From cheastys <@t> svm.vetmed.wisc.edu Mon Dec 1 09:04:17 2008 From: cheastys <@t> svm.vetmed.wisc.edu (Sandra Cheasty) Date: Mon Dec 1 09:04:39 2008 Subject: [Histonet] T Cell and B Cell Markers on Lymph Node Touch Preps Message-ID: Hello, Does anyone have experience with performing Immunocytochemistry (CD3, CD20) on touch preps of canine and feline lymph nodes, particularly the following: Fixation: What reagent(s), what temperature, how long, immediately or wait until air dried, storage post fixing Antigen Retrieval: If formalin is part of the fixative, is there a general rule for HIER on ICC specimens that relates to HIER on paraffin sections of lymph nodes for CD3 and CD20? Thank you so very much. Sandra Cheasty Histology & Necrcopsy Supervisor UW-Madison School of Veterinary Medicine From schaundrawalton <@t> yahoo.com Mon Dec 1 11:57:39 2008 From: schaundrawalton <@t> yahoo.com (Schaundra Walton) Date: Mon Dec 1 11:57:42 2008 Subject: [Histonet] Question RE: HQIP Result Message-ID: <486600.42474.qm@web58905.mail.re1.yahoo.com> We just got our HQIP results and they made a comment regarding the breast tissue H&E slide we submitted.? Can someone please tell me what nuclear bubbling artifact is and what causes it?? ? Thanks! Schaundra Walton BS, HTL (ASCP) Histology Supervisor Swedish American Hospital Rockford, IL From zodiac29 <@t> comcast.net Mon Dec 1 12:10:02 2008 From: zodiac29 <@t> comcast.net (zodiac29@comcast.net) Date: Mon Dec 1 12:10:07 2008 Subject: [Histonet] HALT Message-ID: <120120081810.18797.4934287A000180660000496D2215593414C7CD0C0E070B0196@comcast.net> Hello all, Can anyone tell me where I could buy Halt. It is used to get wrinkle free sections on the floatation bath. Thanks Jenny From tbraud <@t> holyredeemer.com Mon Dec 1 12:13:58 2008 From: tbraud <@t> holyredeemer.com (Terri Braud) Date: Mon Dec 1 12:14:05 2008 Subject: [Histonet] RE: buffy coat preps In-Reply-To: <3a95ff7c00035b12@HolyRedeemer.com> Message-ID: I've done lots of buffy coat preps for IHC using cyto-centrifuge preps, though we always found methanol to be the kiss of death to IHC fixation, but don't have a clue as to why. You'd do better to use 95% Alcohol for at least 10 minutes. Also, by using an alcohol fixative instead of formalin, frequently, much less pretreatment and incubation time is needed. I used to have about 19 Abs worked out for the Ventana with this method. Sadly, the operative word is "used" to. I hope this helps. Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Laboratory Holy Redeemer Hospital and Medical Center 1648 Huntingdon Pike Meadowbrook, PA 19046 (215) 938-3676 phone (215) 938-3689 fax - Message: 2 Date: Sun, 30 Nov 2008 23:54:59 -0500 From: John Kiernan Subject: Re: [Histonet] Buffy coat preparation for immuno T Why go to all the trouble of embedding and sectioning a buffy coat specimen? What's wrong with diluting it in saline and then making smears or (better, if you have the equipment) cyto-centrifuge preparations? You'll get lots of slides. They can be fixed in methanol and the WBC stained with any conventional Romanowsky-Giemsa method, or immunohistochemically. You could even fix in formalin, if there's some special reason to do so. (You have to lower the pH of an ordinary blood stain if the fixative was formaldehyde.) I hope this message is readable; it is sent as plain text. John Kiernan Anatomy, UWO London, Canada = = = --------------------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. From Phyllis_Burnette <@t> bshsi.org Mon Dec 1 12:37:46 2008 From: Phyllis_Burnette <@t> bshsi.org (Burnette, Phyllis) Date: Mon Dec 1 12:36:54 2008 Subject: [Histonet] Question RE: HQIP Result In-Reply-To: <486600.42474.qm@web58905.mail.re1.yahoo.com> References: <486600.42474.qm@web58905.mail.re1.yahoo.com> Message-ID: <2C7750460C3E7545AC11099BE6A7ABD601C994FA@EDC-MAIL-02.ads.bshsi.com> That would be from the primary fixation being alcohol. Phyllis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Schaundra Walton Sent: Monday, December 01, 2008 12:58 PM To: Histonet Subject: [Histonet] Question RE: HQIP Result We just got our HQIP results and they made a comment regarding the breast tissue H&E slide we submitted.? Can someone please tell me what nuclear bubbling artifact is and what causes it?? ? Thanks! Schaundra Walton BS, HTL (ASCP) Histology Supervisor Swedish American Hospital Rockford, IL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. ________________________________________________________________________________________________________ From rjbuesa <@t> yahoo.com Mon Dec 1 14:50:04 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 1 14:50:11 2008 Subject: [Histonet] Question RE: HQIP Result In-Reply-To: <486600.42474.qm@web58905.mail.re1.yahoo.com> Message-ID: <30567.83561.qm@web65708.mail.ac4.yahoo.com> Schaundra: Nuclear bubbling is an artifact caused?when you?oven dry sections that have not been properly drained and above usual temperature. Ren? J. --- On Mon, 12/1/08, Schaundra Walton wrote: From: Schaundra Walton Subject: [Histonet] Question RE: HQIP Result To: "Histonet" Date: Monday, December 1, 2008, 12:57 PM We just got our HQIP results and they made a comment regarding the breast tissue H&E slide we submitted.? Can someone please tell me what nuclear bubbling artifact is and what causes it?? ? Thanks! Schaundra Walton BS, HTL (ASCP) Histology Supervisor Swedish American Hospital Rockford, IL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From steely0511 <@t> yahoo.com Mon Dec 1 14:51:48 2008 From: steely0511 <@t> yahoo.com (John Steel) Date: Mon Dec 1 14:51:52 2008 Subject: [Histonet] EM ListServe Sites Message-ID: <24257.26162.qm@web59709.mail.ac4.yahoo.com> Hi All, Are any of you involved with electron microscopy?? I am interested in EM list services, and would love to hear of what is out there, or even start a list serve of my own?- particularly in the field of cryoTEM as applied to structural biology, bio-polymers, and drug discovery.? I would appreciate any info you could provide - Thanks! Best regards, Jerry From denise.woodward <@t> uconn.edu Mon Dec 1 15:12:00 2008 From: denise.woodward <@t> uconn.edu (Woodward, Denise) Date: Mon Dec 1 15:12:10 2008 Subject: [Histonet] T Cell and B Cell Markers on Lymph Node Touch Preps In-Reply-To: References: Message-ID: <40AC6D73C2B95C4CA21B26B7BF380C4002D0E4A2@EXCHANGED.mgmt.ad.uconn.edu> In my experience, CD20 does not work in canine or feline tissues. Try CD79a instead. I can't comment on the use of touch preps. With FFPE, Dako TRS pH9 for 20 minutes works great. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandra Cheasty Sent: Monday, December 01, 2008 10:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] T Cell and B Cell Markers on Lymph Node Touch Preps Hello, Does anyone have experience with performing Immunocytochemistry (CD3, CD20) on touch preps of canine and feline lymph nodes, particularly the following: Fixation: What reagent(s), what temperature, how long, immediately or wait until air dried, storage post fixing Antigen Retrieval: If formalin is part of the fixative, is there a general rule for HIER on ICC specimens that relates to HIER on paraffin sections of lymph nodes for CD3 and CD20? Thank you so very much. Sandra Cheasty Histology & Necrcopsy Supervisor UW-Madison School of Veterinary Medicine _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stamptrain <@t> yahoo.com Mon Dec 1 15:14:57 2008 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Mon Dec 1 15:15:01 2008 Subject: [Histonet] EM ListServe Sites References: <24257.26162.qm@web59709.mail.ac4.yahoo.com> Message-ID: <844923.92328.qm@web55803.mail.re3.yahoo.com> Jerry: There is a comprehensive listserve at www.microscopy.com.? You don't have to be a member of MSA to use it, but if you are an active electron microscopist (which I was for over 40 years) then the Microscopy Society of America is where you need to be. Roger Moretz, Ph.D. ----- Original Message ---- From: John Steel To: histonet@lists.utsouthwestern.edu Sent: Monday, December 1, 2008 3:51:48 PM Subject: [Histonet] EM ListServe Sites Hi All, Are any of you involved with electron microscopy?? I am interested in EM list services, and would love to hear of what is out there, or even start a list serve of my own?