros and cons of the candidates. If you feel he is making decision which will have a negative impact then your last resort is to discuss it with your lab manager or other superior, they are ultimately responible for you. Good luck Stephen From jcline <@t> wchsys.org Mon May 23 12:04:24 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Who should do the hiring? Message-ID: <000801c55fb9$78a38ce0$1d2a14ac@wchsys.org> I have had the experience of having the pathologists push to hire someone I was still investigating (because we were short staffed). Well, we have multiple problems with this person now. The person that has to check the performance and fill out evals should be the one to pick who gets hired in my opinion. ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From pruegg <@t> ihctech.net Mon May 23 12:31:38 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] looking for Moffit CA Center Investigator Message-ID: <200505231731.j4NHVT1C003922@chip.viawest.net> Pardon the use of this formum for this but I have no other way to contact this person. An investigator from ?Moffit CA Center in ?Tampa Florida called me to request some IHC work on mouse brain tumors. He did not leave his contact info and found me on the HISTONET. He was supposed to email me more details about this work but I have not heard from him. If this person is still on histonet or if someone else knows who this is please have him contact me ASAP at pruegg@ihctech.net Thank you, Patsy From NMargaryan <@t> childrensmemorial.org Mon May 23 13:35:10 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] (no subject) Message-ID: <63B8B599DE283148B92E83C78B32C15DB91A85@cmhexbe2.childrensmemorial.org> Hi Histos! I am having problems with cutting FFPE mouse nipple. Paraffin is cutting well but small tissue just crushing. We can't collect samples. Please help, any suggestion is appreciated. Thanks, Naira Naira V. Margaryan, Ph.D, D.V.M. Research Associate II Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-880-4000/5-6740 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. From dmarsha3 <@t> utmem.edu Mon May 23 13:59:19 2005 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs Message-ID: <104a9911046617.1046617104a991@utnet2.utmem.edu> hi all I am on several listservers and this is definitely the most interesting and the most lively. Between the anchovy gonads and the hairbulbs (and all that has come with that), I'll just say I never delete a subject thread anymore because you just don't know where it will go:-) Happy staining! dm Dana R. Marshall, Ph.D. Research Biologist, VA Medical Center Instructor, UT Health Science Center Memphis, TN 901-523-8990 x7559 ----- Original Message ----- From: Philip Oshel Date: Monday, May 23, 2005 11:53 am Subject: Re: [Histonet] Hair Bulbs > It's Texas, they have open season on *everything*. > > >Are they in season down in Texas? Or do you have open season on > varmints?> > >Joe Nocito wrote: > > > >>Can I just shoot the pathologist first? > >> > >>Joe > >> > >>-----Original Message----- > >>From: histonet-bounces@lists.utsouthwestern.edu > >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of > Rittman,>>Barry R > >>Sent: Saturday, May 21, 2005 6:21 PM > >>To: Joe Nocito; histonet > >>Subject: RE: [Histonet] Hair Bulbs > >> > >> > >>Joe > >>I assume that you are askig about embedding? > >>Can't you treat the hair bulbs as whole specimens, attach them > to a slide > >>(agar perhaps) and then stain the entire hair bulb for fungus? > >>Try this first and then shoot the pathologist. > >>Barry > >> > >>________________________________ > >> > >>From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe > Nocito>>Sent: Sat 5/21/2005 12:02 PM > >>To: histonet > >>Subject: [Histonet] Hair Bulbs > >> > >> > >> > >>Howdy Histoland, > >> I swear one day I'm gonna shoot me a pathologist. We are > diving into the > >>realm of hair pulls by dermatologists to look for fungus. Any > ideas on how > >>to handle these itsy bitsy specimens? Embedding tips? We were > successful on > >>the first specimen, but haven't even come close since then. I'm > pulling my > >>hair out. > >> We are supposed to be a nice little lab that does routine > histology.>>Don't the "reference" part fool ya. Ummmmmm, maybe > some one should have told > >>this pathologist that. > >> As always, thanks in advance for your help and the routine > disclaimer>> "The opinions of the author do not reflect the > opinions of his > >>employers, their lawyers, family members, distant cousins, yada > yada yada. > >> > >>Joe Nocito BS, HT(ASCP)QIHC > >>Histology Manager > >>Pathology Reference Lab > >>San Antonio, TX > > -- > Philip Oshel > Supervisor, BBPIC microscopy facility > Department of Animal Sciences > University of Wisconsin > 1675 Observatory Drive > Madison, WI 53706 > voice: (608) 263-4162 > fax: (608) 262-5157 (dept. fax) > http://www.ansci.wisc.edu/microscopy.htm > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Karen_Skish <@t> rush.edu Mon May 23 14:48:39 2005 From: Karen_Skish <@t> rush.edu (Karen_Skish@rush.edu) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Great research opportunity Message-ID: New research position open. Rush University Medical Center-Alzheimer's Disease Center. Immediate full time opening for research scientist to work on NIA-funded longitudinal clinicopathologic study of motor changes with aging in a stimulating multi-disciplinary academic environment. The project investigates changes in the spinal cord, nerve and muscle and their relationship to motor changes that accompany aging. Position includes working with human tissue using quantitative immunohistochemical techniques, eg. stereology, and routine neuropathology and neuroanatomy. Applicants should have a laboratory background, be able to start-up the laboratory aspect of a new project by ordering equipment and necessary supplies to conduct studies, have experience with tissue based studies, be able to work independently, and have proficiency with computers. Those with experience in neuroscience will be given preference. Post-doctorate or PhD preferred, though MS with 5-years relevant experience may apply. Rank and salary commensurate with experience. Send resume to Julie Schneider MD, Rush Alzheimer's Disease Center, Armour Academic Center, 600 S. Paulina Suite 1022F, Chicago, IL 60612, or email resume to julie_a_schneider@rush.edu Rush is an equal opportunity employer. No recruiters please! Karen M Skish, MS, PA(ASCP)MT Rush Alzheimer's Disease Center Laboratory karen_skish@rush.edu From NLemke <@t> asterand.com Mon May 23 15:08:57 2005 From: NLemke <@t> asterand.com (Nancy Lemke) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] antibody search Message-ID: <56E63440ABF1FC4897947D5BA4CFA44F050355FF@ATL1VEXC005.usdom004.tco.tc> Does anyone know if Cascade Biosciences in Massachusetts has disappeared? I am having trouble finding them online although I can find plenty of references to their antibodies. Does anyone have a favorite phospho antibody vendor? Thanks to all, Nancy Lemke Asterand, Inc. TechOne, Suite 501 440 Burroughs Detroit, MI 48202 This e-mail message, together with any attachments, contains information belonging to Asterand, Inc. that may be confidential, proprietary, copyrighted and/or legally protected or privileged, and is intended for the sole use of the individual or entity named on this message. If you are not the intended recipient, and have received this e-mail message in error, please immediately return this message to its original sender and permanently delete it from your electronic and paper records. Thank You. From DMILES <@t> parknet.pmh.org Mon May 23 15:27:15 2005 From: DMILES <@t> parknet.pmh.org (DARNETTA MILES) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Employment Opportunities Message-ID: Parkland Memorial Hospital Histology Laboratory Registered HT(ASCP) or HTL (ASCP) Duties include: embedding, microtomy, special stains, Immunohistochemistry stains. competitive salary, excellent benefits, excellent working environment. Contact - Darnetta Miles at 214-590-6652 for more information. Darnetta Miles, BS, MT, (ASCP) Anatomic Pathology Manager Parkland Health & Hospital System Phone: 214-590-6652 Fax: 214-590-4474 email: dmiles@parknet.pmh.org This message (including any attachments) is intended only for the use of the addressee(s) and may contain information that is privileged and confidential. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient or an authorized representative of the intended recipient, the use, dissemination or reproduction of this communication is prohibited and may be a violation of federal or state law and regulations. If you have received this communication in error, please destroy all copies of the message and its attachments and notify the sender immediately. The Dallas County Hospital District and its affiliated entities hereby claim all applicable privileges related to this information. From Jackie.O'Connor <@t> abbott.com Mon May 23 15:40:05 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Employment Opportunities Message-ID: It's nice to see where the jobs are - - but sometimes, like this time, you guys don't include where the job is - just the state would be nice. P.S. I'm not looking for a job. I love my job. I love my job. I love my job. "DARNETTA MILES" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/23/2005 03:27 PM To: cc: Subject: [Histonet] Employment Opportunities Parkland Memorial Hospital Histology Laboratory Registered HT(ASCP) or HTL (ASCP) Duties include: embedding, microtomy, special stains, Immunohistochemistry stains. competitive salary, excellent benefits, excellent working environment. Contact - Darnetta Miles at 214-590-6652 for more information. Darnetta Miles, BS, MT, (ASCP) Anatomic Pathology Manager Parkland Health & Hospital System Phone: 214-590-6652 Fax: 214-590-4474 email: dmiles@parknet.pmh.org This message (including any attachments) is intended only for the use of the addressee(s) and may contain information that is privileged and confidential. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient or an authorized representative of the intended recipient, the use, dissemination or reproduction of this communication is prohibited and may be a violation of federal or state law and regulations. If you have received this communication in error, please destroy all copies of the message and its attachments and notify the sender immediately. The Dallas County Hospital District and its affiliated entities hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DMILES <@t> parknet.pmh.org Mon May 23 15:45:22 2005 From: DMILES <@t> parknet.pmh.org (DARNETTA MILES) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Employment Opportunities Message-ID: Sorry.....Dallas Texas >>> "Jackie M O'Connor" 05/23/05 03:40PM >>> It's nice to see where the jobs are - - but sometimes, like this time, you guys don't include where the job is - just the state would be nice. P.S. I'm not looking for a job. I love my job. I love my job. I love my job. "DARNETTA MILES" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/23/2005 03:27 PM To: cc: Subject: [Histonet] Employment Opportunities Parkland Memorial Hospital Histology Laboratory Registered HT(ASCP) or HTL (ASCP) Duties include: embedding, microtomy, special stains, Immunohistochemistry stains. competitive salary, excellent benefits, excellent working environment. Contact - Darnetta Miles at 214-590-6652 for more information. Darnetta Miles, BS, MT, (ASCP) Anatomic Pathology Manager Parkland Health & Hospital System Phone: 214-590-6652 Fax: 214-590-4474 email: dmiles@parknet.pmh.org This message (including any attachments) is intended only for the use of the addressee(s) and may contain information that is privileged and confidential. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient or an authorized representative of the intended recipient, the use, dissemination or reproduction of this communication is prohibited and may be a violation of federal or state law and regulations. If you have received this communication in error, please destroy all copies of the message and its attachments and notify the sender immediately. The Dallas County Hospital District and its affiliated entities hereby claim all applicable privileges related to this information. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From conniegrubaugh <@t> hotmail.com Mon May 23 16:19:34 2005 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs In-Reply-To: Message-ID: I would even take an unpaid vacation to get in on this. Connie

Connie G.




>From: "A. Erickson" <andrae@u.washington.edu>
>To: Paul Bradbury <histology.bc@shaw.ca>
>CC: Joe Nocito <JNocito@Pathreflab.com>,HistoNet Server <histonet@pathology.swmed.edu>
>Subject: Re: [Histonet] Hair Bulbs
>Date: Mon, 23 May 2005 08:47:41 -0700 (PDT)
>
>you could sell tickets! andra erickson
>
>On Sun, 22 May 2005, Paul Bradbury wrote:
>
>>Are they in season down in Texas? Or do you have open season on
>>varmints?
>>
>>Joe Nocito wrote:
>>
>>>Can I just shoot the pathologist first?
>>>
>>>Joe
>>>
>>>-----Original Message-----
>>>From: histonet-bounces@lists.utsouthwestern.edu
>>>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of
>>>Rittman,
>>>Barry R
>>>Sent: Saturday, May 21, 2005 6:21 PM
>>>To: Joe Nocito; histonet
>>>Subject: RE: [Histonet] Hair Bulbs
>>>
>>>
>>>Joe
>>>I assume that you are askig about embedding?
>>>Can't you treat the hair bulbs as whole specimens, attach them to
>>>a slide
>>>(agar perhaps) and then stain the entire hair bulb for fungus?
>>>Try this first and then shoot the pathologist.
>>>Barry
>>>
>>>________________________________
>>>
>>>From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe
>>>Nocito
>>>Sent: Sat 5/21/2005 12:02 PM
>>>To: histonet
>>>Subject: [Histonet] Hair Bulbs
>>>
>>>
>>>
>>>Howdy Histoland,
>>> I swear one day I'm gonna shoot me a pathologist. We are
>>>diving into the
>>>realm of hair pulls by dermatologists to look for fungus. Any
>>>ideas on how
>>>to handle these itsy bitsy specimens? Embedding tips? We were
>>>successful on
>>>the first specimen, but haven't even come close since then. I'm
>>>pulling my
>>>hair out.
>>> We are supposed to be a nice little lab that does routine
>>>histology.
>>>Don't the "reference" part fool ya. Ummmmmm, maybe some one should
>>>have told
>>>this pathologist that.
>>> As always, thanks in advance for your help and the routine
>>>disclaimer
>>> "The opinions of the author do not reflect the opinions of his
>>>employers, their lawyers, family members, distant cousins, yada
>>>yada yada.
>>>
>>>Joe Nocito BS, HT(ASCP)QIHC
>>>Histology Manager
>>>Pathology Reference Lab
>>>San Antonio, TX
>>>_______________________________________________
>>>Histonet mailing list
>>>Histonet@lists.utsouthwestern.edu
>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>>
>>>
>>>
>>>_______________________________________________
>>>Histonet mailing list
>>>Histonet@lists.utsouthwestern.edu
>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>>
>>>
>>>
>>>_______________________________________________
>>>Histonet mailing list
>>>Histonet@lists.utsouthwestern.edu
>>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>>
>>>
>>>
>>
>>
>>
>>_______________________________________________
>>Histonet mailing list
>>Histonet@lists.utsouthwestern.edu
>>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
From froyer <@t> bitstream.net Mon May 23 17:21:58 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Employment Opportunities In-Reply-To: References: Message-ID: <42925786.4070400@bitstream.net> After JKF was tragically shot, I thought everyone knew where Parkland Hospital was... But am I dating myself? DARNETTA MILES wrote: >Sorry.....Dallas Texas > > > >>>>"Jackie M O'Connor" 05/23/05 03:40PM >>>> >>>> >>>> > >It's nice to see where the jobs are - - but sometimes, like this time, >you guys don't include where the job is - just the state would be nice. > > >P.S. I'm not looking for a job. I love my job. I love my job. I >love my job. > > >"DARNETTA MILES" >Sent by: histonet-bounces@lists.utsouthwestern.edu >05/23/2005 03:27 PM > > To: > cc: > Subject: [Histonet] Employment Opportunities > > >Parkland Memorial Hospital >Histology Laboratory >Registered HT(ASCP) or HTL (ASCP) >Duties include: embedding, microtomy, special stains, >Immunohistochemistry stains. >competitive salary, excellent benefits, excellent working environment. >Contact - Darnetta Miles at 214-590-6652 for more information. > >Darnetta Miles, BS, MT, (ASCP) >Anatomic Pathology Manager >Parkland Health & Hospital System >Phone: 214-590-6652 >Fax: 214-590-4474 >email: dmiles@parknet.pmh.org > >This message (including any attachments) is intended only for the use >of the addressee(s) and may contain information that is privileged and >confidential. If you are the intended recipient, further disclosures >are prohibited without proper authorization. If you are not the >intended recipient or an authorized representative of the intended >recipient, the use, dissemination or reproduction of this >communication >is prohibited and may be a violation of federal or state law and >regulations. If you have received this communication in error, please >destroy all copies of the message and its attachments and notify the >sender immediately. The Dallas County Hospital District and its >affiliated entities hereby claim all applicable privileges related to >this information. > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From Frederick.Fifield <@t> sunhealth.org Mon May 23 18:01:22 2005 From: Frederick.Fifield <@t> sunhealth.org (Frederick.Fifield@sunhealth.org) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Fred on APL. Message-ID: I will be out of the office starting 05/23/2005 and will not return until 05/26/2005. I will be out of the lab beginning on 05/23/05. I will respond to your message when I return on Thursday, 05/26/05. From cfavara <@t> niaid.nih.gov Mon May 23 18:08:50 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] (no subject) Message-ID: Maria, Some pertinent questions would be: How are you collecting the sample? What is the processing schedule? I have done some studies here with mouse mammary glands and have collected the gland including the nipple. I have not had any problems with cutting. We shaved the belly being careful around the nipple so as not to injure the tissue. Cut a thin section on either side of the nipple. Process using ETOH 70%-2 hours, 80% -1 hour, 2 95% ETOH - 20 minutes each, 2 100% ETOH 20 minutes each, 3-Propar 30 minutes each, 4 paraffins 30 minutes each. Embedd as you would skin. In my hands most mouse tissue requires some soaking prior to cutting. Be sure the temp of your water bath is not to hot or the fat beneath the skin will just explode. I find the temperature range to be fairly narrow. Hope this is helpful. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Margaryan, Naira [mailto:NMargaryan@childrensmemorial.org] Sent: Monday, May 23, 2005 11:35 AM To: histonet Subject: [Histonet] (no subject) Hi Histos! I am having problems with cutting FFPE mouse nipple. Paraffin is cutting well but small tissue just crushing. We can't collect samples. Please help, any suggestion is appreciated. Thanks, Naira Naira V. Margaryan, Ph.D, D.V.M. Research Associate II Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-880-4000/5-6740 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From NMargaryan <@t> childrensmemorial.org Mon May 23 18:52:19 2005 From: NMargaryan <@t> childrensmemorial.org (Margaryan, Naira) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] RE: mouse mammary glands Message-ID: <63B8B599DE283148B92E83C78B32C15DB91AA5@cmhexbe2.childrensmemorial.org> Thanks a lot for all of your suggestions :-) I appreciate your and our :-) business, Naira -----Original Message----- From: Favara, Cynthia (NIH/NIAID) [mailto:cfavara@niaid.nih.gov] Sent: Monday, May 23, 2005 6:09 PM To: Margaryan, Naira; histonet Subject: RE: [Histonet] (no subject) Maria, Some pertinent questions would be: How are you collecting the sample? What is the processing schedule? I have done some studies here with mouse mammary glands and have collected the gland including the nipple. I have not had any problems with cutting. We shaved the belly being careful around the nipple so as not to injure the tissue. Cut a thin section on either side of the nipple. Process using ETOH 70%-2 hours, 80% -1 hour, 2 95% ETOH - 20 minutes each, 2 100% ETOH 20 minutes each, 3-Propar 30 minutes each, 4 paraffins 30 minutes each. Embedd as you would skin. In my hands most mouse tissue requires some soaking prior to cutting. Be sure the temp of your water bath is not to hot or the fat beneath the skin will just explode. I find the temperature range to be fairly narrow. Hope this is helpful. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Margaryan, Naira [mailto:NMargaryan@childrensmemorial.org] Sent: Monday, May 23, 2005 11:35 AM To: histonet Subject: [Histonet] (no subject) Hi Histos! I am having problems with cutting FFPE mouse nipple. Paraffin is cutting well but small tissue just crushing. We can't collect samples. Please help, any suggestion is appreciated. Thanks, Naira Naira V. Margaryan, Ph.D, D.V.M. Research Associate II Children's Memorial Research Center 2300 Children's Plaza, Box 222 Chicago, IL 60614-3394 Tel: 773-880-4000/5-6740 Fax: 773-755-6594 nmargaryan@childrensmemorial.org For Express Mail: CMRC, Room C.473 2430 N. Halsted Street Chicago, IL 60614-4314 The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The contents of this e-mail message and any attachments are private and confidential communications intended solely for the addressee(s) named in this message. If you are not the intended recipient of this message, please 1) immediately notify the sender by reply e-mail and then delete this message and its attachments and 2) do not read, use, distribute disclose or copy this message and/or any attachments. From cestanecki <@t> ucdavis.edu Mon May 23 19:19:49 2005 From: cestanecki <@t> ucdavis.edu (Catherine Stanecki) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Removing OCT??? Message-ID: <200505240019.j4O0JnNk017314@syrphus.ucdavis.edu> I am new to staining tissues and I recently froze a bunch of tissues in OCT. However, I recently found out that it would be better to have some sample fixed in formalin as well so you can use mutliple staining methods. Is there a way to remove OCT from tissue samples without damaging the tissue, or is the tissue forever stuck in tissue tek once you freeze it back? Thanks! Cathy From katri <@t> cogeco.ca Mon May 23 21:06:01 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Removing OCT??? References: <200505240019.j4O0JnNk017314@syrphus.ucdavis.edu> Message-ID: <007e01c56005$226cdd30$6a9a9618@Katri> Hi Cathy, Just put the OCT embedded tissue in 10% NBF and let come to room temperature, change formalin and let fix appropriate time. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Catherine Stanecki" To: Sent: Monday, May 23, 2005 8:19 PM Subject: [Histonet] Removing OCT??? > > I am new to staining tissues and I recently froze a bunch of tissues in > OCT. However, I recently found out that it would be better to have some > sample fixed in formalin as well so you can use mutliple staining methods. > Is there a way to remove OCT from tissue samples without damaging the > tissue, or is the tissue forever stuck in tissue tek once you freeze it > back? > > Thanks! > Cathy > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doscwk <@t> nus.edu.sg Mon May 23 21:18:27 2005 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Stains for apoptosis Message-ID: Hi, Many thanks to all who responded to my search for the tunel procedure, = especially to Kelly, Edwards, Melissa, Swaram and Andrea. You've = restored my faith in being part of the histonet family. Thanks so much Wai Kam=20 --=20 No virus found in this outgoing message. Checked by AVG Anti-Virus. Version: 7.0.322 / Virus Database: 266.11.15 - Release Date: 5/22/2005 =20 From jnocito <@t> satx.rr.com Mon May 23 22:06:32 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs References: <42914AB9.6010909@shaw.ca> Message-ID: <012201c5600d$972c4770$2423f318@yourxhtr8hvc4p> open season all year long, just like tourist season. We just have restrictions on dove, quail and dear. ----- Original Message ----- From: "Paul Bradbury" To: "Joe Nocito" ; "HistoNet Server" Sent: Sunday, May 22, 2005 10:15 PM Subject: Re: [Histonet] Hair Bulbs > Are they in season down in Texas? Or do you have open season on varmints? > > Joe Nocito wrote: > >>Can I just shoot the pathologist first? >> >>Joe >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rittman, >>Barry R >>Sent: Saturday, May 21, 2005 6:21 PM >>To: Joe Nocito; histonet >>Subject: RE: [Histonet] Hair Bulbs >> >> >>Joe >>I assume that you are askig about embedding? >>Can't you treat the hair bulbs as whole specimens, attach them to a slide >>(agar perhaps) and then stain the entire hair bulb for fungus? >>Try this first and then shoot the pathologist. >>Barry >> >>________________________________ >> >>From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito >>Sent: Sat 5/21/2005 12:02 PM >>To: histonet >>Subject: [Histonet] Hair Bulbs >> >> >> >>Howdy Histoland, >> I swear one day I'm gonna shoot me a pathologist. We are diving into >> the >>realm of hair pulls by dermatologists to look for fungus. Any ideas on how >>to handle these itsy bitsy specimens? Embedding tips? We were successful >>on >>the first specimen, but haven't even come close since then. I'm pulling my >>hair out. >> We are supposed to be a nice little lab that does routine histology. >>Don't the "reference" part fool ya. Ummmmmm, maybe some one should have >>told >>this pathologist that. >> As always, thanks in advance for your help and the routine disclaimer >> "The opinions of the author do not reflect the opinions of his >>employers, their lawyers, family members, distant cousins, yada yada yada. >> >>Joe Nocito BS, HT(ASCP)QIHC >>Histology Manager >>Pathology Reference Lab >>San Antonio, TX >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Mon May 23 22:07:17 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs References: <000001c55f48$b5a99100$7d84080a@wsahs.nsw.gov.au> Message-ID: <012801c5600d$b23a3e50$2423f318@yourxhtr8hvc4p> Bill, can you shoot pathologists down under? Joe ----- Original Message ----- From: "Bill Sinai" To: "histonet (E-mail)" Sent: Sunday, May 22, 2005 10:37 PM Subject: [Histonet] Hair Bulbs > > Joe, > > Our Microbiological/Fungal experts look at scrapings from hairs, skin, > nails > etc. It is almost impossible to orientate hairs to get cuts at full > length. > Many years ago I tried and nearly went insane, and these were in intact > skin. I achieved about one in two hundred success and even then was only > able to get about 5-6 serial sections from each hair. > > > Bill Sinai > Laboratory Manager > Tissue Pathology, ICPMR > Westmead NSW 2145 > Australia > Ph 02 9845 7774 > > > __________________________________________________________________ > > This electronic message and any attachments may be confidential. If you > are not the intended recipient of this message would you please delete the > message and any attachments and advise the sender. Western Sydney > Area Health Services (WSAHS) uses virus scanning software but excludes > any liability for viruses contained in any email or attachment. > > This email may contain privileged and confidential information intended > only for the use of the addressees named above. If you are not the > intended recipient of this email, you are hereby notified that any use, > dissemination, distribution, or reproduction of this email is prohibited. > If > you have received this email in error, please notify WSAHS > immediately. > > Any views expressed in this email are those of the individual sender > except where the sender expressly and with authority states them > to be the views of WSAHS. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Mon May 23 22:09:35 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs References: <20050523115204.17279.qmail@web30404.mail.mud.yahoo.com> Message-ID: <012e01c5600e$03f5dba0$2423f318@yourxhtr8hvc4p> I'm just afraid that I'll be charged with cruelty to an animal. Of course, I'm putting my butt on the line here. I'm sure one day my pathologist will go to conference and my name will pop up about wanting to shoot a pathologist and then I'll be looking for another job. Joe ----- Original Message ----- From: "Stephen Peters M.D." To: Sent: Monday, May 23, 2005 6:52 AM Subject: [Histonet] Hair Bulbs > Joe, > > If you can use frozen sections technique for this chore, I think you might > find it easier to solve your embedding problem using my Precision > Cryoembedding System. If your not familiar with it have a peek at the web > site below. If you do decide to kill your pathologist, let me know , my > residents are always looking for openings. > > Stephen > > > Stephen Peters M.D. > Vice Chairman of Pathology > Hackensack University Medical Center > 201 996 4836 > > Pathology Innovations, LLC > 410 Old Mill Lane, > Wyckoff, NJ 07481 > 201 847 7600 > www.pathologyinnovations.com > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Mon May 23 22:13:05 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs References: <104a9911046617.1046617104a991@utnet2.utmem.edu> Message-ID: <014c01c5600e$81666ff0$2423f318@yourxhtr8hvc4p> where was I? Anchovy gonads. I love anchovies, especially with pepperoni on my pizza. What can I say? It's the Italian in me. Must be getting late. Joe ----- Original Message ----- From: "Dana Marshall" To: "Philip Oshel" Cc: Sent: Monday, May 23, 2005 1:59 PM Subject: Re: [Histonet] Hair Bulbs > hi all > > I am on several listservers and this is definitely the most interesting > and the most lively. Between the anchovy gonads and the hairbulbs (and > all that has come with that), I'll just say I never delete a subject > thread anymore because you just don't know where it will go:-) > Happy staining! > > dm > > Dana R. Marshall, Ph.D. > Research Biologist, VA Medical Center > Instructor, UT Health Science Center > Memphis, TN > 901-523-8990 x7559 > > ----- Original Message ----- > From: Philip Oshel > Date: Monday, May 23, 2005 11:53 am > Subject: Re: [Histonet] Hair Bulbs > >> It's Texas, they have open season on *everything*. >> >> >Are they in season down in Texas? Or do you have open season on >> varmints?