- particularly in the field of cryoTEM as applied to structural biology, bio-polymers, and drug discovery.? I would appreciate any info you could provide - Thanks! Best regards, Jerry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From shive003 <@t> umn.edu Mon Dec 1 15:30:24 2008 From: shive003 <@t> umn.edu (Jan Shivers) Date: Mon Dec 1 15:30:28 2008 Subject: [Histonet] T Cell and B Cell Markers on Lymph Node Touch Preps References: <40AC6D73C2B95C4CA21B26B7BF380C4002D0E4A2@EXCHANGED.mgmt.ad.uconn.edu> Message-ID: Works on canine and feline tissues: CD3; LabVision; cat. # RB-9039; HIER with Dako Target Retrieval high pH 9.0 (or 0.01M citrate buffer, pH 6.0, if used on liver tissue) CD20; LabVision; cat. # RB-9013; no pretreatment necessary. If you have formalin in your fixative, you'll need to do HIER on the CD3 slides... though the high pH will most likely remove some of your cells. If you use alcohol or acetone as your fixative, you won't need to do any HIER. Jan Shivers ----- Original Message ----- From: "Woodward, Denise" To: "Sandra Cheasty" ; Sent: Monday, December 01, 2008 3:12 PM Subject: RE: [Histonet] T Cell and B Cell Markers on Lymph Node Touch Preps In my experience, CD20 does not work in canine or feline tissues. Try CD79a instead. I can't comment on the use of touch preps. With FFPE, Dako TRS pH9 for 20 minutes works great. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sandra Cheasty Sent: Monday, December 01, 2008 10:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] T Cell and B Cell Markers on Lymph Node Touch Preps Hello, Does anyone have experience with performing Immunocytochemistry (CD3, CD20) on touch preps of canine and feline lymph nodes, particularly the following: Fixation: What reagent(s), what temperature, how long, immediately or wait until air dried, storage post fixing Antigen Retrieval: If formalin is part of the fixative, is there a general rule for HIER on ICC specimens that relates to HIER on paraffin sections of lymph nodes for CD3 and CD20? Thank you so very much. Sandra Cheasty Histology & Necrcopsy Supervisor UW-Madison School of Veterinary Medicine _______________________________________________ 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 Herrick.James <@t> mayo.edu Mon Dec 1 16:26:31 2008 From: Herrick.James <@t> mayo.edu (Herrick, James L.) Date: Mon Dec 1 16:26:35 2008 Subject: [Histonet] GMA embedded section attachment to slides Message-ID: <4F820D0A1054E6478FD124A9BE03397271F3B6@MSGEBE35.mfad.mfroot.org> Hi everyone, I have been trying to attach Glycol Methacrylate (GMA) embedded specimens to glass slides and have not been having much luck. So far I have tried gelatin, APES and Histostik. I am using a press and placing them in a 50? C oven for approx. 48 hours. The sections look as if they are attached, until I begin staining. I am pretty sure that the PEG is causing my problem, but have seen an article or two that have attached GMA sections to slides. Does anyone have a protocol that works well with GMA? If so, I would greatly appreciate any help I can get. Thank you much. Jim From ploykasek <@t> phenopath.com Mon Dec 1 17:49:18 2008 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Mon Dec 1 17:49:42 2008 Subject: [Histonet] Expiration date checks Message-ID: Hi all. I am hoping for some clever ideas on the best method to check for expired reagents. We all know that we shouldn't use them, I'm sure we all actively monitor this (right)! I must admit we occasionally have expired reagents - a stain or buffer we haven't used in a while is in a cupboard or fridge & it has expired. What do others do to monitor & prevent this from happening? We have a weekly safety check that spot checks a certain # of reagents for expiration, but we probably have thousands of reagents & don't check every single bottle every week. Thanks for the input. Patti Loykasek This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From Barry.R.Rittman <@t> uth.tmc.edu Mon Dec 1 18:13:26 2008 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Mon Dec 1 18:16:00 2008 Subject: [Histonet] Expiration date checks References: Message-ID: Patti How about just using an adhesive color dot system? Can use one dot to represent the year and second to represent the month. Just a suggestion, its been a long day! Barry. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Patti Loykasek Sent: Mon 12/1/2008 5:49 PM To: histonet Subject: [Histonet] Expiration date checks Hi all. I am hoping for some clever ideas on the best method to check for expired reagents. We all know that we shouldn't use them, I'm sure we all actively monitor this (right)! I must admit we occasionally have expired reagents - a stain or buffer we haven't used in a while is in a cupboard or fridge & it has expired. What do others do to monitor & prevent this from happening? We have a weekly safety check that spot checks a certain # of reagents for expiration, but we probably have thousands of reagents & don't check every single bottle every week. Thanks for the input. Patti Loykasek This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rod.coombe <@t> imvs.sa.gov.au Mon Dec 1 18:27:24 2008 From: rod.coombe <@t> imvs.sa.gov.au (Rod Coombe) Date: Mon Dec 1 18:28:55 2008 Subject: [Histonet] Iso propyl alcohol Message-ID: <005301c95414$bf37bcf0$0267140a@41504n> Hello all, Wondering if anybody has experienced tissue processor problems resulting from long term use of IPA. Rod Coombe Manager Surgical Pathology Frome Road Institute of Medical & Veterinary Science SA Pathology PO Box 14 Rundle Mall Adelaide, South Australia 5000 From steely0511 <@t> yahoo.com Mon Dec 1 19:20:24 2008 From: steely0511 <@t> yahoo.com (John Steel) Date: Mon Dec 1 19:20:29 2008 Subject: [Histonet] Microwave Processors Message-ID: <55063.27940.qm@web59714.mail.ac4.yahoo.com> Hi All, I am very curious about microwave processing technology, as I am told that total processing time is reduced to hours, and could use your candid opinions!? Does anyone have experience with this specific technology?? I understand that there are several units out there, but performance is an issue for me.? If you have such experience, could you share your comments?? How is tissue morphology effected?? Will this technology interfere with current CAP / CLIA/ JCAOH lab guidelines as currently interpreted?? How does this effect histology, special stains, IHC, ISH? Your comments are greatly appreciated! Best regards, Jerry (330) 771-7647 From cdbeads <@t> earthlink.net Mon Dec 1 21:06:46 2008 From: cdbeads <@t> earthlink.net (Daniel) Date: Mon Dec 1 21:06:49 2008 Subject: [Histonet] Free IHC CE course in Seattle Message-ID: <27915183.1228187206484.JavaMail.root@mswamui-backed.atl.sa.earthlink.net> A free continuing education workshop (3 CE units) is being sponsored by Diagnostic BioSystems in cooperation with the Washington State Histotechnology Society this Saturday, the 6th. The title is "IHC Basics and Beyond", presented by Ken Pierce. The workshop will be held at CellNetix: Cellnetix 1124 Columbia St, Suite 200 (206) 386-2676 Date: Saturday, December 6th Time: 9:30 AM ? 12:30 PM Title: IHC: Basics and Beyond Maximum attendance: 60 For more information please write Dan Adams at labdla@vmmc.org or call 206-223-6870. From pieronelva01 <@t> bigpond.com Tue Dec 2 01:52:20 2008 From: pieronelva01 <@t> bigpond.com (Piero Nelva) Date: Tue Dec 2 01:52:29 2008 Subject: [Histonet] protocols on Ventana References: <55063.27940.qm@web59714.mail.ac4.yahoo.com> Message-ID: <3A855683817C405F903BE1FDEA55A252@pentium4> I've deleted the original post, but to the person who was after Ventana protocols for Mismatch Repair proteins, here are ours: MSH2, 1:50, CC1 Mild, 37, 28 MSH6, 1:4000, CC1 Std, 37, 40 plus amplification kit MLH1, Vetnana predilute, CC1 Mild, 42, 28 plus amplification kit PMS2, 1:100, CC1 Mild, 37, 56 plus amplification kit. I can send you some images of our results off list if you like Regards Piero Nelva Anatomical Pathology Monash Medical Centre Australia From heroina <@t> ibiss.bg.ac.yu Tue Dec 2 05:02:44 2008 From: heroina <@t> ibiss.bg.ac.yu (Angelina Subotic) Date: Tue Dec 2 05:02:53 2008 Subject: [Histonet] (no subject) Message-ID: I am trying to find a full procedure for staining with Schiff reagens and naphtol blue black.Does anyone have a protocol they could send me? Any information will be greatly appreciated . Best regards Angelina Subotic From CHRISH <@t> HEALTHCARESCOUTS.COM Tue Dec 2 06:23:27 2008 From: CHRISH <@t> HEALTHCARESCOUTS.COM (Chris Handrahan) Date: Tue Dec 2 06:32:04 2008 Subject: [Histonet] Immediate Need for Histotech in Myrtle Beach, SC Message-ID: The #1 Recruiter for Laboratory/Biotech Specialists Healthcare Scouts is a medical laboratory/biotech recruitment firm that places professionals in full-time, permanent positions throughout the United States. As the leading nationally recognized permanent placement solution for medical laboratory/biotech professionals, Healthcare Scouts has the following histology opening in Myrtle Beach Day shift 8-5 M-F With a brand new urology practice Full time permanent position Looking to hire this week! Referrals With employment opportunities all over the US, we are always looking for qualified and motivated professionals. Recommend a friend or family member, and you could be eligible for up to $1,000 through Healthcare Scouts' referral program. Call us today to learn more! For immediate consideration please contact Chris Handrahan Managing Director of Allied Health Healthcare Scouts 800-708-0605 office 321-231-5427 cell chrish@healthcarescouts.com www.healthcarescouts.com From Susan.Weber2 <@t> va.gov Tue Dec 2 07:48:28 2008 From: Susan.Weber2 <@t> va.gov (Weber, Susan (VHACLE)) Date: Tue Dec 2 07:48:40 2008 Subject: [Histonet] Expiration date checks In-Reply-To: References: Message-ID: <16C83872A53F4346AA9C3A18E3A3AAB903F76E9D@VHAV10MSGA1.v10.med.va.gov> Color coding with sticker "dots" helps a bit. I designate a color for each year (5 yrs/5 colors max) On the top of that years color I put a date (usually 6/08 whatever) and make them easier to spot. (Pardon the pun!)You can find color dots in the office supply catalog, Office Max (or similar) or the craft stores. Happy Holidays To All! Sue Weber -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti Loykasek Sent: Monday, December 01, 2008 6:49 PM To: histonet Subject: [Histonet] Expiration date checks Hi all. I am hoping for some clever ideas on the best method to check for expired reagents. We all know that we shouldn't use them, I'm sure we all actively monitor this (right)! I must admit we occasionally have expired reagents - a stain or buffer we haven't used in a while is in a cupboard or fridge & it has expired. What do others do to monitor & prevent this from happening? We have a weekly safety check that spot checks a certain # of reagents for expiration, but we probably have thousands of reagents & don't check every single bottle every week. Thanks for the input. Patti Loykasek This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Tue Dec 2 07:50:47 2008 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Tue Dec 2 07:51:14 2008 Subject: [Histonet] protocols on Ventana In-Reply-To: <3A855683817C405F903BE1FDEA55A252@pentium4> References: <55063.27940.qm@web59714.mail.ac4.yahoo.com> <3A855683817C405F903BE1FDEA55A252@pentium4> Message-ID: <4934F6E7.2B7F.00C9.0@geisinger.edu> From what company do you order your PMS2, MSH2 and MSH6 concentrates? 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 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! >>> "Piero Nelva" 12/2/2008 2:52 AM >>> I've deleted the original post, but to the person who was after Ventana protocols for Mismatch Repair proteins, here are ours: MSH2, 1:50, CC1 Mild, 37, 28 MSH6, 1:4000, CC1 Std, 37, 40 plus amplification kit MLH1, Vetnana predilute, CC1 Mild, 42, 28 plus amplification kit PMS2, 1:100, CC1 Mild, 37, 56 plus amplification kit. I can send you some images of our results off list if you like Regards Piero Nelva Anatomical Pathology Monash Medical Centre Australia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Bitting, Angela TEL;WORK:570-271-6844 ORG:;Histology EMAIL;WORK;PREF;NGW:AKBITTING@geisinger.edu N:Bitting;Angela END:VCARD From DixonM <@t> vetmed.ufl.edu Tue Dec 2 08:55:03 2008 From: DixonM <@t> vetmed.ufl.edu (MaryAnn Dixon) Date: Tue Dec 2 08:55:10 2008 Subject: [Histonet] antibody Message-ID: <530D827EC657DE418C3572ADD63FCDC3249520@EXGVMCNETWORK.vetmed.ufl.edu> Does anyone out there know where I can find an antibody to buy against Pythium insidiosum? I can't even find it on an antibody search engine!!!!! MaryAnn Dixon BS Biological Scientist Anatomic Pathology UF Veterinary Medical Center (352) 392-2235 Ext. 4517 From sheila_adey <@t> hotmail.com Tue Dec 2 09:04:08 2008 From: sheila_adey <@t> hotmail.com (sheila adey) Date: Tue Dec 2 09:04:22 2008 Subject: [Histonet] Casual Histo tech postition in Port Huron Michigan Message-ID: Hi All, We have a casual histo tech position posted at our facility. If interested please visit the Port Huron Hospital website for further information. Sheila Adey HT MLT Port Huron Hospital Michigan _________________________________________________________________ From renafail <@t> bellsouth.net Tue Dec 2 09:16:38 2008 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Tue Dec 2 09:16:48 2008 Subject: [Histonet] Casual Histo tech postition in Port Huron Michigan In-Reply-To: Message-ID: <120220081516.12979.493551560009D591000032B322230704929B0A02D2089B9A019C04040A0DBF04070E000E020A9D@att.net> Ok , I'll bite. What is a casual histotech? Rena Fail -------------- Original message from sheila adey : -------------- > > Hi All, > We have a casual histo tech position posted at our facility. If interested > please visit the Port Huron Hospital website for further information. > Sheila Adey HT MLT Port Huron Hospital Michigan > _________________________________________________________________ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ktuttle <@t> umm.edu Tue Dec 2 09:18:09 2008 From: ktuttle <@t> umm.edu (Kimberly Tuttle) Date: Tue Dec 2 09:18:42 2008 Subject: [Histonet] Cleaved Caspase-3 In-Reply-To: References: Message-ID: <49350B61.90CE.001A.3@umm.edu> Can anyone recommend control tissue? Thank You Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. From randek <@t> histopathconcepts.com Tue Dec 2 09:18:29 2008 From: randek <@t> histopathconcepts.com (Rande Kline) Date: Tue Dec 2 09:19:46 2008 Subject: [Histonet] Expiration date checks In-Reply-To: <16C83872A53F4346AA9C3A18E3A3AAB903F76E9D@VHAV10MSGA1.v10.med.va.gov> References: <16C83872A53F4346AA9C3A18E3A3AAB903F76E9D@VHAV10MSGA1.v10.med.va.gov> Message-ID: Patty, Chemicals really don't last forever. Changes take place everytime you open the bottle and some just from age even unopened. Salts used in buffers are hygroscopic. They may appear to look okay but they may not be for your application. How long you can use a chemical is dependent on it's application. I think everyone has experienced a solution not working even though you have followed the correct procedure and a repeat has the same outcome. I have wrote a couple of articles a few years back that appeared in Laboratory Medicine Q&A and Microscopy Today on expiration dating of chemicals. The article in Microscopy Today is a little more indepth. I would not mind faxing you a copy. You may want to dig into more than the dot system. I would also suggest disposing chemicals that you may not know the actual age of, if they exist. On Tue, Dec 2, 2008 at 8:48 AM, Weber, Susan (VHACLE) wrote: > Color coding with sticker "dots" helps a bit. I designate a color