> >> >Joe Nocito wrote: >> > >> >>Can I just shoot the pathologist first? >> >> >> >>Joe >> >> >> >>-----Original Message----- >> >>From: histonet-bounces@lists.utsouthwestern.edu >> >>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >> Rittman,>>Barry R >> >>Sent: Saturday, May 21, 2005 6:21 PM >> >>To: Joe Nocito; histonet >> >>Subject: RE: [Histonet] Hair Bulbs >> >> >> >> >> >>Joe >> >>I assume that you are askig about embedding? >> >>Can't you treat the hair bulbs as whole specimens, attach them >> to a slide >> >>(agar perhaps) and then stain the entire hair bulb for fungus? >> >>Try this first and then shoot the pathologist. >> >>Barry >> >> >> >>________________________________ >> >> >> >>From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe >> Nocito>>Sent: Sat 5/21/2005 12:02 PM >> >>To: histonet >> >>Subject: [Histonet] Hair Bulbs >> >> >> >> >> >> >> >>Howdy Histoland, >> >> I swear one day I'm gonna shoot me a pathologist. We are >> diving into the >> >>realm of hair pulls by dermatologists to look for fungus. Any >> ideas on how >> >>to handle these itsy bitsy specimens? Embedding tips? We were >> successful on >> >>the first specimen, but haven't even come close since then. I'm >> pulling my >> >>hair out. >> >> We are supposed to be a nice little lab that does routine >> histology.>>Don't the "reference" part fool ya. Ummmmmm, maybe >> some one should have told >> >>this pathologist that. >> >> As always, thanks in advance for your help and the routine >> disclaimer>> "The opinions of the author do not reflect the >> opinions of his >> >>employers, their lawyers, family members, distant cousins, yada >> yada yada. >> >> >> >>Joe Nocito BS, HT(ASCP)QIHC >> >>Histology Manager >> >>Pathology Reference Lab >> >>San Antonio, TX >> >> -- >> Philip Oshel >> Supervisor, BBPIC microscopy facility >> Department of Animal Sciences >> University of Wisconsin >> 1675 Observatory Drive >> Madison, WI 53706 >> voice: (608) 263-4162 >> fax: (608) 262-5157 (dept. fax) >> http://www.ansci.wisc.edu/microscopy.htm >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jdmd77 <@t> hotmail.com Mon May 23 22:58:17 2005 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs In-Reply-To: <012801c5600d$b23a3e50$2423f318@yourxhtr8hvc4p> Message-ID: >From: "Joe Nocito" >To: "Bill Sinai" ,"histonet (E-mail)" > >Subject: Re: [Histonet] Hair Bulbs >Bill, >can you shoot pathologists down under? >Joe Joe - down under where? >----- Original Message ----- From: "Bill Sinai" > >To: "histonet (E-mail)" >Sent: Sunday, May 22, 2005 10:37 PM >Subject: [Histonet] Hair Bulbs > > >> >>Joe, >> >>Our Microbiological/Fungal experts look at scrapings from hairs, skin, >>nails >>etc. It is almost impossible to orientate hairs to get cuts at full >>length. >>Many years ago I tried and nearly went insane, and these were in intact >>skin. I achieved about one in two hundred success and even then was only >>able to get about 5-6 serial sections from each hair. >> >> >>Bill Sinai >>Laboratory Manager >>Tissue Pathology, ICPMR >>Westmead NSW 2145 >>Australia >>Ph 02 9845 7774 >> >> >>__________________________________________________________________ >> >>This electronic message and any attachments may be confidential. If you >>are not the intended recipient of this message would you please delete the >>message and any attachments and advise the sender. Western Sydney >>Area Health Services (WSAHS) uses virus scanning software but excludes >>any liability for viruses contained in any email or attachment. >> >>This email may contain privileged and confidential information intended >>only for the use of the addressees named above. If you are not the >>intended recipient of this email, you are hereby notified that any use, >>dissemination, distribution, or reproduction of this email is prohibited. >>If >>you have received this email in error, please notify WSAHS >>immediately. >> >>Any views expressed in this email are those of the individual sender >>except where the sender expressly and with authority states them >>to be the views of WSAHS. >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Tue May 24 02:02:57 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] re GFAP localisation in pwax sections Message-ID: <000901c5602e$9d730490$112b5c9f@Carlos> I also use Dako's Z334 anti GFAP. For me, a 1:2000 dilution followed by a std Dako strepABCpx-DAB after TRIS pH10 HIER is optimum. Very good on pwax sections of mouse, rat and chick and .....human! However, as was rightly advised, conditions need to be optimised locally. If you wish, have a look at some pics on this site, p4/5 in Immunohist gallery http://www.immunoportal.com From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 24 02:45:00 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] FW: Who should be doing the hiring? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD195@elht-exch1.xelht.nhs.uk> You and the Manager, keep the Medical Director well away. Hiring requires common sense not an MD. -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:55 AM To: 'histonet@lists.utsouthwester.edu' Subject: Who should be doing the hiring? Hello to all in histoland. I have been finally given another position for a full time tech. My question is, who should ultimately make the decision on who gets the position. Should it be the supervisor, the medical director or the manager? I have been the supervisor here for the last 10 years. I made the decision on who we should hire for the last position that we had open. We now have a new medical director who feels that they should have the ultimate say in the decision. I feel that I should have the last say in the matter due to the fact that I will know what type of person would be best to work in the area, and besides I have to be the one that will supervise this person. The medical director and the manager will not be in the lab for the day to day operations. I'll take any feedback regarding this situation. Thanks Allison Scott CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 24 02:52:17 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Who should be doing the hiring? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD3EC@elht-exch1.xelht.nhs.uk> Apart from the rather patronising view that "The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker", I rather agree with your views. Communication is everything and even though the hiring is the supervisor's ans manager's responsibility, it's best to involve everyone else in the exercise. Run the CV past the Medical Director, can't do any harm, probably won't understand most of it, but DON'T let the Medical Director anywhere near the interviews; they are renown for picking exactly the wrong person!! -----Original Message----- From: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 23 May 2005 18:02 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Who should be doing the hiring? Allison, Pathologist point of view. It is the pathologists job to keep you informed of their histology needs and any new programs, studies, antibodies, techniques, which will require new or additional staffing so that you can adjust your staffing in a timely manner. You should keep track of your changing workload so that you can predict need for additional staff in a timely manner. When it comes time to interview and hire you are best suited to recognize the skill or lack of it in a given candidate and how that person will fill the needs of your department. Checking references should be your job as your knowledge of histology will allow you to best understand the subliminal messages that often are the only negatives you are given. If the pathologist has the interest to call about references I would welcome his help. The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker. When it is time to make a decision I think your impressions and any impression the medical director has made should be discussed by both of you. If you have made a decision on a candidate that you are confident about, I would hope that your medical director would support you in this decision. If you have a medical director who does not care to learn about the candidates as much as he feels the need to control everything in his enviornment then this can lead to problems. Bottom line. You are responsible for the day to day workings of your lab, the working enviornment and quality of the work. You will have to live with the misery of bad hiring decision. You will be the one to feel the heat when mistakes are made or work is repeatedly late. You should be the one to decide who to hire. If you are relitively inexperienced then I would invite other input if it is a tough decision. I would foster a good working relationship with your medical director, keep him informed of your progress and when you have made your decision, discuss it with him and consider his input. If he has faith in you and he is doing his job as Director I hope he would respect your decision and allow you to do the job you were hired to do. If he feels the need to choose a different candidate which you have reservations about then I would discuss your reasoning in a collegeal manner. Explain the pros and cons of the candidates. If you feel he is making decision which will have a negative impact then your last resort is to discuss it with your lab manager or other superior, they are ultimately responible for you. Good luck Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bruyntjes <@t> voeding.tno.nl Tue May 24 04:11:34 2005 From: bruyntjes <@t> voeding.tno.nl (Bruijntjes, J.P.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] (no subject) Message-ID: <3B070848E7C2204F9DEB8BCFD767728004A69575@ntexch1.voeding.tno.nl> Thanks to all who responded to my query about GFAP and rat tissues. Joost Bruijntjes TNO Quality of Life Zeist Holland This e-mail and its contents are subject to the DISCLAIMER at http://www.tno.nl/disclaimer/email.html From ryakay <@t> shands.ufl.edu Tue May 24 06:53:33 2005 From: ryakay <@t> shands.ufl.edu (Kaye Ryan) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] microwave processing Message-ID: We have begun processing tissues using the microwave processor. For the most part, everything we process turns out beautifully but we seem to be having a problem with small endoscopic biopsies. We have begun seeing chatter but it appears to only be in the neoplastic part of the biopsy. I know that normally chatter is due to cutting techniques or over-drying of the tissue but we take the extra steps to ensure the blocks are appropriately cut. Since it seems to be localized to the neoplastic area I was wondering if this could be due to the microwave processing. We occasionally had this problem with routine processing but only occasionally. Everything else cuts very well. Has anyone heard of or experienced similar problems with microwave processing or can you offer some advice or tips as to how to troubleshoot this problem Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 From Jackie.O'Connor <@t> abbott.com Tue May 24 07:02:43 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Employment Opportunities Message-ID: I'm lucky if I remember where to go to work each morning. Ford Royer Sent by: histonet-bounces@lists.utsouthwestern.edu 05/23/2005 05:21 PM To: histonet@lists.utsouthwestern.edu cc: Subject: Re: [Histonet] Employment Opportunities After JKF was tragically shot, I thought everyone knew where Parkland Hospital was... But am I dating myself? DARNETTA MILES wrote: >Sorry.....Dallas Texas > > > >>>>"Jackie M O'Connor" 05/23/05 03:40PM >>>> >>>> >>>> > >It's nice to see where the jobs are - - but sometimes, like this time, >you guys don't include where the job is - just the state would be nice. > > >P.S. I'm not looking for a job. I love my job. I love my job. I >love my job. > > >"DARNETTA MILES" >Sent by: histonet-bounces@lists.utsouthwestern.edu >05/23/2005 03:27 PM > > To: > cc: > Subject: [Histonet] Employment Opportunities > > >Parkland Memorial Hospital >Histology Laboratory >Registered HT(ASCP) or HTL (ASCP) >Duties include: embedding, microtomy, special stains, >Immunohistochemistry stains. >competitive salary, excellent benefits, excellent working environment. >Contact - Darnetta Miles at 214-590-6652 for more information. > >Darnetta Miles, BS, MT, (ASCP) >Anatomic Pathology Manager >Parkland Health & Hospital System >Phone: 214-590-6652 >Fax: 214-590-4474 >email: dmiles@parknet.pmh.org > >This message (including any attachments) is intended only for the use >of the addressee(s) and may contain information that is privileged and >confidential. If you are the intended recipient, further disclosures >are prohibited without proper authorization. If you are not the >intended recipient or an authorized representative of the intended >recipient, the use, dissemination or reproduction of this >communication >is prohibited and may be a violation of federal or state law and >regulations. If you have received this communication in error, please >destroy all copies of the message and its attachments and notify the >sender immediately. The Dallas County Hospital District and its >affiliated entities hereby claim all applicable privileges related to >this information. > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From petepath <@t> yahoo.com Tue May 24 07:27:00 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Who should be doing the hiring? In-Reply-To: 6667 Message-ID: <20050524122700.36701.qmail@web30406.mail.mud.yahoo.com> Roger, I apologize if my remarks were patronizing. I have the great respect for histotechnologists. Our job hinges entirely on the performance of the histology lab. A key role of any supervisor is to maintain staffing and deal with personel issues. It sounds as though Allison has a new director who may not yet have gotten to know his staff well enough to let them do their job without his assistance. Or possibly he is a control freak. My point of view simply outline what I felt was a the role of the supervisor and director in the hiring process. My comment about reviewing the CV did not imply that the supervisor could not do this task equally well. It is simply a role the Director can share in. Unless the director is one of the few pathologists who has taken the time to learn histology anywhere near the level of a technologist, he will not be much help in evaluating skills and experience. This leaves him with the aspects of hiring that we do experience. Recognizing non-technical clues that may prevent hiring of an indevidual who may become a problem. She will develop a better relationship with her director he will more quickly gain confidence in her if she makes an effort now to include him in discussions. When he chills out a bit and the numerous other tasks in his job start wearing on him, there is a strong possibility that his interest "supervising histology" will take a back seat. Hopefully his confidence in Allison will let him leave her to her responsibilities without holding her hand. Stephen "Rogerson Kemlo (ELHT) Pathology" wrote: Apart from the rather patronising view that "The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker", I rather agree with your views. Communication is everything and even though the hiring is the supervisor's ans manager's responsibility, it's best to involve everyone else in the exercise. Run the CV past the Medical Director, can't do any harm, probably won't understand most of it, but DON'T let the Medical Director anywhere near the interviews; they are renown for picking exactly the wrong person!! -----Original Message----- From: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 23 May 2005 18:02 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Who should be doing the hiring? Allison, Pathologist point of view. It is the pathologists job to keep you informed of their histology needs and any new programs, studies, antibodies, techniques, which will require new or additional staffing so that you can adjust your staffing in a timely manner. You should keep track of your changing workload so that you can predict need for additional staff in a timely manner. When it comes time to interview and hire you are best suited to recognize the skill or lack of it in a given candidate and how that person will fill the needs of your department. Checking references should be your job as your knowledge of histology will allow you to best understand the subliminal messages that often are the only negatives you are given. If the pathologist has the interest to call about references I would welcome his help. The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker. When it is time to make a decision I think your impressions and any impression the medical director has made should be discussed by both of you. If you have made a decision on a candidate that you are confident about, I would hope that your medical director would support you in this decision. If you have a medical director who does not care to learn about the candidates as much as he feels the need to control everything in his enviornment then this can lead to problems. Bottom line. You are responsible for the day to day workings of your lab, the working enviornment and quality of the work. You will have to live with the misery of bad hiring decision. You will be the one to feel the heat when mistakes are made or work is repeatedly late. You should be the one to decide who to hire. If you are relitively inexperienced then I would invite other input if it is a tough decision. I would foster a good working relationship with your medical director, keep him informed of your progress and when you have made your decision, discuss it with him and consider his input. If he has faith in you and he is doing his job as Director I hope he would respect your decision and allow you to do the job you were hired to do. If he feels the need to choose a different candidate which you have reservations about then I would discuss your reasoning in a collegeal manner. Explain the pros and cons of the candidates. If you feel he is making decision which will have a negative impact then your last resort is to discuss it with your lab manager or other superior, they are ultimately responible for you. Good luck Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Irene.Campbell <@t> NorthGlasgow.Scot.NHS.UK Tue May 24 07:46:47 2005 From: Irene.Campbell <@t> NorthGlasgow.Scot.NHS.UK (Campbell, Irene) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] MOHS surgery Message-ID: <5D4FCC9BE31DD511B96600508BB9D2CF027DEB3A@northnet-09.northglasgow.scot.nhs.uk> I was wondering if anyone out there has attended the "Fundamentals of Mohs Surgery" training course held in San Diego. Any information about the course would be most helpful. Thanks Irene Irene _________________________________________________________________ The information contained within this e-mail and in any attachments is confidential and may be privileged. If you are not the intended recipient, please destroy this message, delete any copies held on your systems and notify the sender immediately. You should not retain, copy or use this e-mail for any purpose, nor disclose all or any part of its content to any other person. All messages passing through this gateway are checked for viruses but we strongly recommend that you check for viruses using your own virus scanner as NHS Greater Glasgow will not take responsibility for any damage caused as a result of virus infection. From Wanda.Smith <@t> HCAhealthcare.com Tue May 24 07:55:58 2005 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 24 08:01:49 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Who should be doing the hiring? Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD3F3@elht-exch1.xelht.nhs.uk> I did say I agreed with your views, communication is everything, but my point is different jobs should be done by different members of Staff. Supervisors and Managers should hire (and fire) that is their job and they should be trained to do it. An MD confers no skill in management and no innate ability to set on Staff. For example, with respect, my forename is Kemlo not Roger that would have been embarrassing if I had been a job candidate. -----Original Message----- From: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 24 May 2005 13:27 To: Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Who should be doing the hiring? Roger, I apologize if my remarks were patronizing. I have the great respect for histotechnologists. Our job hinges entirely on the performance of the histology lab. A key role of any supervisor is to maintain staffing and deal with personel issues. It sounds as though Allison has a new director who may not yet have gotten to know his staff well enough to let them do their job without his assistance. Or possibly he is a control freak. My point of view simply outline what I felt was a the role of the supervisor and director in the hiring process. My comment about reviewing the CV did not imply that the supervisor could not do this task equally well. It is simply a role the Director can share in. Unless the director is one of the few pathologists who has taken the time to learn histology anywhere near the level of a technologist, he will not be much help in evaluating skills and experience. This leaves him with the aspects of hiring that we do experience. Recognizing non-technical clues that may prevent hiring of an indevidual who may become a problem. She will develop a better relationship with her director he will more quickly gain confidence in her if she makes an effort now to include him in discussions. When he chills out a bit and the numerous other tasks in his job start wearing on him, there is a strong possibility that his interest "supervising histology" will take a back seat. Hopefully his confidence in Allison will let him leave her to her responsibilities without holding her hand. Stephen "Rogerson Kemlo (ELHT) Pathology" wrote: Apart from the rather patronising view that "The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker", I rather agree with your views. Communication is everything and even though the hiring is the supervisor's ans manager's responsibility, it's best to involve everyone else in the exercise. Run the CV past the Medical Director, can't do any harm, probably won't understand most of it, but DON'T let the Medical Director anywhere near the interviews; they are renown for picking exactly the wrong person!! -----Original Message----- From: Stephen Peters M.D. [mailto:petepath@yahoo.com] Sent: 23 May 2005 18:02 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Who should be doing the hiring? Allison, Pathologist point of view. It is the pathologists job to keep you informed of their histology needs and any new programs, studies, antibodies, techniques, which will require new or additional staffing so that you can adjust your staffing in a timely manner. You should keep track of your changing workload so that you can predict need for additional staff in a timely manner. When it comes time to interview and hire you are best suited to recognize the skill or lack of it in a given candidate and how that person will fill the needs of your department. Checking references should be your job as your knowledge of histology will allow you to best understand the subliminal messages that often are the only negatives you are given. If the pathologist has the interest to call about references I would welcome his help. The pathologist experience in reviewing references and recognizing the incongruities of the techs CV may help you to recognizing personal issues that may impact the performance in an otherwise well skilled worker. When it is time to make a decision I think your impressions and any impression the medical director has made should be discussed by both of you. If you have made a decision on a candidate that you are confident about, I would hope that your medical director would support you in this decision. If you have a medical director who does not care to learn about the candidates as much as he feels the need to control everything in his enviornment then this can lead to problems. Bottom line. You are responsible for the day to day workings of your lab, the working enviornment and quality of the work. You will have to live with the misery of bad hiring decision. You will be the one to feel the heat when mistakes are made or work is repeatedly late. You should be the one to decide who to hire. If you are relitively inexperienced then I would invite other input if it is a tough decision. I would foster a good working relationship with your medical director, keep him informed of your progress and when you have made your decision, discuss it with him and consider his input. If he has faith in you and he is doing his job as Director I hope he would respect your decision and allow you to do the job you were hired to do. If he feels the need to choose a different candidate which you have reservations about then I would discuss your reasoning in a collegeal manner. Explain the pros and cons of the candidates. If you feel he is making decision which will have a negative impact then your last resort is to discuss it with your lab manager or other superior, they are ultimately responible for you. Good luck Stephen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 24 08:11:25 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD1A5@elht-exch1.xelht.nhs.uk> In the UK I believe that there are recommendations, for example the number of blocks from prostatic chippings depends on their weight; but a Pathologist is best telling you the College recommendations. -----Original Message----- From: Smith Wanda [mailto:Wanda.Smith@HCAhealthcare.com] Sent: 24 May 2005 13:56 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of Blocks Submitted by PA Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue May 24 08:18:48 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] FW: Who should be doing the hiring? In-Reply-To: Message-ID: Scott, I've always had an integral part of making the decision to hire a tech, but I kept the manager and medical director informed and allowed them to be part of the interview. The lab manager was honest with me and said histology was not her area of expertise, that's why they hired me in the first place. They always deferred to my final decision. I would allow them to meet the candidates just to be courteous, but if you get bad vibes, express them until they see your side. Good luck Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Scott, Allison D Sent: Monday, May 23, 2005 11:00 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] FW: Who should be doing the hiring? -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:58 AM To: 'www.histonet@lists.utsouthwestern.edu' Subject: FW: Who should be doing the hiring? -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:56 AM To: 'www.histonet@lists.utsouthwestern.edu' Subject: FW: Who should be doing the hiring? -----Original Message----- From: Scott, Allison D Sent: Monday, May 23, 2005 10:55 AM To: 'histonet@lists.utsouthwester.edu' Subject: Who should be doing the hiring? Hello to all in histoland. I have been finally given another position for a full time tech. My question is, who should ultimately make the decision on who gets the position. Should it be the supervisor, the medical director or the manager? I have been the supervisor here for the last 10 years. I made the decision on who we should hire for the last position that we had open. We now have a new medical director who feels that they should have the ultimate say in the decision. I feel that I should have the last say in the matter due to the fact that I will know what type of person would be best to work in the area, and besides I have to be the one that will supervise this person. The medical director and the manager will not be in the lab for the day to day operations. I'll take any feedback regarding this situation. Thanks Allison Scott CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. CONFIDENTIALITY NOTICE: If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail and any attachments from your computer system. To the extent the information in this e-mail and any attachments contain protected health information as defined by the Health Insurance Portability and Accountability Act of 1996 ("HIPAA"), PL 104-191; 43 CFR Parts 160 and 164; or Chapter 181, Texas Health and Safety Code, it is confidential and/or privileged. This e-mail may also be confidential and/or privileged under Texas law. The e-mail is for the use of only the individual or entity named above. If you are not the intended recipient, or any authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Tue May 24 08:28:22 2005 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA In-Reply-To: Message-ID: Wanda, as a PA trainee, on a uterine case with tubes, ovaries and fibroids, I was taught that it depended on the number of fibroids and how large. I have submitted up to 24 blocks on a case because I was taught that some of these fibroids might turn out to be leiomyosarcomas. As for any "rules", it's just what your pathologists want to see. Each case is different and each pathologist is different. Trust me, I have 13 pathologists and each have their own way of doing things. Joe Nocito, BS, HT(ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Smith Wanda Sent: Tuesday, May 24, 2005 7:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of Blocks Submitted by PA Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From hmcleod <@t> chempath.uct.ac.za Tue May 24 08:44:11 2005 From: hmcleod <@t> chempath.uct.ac.za (Mcleod) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] CD44 v4 and v7 Message-ID: <42932FAB.5CD6C00D@chempath.uct.ac.za> Dear Histonetters A colleague is having trouble with these 2 antibodies. She has tried hier at ph 6 and 8, enzyme ar and no ar without any success. Her problem is specific stainig amidst lots of non-specific staining and when the dilution is increased the desired signal is lost and she is left with all of the nss. If anybody has successfully worked with these antibodies; any advice / help would be appreciated. She is using Novocastra CD44 v4 and Serotec CD44 v7 (MCA1731) Many many thanks. Heather From kbroomal <@t> NEMOURS.ORG Tue May 24 08:51:42 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Hair Bulbs Message-ID: <6E41111281623B4B8A9AB8F9A7EA3437824318@wlmmsx02.nemours.org> Down Under = Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Julia Dahl Sent: Monday, May 23, 2005 11:58 PM To: jnocito@satx.rr.com; bills@icpmr.wsahs.nsw.gov.au; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Hair Bulbs >From: "Joe Nocito" >To: "Bill Sinai" ,"histonet (E-mail)" > >Subject: Re: [Histonet] Hair Bulbs >Bill, >can you shoot pathologists down under? >Joe Joe - down under where? >----- Original Message ----- From: "Bill Sinai" > >To: "histonet (E-mail)" >Sent: Sunday, May 22, 2005 10:37 PM >Subject: [Histonet] Hair Bulbs > > >> >>Joe, >> >>Our Microbiological/Fungal experts look at scrapings from hairs, skin, >>nails >>etc. It is almost impossible to orientate hairs to get cuts at full >>length. >>Many years ago I tried and nearly went insane, and these were in intact >>skin. I achieved about one in two hundred success and even then was only >>able to get about 5-6 serial sections from each hair. >> >> >>Bill Sinai >>Laboratory Manager >>Tissue Pathology, ICPMR >>Westmead NSW 2145 >>Australia >>Ph 02 9845 7774 >> >> >>__________________________________________________________________ >> >>This electronic message and any attachments may be confidential. If you >>are not the intended recipient of this message would you please delete the >>message and any attachments and advise the sender. Western Sydney >>Area Health Services (WSAHS) uses virus scanning software but excludes >>any liability for viruses contained in any email or attachment. >> >>This email may contain privileged and confidential information intended >>only for the use of the addressees named above. If you are not the >>intended recipient of this email, you are hereby notified that any use, >>dissemination, distribution, or reproduction of this email is prohibited. >>If >>you have received this email in error, please notify WSAHS >>immediately. >> >>Any views expressed in this email are those of the individual sender >>except where the sender expressly and with authority states them >>to be the views of WSAHS. >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KPercival <@t> wyeth.com Tue May 24 09:19:18 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: Hi Wanda, When I worked in clinical histology and the PA put too many blocks through, the pathologists were the ones who talked to her about what and how much to submit. If the pathologists are not complaining about having to read all of the slides that your PA is submitting, then they must be getting what they want. If you feel there are too many blocks being submitted by the PA, then perhaps you should discuss that with your pathologists. See what their response is. The pathologists that I had worked for wanted to read as few slides as possible, so they worked well with the PA to get block submission to a minimun. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> "Smith Wanda" 5/24/2005 8:55:58 AM >>> Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KPercival <@t> wyeth.com Tue May 24 09:23:51 2005 From: KPercival <@t> wyeth.com (Karen Percival) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] microwave processing Message-ID: Kaye, A short soak in soapy water on a frozen cold tray cures all cutting problems associated with biospy specimens, bloody tissues, etc. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> "Kaye Ryan" 5/24/2005 7:53:33 AM >>> We have begun processing tissues using the microwave processor. For the most part, everything we process turns out beautifully but we seem to be having a problem with small endoscopic biopsies. We have begun seeing chatter but it appears to only be in the neoplastic part of the biopsy. I know that normally chatter is due to cutting techniques or over-drying of the tissue but we take the extra steps to ensure the blocks are appropriately cut. Since it seems to be localized to the neoplastic area I was wondering if this could be due to the microwave processing. We occasionally had this problem with routine processing but only occasionally. Everything else cuts very well. Has anyone heard of or experienced similar problems with microwave processing or can you offer some advice or tips as to how to troubleshoot this problem Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Tue May 24 09:38:57 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: Maybe our on-line Pathologists would have a say about this - I recall seeing several texts or reference material on how to dissect different large organs, like a radical prostate, where to take blocks from, and how many should be submitted. I know there are "industry standards". I'm out of the clinical side of pathology now - but yes, I remember it well. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Karen Percival" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/24/2005 09:19 AM To: cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Number of Blocks Submitted by PA Hi Wanda, When I worked in clinical histology and the PA put too many blocks through, the pathologists were the ones who talked to her about what and how much to submit. If the pathologists are not complaining about having to read all of the slides that your PA is submitting, then they must be getting what they want. If you feel there are too many blocks being submitted by the PA, then perhaps you should discuss that with your pathologists. See what their response is. The pathologists that I had worked for wanted to read as few slides as possible, so they worked well with the PA to get block submission to a minimun. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> "Smith Wanda" 5/24/2005 8:55:58 AM >>> Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Tue May 24 10:04:35 2005 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: Whatever the American books say - halve it, and you will sometimes be about right, but mostly, far too fussy. Most cut up protocols now, (and the Brits are getting as bad) generate much noise and little added useful information. Is it really useful: to measure a tumour in 3 planes to take n blocks of a tumour (unless a sarcoma or other rare exceptions) to send yourself batty finding nodes 2mm in diameter to have complicated formulae (which change from time to time) of so many blocks per 10 grammes of tissue etc to ink margins to see where the edge is ? ...and so on (Just head up from 40 odd blocks of a radical prostatectomy specimen, which followed 2 blocks (generous) from each of a pair of 1300 gramme boob tissue - excess to need) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Jackie M O'Connor [mailto:Jackie.O'Connor@abbott.com] Sent: 24 May 2005 15:39 To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Number of Blocks Submitted by PA Maybe our on-line Pathologists would have a say about this - I recall seeing several texts or reference material on how to dissect different large organs, like a radical prostate, where to take blocks from, and how many should be submitted. I know there are "industry standards". I'm out of the clinical side of pathology now - but yes, I remember it well. Jacqueline M. O'Connor HT(ASCP) QIHC Assistant Scientist GPRD Cancer Research Abbott Laboratories, Abbott Park, IL Jackie.OConnor@abbott.com "Karen Percival" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/24/2005 09:19 AM To: cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Number of Blocks Submitted by PA Hi Wanda, When I worked in clinical histology and the PA put too many blocks through, the pathologists were the ones who talked to her about what and how much to submit. If the pathologists are not complaining about having to read all of the slides that your PA is submitting, then they must be getting what they want. If you feel there are too many blocks being submitted by the PA, then perhaps you should discuss that with your pathologists. See what their response is. The pathologists that I had worked for wanted to read as few slides as possible, so they worked well with the PA to get block submission to a minimun. Karen Karen Percival Research Scientist I Wyeth Research 1 Burtt Road G3025 Andover, MA 01810 888-577-1500 x 4058 kpercival @wyeth.com >>> "Smith Wanda" 5/24/2005 8:55:58 AM >>> Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JEllin <@t> yumaregional.org Tue May 24 10:35:06 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] microwave processing Message-ID: I was wondering our Pathologist is wanting to go to this microwave technology and we have budgeted for 2 new processors one microwave and one that it the conventional type. My question would be that how many people out there are doing microwave processing and has anyone seen a discrepancy with immunos or specials due to the microwave processing? Jesus Ellin Yuma Regional Medical Center >>> "Kaye Ryan" 05/24/05 04:53AM >>> We have begun processing tissues using the microwave processor. For the most part, everything we process turns out beautifully but we seem to be having a problem with small endoscopic biopsies. We have begun seeing chatter but it appears to only be in the neoplastic part of the biopsy. I know that normally chatter is due to cutting techniques or over-drying of the tissue but we take the extra steps to ensure the blocks are appropriately cut. Since it seems to be localized to the neoplastic area I was wondering if this could be due to the microwave processing. We occasionally had this problem with routine processing but only occasionally. Everything else cuts very well. Has anyone heard of or experienced similar problems with microwave processing or can you offer some advice or tips as to how to troubleshoot this problem Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Tue May 24 11:00:10 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Removing OCT??? In-Reply-To: <007e01c56005$226cdd30$6a9a9618@Katri> References: <200505240019.j4O0JnNk017314@syrphus.ucdavis.edu> <007e01c56005$226cdd30$6a9a9618@Katri> Message-ID: Hi Katri, I would expect substantial deleterious damage to the cytoarchitecture using this method, as freeze-thaw is generally to be avoided ... are your experiences different? I suggest that Cathy cut frozen sections and post-fix in 10% NBF for 10 min at RT as an alternative. Thanks, Andrea At 10:06 PM -0400 5/23/05, Katri Tuomala wrote: >Hi Cathy, >Just put the OCT embedded tissue in 10% NBF and let come to room >temperature, change formalin and let fix appropriate time. > >Katri > >Katri Tuomala >Hamilton, Ontario, Canada >----- Original Message ----- From: "Catherine Stanecki" > >> >>I am new to staining tissues and I recently froze a bunch of tissues in >>OCT. However, I recently found out that it would be better to have some >>sample fixed in formalin as well so you can use mutliple staining methods. >>Is there a way to remove OCT from tissue samples without damaging the >>tissue, or is the tissue forever stuck in tissue tek once you freeze it back? >> >>Thanks! >>Cathy >> -- From jqb7 <@t> cdc.gov Tue May 24 10:58:15 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Automated special stains Message-ID: Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 From GDawson <@t> dynacaremilwaukee.com Tue May 24 11:01:39 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Automated special stains Message-ID: Dako's Artisan Stainer is great and it has a 7 minute Steiner's that works well. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Tuesday, May 24, 2005 9:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Tue May 24 11:04:47 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Automated special stains Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45DC3@sjhaexc02.sjha.org> We have the DAKO Artisan and are pleased with the company service as well as the stains. GMS is good. We use the Warthin Starry - not sure they have a Steiner. If there would be one change, I would wish that the stains could be started as ordered instead of by batch. Hope this helps! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Tuesday, May 24, 2005 11:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From krat18 <@t> aol.com Tue May 24 11:30:03 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] GOOD markers/pencils for cassettes In-Reply-To: References: Message-ID: <8C72E94F8EEA07D-E44-6A28@mblk-d37.sysops.aol.com> HELP! HELP! HELP! We are having lots of problems with our markers (we're now using StatLab). The writing smears and smudges when we fix tissues in PenFix, from a combination of the grease from the fat and the alcohol in the PenFix. What is everyone using to mark cassettes that will be fixed in alcohol-based fixatives? ANY suggestions are welcome! We cannot at this time buy a cassette marker machine. Are there pencils that work well? Our Tissue Tek pencils used to smear also, but maybe there are better ones? From Heather.A.Harper <@t> pcola.med.navy.mil Tue May 24 11:24:29 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Grossing Message-ID: <807FE48C5A7CC940B973B58D32E701431991BE4E@nhpens-exch1.pcola.med.navy.mil> Currently I am grossing in the small specimens and I do all the miscellaneous stuff, plus all my paper work that a supervisor has to do.. Grossing consumes 2 hrs of my morning and generally do not start until between 8:30-9:00. My shift ends at 2:30. My co-worker works the night shift (10-6) and embeds, cuts and stains (literally). I want to get some opinions. Since I already accession, gross, etc... Is it fair for the pathologists to want me also to do the recuts and special stains? I feel an enormous pressure about this issue and I simply can't understand why some pathologists act like it's so detrimental to be able to view slides and get recuts/special stains the same day. I do not want to be rushed through grossing to accommodate 2 of the 3 pathologists. Am I right or wrong for feeling this way. I need feedback. Heather A. Harper Naval Hospital of Pensacola, FL Supervisor Histology/Morgue From ryakay <@t> shands.ufl.edu Tue May 24 11:47:52 2005 From: ryakay <@t> shands.ufl.edu (Kaye Ryan) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] microwave processing Message-ID: Hi Jesus, We have been doing the microwave processing for a while now and have seen no change in the IHC staining patterns at all. As well, our routine H&E was not altered in any way. The TAT cannot be beat and the cutting of the tissues is much easier. I have only had this "chatter" issue come up since a hospital merged with our lab several months ago. Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 >>> "Jesus Ellin" 05/24/05 11:35 AM >>> I was wondering our Pathologist is wanting to go to this microwave technology and we have budgeted for 2 new processors one microwave and one that it the conventional type. My question would be that how many people out there are doing microwave processing and has anyone seen a discrepancy with immunos or specials due to the microwave processing? Jesus Ellin Yuma Regional Medical Center >>> "Kaye Ryan" 05/24/05 04:53AM >>> We have begun processing tissues using the microwave processor. For the most part, everything we process turns out beautifully but we seem to be having a problem with small endoscopic biopsies. We have begun seeing chatter but it appears to only be in the neoplastic part of the biopsy. I know that normally chatter is due to cutting techniques or over-drying of the tissue but we take the extra steps to ensure the blocks are appropriately cut. Since it seems to be localized to the neoplastic area I was wondering if this could be due to the microwave processing. We occasionally had this problem with routine processing but only occasionally. Everything else cuts very well. Has anyone heard of or experienced similar problems with microwave processing or can you offer some advice or tips as to how to troubleshoot this problem Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Tue May 24 11:58:53 2005 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] freezing of specimens in surgery center Message-ID: <000d01c56081$e0dd14c0$a7d48a80@AMY> Hello Histonetters I have a unique question. We are currently starting to set up procedure for collecting samples from a clinical trial. The clinical trial involves taking multiple synovial biopsies at a surgery center. Since portions of the samples need to be processed for frozen sections we wanted to be able to freeze the specimens at the surgery center via isopentane cooled liquid nitrogen. We really do not want to have to transport the multiple specimens back to the main lab prior to freezing due to the time involved it would probably be 1-2 hours post biopsy before we could freeze the samples. The surgery center is questioning the flammability of the isopentane. Has anyone encountered anything like this? Any suggestions would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From BetStark <@t> aol.com Tue May 24 12:00:58 2005 From: BetStark <@t> aol.com (BetStark@aol.com) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Microm cryostat Message-ID: <2b.73c2dcbf.2fc4b7ca@aol.com> Interested in the pros and cons of the Microm cryostat. We are a small hospital with an active surgical staff considering buying a new cryostat. Any information will be greatly appreciated. Thanks From gcallis <@t> montana.edu Tue May 24 12:41:28 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:07 2005 Subject: There are Re: [Histonet] GOOD markers/pencils for cassettes In-Reply-To: <8C72E94F8EEA07D-E44-6A28@mblk-d37.sysops.aol.com> References: <8C72E94F8EEA07D-E44-6A28@mblk-d37.sysops.aol.com> Message-ID: <6.0.0.22.1.20050524113518.01b27b68@gemini.msu.montana.edu> WE have switched to Cancer Diagnostics Moist Mark Plus, www.cancerdiagnostics.com, Item # MP2100, case of 10 (cheaper too!). Callthem at 877 846-5393 to see if they will send a sample. If not, try the next source below. www.marketlabinc.com, get their 800# they will send you samples to try first before you take the big plunge and buy them to ensure they work in your alcoholic based fixative. We love them! They graciously sent them to me via private email contact, and hopefully looking in on this message. A #2 pencil does the job, just a nice soft lead, but not quite as smeary as the Tissue Tek pencils. Good luck Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue May 24 12:48:24 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:07 2005 Subject: Fwd: Re: [Histonet] freezing of specimens in surgery center Message-ID: <6.0.0.22.1.20050524114813.01b5ccb0@gemini.msu.montana.edu> >Date: Tue, 24 May 2005 11:47:57 -0600 >To: "Elizabeth Chlipala" >From: Gayle Callis >Subject: Re: [Histonet] freezing of specimens in surgery center > >Liz, > >They great cause for concern. They will have to have an explosion proof >freezer for storage. It is highly explosive, flammable. Histologic had >the disaster article on what happened at U of Arkansas at 2 am one morning >written by Bob Skinner. Donna Montague sent me photos also. When >isopentane fumes due to improper strorage in a regular freezer were >exposed to spark of freezer cycling, it blew up and burned the lab, cost >the university $250,000 to get it all squared away and it took out some >very expensive furniture, melted metal, etc. Luckily no one was killed >OR injured. > >At 10:58 AM 5/24/2005, you wrote: >>Hello Histonetters >> >>I have a unique question. We are currently starting to set up procedure >>for collecting samples from a clinical trial. The clinical trial >>involves taking multiple synovial biopsies at a surgery center. Since >>portions of the samples need to be processed for frozen sections we >>wanted to be able to freeze the specimens at the surgery center via >>isopentane cooled liquid nitrogen. We really do not want to have to >>transport the multiple specimens back to the main lab prior to freezing >>due to the time involved it would probably be 1-2 hours post biopsy >>before we could freeze the samples. The surgery center is questioning >>the flammability of the isopentane. Has anyone encountered anything >>like this? Any suggestions would be helpful. Thanks in advance. >> >>Liz >> >>Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC >>Manager >>Premier Laboratory, LLC >>P.O. Box 18592 >>Boulder, Colorado 80308 >>Office: (303) 735-5001 >>Fax: (303) 735-3540 >>liz@premierlab.com >>www.premierlab.com >> >>Ship to Address: >>Premier Laboratory >>University of Colorado >>MCDB, Room A3B40 >>Boulder, Colorado 80309 >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From petepath <@t> yahoo.com Tue May 24 13:07:47 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: <20050524180747.5177.qmail@web30408.mail.mud.yahoo.com> There are a number of excellent references such as the Manual of Surgical Pathology by Susan Lester MD or the index of Rosai's Surgical pathology which offer recommendations on number of sections to be sampled in the common specimens. Depending on the gross findings a pathologist will take additional sections as he sees fit depending on the gross. There are many cases that do not fall into these typical cases and the pathologist or PA will section them as based on their impression of the case. When the pathologist is grossing a case he often has a good idea of what he will need for sections and can be conservative if that is his nature. It is more difficult for the PA to second guess the pathologist so when the PA is unsure how much the pathologist will want they tend to put a bit more through initially so they do not have to go back a second time leading to more work and a delay of the case. The number of blocks will often be inversly proportional to their level of experience of both the PA and the pathologist. A point I try to stress with my group is that even though they may think putting through " more than necessary " may seem like an innocuous practice, in fact overloading the histology staff will stress the staff and can lead to potential mistakes as well as a delay in the work. This will effect the entire department. The same logic applies to ordering of specials and immunos by the less experienced pathologists. If you recognize a trend of increase work coming from a indevidual PA, or pathologist it may be worth discussing it with the medical director to see if their is valid reason to further educate these indeviduals. Stephen From sjchtascp <@t> yahoo.com Tue May 24 13:37:05 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] slide drying Message-ID: <20050524183705.23699.qmail@web90204.mail.scd.yahoo.com> I'm setting up a small Histo lab and using a 60C slide warmer to dry my slides. I've been currently allowing them to air dry vertically for a couple of hours to overnight, then placing the slides on the slide warmer for an hour. About 90% of the time the slides turn out ok, but some slide do show some detaching from the slide. Both charges and non-charged slides with water bath adhesive seem to be involved. I read a past artical on this site not recommending allowing slides to air dry as the section may entrap water? Any suggestions. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! From bmcmahill <@t> incytepathology.com Tue May 24 13:38:27 2005 From: bmcmahill <@t> incytepathology.com (Bonnie J. McMahill) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Automated special stains Message-ID: We've used the Artisan since it was first marketed (since 2000 or so). It does a great job, cost effective, and can vary times/temp according to how you want the stain to look. You do have to yse their reagents, but the costs work out to be very reasonable. The silver stains are great. We use the Warthin Starry for H.pylori and the pathologists love it! Bonnie McMahill Histology Supervisor InCyte Pathology Spokane, WA > -----Original Message----- > From: Bartlett, Jeanine [SMTP:jqb7@cdc.gov] > Sent: Tuesday, May 24, 2005 8:58 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Automated special stains > > Hi everybody: > > I'd like feedback on the automated special stain equipment being used > today. I need something that does all the traditional stains but also > silvers...mainly GMS' and Steiner's. I am most interested in an open > system so as not to be locked into using a particular vendor's reagents > if possible. > > Vendors welcome to email as well. > > Thanks, > Jeanine Bartlett, HT(ASCP) > Centers for Disease Control and Prevention > Infectious Disease Pathology Activity > 1600 Clifton Road, MS/G-32 > Atlanta, GA 30333 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mclarke <@t> allsaintshealthcare.org Tue May 24 13:42:34 2005 From: mclarke <@t> allsaintshealthcare.org (Clarke, Mary) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Good pen/markers Message-ID: <6FE2A453DA437F4D96FAAF363F20ABB20FC845@WFEXBE04.wfsi.priv> We use the Richard-Allan Laboratory Marking Pens for the cassettes and they are great. For slides we use Cancer Diagnostics Moist Mark Plus pen and they are equally as good. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, May 24, 2005 12:02 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 18, Issue 34 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Automated special stains (Bartlett, Jeanine) 2. RE: Automated special stains (Dawson, Glen) 3. RE: Automated special stains (Weems, Joyce) 4. GOOD markers/pencils for cassettes (krat18@aol.com) 5. Grossing (Heather.A.Harper@pcola.med.navy.mil) 6. Re: microwave processing (Kaye Ryan) 7. freezing of specimens in surgery center (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Tue, 24 May 2005 11:58:15 -0400 From: "Bartlett, Jeanine" Subject: [Histonet] Automated special stains To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 ------------------------------ Message: 2 Date: Tue, 24 May 2005 11:01:39 -0500 From: "Dawson, Glen" Subject: RE: [Histonet] Automated special stains To: "Bartlett, Jeanine" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dako's Artisan Stainer is great and it has a 7 minute Steiner's that works well. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Tuesday, May 24, 2005 9:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Tue, 24 May 2005 12:04:47 -0400 From: "Weems, Joyce" Subject: RE: [Histonet] Automated special stains To: "Bartlett, Jeanine" , Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45DC3@sjhaexc02.sjha.org> Content-Type: text/plain; charset="iso-8859-1" We have the DAKO Artisan and are pleased with the company service as well as the stains. GMS is good. We use the Warthin Starry - not sure they have a Steiner. If there would be one change, I would wish that the stains could be started as ordered instead of by batch. Hope this helps! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Tuesday, May 24, 2005 11:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ Message: 4 Date: Tue, 24 May 2005 12:30:03 -0400 From: krat18@aol.com Subject: [Histonet] GOOD markers/pencils for cassettes To: histonet@lists.utsouthwestern.edu Message-ID: <8C72E94F8EEA07D-E44-6A28@mblk-d37.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" HELP! HELP! HELP! We are having lots of problems with our markers (we're now using StatLab). The writing smears and smudges when we fix tissues in PenFix, from a combination of the grease from the fat and the alcohol in the PenFix. What is everyone using to mark cassettes that will be fixed in alcohol-based fixatives? ANY suggestions are welcome! We cannot at this time buy a cassette marker machine. Are there pencils that work well? Our Tissue Tek pencils used to smear also, but maybe there are better ones? ------------------------------ Message: 5 Date: Tue, 24 May 2005 11:24:29 -0500 From: Heather.A.Harper@pcola.med.navy.mil Subject: [Histonet] Grossing To: histonet@lists.utsouthwestern.edu Message-ID: <807FE48C5A7CC940B973B58D32E701431991BE4E@nhpens-exch1.pcola.med.navy.mi l> Content-Type: text/plain Currently I am grossing in the small specimens and I do all the miscellaneous stuff, plus all my paper work that a supervisor has to do.. Grossing consumes 2 hrs of my morning and generally do not start until between 8:30-9:00. My shift ends at 2:30. My co-worker works the night shift (10-6) and embeds, cuts and stains (literally). I want to get some opinions. Since I already accession, gross, etc... Is it fair for the pathologists to want me also to do the recuts and special stains? I feel an enormous pressure about this issue and I simply can't understand why some pathologists act like it's so detrimental to be able to view slides and get recuts/special stains the same day. I do not want to be rushed through grossing to accommodate 2 of the 3 pathologists. Am I right or wrong for feeling this way. I need feedback. Heather A. Harper Naval Hospital of Pensacola, FL Supervisor Histology/Morgue ------------------------------ Message: 6 Date: Tue, 24 May 2005 12:47:52 -0400 From: "Kaye Ryan" Subject: Re: [Histonet] microwave processing To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Jesus, We have been doing the microwave processing for a while now and have seen no change in the IHC staining patterns at all. As well, our routine H&E was not altered in any way. The TAT cannot be beat and the cutting of the tissues is much easier. I have only had this "chatter" issue come up since a hospital merged with our lab several months ago. Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 >>> "Jesus Ellin" 05/24/05 11:35 AM >>> I was wondering our Pathologist is wanting to go to this microwave technology and we have budgeted for 2 new processors one microwave and one that it the conventional type. My question would be that how many people out there are doing microwave processing and has anyone seen a discrepancy with immunos or specials due to the microwave processing? Jesus Ellin Yuma Regional Medical Center >>> "Kaye Ryan" 05/24/05 04:53AM >>> We have begun processing tissues using the microwave processor. For the most part, everything we process turns out beautifully but we seem to be having a problem with small endoscopic biopsies. We have begun seeing chatter but it appears to only be in the neoplastic part of the biopsy. I know that normally chatter is due to cutting techniques or over-drying of the tissue but we take the extra steps to ensure the blocks are appropriately cut. Since it seems to be localized to the neoplastic area I was wondering if this could be due to the microwave processing. We occasionally had this problem with routine processing but only occasionally. Everything else cuts very well. Has anyone heard of or experienced similar problems with microwave processing or can you offer some advice or tips as to how to troubleshoot this problem Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 24 May 2005 10:58:53 -0600 From: "Elizabeth Chlipala" Subject: [Histonet] freezing of specimens in surgery center To: Message-ID: <000d01c56081$e0dd14c0$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Hello Histonetters I have a unique question. We are currently starting to set up procedure for collecting samples from a clinical trial. The clinical trial involves taking multiple synovial biopsies at a surgery center. Since portions of the samples need to be processed for frozen sections we wanted to be able to freeze the specimens at the surgery center via isopentane cooled liquid nitrogen. We really do not want to have to transport the multiple specimens back to the main lab prior to freezing due to the time involved it would probably be 1-2 hours post biopsy before we could freeze the samples. The surgery center is questioning the flammability of the isopentane. Has anyone encountered anything like this? Any suggestions would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 34 **************************************** From gcallis <@t> montana.edu Tue May 24 14:41:48 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Re: slide drying In-Reply-To: <20050524183705.23699.qmail@web90204.mail.scd.yahoo.com> References: <20050524183705.23699.qmail@web90204.mail.scd.yahoo.com> Message-ID: <6.0.0.22.1.20050524133412.01b628b0@gemini.msu.montana.edu> Steve, What tissues are coming off the slides? At 12:37 PM 5/24/2005, you wrote: >I'm setting up a small Histo lab and using a 60C slide warmer to dry my >slides. I've been currently allowing them to air dry vertically for a >couple of hours to overnight, then placing the slides on the slide warmer >for an hour. About 90% of the time the slides turn out ok, but some slide >do show some detaching from the slide. Both charges and non-charged >slides with water bath adhesive seem to be involved. I read a past >artical on this site not recommending allowing slides to air dry as the >section may entrap water? Any suggestions. > >Steve > > > >--------------------------------- >Do You Yahoo!? > Yahoo! Small Business - Try our new Resources site! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From dpahisto <@t> yahoo.com Tue May 24 15:28:56 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Re: Number of blocks Message-ID: <20050524202856.78056.qmail@web33404.mail.mud.yahoo.com> We have the same problem. Our pathologist can do a uterus/tubes/ovaries in 5 blocks and it takes the PA 12. Also our docs can do a placenta in 2 blocks and the PA-4. We have complained. I have created a report comparing like cases and the number of blocks put through and this has gotten me nowhere. So we just cut, and leave it up to the pathologist to complain about how many slides he is getting - then we tell him to talk to the PA. Cindy DuBois Delta Pathology Stockton CA __________________________________ Do you Yahoo!? Yahoo! Small Business - Try our new Resources site http://smallbusiness.yahoo.com/resources/ From Janet.Bonner <@t> FLHOSP.ORG Tue May 24 15:36:37 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] GOOD markers/pencils for cassettes Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB428A@fh2k093.fhmis.net> Statlab pens work great on the slides, but we can't use them on the cassettes. We use the Sanford Sharpie Industrial fine point pen (available from Office Depot)...It has super permanent ink and works fantastically. Janet -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 5/24/2005 12:30 PM Subject: [Histonet] GOOD markers/pencils for cassettes HELP! HELP! HELP! We are having lots of problems with our markers (we're now using StatLab). The writing smears and smudges when we fix tissues in PenFix, from a combination of the grease from the fat and the alcohol in the PenFix. What is everyone using to mark cassettes that will be fixed in alcohol-based fixatives? ANY suggestions are welcome! We cannot at this time buy a cassette marker machine. Are there pencils that work well? Our Tissue Tek pencils used to smear also, but maybe there are better ones? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue May 24 15:40:27 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] freezing of specimens in surgery center References: <000d01c56081$e0dd14c0$a7d48a80@AMY> Message-ID: <4293913B.8F4A9459@uwo.ca> Isopentane is volatile and flammable (to about the same extent as diethyl ether). A safer way for the surgeons to freeze small (2mm) specimens would for you to provide some cryostat chucks packed in dry ice. Small bits of tissue freeze pretty quickly (a few seconds) on contact with metal at -80C. The chucks with mounted specimens can then be put in airtight plastic bags or specimen tubes and kept in a -20C freezer. I've no expperience with synovial tissue. We used to use the above method for muscle. There were usually some ice crystal holes, but they were very small and didn't interfere with fibre typing (ATPase) or measuring the cross sectional areas of the muscle fibres. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Elizabeth Chlipala wrote: > > Hello Histonetters > > I have a unique question. We are currently starting to set up procedure > for collecting samples from a clinical trial. The clinical trial > involves taking multiple synovial biopsies at a surgery center. Since > portions of the samples need to be processed for frozen sections we > wanted to be able to freeze the specimens at the surgery center via > isopentane cooled liquid nitrogen. We really do not want to have to > transport the multiple specimens back to the main lab prior to freezing > due to the time involved it would probably be 1-2 hours post biopsy > before we could freeze the samples. The surgery center is questioning > the flammability of the isopentane. Has anyone encountered anything > like this? Any suggestions would be helpful. Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 From la.sebree <@t> hosp.wisc.edu Tue May 24 15:56:59 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Immunostaining of suspected CJD infected tissue Message-ID: I am wondering if there are any labs out there that will perform immunostains on formic acid treated tissue suspicious for being infected with CJD. Thanks, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From lu_ze <@t> sbcglobal.net Tue May 24 16:17:20 2005 From: lu_ze <@t> sbcglobal.net (Ze Lu) Date: Fri Sep 16 15:25:07 2005 Subject: [Histonet] Fume hood inqury Message-ID: <00ba01c560a5$f8644b80$1302a8c0@OPTIMUM2> Dear all, We are thinking to move to a new lab space but there is no fume hood built in there. Is fume hood required when doing general histological work including tissue process, embedding and staining? By the way, we also have a HPLC system. Anybody know the requirment for HPLC running (not sample preparation which must be done under hood, just HPLC machine). I checked the OSHA web site but can not find useful info. Any helps? Thanks. Ze Lu, Ph.D. Research Scientis Optimum Therapeutics, From cbass <@t> bidmc.harvard.edu Tue May 24 17:54:05 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] HER-2 ICC protocol for breast cancer cell lines Message-ID: <55B639C4-2C99-44CE-8EFC-073C2491CCCE@bidmc.harvard.edu> Hello, I am attempting to stain a couple of breast cancer cell lines for HER-2. Does anyone have a protocol that I could use? I am trying to avoid paraffin embedding as I am not set up to do this. I am using MCF-7, BT474 and MDA-MB-453 cells. Any help would be appreciated. I am relatively new to immunostaining, so even if there is no advice for HER-2 in particular, I would like to hear general staining advice for these cell lines. I think I have to do an antigen retrieval step, but am not sure how to modify the FFPE protocols that I have seen. Thanks, Caroline Bass From alineumann <@t> aol.com Tue May 24 19:24:09 2005 From: alineumann <@t> aol.com (alineumann@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Pathology Assistant books Message-ID: <8C72ED734591789-8E8-20C66@FWM-D41.sysops.aol.com> Does anyone have any suggestions for good Path Asst. textbooks? Thanks From Linitistwo <@t> aol.com Tue May 24 21:37:08 2005 From: Linitistwo <@t> aol.com (Linitistwo@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] dissection table Message-ID: <1ec.3baf925e.2fc53ed4@aol.com> anybody looking for a like new jewitt dissecting station. i have one i want to get rid of. onlu used 2 yrs, like new. gary From jnocito <@t> satx.rr.com Tue May 24 21:49:08 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Automated special stains References: Message-ID: <01ab01c560d4$535e7640$2423f318@yourxhtr8hvc4p> kinda leaves out the Nexus, doesn't it? Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Bartlett, Jeanine" To: Sent: Tuesday, May 24, 2005 10:58 AM Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Tue May 24 21:56:11 2005 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Grossing References: <807FE48C5A7CC940B973B58D32E701431991BE4E@nhpens-exch1.pcola.med.navy.mil> Message-ID: <01c801c560d5$4fe363d0$2423f318@yourxhtr8hvc4p> Heather, it depends on the number of recuts and specials. Before I became a PA trainee, I was doing everything also and it was tough, not going to deny it. I had to prioritize tasks and let the paperwork go until the end of the day or do it while the slides were in the oven or staining. Often, I came in over the weekend to catch up on paperwork. I was told that hiring another person was out of the question, until I ended up in the hospital with heart problems. It got their attention, but I don't recommend it. I would prioritize and do the best you can do. Don't run yourself into the ground, no job is worth sacrificing your health. Trust me, I know. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Tuesday, May 24, 2005 11:24 AM Subject: [Histonet] Grossing > Currently I am grossing in the small specimens and I do all the > miscellaneous stuff, plus all my paper work that a supervisor has to do.. > Grossing consumes 2 hrs of my morning and generally do not start until > between 8:30-9:00. My shift ends at 2:30. My co-worker works the night > shift > (10-6) and embeds, cuts and stains (literally). I want to get some > opinions. > Since I already accession, gross, etc... Is it fair for the pathologists > to > want me also to do the recuts and special stains? I feel an enormous > pressure about this issue and I simply can't understand why some > pathologists act like it's so detrimental to be able to view slides and > get > recuts/special stains the same day. I do not want to be rushed through > grossing to accommodate 2 of the 3 pathologists. Am I right or wrong for > feeling this way. I need feedback. > > > > Heather A. Harper > > Naval Hospital of Pensacola, FL > > Supervisor Histology/Morgue > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kwuny <@t> email.cs.nsw.gov.au Tue May 24 20:08:14 2005 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] CD44 v4 and v7 In-Reply-To: <42932FAB.5CD6C00D@chempath.uct.ac.za> Message-ID: <200505251104108.SM01536@crgcsls814> Hi Heather, I am not sure about CD44 v4 & v7, but I tried CD44 v6 antibody (dil. 1/500) from R&D Systems (Cat No.BRA13). I obtained good results with HIER with EDTA (pH 8) or Citrate buffer. Regards, Young Young Kwun Senior Hospital Scientist Dept of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Phone:61-2-9767 6075 Fax:61-2-9767 8427 Email:kwuny@email.cs.nsw.gov.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mcleod Sent: Tuesday, 24 May 2005 11:44 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CD44 v4 and v7 Dear Histonetters A colleague is having trouble with these 2 antibodies. She has tried hier at ph 6 and 8, enzyme ar and no ar without any success. Her problem is specific stainig amidst lots of non-specific staining and when the dilution is increased the desired signal is lost and she is left with all of the nss. If anybody has successfully worked with these antibodies; any advice / help would be appreciated. She is using Novocastra CD44 v4 and Serotec CD44 v7 (MCA1731) Many many thanks. Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Sydney South West Area Health Service." From ABEE <@t> pml.ac.uk Wed May 25 02:58:38 2005 From: ABEE <@t> pml.ac.uk (Amanda Beesley) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Kulzer histoknives Message-ID: <200505250858.AA293798156@mail.pml.ac.uk> Does anyone know of a company in the UK who sell Kulzer histoknives and holders? regards Amanda Beesley -- Miss Amanda Beesley Cell Biologist/ Ecotoxicologist Plymouth Marine Laboratory Prospect Place West Hoe Plymouth Devon PL4 7PA United Kingdom Direct Dial 01752 633421/ 01752 633486 01752 633100 Website: www.pml.ac.uk Registered Charity No. 1091222 Company No. 4178503 ------------------------------------------------- This e-mail, its content and any file attachments are confidential. If you have received this e-mail in error please do not copy, disclose it to any third party or use the contents or attachments in any way. Please notify the sender by replying to this e-mail or e-mail forinfo@pml.ac.uk and then delete the email without making any copies or using it in any other way. The content of this message may contain personal views which are not the views of Plymouth Marine Laboratory unless specifically stated. Email transmission cannot be guaranteed to be secure as information may be intercepted, corrupted, lost, destroyed, arrive late or incomplete or contain viruses. Plymouth Marine Laboratory accepts no liability for any loss or damage which may be caused by software viruses. -- --- [This E-mail scanned for viruses by Declude Virus] From ABEE <@t> pml.ac.uk Wed May 25 02:59:38 2005 From: ABEE <@t> pml.ac.uk (Amanda Beesley) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Reichert Jung knifemaker Message-ID: <200505250859.AA384893214@mail.pml.ac.uk> We have a Reichert Jung knifemaker that needs to be repaired or replaced. Can anyone tell me where this can be carried out, (preferably the UK). it is for methacrylate sections. If it cannot be repaired, what would you recommend? Regards, Amanda Beesley -- Miss Amanda Beesley Cell Biologist/ Ecotoxicologist Plymouth Marine Laboratory Prospect Place West Hoe Plymouth Devon PL4 7PA United Kingdom Direct Dial 01752 633421/ 01752 633486 01752 633100 Website: www.pml.ac.uk Registered Charity No. 1091222 Company No. 4178503 ------------------------------------------------- This e-mail, its content and any file attachments are confidential. If you have received this e-mail in error please do not copy, disclose it to any third party or use the contents or attachments in any way. Please notify the sender by replying to this e-mail or e-mail forinfo@pml.ac.uk and then delete the email without making any copies or using it in any other way. The content of this message may contain personal views which are not the views of Plymouth Marine Laboratory unless specifically stated. Email transmission cannot be guaranteed to be secure as information may be intercepted, corrupted, lost, destroyed, arrive late or incomplete or contain viruses. Plymouth Marine Laboratory accepts no liability for any loss or damage which may be caused by software viruses. -- --- [This E-mail scanned for viruses by Declude Virus] From Diane.Gladney <@t> se.amedd.army.mil Wed May 25 05:23:31 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Grossing Message-ID: <4D55B2E997EFAE4DA6081DDE100B8302554B9D@amedmlsermc133.amed.ds.army.mil> Heather, I feel your pain. I do grossing of all the small things, also. I am the "Histology Supervisor" but I am the only civilian in my department and the only histotechnologist. I have a military member to embed and help cut blocks in the morning. I do all the special stains, recuts, etc. I also do all the FNA's as we no longer have cytotechnologists (our Pap Smears are shipped out). The pathologist looks at the non-gyn and FNA slides to give the diagnosis. My day starts at 6:00 am and I am suppose to get off of work at 2:45 but that rarely happens (I don't get overtime or comp. time either). I have all the duties of the other lab dept. supervisors but they do not do any bench work. They are also 3 grades higher than me. My paper work has stacked up so high on my desk that I can't even see the surface of my desk. I got told yesterday by our pathologist that he wants me to start keeping up with his QA cases that he sends out for review since our lab QA person said that he doesn't have time to do it!! I showed him my desk and told him that I have 4 SOP's to write, 5 SOP's to revise, tons of other administrative paper work to do (eventually), and a long list of just general lab work to do (filing slides, disposing of waste tissue, etc). His attitude is that I should take this on as an additional duty since it belongs to Histology. I am at my wits end and leave work each day exhausted. I stay behind more than I care and some days I just want to scream. I have tried to talk with the pathologist and the lab manager (both military) but it all falls on deaf ears. It seems that military hospitals everywhere are treating their histo techs this way. If I didn't have 25 1/2 years of federal civilian service, I would leave here for a better job where I am truly appreciated and not worked like a mule. The fact that I am well into my 50's doesn't help me getting hired somewhere else. So I guess that I will just try to ride it out and try to keep my wits about me. Good luck in trying to get your pathologists to see the light. Thanks for letting me vent my frustrations, also. Keep Smiling, Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Tuesday, May 24, 2005 12:24 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Grossing Currently I am grossing in the small specimens and I do all the miscellaneous stuff, plus all my paper work that a supervisor has to do.. Grossing consumes 2 hrs of my morning and generally do not start until between 8:30-9:00. My shift ends at 2:30. My co-worker works the night shift (10-6) and embeds, cuts and stains (literally). I want to get some opinions. Since I already accession, gross, etc... Is it fair for the pathologists to want me also to do the recuts and special stains? I feel an enormous pressure about this issue and I simply can't understand why some pathologists act like it's so detrimental to be able to view slides and get recuts/special stains the same day. I do not want to be rushed through grossing to accommodate 2 of the 3 pathologists. Am I right or wrong for feeling this way. I need feedback. Heather A. Harper Naval Hospital of Pensacola, FL Supervisor Histology/Morgue _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Wed May 25 07:22:02 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Reichert Jung knifemaker Message-ID: Hi Amanda, You might want to try the Leica office in the UK. They should be able a solution. The Reichert Jung label is now part of the Leica-Microsystems company. Leica Microsystems (UK) Ltd Davy Avenue Knowlhill MK5 8LB Bucks Milton Keynes Tel.:+44 1 908 246 246 Fax.:+44 1 908 609992 Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Amanda Beesley" To: Sent by: cc: (bcc: Don Birgerson/USDER/West/Leica) histonet-bounces@lists.utsouth Subject: [Histonet] Reichert Jung knifemaker western.edu 05/25/2005 02:59 AM Please respond to ABEE We have a Reichert Jung knifemaker that needs to be repaired or replaced. Can anyone tell me where this can be carried out, (preferably the UK). it is for methacrylate sections. If it cannot be repaired, what would you recommend? Regards, Amanda Beesley -- Miss Amanda Beesley Cell Biologist/ Ecotoxicologist Plymouth Marine Laboratory Prospect Place West Hoe Plymouth Devon PL4 7PA United Kingdom Direct Dial 01752 633421/ 01752 633486 01752 633100 Website: www.pml.ac.uk Registered Charity No. 1091222 Company No. 4178503 ------------------------------------------------- This e-mail, its content and any file attachments are confidential. If you have received this e-mail in error please do not copy, disclose it to any third party or use the contents or attachments in any way. Please notify the sender by replying to this e-mail or e-mail forinfo@pml.ac.uk and then delete the email without making any copies or using it in any other way. The content of this message may contain personal views which are not the views of Plymouth Marine Laboratory unless specifically stated. Email transmission cannot be guaranteed to be secure as information may be intercepted, corrupted, lost, destroyed, arrive late or incomplete or contain viruses. Plymouth Marine Laboratory accepts no liability for any loss or damage which may be caused by software viruses. -- --- [This E-mail scanned for viruses by Declude Virus] _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From cgfields <@t> lexhealth.org Wed May 25 07:30:34 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Isopentane Message-ID: Yesterday on the Histonet someone referred to an article in the Histologic by Bob Skinner about an accident at the University of Arkansas involving Isopentane. I pulled the Histologic off line (Dec. 1999) but did not find the information. Do you happen to know where the article might be found? The institution I previously worked for uses Isopentane and I would like to send the article to them. Thank you for any information you can give me. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From SSouder <@t> mc.utmck.edu Wed May 25 08:06:07 2005 From: SSouder <@t> mc.utmck.edu (Souder, Starla J.) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] FW: UNSUBSCRIBE Message-ID: <8B799916C6062E4DBB1A6A9E3CD3EFB0A87408@msexch3.mc.utmck.edu> > -----Original Message----- > From: Souder, Starla J. > Sent: Tuesday, May 24, 2005 9:59 AM > To: 'HISTONET@LISTS.UTSOUTHWESTERN.EDU' > Subject: UNSUBSCRIBE > > From mbecker <@t> pathlabinc.com Wed May 25 08:08:45 2005 From: mbecker <@t> pathlabinc.com (Michelle Becker) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Isopentane In-Reply-To: Message-ID: I believe the article you are referring to is on the first page of the May 2003 Histologic entitled "More Than Just a Mandatory Exercise" by Robert Skinner, HTL. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Carole Fields Sent: Wednesday, May 25, 2005 7:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Isopentane Yesterday on the Histonet someone referred to an article in the Histologic by Bob Skinner about an accident at the University of Arkansas involving Isopentane. I pulled the Histologic off line (Dec. 1999) but did not find the information. Do you happen to know where the article might be found? The institution I previously worked for uses Isopentane and I would like to send the article to them. Thank you for any information you can give me. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Wed May 25 08:54:08 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] freezing of specimens in surgery center Message-ID: <5784D843593D874C93E9BADCB87342AB44F9B2@tpiserver03.Coretech-holdings.com> Flash point of Isopentane is -60 degrees F. I think this means a match held over an open surface will cause flames unless the liquid is colder than that. One solution is to keep it chronically colder than that, and relieve the risk. The refrigeration device below is for that purpose. It keeps a cup of isopentane at -80 degrees. It is contained in a metal well. Not much can happen. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=476401&catdesc=Histology+Equipment&CatThreeID=650&CatOneID=4&subcatdesc=Freezing+Devices&idsubcategory=187 Dow Corning makes a silicone liquid, 200 I think, that has the necessary thermal properties, but is none flammable. I have not heard of its use in histology, maybe it blocks staining and water. 3M also makes a nonflammable coolant liquid that might work. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Tuesday, May 24, 2005 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] freezing of specimens in surgery center Hello Histonetters I have a unique question. We are currently starting to set up procedure for collecting samples from a clinical trial. The clinical trial involves taking multiple synovial biopsies at a surgery center. Since portions of the samples need to be processed for frozen sections we wanted to be able to freeze the specimens at the surgery center via isopentane cooled liquid nitrogen. We really do not want to have to transport the multiple specimens back to the main lab prior to freezing due to the time involved it would probably be 1-2 hours post biopsy before we could freeze the samples. The surgery center is questioning the flammability of the isopentane. Has anyone encountered anything like this? Any suggestions would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ja.mitchell <@t> hosp.wisc.edu Wed May 25 09:34:24 2005 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] freezing of specimens in surgery center Message-ID: <583D3E9A1E843445BD54E461D1A2F6F30F83F5D6@uwhis-xchng2.hosp.wisc.edu> "Storage: Keep away from sources of ignition. Keep container closed when not in use. Store in a cool, dry, well-ventilated area away from incompatible substances. Refrigeration has been recommended." The above is directly from a well known company's current MSDS on storage of isopentane. And I know sorting through MSDS's attempting to get a straight answer on on chemical health issues, shelf life, storage can be extremely frustrating. It may be an ignorant question to ask but; Is it not possibly safer to store isopentane (2-Methylbutane) at room temperature vs refrigeration? Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Elizabeth Chlipala Sent: Tuesday, May 24, 2005 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] freezing of specimens in surgery center Hello Histonetters I have a unique question. We are currently starting to set up procedure for collecting samples from a clinical trial. The clinical trial involves taking multiple synovial biopsies at a surgery center. Since portions of the samples need to be processed for frozen sections we wanted to be able to freeze the specimens at the surgery center via isopentane cooled liquid nitrogen. We really do not want to have to transport the multiple specimens back to the main lab prior to freezing due to the time involved it would probably be 1-2 hours post biopsy before we could freeze the samples. The surgery center is questioning the flammability of the isopentane. Has anyone encountered anything like this? Any suggestions would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From froyer <@t> bitstream.net Wed May 25 09:48:22 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Fume hood inqury In-Reply-To: <00ba01c560a5$f8644b80$1302a8c0@OPTIMUM2> References: <00ba01c560a5$f8644b80$1302a8c0@OPTIMUM2> Message-ID: <42949036.9010001@bitstream.net> Any standard HPLC instrument does not need or is required to be in a hood when running. Ford M. Royer, MT(ASCP) Midwest Scienc & Biocenter 6551 Jansen Ave. NE., Ste. 102 Albertville, MN 55301 800-745-4869 Ze Lu wrote: >Dear all, > >We are thinking to move to a new lab space but there is no fume hood built in there. Is fume hood required when doing general histological work including tissue process, embedding and staining? By the way, we also have a HPLC system. Anybody know the requirment for HPLC running (not sample preparation which must be done under hood, just HPLC machine). I checked the OSHA web site but can not find useful info. Any helps? Thanks. > > > >Ze Lu, Ph.D. >Research Scientis >Optimum Therapeutics, >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > From SSouder <@t> mc.utmck.edu Wed May 25 09:50:00 2005 From: SSouder <@t> mc.utmck.edu (Souder, Starla J.) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] unsuscribe Message-ID: <8B799916C6062E4DBB1A6A9E3CD3EFB0A8740B@msexch3.mc.utmck.edu> Please unsuscribe From gcallis <@t> montana.edu Wed May 25 10:15:31 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:08 2005 Subject: Repair Re: [Histonet] Reichert Jung knifemaker In-Reply-To: <200505250859.AA384893214@mail.pml.ac.uk> References: <200505250859.AA384893214@mail.pml.ac.uk> Message-ID: <6.0.0.22.1.20050525090752.01b587b0@gemini.msu.montana.edu> WE sent ours to Energy Beam Sciences - they have a website and may know of a service in UK. Contact this lady for help. Cindy Gaspari Amazing, it was not an expensive repair and timely for an old, well used instrument. At 01:59 AM 5/25/2005, you wrote: >We have a Reichert Jung knifemaker that needs to be repaired or replaced. >Can anyone tell me where this can be carried out, (preferably the UK). it >is for methacrylate sections. >If it cannot be repaired, what would you recommend? > Regards, > Amanda Beesley > >- Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From lelliott <@t> cvh.on.ca Wed May 25 10:36:48 2005 From: lelliott <@t> cvh.on.ca (Lynda Elliott) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] unsubscribe Message-ID: <6A1C183558C13C4EA2B04E9CA97B5A370285C84B@CVH-EXCHANGE.cvh.on.ca> Please unsubscribe Lynda Elliott, MLT, ART Supervisor, Pathology The Credit Valley Hospital 905-813-1100 ext.6332 From mucram11 <@t> comcast.net Wed May 25 10:54:37 2005 From: mucram11 <@t> comcast.net (mucram11@comcast.net) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] unsuscribe Message-ID: <052520051554.24292.42949FBC000A351800005EE42200734748CECE030E9D0C9A03@comcast.net> I am sorry however this happens all of the time and you are given an e-mail the first of every month with directions on how to unsubscribe. Please refer to that information or request it from HistoNet directly by going to addresses below and get the correct method to be dropped from the list. Unsubscribe is not enough anymore. Pam Marcum -------------- Original message -------------- > Please unsuscribe > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Wed May 25 13:25:08 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Fish Eggs-Help Message-ID: <42948AC3.21758.29C403@localhost> Hello Everyone, Who among you have had happy experiences with fixation/processing/sectioning of unfertilized fish eggs?? A graduate student here at the Vet College is trying to section paraffin blocks with Arctic char eggs. The eggs are falling apart on the water bath (or on the way to the waterbath). He only wants to be able to do H&E on (stepped) serial sections. Any advise? Any at all? For insatnce, would frozens be a better option? Is the fixtive choice important, i.e. are fish eggs like eyes- should he be using Bouins? I welcome any and all input. Thanks. Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From Myri37 <@t> aol.com Wed May 25 13:08:07 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] problem with MMA Message-ID: <141.4616cce2.2fc61907@aol.com> Hello histonetters, I am using MMA/ BMA to embed titanium samples on wich we grew cells . The polymerisation step is done at -20?C. Thick sections are made with diamond thread saw and grinded until they reach 50-80 um. Sections are stained with RBS Rapid bone stain) on a hot plate at 56?C. Unfortunately i see on the resulting slides a separation between titanium and cells. can anyone help me understanding what happens and how avoid this separation between titanium and cells. Can it be due to the retraction of BMA/MMA during polymerization ? Do you think the temperature (-20?C) during polymerization has an effect ? In some cases we covered thentitanium with a hydroxyapatite layer before growing cells, but obtained the same results in the end. thanks in advance. best regards. Myriam baali Natural Implant From algranth <@t> u.arizona.edu Wed May 25 13:11:22 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Fish Eggs-Help In-Reply-To: <42948AC3.21758.29C403@localhost> Message-ID: <4.3.2.7.2.20050525105911.00caf080@algranth.inbox.email.arizona.edu> Once I did some frog eggs which are probably similar in that the centers mostly fell out - I did frozens and also paraffin sections. Fortunately the investigator only wanted to observe the walls and not the center yolky part. I found that soaking them and sometimes using a little ammonia water or soapy water helped - I could then get a section or two before I had to resoak. I don't have a processor that uses vacuum and pressure but I was wondering if that would have any better effects on processing? Andi Grantham At 02:25 PM 5/25/2005 -0400, Greg Dobbin wrote: >Hello Everyone, >Who among you have had happy experiences with >fixation/processing/sectioning of unfertilized fish eggs?? > >A graduate student here at the Vet College is trying to section >paraffin blocks with Arctic char eggs. The eggs are falling apart on >the water bath (or on the way to the waterbath). He only wants to be >able to do H&E on (stepped) serial sections. Any advise? Any at >all? > >For insatnce, would frozens be a better option? Is the fixtive choice >important, i.e. are fish eggs like eyes- should he be using Bouins? > >I welcome any and all input. Thanks. >Greg > > >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Greg Dobbin >Research Technologist, >Pathology Lab, >Atlantic Veterinary College, U.P.E.I. >550 University Ave. >Charlottetown, P.E.I. >Canada, C1A 4P3 >Phone: (902)566-0744 >Fax: (902)566-0851 >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Happiness is a journey, not a destination. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From pruegg <@t> ihctech.net Wed May 25 13:13:09 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] NADH stain Message-ID: <200505251813.j4PID81C027537@chip.viawest.net> A pathologist is asking for a stain, possibly IHC he is calling NADH, does this mean anything to anyone? Patsy From pruegg <@t> ihctech.net Wed May 25 13:17:43 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] think we have the NADH/NADPH request solved Message-ID: <200505251817.j4PIHg1C028846@chip.viawest.net> NADH ? a peroxidase method for blood smears (is this like myleoperoxidase for endogenous peroxidase?) NADPH ? NitroBlue Tetrazolium ?sp From gcallis <@t> montana.edu Wed May 25 13:22:27 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:08 2005 Subject: isopentane aka 2 methyl butane storage RE: [Histonet] freezing of specimens in surgery center In-Reply-To: <583D3E9A1E843445BD54E461D1A2F6F30F83F5D6@uwhis-xchng2.hosp .wisc.edu> References: <583D3E9A1E843445BD54E461D1A2F6F30F83F5D6@uwhis-xchng2.hosp.wisc.edu> Message-ID: <6.0.0.22.1.20050525120847.01b6cff0@gemini.msu.montana.edu> Dear Jean, Not an ignorant question, and that is exactly what we do, RT storage isopentane and hexane. The room is well ventilated, and the 1 liter bottle size is all we have around at any one time is placed inside a plastic container to cushion the bottle from bumping into another bottle or hard surface. No one here is about to invest in an explosion proof freezer, refrigerator, or expensive freezing devices (although that would be nice!) and storage in a regular freezer/refrigerator is out of the question. Amazing, and with pure blind luck, I stored isopentane for years in a regular refrigerator/freezer without blowing up the lab. I would not take that chance now. What we are trying to do is eliminate isopentane completely as a snap freezing method and going to a petri dish floating in Liquid nitrogen (with dish supported by a metal block or rack). Ventilation is still important, snap freezing is excellent, without freezing artifact and we can do many blocks at a dissection session. This is a cheap, easy setup as long as one has liquid nitrogen in the lab - we always have it around - fortunately. At 08:34 AM 5/25/2005, you wrote: >"Storage: Keep away from sources of ignition. Keep container closed when >not in use. Store in a cool, dry, well-ventilated area away from >incompatible substances. Refrigeration has been recommended." > >The above is directly from a well known company's current MSDS on >storage of isopentane. And I know sorting through MSDS's attempting to >get a straight answer on on chemical health issues, shelf life, storage >can be extremely frustrating. > >It may be an ignorant question to ask but; Is it not possibly safer to >store isopentane (2-Methylbutane) at room temperature vs refrigeration? > >Jean Mitchell, BS, HT (ASCP) >University of Wisconsin Hospital & Clinics >Neuromuscular Laboratory >Madison, WI > > > > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >Elizabeth Chlipala >Sent: Tuesday, May 24, 2005 11:59 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] freezing of specimens in surgery center > >Hello Histonetters > >I have a unique question. We are currently starting to set up procedure >for collecting samples from a clinical trial. The clinical trial >involves taking multiple synovial biopsies at a surgery center. Since >portions of the samples need to be processed for frozen sections we >wanted to be able to freeze the specimens at the surgery center via >isopentane cooled liquid nitrogen. We really do not want to have to >transport the multiple specimens back to the main lab prior to freezing >due to the time involved it would probably be 1-2 hours post biopsy >before we could freeze the samples. The surgery center is questioning >the flammability of the isopentane. Has anyone encountered anything >like this? Any suggestions would be helpful. Thanks in advance. > >Liz > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC >P.O. Box 18592 Boulder, Colorado 80308 >Office: (303) 735-5001 >Fax: (303) 735-3540 >liz@premierlab.com >www.premierlab.com > >Ship to Address: >Premier Laboratory >University of Colorado >MCDB, Room A3B40 >Boulder, Colorado 80309 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From juan.gutierrez <@t> christushealth.org Wed May 25 13:24:18 2005 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] NADH stain Message-ID: The only NADH that I am aware of is the histochemistry stain done on frozen muscle sections. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, May 25, 2005 1:13 PM To: 'Histonet' Subject: [Histonet] NADH stain A pathologist is asking for a stain, possibly IHC he is calling NADH, does this mean anything to anyone? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From emerald_lake77 <@t> yahoo.com Wed May 25 13:48:29 2005 From: emerald_lake77 <@t> yahoo.com (- -) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Anti-beta Galactosidase - General info Message-ID: <20050525184829.40586.qmail@web31711.mail.mud.yahoo.com> Hello Histonetters! I might be working with anti beta galactosidase anitbodies in the near future. I have no idea about any that work or how they work. Can anybody give me some information regarding these antibodies? Which work? If they should be used on frozen samples or paraffin sections? Any info will be great! Thank you! __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Myri37 <@t> aol.com Wed May 25 13:51:41 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] (sans sujet) Message-ID: <86.28d42987.2fc6233d@aol.com> BMA is butylmethacrylate, i make a mixture of Methylmethacrylate 60ml, butylmethacrylate 35 ml, methylbenzoate 5ml, and 1.2 ml PEG 400. i Fix samples in paraformaldehyde 4%, then dehydrate in ethanol, clear in xylene, then in : MMA 1 : mixture as below. MMA2 : mixture + 04% of PBO (peroxyde benzoyle dried) MMA3 : mixture + 0.8 % of PBO. all these stpes at 4?C. then polymerisation MMA3 + N N dimethyltolidine at -20?C. titanium samples are disks of 1cm od diameter. thanks Myriam Baali From Myri37 <@t> aol.com Wed May 25 13:54:02 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] problem with MMA Message-ID: <1a0.348c260c.2fc623ca@aol.com> BMA is butylmethacrylate, i make a mixture of Methylmethacrylate 60ml, butylmethacrylate 35 ml, methylbenzoate 5ml, and 1.2 ml PEG 400. i Fix samples in paraformaldehyde 4%, then dehydrate in ethanol, clear in xylene, then in : MMA 1 : mixture as below. MMA2 : mixture + 04% of PBO (peroxyde benzoyle dried) MMA3 : mixture + 0.8 % of PBO. all these stpes at 4?C. then polymerisation MMA3 + N N dimethyltoluidine at -20?C. titanium samples are disks of 1cm diameter. thanks Myriam Baali From kbroomal <@t> NEMOURS.ORG Wed May 25 13:54:43 2005 From: kbroomal <@t> NEMOURS.ORG (Kristen Broomall) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] think we have the NADH/NADPH request solved Message-ID: <6E41111281623B4B8A9AB8F9A7EA343782431F@wlmmsx02.nemours.org> NADH is nicotinamide adenine dinucleotide reductase, a muscle enzyme histochemistry stain. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, May 25, 2005 2:18 PM To: 'Histonet' Subject: [Histonet] think we have the NADH/NADPH request solved NADH ? a peroxidase method for blood smears (is this like myleoperoxidase for endogenous peroxidase?) NADPH ? NitroBlue Tetrazolium ?sp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From weneng2004 <@t> yahoo.com Wed May 25 14:00:23 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] H & E staining on frozen sections Message-ID: <20050525190024.53869.qmail@web53407.mail.yahoo.com> Dear histonetters, Long time ago I used a H&E stain solution on frozen sections. I remember it's a one step staining. But I couldn't find it now. Does anybody know where I can order it? Thanks in advance!\ Wen --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! From mcauliff <@t> umdnj.edu Wed May 25 14:04:44 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] NADH stain In-Reply-To: <200505251813.j4PID81C027537@chip.viawest.net> References: <200505251813.j4PID81C027537@chip.viawest.net> Message-ID: <4294CC4C.8040202@umdnj.edu> NADH is Nicotine Adenine Dinucleotide Phosphate, it is a diaphorase (catalyzes the reoxidation of reduced coenzymes). Old name is TPN. I suggest looking at a histochemisty text, John Kiernan's book is excellent. You can do the rxn on formalin fixed frozen sections (not wax). Geoff Patsy Ruegg wrote: >A pathologist is asking for a stain, possibly IHC he is calling NADH, does >this mean anything to anyone? >Patsy >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From dw18 <@t> uchicago.edu Wed May 25 14:08:53 2005 From: dw18 <@t> uchicago.edu (David A. Wright) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Re: Sharpies, GOOD markers for cassettes & pencils Message-ID: <58b4342.2201739.822fd00@m4500-00.uchicago.edu> Hi I agree with Janet Bonner. Sanford Sharpie Industrial Extra Fine Point permanent markers (Sanford #13801) make excellent lab labeling pens for use in difficult environments. (I also use the Industrial "fat" sharpies too). You can easily get them from most office supply companies. I use them for everything - microfuge tubes, glassware, labeling microslides... which is how I came to find them. I needed something better than pencil (which I had been brought up to use as the "best" solvent resistant frosted slide marker) when I began using SuperFrost Plus slides. Pencil on slides was was a recent discussion topic on Histonet. I meant to write at the time that with charged glass, as with Superfrost Plus or DIY APES, fine graphite particles produced by pencil writing are attracted to the glass to give an ugly background speckling, especially at the label end. These pens solved my problem. Depending on the surface (non)porosity & (non)"wickability", you can produce really quite small writing. I routinely use the pens to write a dozen characters or so along the edge of an APES coverglass used for processing those frozen sections which I plan to prepare with aqueous mountant (Moewiol) and then make permanent with resin using John Kiernan's recommended "double coverslip sandwich on a slide" technique (see the Archives! [My title, not his - is there an easier name for this?]) I should point out the pens are "solvent resistant" in that the writing is still very easy to read after solvent/steam exposure. A small amount of dye does however leach out, just on first contact with a strong solvent, so that my first clearing bath of Xylene/Citrisolv does turn yellow-brown. I did worry at first that this might dye unstained tissue but the next bath presumably removes it and I haven't seen any problems. All the best -David ================================================= Message: 10 Date: Tue, 24 May 2005 16:36:37 -0400 From: "Bonner, Janet" Subject: RE: [Histonet] GOOD markers/pencils for cassettes To: "'krat18@aol.com '" , Statlab pens work great on the slides, but we can't use them on the cassettes. We use the Sanford Sharpie Industrial fine point pen (available from Office Depot)...It has super permanent ink and works fantastically. -Janet David A. Wright, Ph.D. University of Chicago Section of Neurosurgery, MC3026 5841 S. Maryland Ave Chicago, IL 60637 USA [W] 773-834-1485 (Office) SBRI Rm J325 773-834-0395 (Lab) SBRI Rm J338 [H] 773-643-6976 Pager (@773-753-1880) #9776 Text page 120 char E-mail 7738456834@myairmail.com or web: http://www.myairmail.com to 7738456834 FAX (Neurosurgery) 773.702.3518 Web FAX & personal voicemail: (815)642-9630 ================================================ Does 2+2=5 for large values of 2? From northma <@t> ohsu.edu Wed May 25 14:16:49 2005 From: northma <@t> ohsu.edu (Mary North) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] NADH stain Message-ID: The NADH stain is routinely requested for skeletal muscle enzyme histochemistry, performed on frozen sections. The technique is available in the AFIP's Advanced Laboratory Methods in Histology and Pathology or I can email you personally with the protocol we use in this lab. Mary North, HT(ASCP), HTL Neuromuscular Laboratory Oregon Health & Science University Portland, OR >>> "Patsy Ruegg" 05/25/05 11:13 AM >>> A pathologist is asking for a stain, possibly IHC he is calling NADH, does this mean anything to anyone? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sbreeden <@t> nmda.nmsu.edu Wed May 25 14:34:06 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Looking for Donna Willis Message-ID: I'd like to get in touch with Donna Willis (TX); please email me regarding TX State Society. Thanks! Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From mfrei <@t> sial.com Wed May 25 14:40:10 2005 From: mfrei <@t> sial.com (Mark Frei) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] think we have the NADH/NADPH request solved Message-ID: NADH is Nicotinamide adenine dinucleotide, reduced form. It is considered a coenzyme, not an enzyme. It participates in enzymatic reactions by donating electrons. Mark Frei MT(ASCP) "Kristen Broomall" Sent by: histonet-bounces@lists.utsouthwestern.edu 05/25/2005 01:54 PM To: "'Patsy Ruegg'" , "'Histonet'" cc: Subject: RE: [Histonet] think we have the NADH/NADPH request solved NADH is nicotinamide adenine dinucleotide reductase, a muscle enzyme histochemistry stain. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, May 25, 2005 2:18 PM To: 'Histonet' Subject: [Histonet] think we have the NADH/NADPH request solved NADH ? a peroxidase method for blood smears (is this like myleoperoxidase for endogenous peroxidase?) NADPH ? NitroBlue Tetrazolium ?sp _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nmuvarak <@t> facstaff.wisc.edu Wed May 25 17:13:03 2005 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:25:08 2005 Subject: isopentane aka 2 methyl butane storage RE: [Histonet] freezing of specimens in surgery center Message-ID: <122f5812339f.12339f122f58@wiscmail.wisc.edu> We have stored isopentane at RT under the fume hood for an entire year. Then when the department got an explosion-proof frig, that's where I started storing it. As for freezing, I personally found that freezing on the petri dish without isopentane is not as good... blocks tended to crack if frozen too quickly without isopentane, but it may work for others. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 ----- Original Message ----- From: Gayle Callis Date: Wednesday, May 25, 2005 1:22 pm Subject: isopentane aka 2 methyl butane storage RE: [Histonet] freezing of specimens in surgery center > Dear Jean, > > Not an ignorant question, and that is exactly what we do, RT > storage > isopentane and hexane. The room is well ventilated, and the 1 > liter bottle > size is all we have around at any one time is placed inside a > plastic > container to cushion the bottle from bumping into another bottle > or hard > surface. > > No one here is about to invest in an explosion proof freezer, > refrigerator, > or expensive freezing devices (although that would be nice!) and > storage in > a regular freezer/refrigerator is out of the question. Amazing, > and with > pure blind luck, I stored isopentane for years in a regular > refrigerator/freezer without blowing up the lab. I would not take > that > chance now. > > What we are trying to do is eliminate isopentane completely as a > snap > freezing method and going to a petri dish floating in Liquid > nitrogen (with > dish supported by a metal block or rack). Ventilation is still > important, > snap freezing is excellent, without freezing artifact and we can > do many > blocks at a dissection session. This is a cheap, easy setup as > long as one > has liquid nitrogen in the lab - we always have it around - > fortunately. > > At 08:34 AM 5/25/2005, you wrote: > >"Storage: Keep away from sources of ignition. Keep container > closed when > >not in use. Store in a cool, dry, well-ventilated area away from > >incompatible substances. Refrigeration has been recommended." > > > >The above is directly from a well known company's current MSDS on > >storage of isopentane. And I know sorting through MSDS's > attempting to > >get a straight answer on on chemical health issues, shelf life, > storage>can be extremely frustrating. > > > >It may be an ignorant question to ask but; Is it not possibly > safer to > >store isopentane (2-Methylbutane) at room temperature vs > refrigeration?> > >Jean Mitchell, BS, HT (ASCP) > >University of Wisconsin Hospital & Clinics > >Neuromuscular Laboratory > >Madison, WI > > > > > > > > > > > > > > > >-----Original Message----- > >From: histonet-bounces@lists.utsouthwestern.edu > >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > >Elizabeth Chlipala > >Sent: Tuesday, May 24, 2005 11:59 AM > >To: histonet@lists.utsouthwestern.edu > >Subject: [Histonet] freezing of specimens in surgery center > > > >Hello Histonetters > > > >I have a unique question. We are currently starting to set up > procedure>for collecting samples from a clinical trial. The > clinical trial > >involves taking multiple synovial biopsies at a surgery center. > Since>portions of the samples need to be processed for frozen > sections we > >wanted to be able to freeze the specimens at the surgery center via > >isopentane cooled liquid nitrogen. We really do not want to have to > >transport the multiple specimens back to the main lab prior to > freezing>due to the time involved it would probably be 1-2 hours > post biopsy > >before we could freeze the samples. The surgery center is > questioning>the flammability of the isopentane. Has anyone > encountered anything > >like this? Any suggestions would be helpful. Thanks in advance. > > > >Liz > > > >Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier > Laboratory, LLC > >P.O. Box 18592 Boulder, Colorado 80308 > >Office: (303) 735-5001 > >Fax: (303) 735-3540 > >liz@premierlab.com > >www.premierlab.com > > > >Ship to Address: > >Premier Laboratory > >University of Colorado > >MCDB, Room A3B40 > >Boulder, Colorado 80309 > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 (lab with voice mail) > 406 994-4303 (FAX) > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From katri <@t> cogeco.ca Wed May 25 19:20:30 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Removing OCT??? References: <200505240019.j4O0JnNk017314@syrphus.ucdavis.edu><007e01c56005$226cdd30$6a9a9618@Katri> Message-ID: <005401c56188$b9af5810$6a9a9618@Katri> Andrea, We sometimes end up processing to paraffin the frozen needle biopsies of kidney, after IF (immunofluorescence) has been completed (original paraffin blocks did not have glomeruli) and they seem to turn out OK. Maybe it depends a lot on type of tissue and what you are going to do with it after. Katri ----- Original Message ----- From: "Andrea T. Hooper" To: "Katri Tuomala" ; ; "Catherine Stanecki" Sent: Tuesday, May 24, 2005 12:00 PM Subject: Re: [Histonet] Removing OCT??? > Hi Katri, > > I would expect substantial deleterious damage to the cytoarchitecture > using this method, as freeze-thaw is generally to be avoided ... are your > experiences different? I suggest that Cathy cut frozen sections and > post-fix in 10% NBF for 10 min at RT as an alternative. > > Thanks, > Andrea > > > At 10:06 PM -0400 5/23/05, Katri Tuomala wrote: >>Hi Cathy, >>Just put the OCT embedded tissue in 10% NBF and let come to room >>temperature, change formalin and let fix appropriate time. >> >>Katri >> >>Katri Tuomala >>Hamilton, Ontario, Canada >>----- Original Message ----- From: "Catherine Stanecki" >> >>> >>>I am new to staining tissues and I recently froze a bunch of tissues in >>>OCT. However, I recently found out that it would be better to have some >>>sample fixed in formalin as well so you can use mutliple staining >>>methods. >>>Is there a way to remove OCT from tissue samples without damaging the >>>tissue, or is the tissue forever stuck in tissue tek once you freeze it >>>back? >>> >>>Thanks! >>>Cathy >>> > > -- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Megan.Clarke <@t> hnehealth.nsw.gov.au Wed May 25 21:35:46 2005 From: Megan.Clarke <@t> hnehealth.nsw.gov.au (Megan Clarke) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] killing bugs Message-ID: why does 70% alcohol kill bugs better than 100% alcohol ? From n.cragg <@t> epistem.co.uk Thu May 26 04:26:48 2005 From: n.cragg <@t> epistem.co.uk (ncragg) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Antibodies for FACS and IHC Message-ID: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> Hello, Please can anyone help? How are antibodies produced for different applications different? Is it possible to use antibodies that are currently being used for IHC, for FACS analysis? We currently have unconjugated primary antibodies which we use for IHC, could we use a fluorescent secondary antibody to then use it for FACS. If I wanted to source a new antibody that could be used for IHC (probably frozen sections) and FACS - do I need to be sure it's been tested in both applications or can I make a judgement based on it's performance in just one of the applications? Is it best to source a fluorescent-conjugated primary? Or will this mean there won't be enough amplification for the IHC application? Sorry for all the questions but I'm venturing into new territory!! Thank you in advance, Nic Nicola Cragg BSc Epistem Ltd Manchester, UK From Sjohnso616 <@t> aol.com Thu May 26 04:49:45 2005 From: Sjohnso616 <@t> aol.com (Sjohnso616@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Second Post of Position in Fl Message-ID: In a message dated 5/23/05 1:01:19 P.M. Eastern Daylight Time, histonet-request@lists.utsouthwestern.edu writes: Message: 5 Date: Mon, 23 May 2005 04:32:54 EDT From: Sjohnso616@aol.com Subject: [Histonet] Position available In Southwest Fla To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="US-ASCII" Hi All, Sarasota Pathology in Southwest Florida is looking to hire a HT or HTL. Our successful candidate will have strong skills in embedding, cutting and special stains. We offer excellent salary and benefits as well as an abundance of SUN. Please forward your resume to: Nancy Day 2001 Webber St Sarasota Florida 34239-5737 Fax 941-362-8992 Phone: 941 362-8900 From pex0220 <@t> yahoo.com.cn Thu May 26 05:28:23 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] antigen retrieval! Message-ID: <20050526102823.6368.qmail@web15506.mail.cnb.yahoo.com> Hello, all, I want to use HIER on bone sections, but I would choose a more gentle method such as steaming rather than boiling. If you have some experience in it, Can you do me a favor? Thank you! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From lpwenk <@t> sbcglobal.net Thu May 26 05:46:36 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] killing bugs In-Reply-To: Message-ID: <1095018647-217737973@pathology.swmed.edu> By "bugs" - do you mean micro-organisms or insects? With this group, it could be either. Second part of my question - "who" is it that is saying that 70% is killing "bugs" better than 100%? And how and why are you trying to kill them? Disinfecting a countertop? Fixing them? Sorry, need a little bit more information before I can suggest an explanation for your question. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Megan Clarke Sent: Wednesday, May 25, 2005 10:36 PM To: histonet@pathology.swmed.edu Subject: [Histonet] killing bugs why does 70% alcohol kill bugs better than 100% alcohol ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Thu May 26 05:50:33 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Isopentane In-Reply-To: Message-ID: Go to Histo-Logic (under Sakura's webpage) http://www.sakura-americas.com/histologic/menu.html Click on Archive issues Then click on Index I believe the article is under Safety, within the last 2-3 years. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carole Fields Sent: Wednesday, May 25, 2005 8:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Isopentane Yesterday on the Histonet someone referred to an article in the Histologic by Bob Skinner about an accident at the University of Arkansas involving Isopentane. I pulled the Histologic off line (Dec. 1999) but did not find the information. Do you happen to know where the article might be found? The institution I previously worked for uses Isopentane and I would like to send the article to them. Thank you for any information you can give me. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Thu May 26 05:53:25 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] H & E staining on frozen sections In-Reply-To: <20050525190024.53869.qmail@web53407.mail.yahoo.com> Message-ID: H&E is always a 2 step stain (Hematoxylin in one step, Eosin in another, with lots of other solutions between). Any chance you were doing a Giemsa? Or a Paragon - A basic fuchsin/toluidine blue combination? These are both one step. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wen eng Sent: Wednesday, May 25, 2005 3:00 PM To: histonet Subject: [Histonet] H & E staining on frozen sections Dear histonetters, Long time ago I used a H&E stain solution on frozen sections. I remember it's a one step staining. But I couldn't find it now. Does anybody know where I can order it? Thanks in advance!