for > each year (5 yrs/5 colors max) On the top of that years color I put a > date (usually 6/08 whatever) and make them easier to spot. (Pardon the > pun!)You can find color dots in the office supply catalog, Office Max > (or similar) or the craft stores. Happy Holidays To All! > > Sue Weber > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti > Loykasek > Sent: Monday, December 01, 2008 6:49 PM > To: histonet > Subject: [Histonet] Expiration date checks > > Hi all. I am hoping for some clever ideas on the best method to check > for > expired reagents. We all know that we shouldn't use them, I'm sure we > all > actively monitor this (right)! I must admit we occasionally have expired > reagents - a stain or buffer we haven't used in a while is in a cupboard > or > fridge & it has expired. What do others do to monitor & prevent this > from > happening? We have a weekly safety check that spot checks a certain # of > reagents for expiration, but we probably have thousands of reagents & > don't > check every single bottle every week. Thanks for the input. > > Patti Loykasek > > > > This e-mail message, including any attachments, is for the sole use of > the > intended recipients and may contain privileged information. Any > unauthorized > review, use, disclosure or distribution is prohibited. If you are not > the intended > recipient, please contact the sender by e-mail and destroy all copies of > the > original message, or you may call PhenoPath Laboratories, Seattle, WA > U.S.A. > at (206) 374-9000. > > > _______________________________________________ > 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 > -- Rande Kline, HT (ASCP) Histopathology Lab Concepts P-609-744-0803 F-609-939-0270 randek@histopathconcepts.com From lblazek <@t> digestivespecialists.com Tue Dec 2 09:24:08 2008 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Tue Dec 2 09:23:00 2008 Subject: [Histonet] Casual Histo tech postition in Port Huron Michigan In-Reply-To: <120220081516.12979.493551560009D591000032B322230704929B0A02D2089B9A019C04040A0DBF04070E000E020A9D@att.net> References: <120220081516.12979.493551560009D591000032B322230704929B0A02D2089B9A019C04040A0DBF04070E000E020A9D@att.net> Message-ID: <5A2BD13465E061429D6455C8D6B40E3907C05DF23C@IBMB7Exchange.digestivespecialists.com> You work only when you feel like it????????? And in jeans and t shirt. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of renafail@bellsouth.net Sent: Tuesday, December 02, 2008 10:17 AM To: sheila adey; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Casual Histo tech postition in Port Huron Michigan Ok , I'll bite. What is a casual histotech? Rena Fail -------------- Original message from sheila adey : -------------- > > Hi All, > We have a casual histo tech position posted at our facility. If interested > please visit the Port Huron Hospital website for further information. > Sheila Adey HT MLT Port Huron Hospital Michigan > _________________________________________________________________ > > _______________________________________________ > 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 slappycraw <@t> yahoo.com Tue Dec 2 09:32:27 2008 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Tue Dec 2 09:32:38 2008 Subject: [Histonet] Cleaved Caspase-3 References: <49350B61.90CE.001A.3@umm.edu> Message-ID: <855588.59037.qm@web53610.mail.re2.yahoo.com> Tumor tissue works well. ? Larry A. Woody Seattle, Wa. ________________________________ From: Kimberly Tuttle To: histonet@lists.utsouthwestern.edu Sent: Tuesday, December 2, 2008 7:18:09 AM Subject: [Histonet] Cleaved Caspase-3 Can anyone recommend control tissue? Thank You Kimberly C. Tuttle? HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jdhisto <@t> yahoo.com Tue Dec 2 09:41:56 2008 From: jdhisto <@t> yahoo.com (JD) Date: Tue Dec 2 09:42:09 2008 Subject: [Histonet] Casual Histotech Message-ID: My thoughts exactly. Casual histotech? From jqb7 <@t> cdc.gov Tue Dec 2 09:43:35 2008 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Tue Dec 2 09:43:53 2008 Subject: [Histonet] Casual Histotech In-Reply-To: <652B88F04HG494005-01@EMF> References: <652B88F04HG494005-01@EMF> Message-ID: <1CE1847DFEA0A647B1CCDE4108EA60A70208C5A8@LTA3VS011.ees.hhs.gov> As opposed to the rest of us...you know, the formal ones. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JD Sent: Tuesday, December 02, 2008 10:42 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Casual Histotech My thoughts exactly. Casual histotech?_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWeems <@t> sjha.org Tue Dec 2 09:46:58 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Tue Dec 2 09:47:11 2008 Subject: [Histonet] Casual Histotech In-Reply-To: <1CE1847DFEA0A647B1CCDE4108EA60A70208C5A8@LTA3VS011.ees.hhs.gov> References: <652B88F04HG494005-01@EMF> <1CE1847DFEA0A647B1CCDE4108EA60A70208C5A8@LTA3VS011.ees.hhs.gov> Message-ID: <5D64396A0D4A5346BEBC759022AAEAA516A407@ITSSSXM01V6.one.ads.che.org> Just like formal alcohol... We wear our tuxedos... :>) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bartlett, Jeanine (CDC/CCID/NCZVED) Sent: Tuesday, December 02, 2008 10:44 AM To: JD; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Casual Histotech As opposed to the rest of us...you know, the formal ones. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JD Sent: Tuesday, December 02, 2008 10:42 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Casual Histotech My thoughts exactly. Casual histotech?_______________________________________________ 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: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From anh2006 <@t> med.cornell.edu Tue Dec 2 09:42:39 2008 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Tue Dec 2 09:49:40 2008 Subject: [Histonet] Cleaved Caspase-3 Message-ID: <1136679857-1228232966-cardhu_decombobulator_blackberry.rim.net-946880122-@bxe173.bisx.prod.on.blackberry> Tonsil is great. Also tumor and spleen can work. ------Original Message------ From: Kimberly Tuttle Sender: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: Dec 2, 2008 10:18 AM Subject: [Histonet] Cleaved Caspase-3 Can anyone recommend control tissue? Thank You Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Tue Dec 2 09:56:45 2008 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Dec 2 09:57:10 2008 Subject: AW: [Histonet] Question RE: HQIP Result In-Reply-To: <486600.42474.qm@web58905.mail.re1.yahoo.com> References: <486600.42474.qm@web58905.mail.re1.yahoo.com> Message-ID: <4310F95DCE954C5C92DFB73A61DBA87B@dielangs.at> Schaundra, In the following publication Fox describes bubbles that occur in cultered cells after 5-30 min after the addition of formaldehyd solution. Fox Cecil H ua., Formaldehyd Fixation; J. of. Histochemistry and Cytochemistry 33(8):845-853, 1985 So bubbles could be a sign of very short fixation. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Schaundra Walton Gesendet: Montag, 01. Dezember 2008 18:58 An: Histonet Betreff: [Histonet] Question RE: HQIP Result We just got our HQIP results and they made a comment regarding the breast tissue H&E slide we submitted.? Can someone please tell me what nuclear bubbling artifact is and what causes it?? ? Thanks! Schaundra Walton BS, HTL (ASCP) Histology Supervisor Swedish American Hospital Rockford, IL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Tue Dec 2 10:32:50 2008 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Tue Dec 2 10:33:24 2008 Subject: [Histonet] Casual Histo tech postition in Port Huron Michigan References: Message-ID: <5F31F38C96781A4FBE3196EBC22D47807F2852@fhosxchmb006.ADVENTISTCORP.NET> Is this Part-time? Janet L. Bonner, HTL (ASCP) Pathology Laboratory Florida Hospital Winter Park janet.bonner@FLHOSP.org 407-646-7559 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of sheila adey Sent: Tue 12/2/2008 10:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Casual Histo tech postition in Port Huron Michigan Hi All, We have a casual histo tech position posted at our facility. If interested please visit the Port Huron Hospital website for further information. Sheila Adey HT MLT Port Huron Hospital Michigan _________________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ======================================================= The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ======================================================= From fernandom.munoz <@t> usc.es Tue Dec 2 10:33:20 2008 From: fernandom.munoz <@t> usc.es (fernandom.munoz@usc.es) Date: Tue Dec 2 10:33:27 2008 Subject: [Histonet] Re: GMA embedded section attachment to slides Message-ID: <20081202173320.xahxe8snwg0sookk@correoweb.usc.es> Hi James: We work with technovit 7200 VLC and use Silicoup (kulzer) as pretreatment for the glass slides. The blocks must be completely dry (I leave one day drying in the lab without working on it). Fernando Mu?oz From tanisha.mcknight <@t> covance.com Tue Dec 2 10:33:22 2008 From: tanisha.mcknight <@t> covance.com (McKnight, Tanisha) Date: Tue Dec 2 10:33:36 2008 Subject: [Histonet] Best Tissue for H&E Control Message-ID: <816E3C72F855F14985FC31D7C963AE6F0BF84231@indexch03.ent.covance.com> Hello All: Can I get some of you to share with me, what types of tissue you use to assess the quality of your H&E? Or what tissue type do you believe to be best for assessing the quality of an H&E? Thanks, Tanisha Neely, HT(ASCP) AP-Histology/Specimen Management Covance CLS, Indianapolis ----------------------------------------------------- Confidentiality Notice: This e-mail transmission may contain confidential or legally privileged information that is intended only for the individual or entity named in the e-mail address. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or reliance upon the contents of this e-mail is strictly prohibited. If you have received this e-mail transmission in error, please reply to the sender, so that we can arrange for proper delivery, and then please delete the message from your inbox. Thank you. From dholmes <@t> anatomy.umsmed.edu Tue Dec 2 10:44:17 2008 From: dholmes <@t> anatomy.umsmed.edu (Dianne Holmes) Date: Tue Dec 2 10:45:01 2008 Subject: [Histonet] Histo humor :-) Message-ID: <4935118102000082000308E8@GWIA1.umsmed.edu> Not only do I get valuable info from this group but histologists have a GREAT sense of humor in common !! I loved the 'casual histo tech' volley!! Individuals who have received this information in error or are not authorized to receive it must promptly return or dispose of the information and notify the sender. Those individuals are hereby notified that they are strictly prohibited from reviewing, forwarding, printing, copying, distributing or using this information in any way. From Phyllis_Burnette <@t> bshsi.org Tue Dec 2 10:54:13 2008 From: Phyllis_Burnette <@t> bshsi.org (Burnette, Phyllis) Date: Tue Dec 2 10:53:27 2008 Subject: [Histonet] Question RE: HQIP Result In-Reply-To: <4310F95DCE954C5C92DFB73A61DBA87B@dielangs.at> References: <486600.42474.qm@web58905.mail.re1.yahoo.com> <4310F95DCE954C5C92DFB73A61DBA87B@dielangs.at> Message-ID: <2C7750460C3E7545AC11099BE6A7ABD601C99506@EDC-MAIL-02.ads.bshsi.com> When responding about the primary fixative being alcohol, I was also referencing HQIP Participant Summary from 2007 which states "Cell shrinkage and nuclear bubbling, artifacts induced by aggressive processing schedules with resulting primary fixation largely in alcohols". (It's weird, I do actually read these things!) I know for us this was endometrial tissue/ small biopsies that was evaluated and the infiltration of fixative was not long enough (even core breast bx's are now recommended to be included in the no less than 6 hour fixative time frame for Her2 testing). So looking at your fixation processing times may be of help. This is just what has helped the quality of our tissue sections in our lab as we tend to rush these small biopsies through. Fyi, Phyllis I do have a question though, now that I'm on the subject of the HQIP surveys. Did anyone read the comment on the AFB staining from HQIP-B 2007 final critique pg. 18?? I keep waiting for some further feed back about IHC for AFB. They mention IHC sensitivity is 74%-100% whereas the traditional special is only 36%-44% (they made it sound like labs are doing this IHC routinely). Their reference is to the Indian Journal of Tuberculosis. I have called all of our major vendors and they are not aware of a monoclonal antibody to 38k da protein of mycobacterium tuberculosis complex. I just don't understand why CAP would recommend something that I cannot find readily available on the market. Any comments?? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Tuesday, December 02, 2008 10:57 AM To: schaundrawalton@yahoo.com Cc: histonet@lists.utsouthwestern.edu Subject: AW: [Histonet] Question RE: HQIP Result Schaundra, In the following publication Fox describes bubbles that occur in cultered cells after 5-30 min after the addition of formaldehyd solution. Fox Cecil H ua., Formaldehyd Fixation; J. of. Histochemistry and Cytochemistry 33(8):845-853, 1985 So bubbles could be a sign of very short fixation. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Schaundra Walton Gesendet: Montag, 01. Dezember 2008 18:58 An: Histonet Betreff: [Histonet] Question RE: HQIP Result We just got our HQIP results and they made a comment regarding the breast tissue H&E slide we submitted.? Can someone please tell me what nuclear bubbling artifact is and what causes it?? ? Thanks! Schaundra Walton BS, HTL (ASCP) Histology Supervisor Swedish American Hospital Rockford, IL _______________________________________________ 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 in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone, and permanently delete the original e-mail, attachment(s), and any copies. ________________________________________________________________________________________________________ From Shirley.Chu <@t> moldev.com Tue Dec 2 11:02:58 2008 From: Shirley.Chu <@t> moldev.com (Chu, Shirley) Date: Tue Dec 2 11:03:21 2008 Subject: [Histonet] Rescheduling of previously cancelled LCM Webinar Message-ID: The cancelled webinar that was scheduled for Nov 24th has been rescheduled for Friday, Dec 5th at 10AM PST or 1PM EST. The speaker for this webinar will be Charmain Pietersen, PhD., Laboratory for Structural and Molecular Neuroscience, McLean Hospital. Her presentation is titled: "Gene expression analysis in homogenous single cell populations in the superior temporal gyrus in postmortem brains from subjects with schizophrenia". This webinar is free to all registrants. For further information and/or to register for this webinar, please connect to the following website: http://www.moleculardevices.com/pages/lcm_webinar_11.2008.html. Please note anyone that had previously registered for this webinar do not need to register again. Please use the log in information that was forwarded to you upon registering to log into the webinar this friday. Shirley Chu Application Scientist, Arcturus LCM Products Molecular Devices (now a part of MDS Analytical Technologies) 408-747-3765 | www.moleculardevices.com From Shirley.Chu <@t> moldev.com Tue Dec 2 11:12:12 2008 From: Shirley.Chu <@t> moldev.com (Chu, Shirley) Date: Tue Dec 2 11:12:27 2008 Subject: [Histonet] Update - Rescheduling of previously cancelled LCM Webinar Message-ID: My apologies to all, but the link that I provided below for registration is incorrect, please use this link to register for this webinar: http://www.moleculardevices.com/pages/lcm_webinar_12.2008.html ________________________________ From: Chu, Shirley Sent: Tuesday, December 02, 2008 9:03 AM To: 'histonet@lists.utsouthwestern.edu' Cc: 'NYSHistotech' Subject: Rescheduling of previously cancelled LCM Webinar The cancelled webinar that was scheduled for Nov 24th has been rescheduled for Friday, Dec 5th at 10AM PST or 1PM EST. The speaker for this webinar will be Charmain Pietersen, PhD., Laboratory for Structural and Molecular Neuroscience, McLean Hospital. Her presentation is titled: "Gene expression analysis in homogenous single cell populations in the superior temporal gyrus in postmortem brains from subjects with schizophrenia". This webinar is free to all registrants. For further information and/or to register for this webinar, please connect to the following website: http://www.moleculardevices.com/pages/lcm_webinar_11.2008.html. Please note anyone that had previously registered for this webinar do not need to register again. Please use the log in information that was forwarded to you upon registering to log into the webinar this friday. Shirley Chu Application Scientist, Arcturus LCM Products Molecular Devices (now a part of MDS Analytical Technologies) 408-747-3765 | www.moleculardevices.com From CBark <@t> memorialcare.org Tue Dec 2 12:08:27 2008 From: CBark <@t> memorialcare.org (Christine Bark) Date: Tue Dec 2 12:08:45 2008 Subject: [Histonet] RE: Halt In-Reply-To: <652B8AEE1V8454816-01@emf1.memorialcare.org> Message-ID: You can get it from Poly Scientific www.polyrnd.com Christine Bark HT(ASCP) CM Lead Histotech, Pathology Saddleback Memorial Medical Center 949-452-3548 cbark@memorialcare.org Hello all, Can anyone tell me where I could buy Halt. It is used to get wrinkle free sections on the floatation bath. Thanks Jenny ______________________________________________________________________________ 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 Dorothy.L.Webb <@t> HealthPartners.Com Tue Dec 2 12:09:50 2008 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Tue Dec 2 12:09:59 2008 Subject: [Histonet] Control tissue Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635B53@hpes1.HealthPartners.int> The best control is healthy skin with the connective tissue in place but a tonsil is also great for checking how well your lymph tissue is staining. Tonsil should be cut thinner just like the lymph nodes so you should use the tonsil when staining lymph nodes. Make sure the tonsil has connective tissue in it so you can see the eosin staining in the collagen, blood vessels, cytoplasm, and RBC's. Another good tissue is cervix with the glands. The glandular area gives you the nice hematoxylin staining and the muscle and collagen fibers give you a nice contrast on the eosin staining plus you can always get cervix. I would stay away from the appendix since there is usually a lot of necrosis and the placenta is a little to bloody so the eosin is always some what overwhelming for a good evaluation. This is information I was given when posing the question years ago!! Dorothy Webb, HT Regions Histology Technical Supervisor 651-254-2962 ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From KKay <@t> chr.ab.ca Tue Dec 2 12:12:07 2008 From: KKay <@t> chr.ab.ca (Kay, Karen) Date: Tue Dec 2 12:12:13 2008 Subject: [Histonet] RE: Histonet Digest, Vol 61, Issue 2 H&E CONTROL TISSUE In-Reply-To: Message-ID: <9C0BD812BAB0BA4DB7E7FD4F5FA3CAA80A494DBF@exbe.chr.ab.ca> Hello Tanisha, We feel that no one tissue is suitable on its own, therefore we create a multi-tissue control block for our H&E stains. The tissues included are liver, skin, appendix, and placenta. This enables us to assess the stain from a number of different perspectives according to the tissue types included. We use small portions of each type, embedded into a rectangular shaped mold. Hope this helps you Karen J Kay, MLT Pathology Supervisor Chinook Health Laboratory Chinook Regional Hospital 960 - 19 st South Lethbridge, Alberta, Canada T1J 1W5 (403) 388 - 6061 (Phone) (403) 388 - 6067 (Fax) Message: 8 Date: Tue, 2 Dec 2008 11:33:22 -0500 From: "McKnight, Tanisha" Subject: [Histonet] Best Tissue for H&E Control To: histonet@lists.utsouthwestern.edu Message-ID: <816E3C72F855F14985FC31D7C963AE6F0BF84231@indexch03.ent.covance.com> Content-Type: text/plain; charset="us-ascii" Hello All: Can I get some of you to share with me, what types of tissue you use to assess the quality of your H&E? Or what tissue type do you believe to be best for assessing the quality of an H&E? Thanks, Tanisha Neely, HT(ASCP) AP-Histology/Specimen Management Covance CLS, Indianapolis This communication is intended for the use of the recipient to which it is addressed, and may contain confidential, personal and or privileged information. Please contact us immediately if you are not the intended recipient. Do not copy, distribute or take action relying on it. Any communication received in error, or subsequent reply, should be deleted or destroyed. From histonetalias <@t> gmail.com Tue Dec 2 13:19:21 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Tue Dec 2 13:19:32 2008 Subject: [Histonet] Ventana Benchmark XTs In-Reply-To: References: <2DFAEEFF192A9141ABACFF88BE613BF30BBC56@EXMBXC1.crha.bewell.ca> <4b6c85510811281151w4aec5437ybf85920655ab9def@mail.gmail.com> <5b6eb13e0811281210j7fd2f0d5w1c3283bd9bdead11@mail.gmail.com> <4b6c85510811290803v7e52528bw2aee194670077f0@mail.gmail.com> Message-ID: <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> Mr. Della Speranza, I respect what you have done for the histology community and we have had pleasant conversations in the past but you can not speak for the Histonet members. You act as if I am hiding my identity to create havoc and disruption. I have not done this nor will I do this. I have been on histonet for many years and I will continue to do so. As I explained in my previous posts my current employer does not want any opinions, help, or questions associated with the laboratory. That is the only reason that I have chosen to remain anonymous. Would it make anyone feel better if I made up a fake name and lab as my signature? I will continue to post and speak freely without breaking any rules of this listserver. If I respond or ask a question, those who find my years of experience not "credible" then I apologize. On Mon, Dec 1, 2008 at 11:21 AM, Della Speranza, Vinnie wrote: > I'm afraid your opinion will not be credible as long as you insist on > remaining anonymous. It has nothing to do with whether you like or dislike a > particular product or instrument. Histonet members appreciate the > responsibility that comes with posting one's views or knowledge by > identifying themselves accordingly. In my opinion anyone who insists on > remaining anonymous should not be permitted to participate on the list. > > > Vinnie Della Speranza > Manager for Anatomic Pathology Services > Medical University of South Carolina > 165 Ashley Avenue Suite 309 > Charleston, South Carolina 29425 > Tel: (843) 792-6353 > Fax: (843) 792-8974 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto: > histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias > Sent: Saturday, November 29, 2008 11:03 AM > To: Mark Tarango > Cc: histonet@lists.utsouthwestern.edu; Martin Trotter; Tracey Lenek; > Burton, Lynn; Joanna Bartczak-McKay > Subject: Re: [Histonet] Ventana Benchmark XTs > > Sorry I don't work for Ventana and I have no beef with them. It > may disappoint some that I have not had a bad experience. So my opinion > does > not count because I have had a different experience with my XT than you? I > use the name of The Unknown HT so I can express my opinions freely without > my employer receiving phone calls because certain people don't like my > opinions. > > On Fri, Nov 28, 2008 at 3:10 PM, Mark Tarango > wrote: > > > I don't know how much the opinion of "The Unknown HT(ASCP)" counts. You > > aren't with Ventana I hope. > > > > Mark > > > > On Fri, Nov 28, 2008 at 11:51 AM, Histonet Alias < > histonetalias@gmail.com>wrote: > > > >> They are switching to a more reliable manufacturer. They are being > >> proactive > >> and switching them before they fail down the road. My XT has been a > >> workhorse but we know certain peoples opinions about the company on > here. > >> I > >> will take it any day over the rest. > >> > >> On Wed, Nov 26, 2008 at 5:29 PM, Burton, Lynn >> >wrote: > >> > >> > We have had some