\ Wen --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Thu May 26 06:46:26 2005 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] killing bugs Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB4291@fh2k093.fhmis.net> Since bugs are water-based, the aqueous properties of the 70% allow for faster and greater replacement of the existing water by the 70% EtOH. @:) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@pathology.swmed.edu Sent: 5/25/2005 10:35 PM Subject: [Histonet] killing bugs why does 70% alcohol kill bugs better than 100% alcohol ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dobbin <@t> upei.ca Thu May 26 08:42:19 2005 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Fish Eggs-Thank you!! Message-ID: <429599FB.6403.29A8FB@localhost> Hello Everyone, A big thank you to the 6 people to who took the time to offer very helpful suugestions to deal with fish egg histo techniques. The graduate student is VERY grateful! Five of the six replies were not copied to the group, so if anyone else is interested in having this information, contact me directly and I will share the responses. Thanks again. Have a nice day everyone. Greg ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Greg Dobbin Research Technologist, Pathology Lab, Atlantic Veterinary College, U.P.E.I. 550 University Ave. Charlottetown, P.E.I. Canada, C1A 4P3 Phone: (902)566-0744 Fax: (902)566-0851 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Happiness is a journey, not a destination. From louise.renton <@t> gmail.com Thu May 26 07:49:47 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] gallon to liter conversion Message-ID: hello all, I have 2 sachets of powder that can be made into 1 gallon of formalin. What converting factor do I use? Given that this powder is from a company in Stanwood, WA, should I use American gallons = 3.79 liter or Imperial gallons = 4.41litre? Does it make any difference in the long run? I have contacted the company, but am still waiting for a reply. Many thanks -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From Barry.R.Rittman <@t> uth.tmc.edu Thu May 26 08:14:44 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] gallon to liter conversion Message-ID: <566FB0B522443D43AF02D2ADBE35A6F0019AE0B3@UTHEVS3.mail.uthouston.edu> Louise I think that you should wait for a response from the company as the measure that you use will, I assume, depend primarily on the market for which the company marketed the material. If this is to make buffered formalin then it probably doesn't make a whole lot of difference as long as you check the pH. The final concentration of formalin is usually acceptable within a range. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise renton Sent: Thursday, May 26, 2005 7:50 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] gallon to liter conversion hello all, I have 2 sachets of powder that can be made into 1 gallon of formalin. What converting factor do I use? Given that this powder is from a company in Stanwood, WA, should I use American gallons = 3.79 liter or Imperial gallons = 4.41litre? Does it make any difference in the long run? I have contacted the company, but am still waiting for a reply. Many thanks -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From info <@t> instrumedics.com Thu May 26 09:43:27 2005 From: info <@t> instrumedics.com (Instrumedics) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] freezing tissue Message-ID: <015901c56201$4b5750b0$6401a8c0@INSTRUMEDICS22> Many years ago we stored isopentane in our lab refrigerator and lo and behold we came in one morning to find that the refrigerator was a mess because of a fire due to an spark that ignited the isopentane. Now I'm gun shy and do not have isopentane in the lab. We developed the Gentle Jane method which only uses liquid nitrogen, which is safe with proper ventilation. We found that immersion in LN2 had problems; a vapor envelop and also cracking of the tissue. The Gentle Jane method chills a heat extractor to LN2 temperature and then contacts the tissue and embedding medium. The tissue is snap frozen in 8-10 seconds. The low temperature of the LN2 (-196C) and the thermal exchange the heat extractor provides, produce small ice crystals which are not damaging to the cells,. The freezing artifact is minimized See the Gentle Jane method on www.instrumedics.com Bernice From marco.prunotto <@t> medecine.unige.ch Thu May 26 09:55:35 2005 From: marco.prunotto <@t> medecine.unige.ch (Marco Prunotto) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] macrophages for the dog In-Reply-To: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> References: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> Message-ID: <4295E367.7070506@medecine.unige.ch> Dear all, have you got any idea if the DAKO CD68 (clone KP1) works in the dog? Have you got any idea for a good marker for macrophages in the dog? thanks a lot marco --------------------------------- Marco Prunotto, PhD Dept. of Pathology & Immunology, CMU 1, rue michel-Servet 1211 Geneva 4, Switzerland Tel: +41 22 3795763 Fax: +41 22 3795746 From scoop <@t> mail.nih.gov Thu May 26 09:58:32 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Antibodies for FACS and IHC In-Reply-To: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> References: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> Message-ID: You can use antibodies that are routinely used for FACS for IHC and visa versa. Most FACS usable antibodies I have tried that work with formalin fixed cells on FACS (eg. you fix the cells first) will work at least on frozen sections, sometimes on FFPE sections. Sometimes antibodies that work on one method have too much background or not enough signal on the other method. You don't need to have a fluorescence tagged primary for FACS and sometimes it helps to increase your signal by using a secondary antibody. Sometimes a fluorescent primary is enough signal for immunofluorescence and sometimes not, but you can always use a fluorescent secondary (tagged with the same fluor) with a tagged primary. But, I don't have any hard and fast rules (other more experienced people may) I just try it and work it out for each antibody as the need arises. Sharon >Hello, > >Please can anyone help? How are antibodies produced for different >applications different? Is it possible to use antibodies that are >currently being used for IHC, for FACS analysis? We currently have >unconjugated primary antibodies which we use for IHC, could we use a >fluorescent secondary antibody to then use it for FACS. If I wanted to >source a new antibody that could be used for IHC (probably frozen sections) >and FACS - do I need to be sure it's been tested in both applications or >can I make a judgement based on it's performance in just one of the >applications? Is it best to source a fluorescent-conjugated primary? Or >will this mean there won't be enough amplification for the IHC application? > Sorry for all the questions but I'm venturing into new territory!! > >Thank you in advance, > >Nic > >Nicola Cragg BSc >Epistem Ltd >Manchester, UK > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From lsallen <@t> lexhealth.org Thu May 26 10:03:30 2005 From: lsallen <@t> lexhealth.org (Lawrence Allen) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] (no subject) Message-ID: WE ARE HAVING PROBLEMS WITH IHC BACKGROUND STAINING. WE ARE USING VANTANA REAGENTS, IS ANY BODY HAS OR HAVE THIS PROBLEM. THANK YOU, LAWRENCE ALLEN _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From billingconsultants <@t> yahoo.com Thu May 26 10:04:26 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Re: Immuno & Special Stainers Attn Vendors Message-ID: <20050526150426.36918.qmail@web54208.mail.yahoo.com> Hi everyone, I have a client interested in puchasing an immuno stainer and special stainers. Are there any out there that do both on one machine? Refurbished and used equipment are also a possibility. Any assistance you can provide would be greatly appreciated. Louri __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From pmarcum <@t> vet.upenn.edu Thu May 26 10:18:33 2005 From: pmarcum <@t> vet.upenn.edu (pmarcum@vet.upenn.edu) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: <1117120713.4295e8c9f1de0@imp.vet.upenn.edu> Lawrence, It will require knowing which antibodies you are using and are they pre-dilutes or did you titer them. Backgound is to general for real assistance. Have you spoken to Ventana Tech Service if it is all of your antibodies to see if you have an instrutment problem. Pam Marcum Quoting Lawrence Allen : > WE ARE HAVING PROBLEMS WITH IHC BACKGROUND STAINING. > WE ARE USING VANTANA REAGENTS, IS ANY BODY HAS OR HAVE THIS PROBLEM. > > > THANK YOU, > LAWRENCE ALLEN > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RizoC <@t> chi.osu.edu Thu May 26 10:41:34 2005 From: RizoC <@t> chi.osu.edu (Rizo, Christian) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] EM Tech position Message-ID: <979FF5962E234F45B06CF0DB7C1AABB201FEA70E@chi2k3ms01.columbuschildrens.net> My name is Christian M. Rizo. I am the Manager of the Anatomic Pathology Lab at Children's Hospital in Columbus Ohio. The reason for this e-mail is that I wanted to inform you that we have an opening for an EM Technologist at our institution. There is a first shift (8:00 A.M. - 4:30 P.M.) Electron Microscopy Technologist position in the Anatomic Pathology Laboratory that is immediately available. The position is a full-time position, working Monday through Friday. Applicants must be a B.S. degree. Preference is given to candidates with some electron microscopy experience or M.S. degree candidate. This position is responsible for varied laboratory procedures, including specimen processing, electron microtomy, immunogold labeling and utilization of electron microscope. Duties may change depending on workload. Applicants should have moderate computer skills. Good customer relations, teamwork, dependability and communication are a must for successful candidates. Children's Hospital Columbus Founded by a determined group of women in 1892, Children's Hospital began as a local charity to serve a dozen very ill children. Throughout the following century, this tiny community-funded mission matured into a health care system that today spans the Midwest as one of its preferred providers of pediatric health care. Columbus Children's today is ranked as one of the nation's ten largest children's hospitals and pediatric research centers. Our hospital campus has nearly 700,000 patient visits every year. Located just minutes from downtown Columbus, Children's Hospital is one of the nation's most progressive and sophisticated health care institutions. Specialty areas include * Surgery * Heart transplant/cardiac care * Cancer * Trauma * Rehabilitation * Dialysis * Bone Marrow Transplant * Neurosciences * Research Children's is also the region's only pediatric Level 1 Trauma Center. With over 1.5-million square feet of interior space, the campus at Children's Hospital is vast. * The main hospital is home to the emergency department (one of the busiest pediatric emergency departments in the nation), surgery suites, interventional radiology and all inpatient units. * In August 2005, a new clinical expansion will be established which includes 14 additional state-of-the-art operating suites (which include intra-operative MRI and robotic technologies), new space for the Children's Heart Center to include catheterization laboratories and an additional 28-bed neonatal intensive care unit as well as integrated clinical laboratory services Thank you for your time. Please forward the e-mail to personnel who you think will benefit from it. Christian M. Rizo MBA, HTL (ASCP) Manager, Anatomic Pathology Childrens Hospital 700 Childrens Drive Columbus, Ohio 43205 614.722.5465 RizoC@chi.osu.edu ----------------------------------------- Confidentiality Notice: This e-mail message, from Children's Hospital, Columbus, Ohio, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. The recipient is responsible to maintain the confidentiality of this information and to use the information only for authorized purposes pursuant to Children's Hospital's confidentiality policies. If you are not the intended recipient (or authorized to receive information for the intended recipient), you are hereby notified that any review, use, disclosure, distribution, copying, printing, or action taken in reliance on the contents of this e-mail is strictly prohibited. If you have received this communication in error, please notify us immediately by reply e-mail and destroy all copies of the original message. Thank you. From gcallis <@t> montana.edu Thu May 26 10:51:43 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Long reply to Re: Antibodies for FACS and IHC In-Reply-To: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> References: <01C561DD.6CDE6CB0.n.cragg@epistem.co.uk> Message-ID: <6.0.0.22.1.20050526090528.01b758e8@gemini.msu.montana.edu> First of all what species tissues/cells are you going to be working with, mouse? Rat? Human? We use the same antibodies for both FACS and IHC. First of all, when you are purchasing anitibodies, as from BD Pharmingen, they tell you what applications are recommended in their catalogs - little abbreviations. Be sure to access the specification sheets on their websites, these are filled with info, and even references to publications for any given antibody. You have to do a bit of sleuthing - a bit of work but fun too. Some of our combinations are For FACS 1. purified antibody ( we also use monoclonal antibody supernates from cell lines producing antibodies, grown up in house) secondary antibody conjugated to fluorophore 2. although infrequent FACS, but a possibility Purified antibody Secondary antibody-biotin Strepavidn-Alexa dye conjugate For IHC 1. Purified antibody Secondary antibody -Biotin Strepavdin-HRP or AP Chromogen 2. Purified antibody Secondary antibody-HRP or AP Chromogen Our favorite is and works very well murine IHC when the primary is a Rat antiMouse monoclonal . 1. For FACS and fluorescence microscopy Biotinylated primary Strepavidin -Alexa fluorophore conjugate (499 = FITC, 555 = rhodamine) for FACS or fluorescence microscopy OR 2. For IHC, use Strepavidin-HRP for peroxidase IHC or Strepavidin-AP if desired. Chromogen a. For FACS Primary antibody-conjugated to FITC - can use this alone for FACS b. For IHC Primary conj FITC Rabbit antiFITC (DAKO) Donkey antiRabbit-HRP Chromogen If the primary is conjugated to PE, you can even buy antiPE and do IHC. Most companies have recommended dilutions of their antibody (ug/ml) for FACS, however for IHC you would have to do a dilution panel to determine your working concentration for any given fixative, paraffin processing, retrieval etc. Most companies will test antibodies for frozen section use, but not always for paraffin embedded tissues. Dilution panel target concentration can start at 10 ug/ml, and then you dilute out. Basically you will do the testing based on what they already tell you, be prepared to do some work. Best sources of fluorescent secondaries - some companies that are excellent, and there are others. It is advisable that the secondary antibody be adsorbed against the species being stained. If you use a biotinylated primary the negative control must be host of primary IgG or isotype matched IgG that is also biotinylated. You can purchase all different species IgG conjugated to biotin, FITC, etc, etc from Jackson and other sources. Southern Biotechnology - excellent antibodies for murine work. Jackson ImmunoResearch Rockland BD PharminGen If you want to try secondary antibodies conjugated to Alexa fluorophores, Molecular Probes, they are available, we do not use these OR you can buy an Alexa fluor conjugation kit and make your own secondaries, particularly if you have a company preference for buying secondary antibodies. One thing to keep in mind, there are now primary antibodies conjugated to Alexa fluorophores, and you probably would have to buy antiAlexaFluor secondary from Molecular Probes, I have not done this. I have never had problems using primary antibodies for FACS and then IHC application. You will have optimize your staining kit/system for the antibody you are using. Just do the dilution panel on the primary, as kits are often very standardized to help you out there. We have a protocol for biotinylated Rat antiMouse CD4, (0.5mg/) diluted out 1:15,000 and fading at 1:20,000 for 30 min Strepavidin-HRP 1:500 dilution of 0.5mg/ml, for 20 min DAKO DAB+, followed by their DAB enhancer. There was some special blocking done prior to staining, and not the same as some else might do it. If someone else was using this antibody as we do, their Strepavidin-HRP may not be as effective as mine, nor will someone elses DAB chromogen. And they may not have the same working dilution as we used for the primary. You can go to company websites which provide staining protocols on how THEY do their frozen section work, etc and tweak the method to suit your needs. There are basic rules but no hard and fast rules on how you do IHC with antibodies used for FACS, Histonet abounds with answers, so if you have a particular antibody in mind, use Histonet Archives for a search www.histosearch.org, and if you don't find it, ask. Use publications also, they should have information in methods and materials, invaluable to see how others have done it, plus it usually provides sources of antibodies/secondaries, etc all reagents used for both FACS and IHC. I suggest you get the Molecular Probes 10th Edition handbook, they will send it to you or you can access it online from their website. This book is very educational for fluorophore usage and also just understanding some of the chemistry of these fluorescent dyes. Good luck, sorry this was so long 03:26 AM 5/26/2005, you wrote: >Hello, > >Please can anyone help? How are antibodies produced for different >applications different? Is it possible to use antibodies that are >currently being used for IHC, for FACS analysis? We currently have >unconjugated primary antibodies which we use for IHC, could we use a >fluorescent secondary antibody to then use it for FACS. If I wanted to >source a new antibody that could be used for IHC (probably frozen sections) >and FACS - do I need to be sure it's been tested in both applications or >can I make a judgement based on it's performance in just one of the >applications? Is it best to source a fluorescent-conjugated primary? Or >will this mean there won't be enough amplification for the IHC application? > Sorry for all the questions but I'm venturing into new territory!! >Thank you in advance, >Nicola Cragg BSc >Epistem Ltd >Manchester, UK Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From clarissabush <@t> sbcglobal.net Thu May 26 11:39:54 2005 From: clarissabush <@t> sbcglobal.net (CM Bush) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Will brain tissue shrink even more if stored in 70% alcohol prior to processing? Message-ID: <20050526163954.4847.qmail@web80314.mail.yahoo.com> Dear Histonet, Hello, here is my first post to the list, thank you in advance for your help. Summary: Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) Maybe this is a little bit long...thank you very much for your time. CM Bush From lillinda70 <@t> aol.com Thu May 26 11:50:10 2005 From: lillinda70 <@t> aol.com (lillinda70@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] help In-Reply-To: <200505241300.71942935db240@rly-xi06.mx.aol.com> References: <200505241300.71942935db240@rly-xi06.mx.aol.com> Message-ID: <8C7302A1CFF6034-C48-2760F@mblk-r37.sysops.aol.com> I would like to be on the Histonet message list instead of the Histonet Digest. -----Original Message----- From: histonet-request@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: Tue, 24 May 2005 1:01:31 PM Eastern Daylight Time Subject: Histonet Digest, Vol 18, Issue 34 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. Automated special stains (Bartlett, Jeanine) 2. RE: Automated special stains (Dawson, Glen) 3. RE: Automated special stains (Weems, Joyce) 4. GOOD markers/pencils for cassettes (krat18@aol.com) 5. Grossing (Heather.A.Harper@pcola.med.navy.mil) 6. Re: microwave processing (Kaye Ryan) 7. freezing of specimens in surgery center (Elizabeth Chlipala) ---------------------------------------------------------------------- Message: 1 Date: Tue, 24 May 2005 11:58:15 -0400 From: "Bartlett, Jeanine" Subject: [Histonet] Automated special stains To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 ------------------------------ Message: 2 Date: Tue, 24 May 2005 11:01:39 -0500 From: "Dawson, Glen" Subject: RE: [Histonet] Automated special stains To: "Bartlett, Jeanine" , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" Dako's Artisan Stainer is great and it has a 7 minute Steiner's that works well. Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Tuesday, May 24, 2005 9:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Tue, 24 May 2005 12:04:47 -0400 From: "Weems, Joyce" Subject: RE: [Histonet] Automated special stains To: "Bartlett, Jeanine" , Message-ID: <83AACDB0810528418AA106F9AE9B7F7EA45DC3@sjhaexc02.sjha.org> Content-Type: text/plain; charset="iso-8859-1" We have the DAKO Artisan and are pleased with the company service as well as the stains. GMS is good. We use the Warthin Starry - not sure they have a Steiner. If there would be one change, I would wish that the stains could be started as ordered instead of by batch. Hope this helps! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 404-851-7376 - Phone 404-851-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine Sent: Tuesday, May 24, 2005 11:58 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Automated special stains Hi everybody: I'd like feedback on the automated special stain equipment being used today. I need something that does all the traditional stains but also silvers...mainly GMS' and Steiner's. I am most interested in an open system so as not to be locked into using a particular vendor's reagents if possible. Vendors welcome to email as well. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ Message: 4 Date: Tue, 24 May 2005 12:30:03 -0400 From: krat18@aol.com Subject: [Histonet] GOOD markers/pencils for cassettes To: histonet@lists.utsouthwestern.edu Message-ID: <8C72E94F8EEA07D-E44-6A28@mblk-d37.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" HELP! HELP! HELP! We are having lots of problems with our markers (we're now using StatLab). The writing smears and smudges when we fix tissues in PenFix, from a combination of the grease from the fat and the alcohol in the PenFix. What is everyone using to mark cassettes that will be fixed in alcohol-based fixatives? ANY suggestions are welcome! We cannot at this time buy a cassette marker machine. Are there pencils that work well? Our Tissue Tek pencils used to smear also, but maybe there are better ones? ------------------------------ Message: 5 Date: Tue, 24 May 2005 11:24:29 -0500 From: Heather.A.Harper@pcola.med.navy.mil Subject: [Histonet] Grossing To: histonet@lists.utsouthwestern.edu Message-ID: <807FE48C5A7CC940B973B58D32E701431991BE4E@nhpens-exch1.pcola.med.navy.mil> Content-Type: text/plain Currently I am grossing in the small specimens and I do all the miscellaneous stuff, plus all my paper work that a supervisor has to do.. Grossing consumes 2 hrs of my morning and generally do not start until between 8:30-9:00. My shift ends at 2:30. My co-worker works the night shift (10-6) and embeds, cuts and stains (literally). I want to get some opinions. Since I already accession, gross, etc... Is it fair for the pathologists to want me also to do the recuts and special stains? I feel an enormous pressure about this issue and I simply can't understand why some pathologists act like it's so detrimental to be able to view slides and get recuts/special stains the same day. I do not want to be rushed through grossing to accommodate 2 of the 3 pathologists. Am I right or wrong for feeling this way. I need feedback. Heather A. Harper Naval Hospital of Pensacola, FL Supervisor Histology/Morgue ------------------------------ Message: 6 Date: Tue, 24 May 2005 12:47:52 -0400 From: "Kaye Ryan" Subject: Re: [Histonet] microwave processing To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Jesus, We have been doing the microwave processing for a while now and have seen no change in the IHC staining patterns at all. As well, our routine H&E was not altered in any way. The TAT cannot be beat and the cutting of the tissues is much easier. I have only had this "chatter" issue come up since a hospital merged with our lab several months ago. Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 >>> "Jesus Ellin" 05/24/05 11:35 AM >>> I was wondering our Pathologist is wanting to go to this microwave technology and we have budgeted for 2 new processors one microwave and one that it the conventional type. My question would be that how many people out there are doing microwave processing and has anyone seen a discrepancy with immunos or specials due to the microwave processing? Jesus Ellin Yuma Regional Medical Center >>> "Kaye Ryan" 05/24/05 04:53AM >>> We have begun processing tissues using the microwave processor. For the most part, everything we process turns out beautifully but we seem to be having a problem with small endoscopic biopsies. We have begun seeing chatter but it appears to only be in the neoplastic part of the biopsy. I know that normally chatter is due to cutting techniques or over-drying of the tissue but we take the extra steps to ensure the blocks are appropriately cut. Since it seems to be localized to the neoplastic area I was wondering if this could be due to the microwave processing. We occasionally had this problem with routine processing but only occasionally. Everything else cuts very well. Has anyone heard of or experienced similar problems with microwave processing or can you offer some advice or tips as to how to troubleshoot this problem Kaye Ryan Histology Manager/Educational Coordinator Rocky Point Laboratories 265-0111, 72093 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Tue, 24 May 2005 10:58:53 -0600 From: "Elizabeth Chlipala" Subject: [Histonet] freezing of specimens in surgery center To: Message-ID: <000d01c56081$e0dd14c0$a7d48a80@AMY> Content-Type: text/plain; charset="us-ascii" Hello Histonetters I have a unique question. We are currently starting to set up procedure for collecting samples from a clinical trial. The clinical trial involves taking multiple synovial biopsies at a surgery center. Since portions of the samples need to be processed for frozen sections we wanted to be able to freeze the specimens at the surgery center via isopentane cooled liquid nitrogen. We really do not want to have to transport the multiple specimens back to the main lab prior to freezing due to the time involved it would probably be 1-2 hours post biopsy before we could freeze the samples. The surgery center is questioning the flammability of the isopentane. Has anyone encountered anything like this? Any suggestions would be helpful. Thanks in advance. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 18, Issue 34 **************************************** From David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu May 26 12:02:05 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] slide drying Message-ID: Has anyone suggested that the sections need melting on rather than just drying? or have you done this in the process described. Shaking trapped water off before drying and then melting may be an issue if you have trapped amounts of water under the sections. Whether melted or just dried on. David Christie Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: 24 May 2005 19:37 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] slide drying I'm setting up a small Histo lab and using a 60C slide warmer to dry my slides. I've been currently allowing them to air dry vertically for a couple of hours to overnight, then placing the slides on the slide warmer for an hour. About 90% of the time the slides turn out ok, but some slide do show some detaching from the slide. Both charges and non-charged slides with water bath adhesive seem to be involved. I read a past artical on this site not recommending allowing slides to air dry as the section may entrap water? Any suggestions. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Patricia.Traphagan <@t> ParkNicollet.com Wed May 25 12:21:34 2005 From: Patricia.Traphagan <@t> ParkNicollet.com (Traphagan, Patricia) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] EGFR Message-ID: We have been trying to achieve good staining using this antibody. We use the Dako EGFR kit. Dako's control that comes with the kit, turns out beautifully, which tells us that the stain procedure is correct. Our problem lies with our internal control. The pathologist are just not happy with it. According to them we have: Over staining of the smooth muscle, normal epithelial tissue does not stain correctly, and have un-uniform staining of the membranes. Can anyone tell me what this means? We use 10% NBF, fixation is probably no more than 8 hours. Our processing run is about 12 hours for large tissue specimens, and we use an alcoholic formalin in the second station, which then goes to 70% ETOH and the graded series of alcohols, xylenes, paraffin. We do not put the cut tissue in the oven, we let it air-dry overnight, possibly two nights. We are at a total loss. Any suggestions. We really need HELP!!!!! From pex0220 <@t> yahoo.com.cn Thu May 26 13:56:19 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Take some pictures after the skeletal staining Message-ID: <20050526185619.60674.qmail@web15506.mail.cnb.yahoo.com> Hello, all, I am doing the skeletal staining in mouse embryo. Now I finish staining, but I do not know how to take some pictures. If you have some experience, can you do me a favor? Thank you! Guofeng --------------------------------- Do You Yahoo!? ×¢²áÊÀ½çÒ»Á÷Æ·ÖʵÄÑÅ»¢Ãâ·ÑµçÓÊ From AnthonyH <@t> chw.edu.au Thu May 26 18:25:15 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] NADH stain Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E266@simba.kids> Patsy, The NADH probably refers to the enzymatic demonstration of Nicotinic Acid Adenine Dinucleotide Dehydrogenase. It is used for the demonstration of mithochondria especially in frozen sections of muscle biopsies. The principle of the stain: The oxidation of NADH can be catalysed by flavin enzymes (formerly called diaphorases). NADH-TR is a flavin enzyme which takes up hydrogen reversibly and transfers it to the acceptor, a tetrazolium salt. The tetrazolium is converted to purple-blue formazan pigment marking the site of enzyme activity. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Thursday, 26 May 2005 4:13 AM To: 'Histonet' Subject: [Histonet] NADH stain A pathologist is asking for a stain, possibly IHC he is calling NADH, does this mean anything to anyone? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Thu May 26 18:50:01 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] H & E staining on frozen sections Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3741269E267@simba.kids> Alternatively, you could try Kiernan's Chromoxane cyanine R. This solution stains in 3 minutes and gives a H&E picture. It is an acidic aqueous solution containing Chromoxane cyanine R (CI 43820) and ferric chloride. Reference in his book. Another technique is to do a Diff quik on ethanol fixed frozen sections. This will also give a H&E like picture. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Lee & Peggy Wenk [mailto:lpwenk@sbcglobal.net] Sent: Thursday, 26 May 2005 8:53 PM To: 'wen eng'; 'histonet' Subject: RE: [Histonet] H & E staining on frozen sections H&E is always a 2 step stain (Hematoxylin in one step, Eosin in another, with lots of other solutions between). Any chance you were doing a Giemsa? Or a Paragon - A basic fuchsin/toluidine blue combination? These are both one step. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 Lee & Peggy Wenk -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of wen eng Sent: Wednesday, May 25, 2005 3:00 PM To: histonet Subject: [Histonet] H & E staining on frozen sections Dear histonetters, Long time ago I used a H&E stain solution on frozen sections. I remember it's a one step staining. But I couldn't find it now. Does anybody know where I can order it? Thanks in advance!\ Wen --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From kappeler <@t> patho.unibe.ch Fri May 27 00:46:37 2005 From: kappeler <@t> patho.unibe.ch (Andi Kappeler) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] EGFR References: Message-ID: <003201c5627f$87730d40$27955c82@patho.unibe.ch> Hi Patricia I assume you are talking about staining of colorectal adenocarcinoma - that is what the DakoCytomation EGFR pharmDx kit is most frequently used for. A few comments: - Smooth muscle can stain positively for EGFR (in a low percentage of cases) when using the DakoCytomation kit. - not all "norma epithelia" are EGFR positive. If you look at colon, may be half of the samples will show staining of the normal epithelium. - I would love to see "uniform staining" of the membranes... This is limited to a certain percentage of tumors. The majority of cases show focal or heterogeneous staining of the tumor; often the majority of tumor cells have partial membrane reactivity, most tumors also show cytoplasmic staining. - 8 hours fixation (room temperature, without microwave) may be somewhat short, if your samples are more than just small biopsies. ... and a few suggestions: The DakoCytomation protocol asks to place slides in water before the application of Proteinase K (and also beforte the peroxidase block). We found this to be the cause for uneven digestion and have replaced the water with wash buffer from the kit (contains detergents --> better distribution of ProtK in subsequent step) (the FDA is faaaar away from Switzerland, and we care more about our patients than about the FDA...). Comparative staining of slides with water or with wash buffer in this particular step revealed no difference - except for a more even staining in particular of the border areas of tissue samples on those slides that went through wash buffer. We do a CK8 (e.g. clone CAM 5.2) staining for all cases that are stained for EGFR. This clearly demonstrates the carcinoma cells and helps to identify small groups of cells in the invasive part of the tumor. Particularly useful, if you have one of those cases where smooth muscle and/or connective tissue show some reactivity for EGFR. As an in house control we use cases of liver metastasis of colorectal adenocarcinoma, where the adenocarcinoma is heterogeneously 2+ positive (I don't want a 'retina blaster' as positive control) and where the hepatocytes can serve as strong positive control. If you don't have a copy of the DakoCytomation 'EGFR pharmDx Interpretation Manual' (Nr. 08052), call DakoCytomation and tell them to send you a few copies - and then force your pathologists to put their noses in it, it may help them understand that not all tumors have 3+ complete membrane staining in 110% of all tumor cells. Hope this helps (and let time work for you: EGFR testing will go as it has come, just like many other tests ...) Andi Kappeler Institute of Pathology, University of Bern, Switzerland ----- Original Message ----- From: "Traphagan, Patricia" To: Sent: Wednesday, May 25, 2005 7:21 PM Subject: [Histonet] EGFR > We have been trying to achieve good staining using this antibody. We use > the Dako EGFR kit. Dako's control that comes with the kit, turns out > beautifully, which tells us that the stain procedure is correct. Our > problem lies with our internal control. The pathologist are just not > happy with it. According to them we have: Over staining of the smooth > muscle, normal epithelial tissue does not stain correctly, and have > un-uniform staining of the membranes. Can anyone tell me what this > means? We use 10% NBF, fixation is probably no more than 8 hours. Our > processing run is about 12 hours for large tissue specimens, and we use > an alcoholic formalin in the second station, which then goes to 70% ETOH > and the graded series of alcohols, xylenes, paraffin. We do not put the > cut tissue in the oven, we let it air-dry overnight, possibly two > nights. We are at a total loss. Any suggestions. We really need > HELP!!!!! > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Myri37 <@t> aol.com Fri May 27 03:46:13 2005 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] substitute for xylene Message-ID: <5F0A3586.0C350B2F.0005167B@aol.com> Hello everyone i have some samples made of polystyrene and i want to embed them in paraffin or MMA. But xylene and toluene attacks polystyrene. DO you Know another clearing agent to replace Xylene ? Thank you very much in advance. Myriam baali natural implant From katri <@t> cogeco.ca Fri May 27 07:59:03 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] EGFR References: <003201c5627f$87730d40$27955c82@patho.unibe.ch> Message-ID: <000601c562bb$dc659b60$6a9a9618@Katri> Andi, I loved your response to Patricia. Very informative and down to earth... and helpful to me also, since I am working with EGFR, although, not with the Dako Kit... Thank You. Katri ----- Original Message ----- From: "Andi Kappeler" To: "Traphagan, Patricia" ; "Histonet" Sent: Friday, May 27, 2005 1:46 AM Subject: Re: [Histonet] EGFR > Hi Patricia > > I assume you are talking about staining of colorectal adenocarcinoma - > that is what the DakoCytomation EGFR pharmDx kit is most frequently used > for. A few comments: > - Smooth muscle can stain positively for EGFR (in a low percentage of > cases) when using the DakoCytomation kit. > - not all "norma epithelia" are EGFR positive. If you look at colon, may > be half of the samples will show staining of the normal epithelium. > - I would love to see "uniform staining" of the membranes... This is > limited to a certain percentage of tumors. The majority of cases show > focal or heterogeneous staining of the tumor; often the majority of tumor > cells have partial membrane reactivity, most tumors also show cytoplasmic > staining. > - 8 hours fixation (room temperature, without microwave) may be somewhat > short, if your samples are more than just small biopsies. > > ... and a few suggestions: > The DakoCytomation protocol asks to place slides in water before the > application of Proteinase K (and also beforte the peroxidase block). We > found this to be the cause for uneven digestion and have replaced the > water with wash buffer from the kit (contains detergents --> better > distribution of ProtK in subsequent step) (the FDA is faaaar away from > Switzerland, and we care more about our patients than about the FDA...). > Comparative staining of slides with water or with wash buffer in this > particular step revealed no difference - except for a more even staining > in particular of the border areas of tissue samples on those slides that > went through wash buffer. > We do a CK8 (e.g. clone CAM 5.2) staining for all cases that are stained > for EGFR. This clearly demonstrates the carcinoma cells and helps to > identify small groups of cells in the invasive part of the tumor. > Particularly useful, if you have one of those cases where smooth muscle > and/or connective tissue show some reactivity for EGFR. > As an in house control we use cases of liver metastasis of colorectal > adenocarcinoma, where the adenocarcinoma is heterogeneously 2+ positive (I > don't want a 'retina blaster' as positive control) and where the > hepatocytes can serve as strong positive control. > If you don't have a copy of the DakoCytomation 'EGFR pharmDx > Interpretation Manual' (Nr. 08052), call DakoCytomation and tell them to > send you a few copies - and then force your pathologists to put their > noses in it, it may help them understand that not all tumors have 3+ > complete membrane staining in 110% of all tumor cells. > Hope this helps (and let time work for you: EGFR testing will go as it has > come, just like many other tests ...) > > Andi Kappeler > Institute of Pathology, University of Bern, Switzerland > > ----- Original Message ----- > From: "Traphagan, Patricia" > To: > Sent: Wednesday, May 25, 2005 7:21 PM > Subject: [Histonet] EGFR > > >> We have been trying to achieve good staining using this antibody. We use >> the Dako EGFR kit. Dako's control that comes with the kit, turns out >> beautifully, which tells us that the stain procedure is correct. Our >> problem lies with our internal control. The pathologist are just not >> happy with it. According to them we have: Over staining of the smooth >> muscle, normal epithelial tissue does not stain correctly, and have >> un-uniform staining of the membranes. Can anyone tell me what this >> means? We use 10% NBF, fixation is probably no more than 8 hours. Our >> processing run is about 12 hours for large tissue specimens, and we use >> an alcoholic formalin in the second station, which then goes to 70% ETOH >> and the graded series of alcohols, xylenes, paraffin. We do not put the >> cut tissue in the oven, we let it air-dry overnight, possibly two >> nights. We are at a total loss. Any suggestions. We really need >> HELP!!!!! >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Fri May 27 08:04:58 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] substitute for xylene Message-ID: Hi Myriam, The June 2000 issue of Journal of Histotechnology (volume 23, number 2) has an article dealing with using mineral oil inplace of xylene for processing. Many years ago I used peanut oil in processing catheters removed from dog hearts (to examine the surrounding tissue for something, though I don't remember what). If you don't have access to the JOH article I could fax/mail it to you. Fred >>> 05/27/05 04:46AM >>> Hello everyone i have some samples made of polystyrene and i want to embed them in paraffin or MMA. But xylene and toluene attacks polystyrene. DO you Know another clearing agent to replace Xylene ? Thank you very much in advance. Myriam baali natural implant _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ryaskovich <@t> dir.nidcr.nih.gov Fri May 27 08:18:17 2005 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] substitute for xylene Message-ID: <8F3AB322628548428A992EFB0E80F5D315D63155@nihexchange8.nih.gov> Myriam, If your samples are just the polystyrene without tissue just use the MMA and hand process. You really don't need any xylene. I have done it before with no problems. Ruth Yaskovich National Institutes of Health N.I.D.C.R. Pain Neurosensory Mechanism Branch Drug Discovery > ---------- > From: Myri37@aol.com > Sent: Friday, May 27, 2005 3:46 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] substitute for xylene > > Hello everyone > > i have some samples made of polystyrene and i want to embed them in > paraffin or MMA. But xylene and toluene attacks polystyrene. DO you Know > another clearing agent to replace Xylene ? > Thank you very much in advance. > > Myriam baali > natural implant > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Jan.Minshew <@t> leica-microsystems.com Fri May 27 08:56:28 2005 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] substitute for xylene Message-ID: Hi Myriam and Ruth, If you have access to a vibrating microtome, you might be able to section the polystyrene without embedding it. Best wishes, Jan "Yaskovich, Ruth A (NIH/NIDCR)" To: histonet@pathology.swmed.edu, "'Myri37@aol.com'" Sent by: cc: (bcc: Jan Minshew/USDER/West/Leica) histonet-bounces@lists.utsouth Subject: RE: [Histonet] substitute for xylene western.edu 05/27/2005 08:18 AM Myriam, If your samples are just the polystyrene without tissue just use the MMA and hand process. You really don't need any xylene. I have done it before with no problems. Ruth Yaskovich National Institutes of Health N.I.D.C.R. Pain Neurosensory Mechanism Branch Drug Discovery > ---------- > From: Myri37@aol.com > Sent: Friday, May 27, 2005 3:46 AM > To: histonet@pathology.swmed.edu > Subject: [Histonet] substitute for xylene > > Hello everyone > > i have some samples made of polystyrene and i want to embed them in > paraffin or MMA. But xylene and toluene attacks polystyrene. DO you Know > another clearing agent to replace Xylene ? > Thank you very much in advance. > > Myriam baali > natural implant > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _____________________________________________________________________ This e-mail has been scanned for viruses by MCI's Internet Managed Scanning Services - powered by MessageLabs. For further information visit http://www.mci.com From jkiernan <@t> uwo.ca Fri May 27 10:46:02 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Will brain tissue shrink even more if stored in 70% alcoholprior to processing? References: <20050526163954.4847.qmail@web80314.mail.yahoo.com> Message-ID: <429740B9.90F1F9F6@uwo.ca> All tissues, including brain, shrink when their water is replaced by other liquids and then by wax. Graded alcohols will slow down the shrinkage but will not prevent it. The lower alcohols (weaker than 70%) also usefully remove alcohol-insoluble inorganic solutes, such as sodium phosphate derived from the fixative. In the case of liver, formaldehyde fixation reduces the volume to 99% of the original. The volume is 80% after dehydration to 100% ethanol, and 70% after clearing and infiltration with paraffin. [Data from JR Baker's "Principles of Biological Microtechnique" (1958) p.76, citing work by W.Berg (1908) that was quoted in K.Tellyesniczky's article in R. Krause's "Enzyklopaedie der mikroskopischen Technik", Vol 2 (1926). Krause and Baker are great classics in this field.] The reason for storage in 70% alcohol is that this liquid is generally considered to protect specimens from autolysis and attack by microorganisms without making the tissue unduly hard. If the original dimensions of your CNS specimens are important data for your investigation, you should make linear measurements before starting to dehydrate and after the sections are mounted and stained, to obtain a fudge factor for converting measurements to those of the wet specimen. This factor will not take into account differential shrinkage of cells, which is a prominent artifact in paraffin sections of formaldehyde-fixed CNS. The empty spaces seen around large neurons (and often also around capillaries) form during dehydration, clearing and infiltration - probably mostly in the last step, because they are not usually seen in sections of plastic-embedded brain. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ CM Bush wrote: > > Dear Histonet, > > Hello, here is my first post to the list, thank you in advance for your help. > > Summary: > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > Maybe this is a little bit long...thank you very much for your time. > > CM Bush --------- From ddeibler <@t> centexpathlab.com Fri May 27 10:46:04 2005 From: ddeibler <@t> centexpathlab.com (David Deibler) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Employment Opportunity Message-ID: Central Texas Pathology Laboratory ( Waco, Texas) has an immediate opening for an immuno-tech. Qualified applicants will be registered HT(ASCP) and have minimum 5 years post registry experience in an anatomic pathology laboratory. Previous IHC experience is preferred. We have an expanding state-of-the-art IHC department with two Dako Immunostainers. CTPL offers a great benefit package including paid holidays and sick time. Please contact Cindy Bulmer HT(ASCP),QIHC IHC supervisor at 254-752-9621 ext 247 Or David Deibler HT(ASCP) Histology supervisor at 254-752-9621 ext 219 or ddeibler@centexpathlab.com From krat18 <@t> aol.com Fri May 27 10:45:33 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] substitute for xylene In-Reply-To: <5F0A3586.0C350B2F.0005167B@aol.com> References: <5F0A3586.0C350B2F.0005167B@aol.com> Message-ID: <8C730EA40A56691-81C-22AF3@mblk-r29.sysops.aol.com> Hi, Myriam, CBG Biotech (a company that does solvent recycling) has a Formula 83 clearing solvent that they say replaces xylene. Our hospital has received a sample of it, but we have not yet tried it (I think it has naphtha but not sure). Their email address is cbgbiotech.com, if you want to check it out. Hope this will work for you. Karen Raterman -----Original Message----- From: Myri37@aol.com To: histonet@pathology.swmed.edu Sent: Fri, 27 May 2005 04:46:13 -0400 Subject: [Histonet] substitute for xylene Hello everyone i have some samples made of polystyrene and i want to embed them in paraffin or MMA. But xylene and toluene attacks polystyrene. DO you Know another clearing agent to replace Xylene ? Thank you very much in advance. Myriam baali natural implant _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dpahisto <@t> yahoo.com Fri May 27 10:59:20 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] California Grossing Requirements Message-ID: <20050527155920.91269.qmail@web33401.mail.mud.yahoo.com> The person in our lab who grosses most of the surgical specimens is not a certified pathology assistant (or an HT). At our last CLIA inspection the inspector asked us for the "PA's" certification documentation. I deferred the question to the "PA" themselves. She managed to change the subject quite quickly and the topic was not brought up again. Now our grossing person is thinking that she is not qualified and should not be grossing according to government regulations. I have not been able to find anything anywhere (google search) stating who can gross or not gross tissue specimens. Can anyone help me with this question? Cindy DuBois __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From dellav <@t> musc.edu Fri May 27 12:54:13 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Will brain tissue shrink even more if stored in 70%alcoholprior to processing? Message-ID: You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. you can access the article by selecting the link for HistoLogic at www.sakuraus.com CM Bush wrote: > > Dear Histonet, > > Hello, here is my first post to the list, thank you in advance for your help. > > Summary: > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > Maybe this is a little bit long...thank you very much for your time. > > CM Bush --------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 From FlemingT <@t> uthscsa.edu Fri May 27 13:06:40 2005 From: FlemingT <@t> uthscsa.edu (Fleming, Tiffany M) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Double Labeling Same Species Message-ID: Hello, I do a lot of double-labeling- but my anti-bodies are from different species (i.e; Cfos (rabbit) Oxytocin (mouse) FosB (goat)), with fantastic results. I am attempting to stain mice brain tissue for a different P.I with : Cfos(rabbit) with DAB GFP (rabbit) with CY3 AVP (rabbit) with Alexa 488 I did a trial run with Cfos (rabbit) cy3 GFP (rabbit) 488 I am having a terrible time with cross staining/ cross reaction. The professor has no problem staining with GFP and AVP (both rabbit). I am using a combination of his protocol with my cfos protocol. Any suggestions? Tiffany M. Fleming Cunningham Lab 225C UTHSCSA Pharmacology Department 7703 Floyd Curl Dr San Antonio TX 78229-3900 lab (210) 567-4180 (210) 567- 4273 fax 210-567-4300 From weneng2004 <@t> yahoo.com Fri May 27 13:33:08 2005 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] blocking Message-ID: <20050527183308.63692.qmail@web53403.mail.yahoo.com> Hello, I need your advice on blocking. The animal was injected with human antibody. After the tissue are havested I have to do IHC using the same human antibody. I know the secondary antibody will pick up the antibody that is injected as well as the one I applied as primary antibody in IHC. I hope I said it clearly. So please tell me what kind of blocking I can use to block the injected human antibody? And how? Will normal human serum work? Any input will be highly appreciated! Wen __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From lowman034 <@t> yahoo.com Fri May 27 14:03:20 2005 From: lowman034 <@t> yahoo.com (Dave Low) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] California Grossing Requirements In-Reply-To: 6667 Message-ID: <20050527190320.57492.qmail@web32004.mail.mud.yahoo.com> Cindy, This is the new requirement for the APR 2005 CAP#: ANP.11610 Phase II N/A YES NO If specimens are dissected by individuals other than a pathologist or pathology resident, do such individuals qualify as high complexity testing personnel under CLIA-88 regulations? NOTE: The laboratory director may delegate the dissection of specimens to non-pathologist individuals; these individuals must be qualified as high complexity testing personnel under CLIA-88 regulations. The minimum training/experience required of such personnel is: 1. An earned associate degree in a laboratory science or medical laboratory technology, obtained from an accredited institution, OR 2. Education/training equivalent to the above that includes at least 60 semester hours or equivalent from an accredited institution. This education must include 24 semester hours of medical laboratory technology courses, OR 24 semester hours of science courses that includes 6 semester hours of chemistry, 6 semester hours of biology, and 12 semester hours of chemistry, biology or medical laboratory technology in any combination. In addition, the individual must have laboratory training including either completion of a clinical laboratory training program approved or accredited by the ABHES, the CAHEA, or other organization approved by HHS (note that this training may be included in the 60 semester hours listed above), OR at least 3 months documented laboratory training in each specialty in which the individual performs high complexity testing. In addition, the CLIA-88 regulations include exceptions for grandfathered individuals; these regulations (42CFR493.1489 and 1491) may be found at http://www.phppo.cdc.gov/clia/regs/subpart_m.aspx#493.1487 This checklist question applies only to laboratories subject to CLIA-88. D. Low --- Cindy DuBois wrote: > The person in our lab who grosses most of the > surgical specimens is not a certified pathology > assistant (or an HT). At our last CLIA inspection > the inspector asked us for the "PA's" certification > documentation. I deferred the question to the "PA" > themselves. She managed to change the subject quite > quickly and the topic was not brought up again. Now > our grossing person is thinking that she is not > qualified and should not be grossing according to > government regulations. I have not been able to > find anything anywhere (google search) stating who > can gross or not gross tissue specimens. > > Can anyone help me with this question? > > Cindy DuBois > > > > __________________________________________________ > Do You Yahoo!? > Tired of spam? Yahoo! Mail has the best spam > protection around > http://mail.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From sjchtascp <@t> yahoo.com Fri May 27 14:32:00 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] gram stain control Message-ID: <20050527193200.41795.qmail@web90202.mail.scd.yahoo.com> I'm looking for a good gram stain control with both gram +/-. Any good suppliers out there. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! From Jenny-Oblander <@t> omrf.ouhsc.edu Fri May 27 14:43:06 2005 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] gram stain control Message-ID: Slim Jim's -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: Friday, May 27, 2005 2:32 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] gram stain control I'm looking for a good gram stain control with both gram +/-. Any good suppliers out there. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rachael_Emerson <@t> URMC.Rochester.edu Fri May 27 14:51:24 2005 From: Rachael_Emerson <@t> URMC.Rochester.edu (Emerson, Rachael) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] frozen sections Message-ID: Hello. I am in need of some help with trying to freeze mouse embryos for frozen sectioning. I am dissecting mouse embryos out in PBS (Embryonic day 8.5-14.5) and freezing them in VWR's Neg 50 Embedding Media. Initially I put a small amount of Neg 50 in the bottom of a Polyscience Plastic Peel-Away Mold, oriented the embryo, and then added more Neg 50. I then put the molds in the -80 Freezer and let them solidify. They seem to cut OK, but the morphology was terrible. Not so much in the E14.5, but other embryos looked very degraded. Next, I tried the same procedure but froze them by floating the mold in ethanol with dry ice. Still the morphology was terrible. I tried to freeze the embryos directly and then embed them, but they just turned to mush. On my final attempt I repeated the same procedure, but froze the mold by holding the bottom in liquid nitrogen. The block froze in seconds, but when I took them it out of the mold it was cracked and very hard to cut. The few sections I did manage were terrible and you could see the ice crystals in the embryos. I would really appreciate any advice or suggestions you have to offer. Thank you Rachael Emerson From jqb7 <@t> cdc.gov Fri May 27 14:51:46 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] gram stain control Message-ID: I heard that they weren't good for BOTH gram +/-...not true? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jenny Oblander Sent: Fri 5/27/2005 3:43 PM To: 'Steven Coakley'; Histonet@lists.utsouthwestern.edu Cc: Subject: RE: [Histonet] gram stain control Slim Jim's -----Original Message----- From: Steven Coakley [mailto:sjchtascp@yahoo.com] Sent: Friday, May 27, 2005 2:32 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] gram stain control I'm looking for a good gram stain control with both gram +/-. Any good suppliers out there. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From djohnson14 <@t> hotmail.com Fri May 27 14:57:45 2005 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] gram stain control In-Reply-To: Message-ID: Ha ha, thats a good one Mercedes Medical has a pretty good one. Give us a call Dave JOhnson Mercedes Medical 800-331-2716 >From: "Bartlett, Jeanine" >To: "Jenny Oblander" ,"Steven Coakley" >, >Subject: RE: [Histonet] gram stain control >Date: Fri, 27 May 2005 15:51:46 -0400 > >I heard that they weren't good for BOTH gram +/-...not true? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jenny >Oblander > Sent: Fri 5/27/2005 3:43 PM > To: 'Steven Coakley'; Histonet@lists.utsouthwestern.edu > Cc: > Subject: RE: [Histonet] gram stain control > > > > Slim Jim's > > > -----Original Message----- > From: Steven Coakley [mailto:sjchtascp@yahoo.com] > Sent: Friday, May 27, 2005 2:32 PM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] gram stain control > > I'm looking for a good gram stain control with both gram +/-. Any good > suppliers out there. > > Steve > > > --------------------------------- > Do You Yahoo!? > Yahoo! Small Business - Try our new Resources site! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From david.kinsley <@t> spcorp.com Fri May 27 14:59:24 2005 From: david.kinsley <@t> spcorp.com (Kinsley, David) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] frozen sections Message-ID: Hi Rachael, I use OCT embedding compound supplied by Sakura/Tissue Tek. I put a small amount in the bottom of a plastic embedding mold, orientate the specimen, fill the mold with OCT and then immerse the entire mold into a slurry of 2-methyl Butane (isopentane) in dry ice. With the technique you can keep the sample immersed a few minutes (2-3)without it cracking. It is best to immerse the sample, as it will freeze from all sides rather than from the bottom up as a floating technique would do. Liquid Nitrogen freezes quickly, but you have to be very careful as the blocks tend to crack very often. Freezing in the -80 is too slow and ice crystal will form in your samples. Good luck Dave -----Original Message----- From: Emerson, Rachael [mailto:Rachael_Emerson@URMC.Rochester.edu] Sent: Friday, May 27, 2005 3:51 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] frozen sections Hello. I am in need of some help with trying to freeze mouse embryos for frozen sectioning. I am dissecting mouse embryos out in PBS (Embryonic day 8.5-14.5) and freezing them in VWR's Neg 50 Embedding Media. Initially I put a small amount of Neg 50 in the bottom of a Polyscience Plastic Peel-Away Mold, oriented the embryo, and then added more Neg 50. I then put the molds in the -80 Freezer and let them solidify. They seem to cut OK, but the morphology was terrible. Not so much in the E14.5, but other embryos looked very degraded. Next, I tried the same procedure but froze them by floating the mold in ethanol with dry ice. Still the morphology was terrible. I tried to freeze the embryos directly and then embed them, but they just turned to mush. On my final attempt I repeated the same procedure, but froze the mold by holding the bottom in liquid nitrogen. The block froze in seconds, but when I took them it out of the mold it was cracked and very hard to cut. The few sections I did manage were terrible and you could see the ice crystals in the embryos. I would really appreciate any advice or suggestions you have to offer. Thank you Rachael Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From dellav <@t> musc.edu Fri May 27 15:09:34 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] frozen sections Message-ID: There are two very good articles discussing preparing tissues for frozen cryotomy by Gayle Callis in the Spring and Autumn 2004 issues of HistoLogic. you can access them by using the HistoLogic link at www.sakuraus.com Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Emerson, Rachael" 05/27/05 03:51PM >>> Hello. I am in need of some help with trying to freeze mouse embryos for frozen sectioning. I am dissecting mouse embryos out in PBS (Embryonic day 8.5-14.5) and freezing them in VWR's Neg 50 Embedding Media. Initially I put a small amount of Neg 50 in the bottom of a Polyscience Plastic Peel-Away Mold, oriented the embryo, and then added more Neg 50. I then put the molds in the -80 Freezer and let them solidify. They seem to cut OK, but the morphology was terrible. Not so much in the E14.5, but other embryos looked very degraded. Next, I tried the same procedure but froze them by floating the mold in ethanol with dry ice. Still the morphology was terrible. I tried to freeze the embryos directly and then embed them, but they just turned to mush. On my final attempt I repeated the same procedure, but froze the mold by holding the bottom in liquid nitrogen. The block froze in seconds, but when I took them it out of the mold it was cracked and very hard to cut. The few sections I did manage were terrible and you could see the ice crystals in the embryos. I would really appreciate any advice or suggestions you have to offer. Thank you Rachael Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sharon.willman <@t> bms.com Fri May 27 15:20:49 2005 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Sep 16 15:25:08 2005 Subject: [Histonet] Hostonet Mallory Heidenhain stain Message-ID: <42978121.5050807@bms.com> Hi, I am needing information on the Mallory Heidenhain staining procedure regarding washing out of the slides. The tissues have been fixed in 10% formalin and processed through a routine processing schedule. The slides are stained with the Mallory Heidenhain stain for 5 minutes and washed gently in running tap water for 30 seconds. They are dehydrated, cleared and then coverslipped. If anyone has experience in this stain, I would appreciate any hints that you may be willing to share. Thanks in advance. Sharon Willman From jhnspam <@t> aol.com Fri May 27 15:24:48 2005 From: jhnspam <@t> aol.com (jhnspam@aol.com) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] How to prevent specimens from freezing Message-ID: Does anyone know what can be done to prevent specimens from freezing when put outside in a courier pick up box during the cold winter months? The specimen has a freeze looking artifact after processing on a conventional processor. Thanks for any information. Pam From jkiernan <@t> uwo.ca Fri May 27 15:27:49 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Will brain tissue shrink even more if stored in70%alcoholprior to processing? References: Message-ID: <429782C5.4BB0F302@uwo.ca> In the HistoLogic May 2000 paper the duration of the initial formaldehyde fixation is not stated; only that the brains were "previously well fixed in neutral buffered formalin." Frequently specimens are not fixed for long enough to make them resistant to bad effects of solvents etc. It's also seems from the paper that the test pieces of brain were passed directly from the buffered fixative into 70% alcohol. This causes precipitation of sodium phosphate in the tissue (see Freida Carson 1996 "Histotechnology" 2nd edn, p.27). The control specimens in the HistoLogic paper were passed from buffered formalin to 60% ethanol (which safely extracts the phosphate buffer salts). Could the holes in the white matter be made by buffer salt crystals rather than forming slowly during storage in 70% ethanol? Just a thought! -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Vinnie Della Speranza wrote: > > You may wish to look at an article authored by Jane Chladny, published in HistoLogic, May 2000, entitled "Artifact in Tissues Held in 70% Ethanol" > > the author reported the appearance of 'vacuole' artifact in nervous tissues observed in a variety of mammalian tissues after storage in 70% ethanol. > > you can access the article by selecting the link for HistoLogic at www.sakuraus.com > > CM Bush wrote: > > > > Dear Histonet, > > > > Hello, here is my first post to the list, thank you in advance for your help. > > > > Summary: > > Having problems with mouse and human brain tissue shrinkage durning processing, but would like to better preserve antigens, concerned storage of tissue 70% ethanol will shrink tissue even further: > > > > We have lots of brain specimens, stored in formalin for a long time (months up to 10 years), both human and mouse. We perform immunohistochemistry on paraffin embedded brain. Of course, there is the massive over fixation. Also the tissue shrinks a lot during processing. > > > > In my last lab, after brain tissue had been fixed in formalin for 7 days we would store the tissue in 70% ethanol. I'd like to start storing the brain tissue I work with now in 70%, but I'm worried about the tissue possibly shinking too much, as the tissue already seems to shrink by greater than 60% after processing. > > > > (Previously, we would gradually increase the alcohols, 30% for 1 hour, then 50% for an hour in a bucket, room temperature on a rocker table, then put the cassettes into a bucket of 70% and store at room temp or 4C- does this process help any with controling the shrinkage factor?) > > > > Maybe this is a little bit long...thank you very much for your time. > > > > CM Bush > --------- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Vinnie Della Speranza > Manager for Anatomic Pathology Services > Medical University of South Carolina > 165 Ashley Avenue Suite 309 > Charleston, SC 29425 > Ph: 843-792-6353 > fax: 843-792-8974 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri May 27 15:32:12 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] How to prevent specimens from freezing Message-ID: I don't have your answer, but does anyone know how to keep a bull from charging? Hide his credit cards. (I'm sorry, . . . . . I had to). jhnspam@aol.com Sent by: histonet-bounces@lists.utsouthwestern.edu 05/27/2005 03:24 PM To: histonet@pathology.swmed.edu cc: Subject: [Histonet] How to prevent specimens from freezing Does anyone know what can be done to prevent specimens from freezing when put outside in a courier pick up box during the cold winter months? The specimen has a freeze looking artifact after processing on a conventional processor. Thanks for any information. Pam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri May 27 15:34:35 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Will brain tissue shrink even more if stored in70%alcoholprior to processing? Message-ID: Is this the reason for hangovers? From jkiernan <@t> uwo.ca Fri May 27 15:51:01 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Hostonet Mallory Heidenhain stain References: <42978121.5050807@bms.com> Message-ID: <42978835.B5D6C4D7@uwo.ca> You'll need to provide more details. Mallory's and Heidenhain's methods are quite different, and both require visual checking after the different steps. This is particularly important with Heidenhain's AZAN, which has two critical differentiations in the procedure. Whatever method you're using, the cause of weak or uneven staining could lie with the fixation of the tissue or the washing and dehydration of the sections after any or all stages of the staining procedure. Fixation. Simple formaldehyde fixation is not the best for trichrome methods. Treating the hydrated slides with picric acid for a few hours (followed by washing in water) before staining improves the result. Your note implies a single staining solution - one of the one-step trichromes like Gomori's or Cason's. Such methods can work well, but the user has no control over the actions of the three anionic dyes and the PTA (or PMA). Mallory and Heidenhain must be turning in their graves, having their names attached to a one-step trichrome - if indeed that is the case. Washing & dehydration. Never wash in tap water after any step of any trichrome method. If it's neutral or slightly alkaline the water will extract the dyes and leave a messy result. The rinses should be in water that has been slightly acidified. (I put about 5ml of 5% acetic acid in about 100 ml of tap water; no need to measure it!) After the last rinse in acidified water, take the slides into the first of 3 changes of new 100% alcohol. Avoid "graded" alcohols because water-alcohol mixtures can extract the dyes. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Sharon E Willman wrote: > > Hi, > > I am needing information on the Mallory Heidenhain staining procedure > regarding washing out of the slides. The tissues have been fixed in 10% > formalin and processed through a routine processing schedule. The > slides are stained with the Mallory Heidenhain stain for 5 minutes and > washed gently in running tap water for 30 seconds. They are dehydrated, > cleared and then coverslipped. > > If anyone has experience in this stain, I would appreciate any hints > that you may be willing to share. > > Thanks in advance. > > Sharon Willman > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dndsomi <@t> aol.com Fri May 27 16:19:37 2005 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] substitute for xylene Message-ID: <1a8.38a9a840.2fc8e8e9@aol.com> We switched to Formula 83 a few months ago and it works really well for us. DDietz, HT From sjchtascp <@t> yahoo.com Fri May 27 19:29:15 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] gram control Message-ID: <20050528002915.25997.qmail@web90202.mail.scd.yahoo.com> Thanks Dave, but unable to open. --------------------------------- Yahoo! Mail Stay connected, organized, and protected. Take the tour From themagoos <@t> rushmore.com Fri May 27 20:11:14 2005 From: themagoos <@t> rushmore.com (Jason and Heather) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] gram stain control References: <20050527193200.41795.qmail@web90202.mail.scd.yahoo.com> Message-ID: <004401c56322$275e5a60$b3fc9fd1@magoo> We use Necomer Supply. Jason ----- Original Message ----- From: "Steven Coakley" To: Sent: Friday, May 27, 2005 1:32 PM Subject: [Histonet] gram stain control > I'm looking for a good gram stain control with both gram +/-. Any good > suppliers out there. > > Steve > > > --------------------------------- > Do You Yahoo!? > Yahoo! Small Business - Try our new Resources site! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gu.lang <@t> gmx.at Sat May 28 01:45:29 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Fri Sep 16 15:25:09 2005 Subject: AW: [Histonet] frozen sections In-Reply-To: Message-ID: This method for freezing sounds good but rather time-consuming. I would like to know, how surgery-labs handle tissue that is to asservate (perhaps from native lymphnodes or other interesting things sent in for frozen-sections). We put a small piece of tissue in a plastic-tube and let it sink into the liquid nitrogen. There we store it, untill the tissue is either sent to another lab or thrown away. Is this a usual method? Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Vinnie Della Speranza Gesendet: Freitag, 27. Mai 2005 22:10 An: histonet@lists.utsouthwestern.edu; Rachael_Emerson@URMC.Rochester.edu Betreff: Re: [Histonet] frozen sections There are two very good articles discussing preparing tissues for frozen cryotomy by Gayle Callis in the Spring and Autumn 2004 issues of HistoLogic. you can access them by using the HistoLogic link at www.sakuraus.com Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, SC 29425 Ph: 843-792-6353 fax: 843-792-8974 >>> "Emerson, Rachael" 05/27/05 03:51PM >>> Hello. I am in need of some help with trying to freeze mouse embryos for frozen sectioning. I am dissecting mouse embryos out in PBS (Embryonic day 8.5-14.5) and freezing them in VWR's Neg 50 Embedding Media. Initially I put a small amount of Neg 50 in the bottom of a Polyscience Plastic Peel-Away Mold, oriented the embryo, and then added more Neg 50. I then put the molds in the -80 Freezer and let them solidify. They seem to cut OK, but the morphology was terrible. Not so much in the E14.5, but other embryos looked very degraded. Next, I tried the same procedure but froze them by floating the mold in ethanol with dry ice. Still the morphology was terrible. I tried to freeze the embryos directly and then embed them, but they just turned to mush. On my final attempt I repeated the same procedure, but froze the mold by holding the bottom in liquid nitrogen. The block froze in seconds, but when I took them it out of the mold it was cracked and very hard to cut. The few sections I did manage were terrible and you could see the ice crystals in the embryos. I would really appreciate any advice or suggestions you have to offer. Thank you Rachael Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Krat18 <@t> aol.com Sat May 28 18:46:34 2005 From: Krat18 <@t> aol.com (Krat18@aol.com) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] gram stain control Message-ID: In a message dated 5/27/2005 8:19:46 P.M. Central Standard Time, themagoos@rushmore.com writes: We use Necomer Supply. Jason Ditto Karen Raterman _Karen_Raterman@ssmhc.com_ (mailto:Karen_Raterman@ssmhc.com) From louise.renton <@t> gmail.com Mon May 30 02:27:29 2005 From: louise.renton <@t> gmail.com (louise renton) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] How to prevent specimens from freezing In-Reply-To: References: Message-ID: Would a polystyrene or cooler box be of any use? You know the sort use to keep your beers cool in summer? On 5/27/05, jhnspam@aol.com wrote: > Does anyone know what can be done to prevent specimens from freezing when > put outside in a courier pick up box during the cold winter months? The specimen > has a freeze looking artifact after processing on a conventional processor. > Thanks for any information. > > Pam > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "....I know who I am. No-one else knows who I am. If I was a giraffe, and someone said I was a snake, I'd think no, actually I'm a giraffe" Richard Gere From Jacqueline.Malam <@t> rli.mbht.nhs.uk Tue May 31 04:36:09 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Gram control Message-ID: I have used stilton blue cheese in the past and it works quite well ! Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Kemlo.Rogerson <@t> elht.nhs.uk Tue May 31 04:51:05 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Gram control Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9E5AD1F1@elht-exch1.xelht.nhs.uk> Nicked from Tony's sarnies? -----Original Message----- From: Malam Jacqueline (Morecambe Bay Hospitals NHS Trust) Sent: 31 May 2005 10:36 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Gram control I have used stilton blue cheese in the past and it works quite well ! Jacqui Malam Royal Lancaster Infirmary England DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Tue May 31 07:34:05 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] slide drying Message-ID: <5D2189E74151CC42BEC02906BA8996322B90B9@exchsrv01.barrynet.barry.edu> I have tried melting the wax. If the wax actually melts (becomes liquid), the sections are likely to warp. Drying the sections for 48 hours just below the melting point of the wax has worked best for me. I suspect that the longer warming time denatures the albumin and makes it a better adhesive. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Edmondson David (RBV) NHS Christie Tr Sent: Thursday, May 26, 2005 1:02 PM To: Steven Coakley; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] slide drying Has anyone suggested that the sections need melting on rather than just drying? or have you done this in the process described. Shaking trapped water off before drying and then melting may be an issue if you have trapped amounts of water under the sections. Whether melted or just dried on. David Christie Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Steven Coakley Sent: 24 May 2005 19:37 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] slide drying I'm setting up a small Histo lab and using a 60C slide warmer to dry my slides. I've been currently allowing them to air dry vertically for a couple of hours to overnight, then placing the slides on the slide warmer for an hour. About 90% of the time the slides turn out ok, but some slide do show some detaching from the slide. Both charges and non-charged slides with water bath adhesive seem to be involved. I read a past artical on this site not recommending allowing slides to air dry as the section may entrap water? Any suggestions. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From jessgrocki <@t> yahoo.com Sun May 29 10:50:04 2005 From: jessgrocki <@t> yahoo.com (Jessica Piche) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] New Mc Letchies Trichrome Kit Message-ID: <20050529155004.38106.qmail@web81710.mail.yahoo.com> Hi All, We're looking into a new trichrome stain and are wondering if anyone has tried or is now using the New Mc Letchie's Trichrome kit (New Comers Supply)? And what does everyone who is using it think about it? Any feedback would be appreciated. Thanks, Jessica Piche-Grocki, HT From brett_connolly <@t> merck.com Tue May 31 08:23:27 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Bielschowsky's on frozens Message-ID: Has anyone performed Bielschowsky's silver staining on frozen sections? I have some 10 micron unfixed, frozen sections in the freezer and would like some guidance on fixation and any possible adjustments to the paraffin protocol. Thanks, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From jqb7 <@t> cdc.gov Tue May 31 08:33:32 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] New Mc Letchies Trichrome Kit Message-ID: We just received our free stained slides and technical memo from Newcomer. The pathologist's all love it so I plan to order it soon. I'll let you know how it works for me. Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road MS/G32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jessica Piche Sent: Sunday, May 29, 2005 11:50 AM To: histonet@pathology.swmed.edu Subject: [Histonet] New Mc Letchies Trichrome Kit Hi All, We're looking into a new trichrome stain and are wondering if anyone has tried or is now using the New Mc Letchie's Trichrome kit (New Comers Supply)? And what does everyone who is using it think about it? Any feedback would be appreciated. Thanks, Jessica Piche-Grocki, HT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From susan.wells <@t> bms.com Tue May 31 08:38:33 2005 From: susan.wells <@t> bms.com (Susan Q Wells) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] perkinje cell staining Message-ID: <429C68D9.3060907@bms.com> Can anyone offer insight into why an antibody known to be expressed on activated T cells is lighting up perkinje cells in frozen human cerebellum sections ????? I'm using a pretty standard staining scenario,avidin/ biotin block, Fc block, biotinylated primary, streptavidin HRP, DAB,. Thanks in advance, Susan Wells From dpahisto <@t> yahoo.com Tue May 31 09:19:53 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Stain Racks Message-ID: <20050531141953.33260.qmail@web33410.mail.mud.yahoo.com> Does anyone know where I can purchase slide racks for the TissueTek DRS ? We currently run about 250 slides a day and only have 6 racks (one which is broken). Sakura is not currently manufacturing them due to a production problem. Any suggestions is greatly appreciated, Cindy Dubois Delta Pathology Assoc. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From dsnider <@t> shrinenet.org Tue May 31 09:46:43 2005 From: dsnider <@t> shrinenet.org (Snider, Deanna) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] cutting GMA blocks Message-ID: Dear histonetters, I hope you can help!! Thanks in advance. I have learned to process and embed GMA specimens with no problem, BUT the cutting is a nigthmare! For one I can hardly see them, and 2, they are of the absolute worst quality- totally unacceptable to me! There are compressions, wrinkles, folds- you name it! By far the worst work I have done since learning in 1985! Does anybody have any hands on hints or tips to get a nice smooth quality section from GMA's? Deanna Snider HT ASCP Shriners Hospital for Children Research Dept. Cincinnatti, Oh 513-872-6388 CONFIDENTIALITY NOTICE: This e-mail communication and any attachments may contain confidential and privileged information for the use of the designated recipients. If you are not the intended recipient, (or authorized to receive for the recipient) you are hereby notified that you have received this communication in error and that any review, disclosure, dissemination, distribution or copying of it or its contents is prohibited. If you have received this communication in error, please destroy all copies of this communication and any attachments and contact the sender by reply e-mail or telephone (813) 281-0300. From contact <@t> excaliburpathology.com Tue May 31 11:05:41 2005 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] cutting GMA sections Message-ID: <20050531160541.6007.qmail@web50310.mail.yahoo.com> Try floating the GMA sections in a room temp waterbath filled with weak ammonia water. Paula Pierce, HTL(ASCP)HT Excalibur Pathology, Inc. 630 N. Broadway Moore, OK 73160 405-759-3953 contact@excaliburpathology.com www.excaliburpathology.com From doninn <@t> mail.nih.gov Tue May 31 10:54:41 2005 From: doninn <@t> mail.nih.gov (Donin, Nick (NIH/NCI)) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] mouse endothelial cells in brain Message-ID: <16A0583FB1644E4DB8C0A0265028B6FD02B4E725@nihexchange13.nih.gov> Histonetters, This is my first posting, so I'm excited that someone out there might be able to help me. I am trying to stain endothelial cells in brain tumors in formalin fixed paraffin embedded mouse tissues. I have tried several antibodies, and none of them seem to work. I am consistently getting high background and no true staining. I have used the following antibodies: BD Pharmigen anti-mouse CD31 (PECAM-1) cat #557355 Santa Cruz CD31 (PECAM-1) cat # sc-1506 (I have seen postings that this works, but my protocol didn't produce good results) Cedarlane mouse CD31 cat#CL8930 AP Cymbus Biotechnology anti-ms CD31 cat# CBL1337 The protocol I use is shown below. I'm confident that out of these antibodies, at least one will be able to stain the endothelial cells in these tumors, but I think that my protocol may be the problem. Could someone possibly suggest and antibody or protocol that would work for me? Thanks so much for your help, much appreciated. Nick Donin CRTA Neuro-Oncology Branch National Cancer Institute National Institutes of Health 9000 Rockville Pike Building 35, Room 2B-203 Bethesda, MD 20892 From sjchtascp <@t> yahoo.com Tue May 31 11:39:13 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] new trichrome stain Message-ID: <20050531163913.30668.qmail@web90209.mail.scd.yahoo.com> I'd certainly do some comparitive studies and ask for a sample. Gomori's, is very easy to make up yourself. Even the componets can be adjusted to render more sutable staining intensities depending on what your Pathologist preferes. Steve --------------------------------- Do You Yahoo!? Yahoo! Small Business - Try our new Resources site! From sbreeden <@t> nmda.nmsu.edu Tue May 31 13:33:01 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Speaking of Gram's... Message-ID: I'm having trouble with my Gram's in that my positive bacteria are losing color. So, I'm wondering what the exact differentiation time should be for the step after the Gram's Iodine (I'm using the Sigma kit)? The instructions are not precise, so I'm thinkin' maybe I'm over differentiating or under-Safranin-ing or aliens (this is the State that's home to ROSWELL...) have taken over my kit. Anyone have anything a bit more specific? I'd appreciate any help... Sara A. Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From sharon.osborn <@t> dnax.org Tue May 31 14:41:02 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Freezing muscle biopsies Message-ID: <29B25753F6B1D51196110002A589D44402397FB8@PALMSG30.us.schp.com> Liz, Our researchers often freeze tissue for later study. They use the plastic disposable embedding molds. These can be used with the dry ice as John Kiernan suggested. There is a thin layer of OCT placed in bottom of mold then the tissue is oriented in this with additional OCT to fill the mold. The mold can be labeled with a permanent felt tip pen. Each specimen is then placed into the labeled sterile sampling bags, packed in dry ice and shipped to respective location. We have experienced very little distortion of the tissue due to ice crystals, etc. The testing has worked out okay. The main tissues were liver, kidney, lung. You may wish to experiment with some tissues before actually tackling the research samples to get the right combination for you. Sharon Osborn DNAX ScheringPlough Biopharma Palo Alto, CA Hello Histonetters > > I have a unique question. We are currently starting to set up > procedure for collecting samples from a clinical trial. The clinical > trial involves taking multiple synovial biopsies at a surgery center. > Since portions of the samples need to be processed for frozen sections > we wanted to be able to freeze the specimens at the surgery center via > isopentane cooled liquid nitrogen. We really do not want to have to > transport the multiple specimens back to the main lab prior to > freezing due to the time involved it would probably be 1-2 hours post > biopsy before we could freeze the samples. The surgery center is > questioning the flammability of the isopentane. Has anyone > encountered anything like this? Any suggestions would be helpful. > Thanks in advance. > > Liz > > Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC > Manager > Premier Laboratory, LLC > P.O. Box 18592 > Boulder, Colorado 80308 > Office: (303) 735-5001 > Fax: (303) 735-3540 > liz@premierlab.com > www.premierlab.com > > Ship to Address: > Premier Laboratory > University of Colorado > MCDB, Room A3B40 > Boulder, Colorado 80309 ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From Guy.Mitchell <@t> carolinashealthcare.org Tue May 31 14:42:59 2005 From: Guy.Mitchell <@t> carolinashealthcare.org (Mitchell, Guy) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Enzyme Staining and Billing for Controls Message-ID: <4DCF5CA56B4AA04ABC383DE358BAD3820289100D@dcr-xchg-04.carolinas.org> A question has come up concerning charges for muscle enzyme stains. We stain for Adenylate Deaminase and Phosphofructokinase as well as negatives for each. I assume they should be treated as controls without a charge but others disagree. Guy W. Mitchell (704) 379-5984 (704) 379-6144 Fax ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. From Charles.Embrey <@t> carle.com Tue May 31 15:42:58 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Number of Blocks Submitted by PA Message-ID: I have been gone for a week and just got a chance to check my e-mails and found this question interesting. First, have you asked the PA? I know it seems like a simple question but you would be surprised at all the misunderstanding that can be cleared up with a simple question asked? Please know that I have NEVER met a pathologist that will gladly read more slides than he thinks are necessary for a good diagnosis on a routine basis. I gross for seven pathologists and it is tricky to satisfy all at the same time. Luckily, my pathologists and I here have great communication and we manage to keep the block count, as well a gross recut number down. As far as "rules of thumb" we do have certain guidelines that are basically thought of as "standards of care". Without knowing other details the best advice I could give is to ask the PA. Charles Embrey PA(ASCP) Urbana IL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith Wanda Sent: Tuesday, May 24, 2005 7:56 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Number of Blocks Submitted by PA Good Morning Histonetters, I have a question for you this morning...Are there written standards or "Rules of Thumb" regarding number of blocks that should be submitted on certain tissues? The reason I ask is, "it seems to me" that sometimes the PA submits an extreme number of blocks on some specimens that could be accomplished with less. I'm not advocating cutting corners or not performing good medical practice, I just need to know. An example would be a uterus w/ fibroids, tubes and ovaries which we have gotten up to 24 blocks on. I'm not a PA, but it's interesting to me that when the Pathologist cut, we get alot less blocks. Please advise. Thanks, Wanda > Wanda G. Smith, HTL/HT(ASCP) > Pathology Supervisor > Trident Medical Center Laboratory Services > *9330 Medical Plaza Drive, Charleston, SC 29406 > *843-797-4586 *fax 843-797-4296 > *wanda.smith@hcahealthcare.com > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tony_Reilly <@t> health.qld.gov.au Tue May 31 23:13:15 2005 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Fri Sep 16 15:25:09 2005 Subject: [Histonet] Aquaporin immunohistochemistry Message-ID: Hello All I have been requested to be involved in a research study demonstrating Aquaporin 1 (AQ1, AQP1) using IHC. To date I have found 2 antibodies available from Chemicon. My questions are: 1. Are there other commercial suppliers of this Ab. 2. If not, which of the 2 from Chemicon is the preferred option. 3. Do you realyy need to fix with paraformaldehyde as stated in most methods, and why? i would appreciate any advice forwarded. regards Tony Reilly Chief Scientist Anatomical Pathology Northside Pathology Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au *********************************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is prohibited. It may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone or by return email. You should also delete this email and destroy any hard copies produced. ***********************************************************************************