trouble but they are planning to replace them. We > >> received > >> > a letter that they had changed vendors for parts and found they need > to > >> > switch back. They did replace one section earlier when it stopped > >> working > >> > completely. > >> > > >> > Lynn Burton > >> > Lab Assoc. I > >> > Animal Disease Lab > >> > Galesburg, Il 61401 > >> > > >> > ________________________________ > >> > > >> > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tracey > >> Lenek > >> > Sent: Wed 11/26/2008 10:59 AM > >> > To: histonet@lists.utsouthwestern.edu > >> > Cc: Martin Trotter; Joanna Bartczak-McKay > >> > Subject: [Histonet] Ventana Benchmark XTs > >> > > >> > > >> > > >> > Hi, > >> > > >> > We have had on-going thermal slide pad issues with our XTs since they > >> were > >> > installed > >> > in February. The slide tray assemblies have been replaced not once but > >> > twice and we are still having inconsistent > >> > results with the temp verifier slides. Has anyone experienced the > same > >> > issues with this > >> > instrumentation? > >> > > >> > Thanks > >> > Tracey Lenek > >> > Tech III - Anatomic Pathology > >> > Calgary Laboratory Services > >> > 403-770-3588 > >> > > >> > ________________________________ > >> > This message and any attached documents are only for the use of the > >> > intended recipient(s), are confidential and may contain privileged > >> > information. Any unauthorized review, use, retransmission, or other > >> > disclosure is strictly prohibited. If you have received this message > in > >> > error, please notify the sender immediately, and then delete the > >> original > >> > message. Thank you. > >> > _______________________________________________ > >> > 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 Unknown HT(ASCP) > >> _______________________________________________ > >> Histonet mailing list > >> Histonet@lists.utsouthwestern.edu > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >> > > > > > > > -- > The Unknown HT(ASCP) > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- The Unknown HT(ASCP) From histonetalias <@t> gmail.com Tue Dec 2 13:21:53 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Tue Dec 2 13:22:02 2008 Subject: [Histonet] Best Tissue for H&E Control In-Reply-To: <816E3C72F855F14985FC31D7C963AE6F0BF84231@indexch03.ent.covance.com> References: <816E3C72F855F14985FC31D7C963AE6F0BF84231@indexch03.ent.covance.com> Message-ID: <4b6c85510812021121l3445be59l2d489ea106e237be@mail.gmail.com> Hi Tanisha, We use an appendix for our H&E qc. On Tue, Dec 2, 2008 at 11:33 AM, McKnight, Tanisha < tanisha.mcknight@covance.com> wrote: > Hello All: > > Can I get some of you to share with me, what types of tissue you use to > assess the quality of your H&E? Or what tissue type do you believe to be > best for assessing the quality of an H&E? > > Thanks, > > Tanisha Neely, HT(ASCP) > AP-Histology/Specimen Management > Covance CLS, Indianapolis > > > > > ----------------------------------------------------- > Confidentiality Notice: This e-mail transmission > may contain confidential or legally privileged > information that is intended only for the individual > or entity named in the e-mail address. If you are not > the intended recipient, you are hereby notified that > any disclosure, copying, distribution, or reliance > upon the contents of this e-mail is strictly prohibited. > > If you have received this e-mail transmission in error, > please reply to the sender, so that we can arrange > for proper delivery, and then please delete the message > from your inbox. Thank you. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- The Unknown HT(ASCP) From histonetalias <@t> gmail.com Tue Dec 2 13:24:44 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Tue Dec 2 13:24:56 2008 Subject: [Histonet] Expiration date checks In-Reply-To: References: Message-ID: <4b6c85510812021124p6b96d961j964be823fc254a19@mail.gmail.com> I actually have an excel spread sheet of expiration dates. I also place those dates into the outlook calender and I receive a notification a week before the expiration. On Mon, Dec 1, 2008 at 6:49 PM, Patti Loykasek wrote: > Hi all. I am hoping for some clever ideas on the best method to check for > expired reagents. We all know that we shouldn't use them, I'm sure we all > actively monitor this (right)! I must admit we occasionally have expired > reagents - a stain or buffer we haven't used in a while is in a cupboard or > fridge & it has expired. What do others do to monitor & prevent this from > happening? We have a weekly safety check that spot checks a certain # of > reagents for expiration, but we probably have thousands of reagents & don't > check every single bottle every week. Thanks for the input. > > Patti Loykasek > > > > This e-mail message, including any attachments, is for the sole use of the > intended recipients and may contain privileged information. Any > unauthorized > review, use, disclosure or distribution is prohibited. If you are not the > intended > recipient, please contact the sender by e-mail and destroy all copies of > the > original message, or you may call PhenoPath Laboratories, Seattle, WA > U.S.A. > at (206) 374-9000. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- The Unknown HT(ASCP) From Kristopher.Kalleberg <@t> unilever.com Tue Dec 2 14:03:29 2008 From: Kristopher.Kalleberg <@t> unilever.com (Kalleberg, Kristopher) Date: Tue Dec 2 14:03:39 2008 Subject: [Histonet] Fontana Masson Message-ID: <0E6BC087F70F9C47ACFF2C203D6E329C052FF1BD@NTRSEVS30002.s3.ms.unilever.com> All, Does anyone have a reference for me to read up on the chemistry involved in Fontana Masson staining of melanin? I am having a hard time finding in depth information on exactly how FM stain stains the melanosmes and the chemistry and reactions involved. Any information, papers, or references would be greatly appreciated. Thank you in advance. Kris Kalleberg Research Scientist Unilever R&D 40 Merritt Blvd. Trumbull, CT 06611 (203) 381-5765 From marktarango <@t> gmail.com Tue Dec 2 14:24:51 2008 From: marktarango <@t> gmail.com (Mark Tarango) Date: Tue Dec 2 14:25:03 2008 Subject: [Histonet] Ventana Benchmark XTs In-Reply-To: <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> References: <2DFAEEFF192A9141ABACFF88BE613BF30BBC56@EXMBXC1.crha.bewell.ca> <4b6c85510811281151w4aec5437ybf85920655ab9def@mail.gmail.com> <5b6eb13e0811281210j7fd2f0d5w1c3283bd9bdead11@mail.gmail.com> <4b6c85510811290803v7e52528bw2aee194670077f0@mail.gmail.com> <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> Message-ID: <5b6eb13e0812021224k1be955f9l4341e202b5ca4d3@mail.gmail.com> Can't you just post without saying where exactly you work? I don't mention where I'm working these days. Does your employer own you? ...or maybe you're some famous histotech and you get paid for endorsements? lol Mark On Tue, Dec 2, 2008 at 11:19 AM, Histonet Alias wrote: > Mr. Della Speranza, I respect what you have done for the histology > community and we have had pleasant conversations in the past but you can not > speak for the Histonet members. You act as if I am hiding my identity to > create havoc and disruption. I have not done this nor will I do this. I have > been on histonet for many years and I will continue to do so. As I explained > in my previous posts my current employer does not want any opinions, help, > or questions associated with the laboratory. That is the only reason that I > have chosen to remain anonymous. Would it make anyone feel better if I made > up a fake name and lab as my signature? I will continue to post and speak > freely without breaking any rules of this listserver. If I respond or ask a > question, those who find my years of experience not "credible" then > I apologize. > > > On Mon, Dec 1, 2008 at 11:21 AM, Della Speranza, Vinnie wrote: > >> I'm afraid your opinion will not be credible as long as you insist on >> remaining anonymous. It has nothing to do with whether you like or dislike a >> particular product or instrument. Histonet members appreciate the >> responsibility that comes with posting one's views or knowledge by >> identifying themselves accordingly. In my opinion anyone who insists on >> remaining anonymous should not be permitted to participate on the list. >> >> >> Vinnie Della Speranza >> Manager for Anatomic Pathology Services >> Medical University of South Carolina >> 165 Ashley Avenue Suite 309 >> Charleston, South Carolina 29425 >> Tel: (843) 792-6353 >> Fax: (843) 792-8974 >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias >> Sent: Saturday, November 29, 2008 11:03 AM >> To: Mark Tarango >> Cc: histonet@lists.utsouthwestern.edu; Martin Trotter; Tracey Lenek; >> Burton, Lynn; Joanna Bartczak-McKay >> Subject: Re: [Histonet] Ventana Benchmark XTs >> >> Sorry I don't work for Ventana and I have no beef with them. It >> may disappoint some that I have not had a bad experience. So my opinion >> does >> not count because I have had a different experience with my XT than you? I >> use the name of The Unknown HT so I can express my opinions freely without >> my employer receiving phone calls because certain people don't like my >> opinions. >> >> On Fri, Nov 28, 2008 at 3:10 PM, Mark Tarango >> wrote: >> >> > I don't know how much the opinion of "The Unknown HT(ASCP)" counts. You >> > aren't with Ventana I hope. >> > >> > Mark >> > >> > On Fri, Nov 28, 2008 at 11:51 AM, Histonet Alias < >> histonetalias@gmail.com>wrote: >> > >> >> They are switching to a more reliable manufacturer. They are being >> >> proactive >> >> and switching them before they fail down the road. My XT has been a >> >> workhorse but we know certain peoples opinions about the company on >> here. >> >> I >> >> will take it any day over the rest. >> >> >> >> On Wed, Nov 26, 2008 at 5:29 PM, Burton, Lynn < >> Lynn.Burton@illinois.gov >> >> >wrote: >> >> >> >> > We have had some trouble but they are planning to replace them. We >> >> received >> >> > a letter that they had changed vendors for parts and found they need >> to >> >> > switch back. They did replace one section earlier when it stopped >> >> working >> >> > completely. >> >> > >> >> > Lynn Burton >> >> > Lab Assoc. I >> >> > Animal Disease Lab >> >> > Galesburg, Il 61401 >> >> > >> >> > ________________________________ >> >> > >> >> > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tracey >> >> Lenek >> >> > Sent: Wed 11/26/2008 10:59 AM >> >> > To: histonet@lists.utsouthwestern.edu >> >> > Cc: Martin Trotter; Joanna Bartczak-McKay >> >> > Subject: [Histonet] Ventana Benchmark XTs >> >> > >> >> > >> >> > >> >> > Hi, >> >> > >> >> > We have had on-going thermal slide pad issues with our XTs since they >> >> were >> >> > installed >> >> > in February. The slide tray assemblies have been replaced not once >> but >> >> > twice and we are still having inconsistent >> >> > results with the temp verifier slides. Has anyone experienced the >> same >> >> > issues with this >> >> > instrumentation? >> >> > >> >> > Thanks >> >> > Tracey Lenek >> >> > Tech III - Anatomic Pathology >> >> > Calgary Laboratory Services >> >> > 403-770-3588 >> >> > >> >> > ________________________________ >> >> > This message and any attached documents are only for the use of the >> >> > intended recipient(s), are confidential and may contain privileged >> >> > information. Any unauthorized review, use, retransmission, or other >> >> > disclosure is strictly prohibited. If you have received this message >> in >> >> > error, please notify the sender immediately, and then delete the >> >> original >> >> > message. Thank you. >> >> > _______________________________________________ >> >> > 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 Unknown HT(ASCP) >> >> _______________________________________________ >> >> Histonet mailing list >> >> Histonet@lists.utsouthwestern.edu >> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > >> > >> >> >> -- >> The Unknown HT(ASCP) >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > The Unknown HT(ASCP) > From rjbuesa <@t> yahoo.com Tue Dec 2 14:52:11 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 2 14:52:22 2008 Subject: [Histonet] Ventana Benchmark XTs In-Reply-To: <5b6eb13e0812021224k1be955f9l4341e202b5ca4d3@mail.gmail.com> Message-ID: <946428.46996.qm@web65706.mail.ac4.yahoo.com> This is getting somewhat confusing BUT your initial commentary fully endorsing Ventana OVER ANY OTHER, is the one that raised some suspicion because there are other autostainers generally accepted as MUCH BETTER than Ventana or put in some other way, NO OTHER autostainer has generated so many bad reports. Either you work for Verntana or not I do not care, but I think that if your boss restricts so much somebody with the many years of experience you claim to have, either you have a problem with your boss, or you should think in working some other place before retiring (after so many years of work in histology). That is my honest and not anonymous opinion Ren? J. --- On Tue, 12/2/08, Mark Tarango wrote: From: Mark Tarango Subject: Re: [Histonet] Ventana Benchmark XTs To: "Histonet Alias" Cc: "histonet@lists.utsouthwestern.edu" , "Tracey Lenek" , "Burton, Lynn" , "Joanna Bartczak-McKay" , "Martin Trotter" Date: Tuesday, December 2, 2008, 3:24 PM Can't you just post without saying where exactly you work? I don't mention where I'm working these days. Does your employer own you? ...or maybe you're some famous histotech and you get paid for endorsements? lol Mark On Tue, Dec 2, 2008 at 11:19 AM, Histonet Alias wrote: > Mr. Della Speranza, I respect what you have done for the histology > community and we have had pleasant conversations in the past but you can not > speak for the Histonet members. You act as if I am hiding my identity to > create havoc and disruption. I have not done this nor will I do this. I have > been on histonet for many years and I will continue to do so. As I explained > in my previous posts my current employer does not want any opinions, help, > or questions associated with the laboratory. That is the only reason that I > have chosen to remain anonymous. Would it make anyone feel better if I made > up a fake name and lab as my signature? I will continue to post and speak > freely without breaking any rules of this listserver. If I respond or ask a > question, those who find my years of experience not "credible" then > I apologize. > > > On Mon, Dec 1, 2008 at 11:21 AM, Della Speranza, Vinnie wrote: > >> I'm afraid your opinion will not be credible as long as you insist on >> remaining anonymous. It has nothing to do with whether you like or dislike a >> particular product or instrument. Histonet members appreciate the >> responsibility that comes with posting one's views or knowledge by >> identifying themselves accordingly. In my opinion anyone who insists on >> remaining anonymous should not be permitted to participate on the list. >> >> >> Vinnie Della Speranza >> Manager for Anatomic Pathology Services >> Medical University of South Carolina >> 165 Ashley Avenue Suite 309 >> Charleston, South Carolina 29425 >> Tel: (843) 792-6353 >> Fax: (843) 792-8974 >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias >> Sent: Saturday, November 29, 2008 11:03 AM >> To: Mark Tarango >> Cc: histonet@lists.utsouthwestern.edu; Martin Trotter; Tracey Lenek; >> Burton, Lynn; Joanna Bartczak-McKay >> Subject: Re: [Histonet] Ventana Benchmark XTs >> >> Sorry I don't work for Ventana and I have no beef with them. It >> may disappoint some that I have not had a bad experience. So my opinion >> does >> not count because I have had a different experience with my XT than you? I >> use the name of The Unknown HT so I can express my opinions freely without >> my employer receiving phone calls because certain people don't like my >> opinions. >> >> On Fri, Nov 28, 2008 at 3:10 PM, Mark Tarango >> wrote: >> >> > I don't know how much the opinion of "The Unknown HT(ASCP)" counts. You >> > aren't with Ventana I hope. >> > >> > Mark >> > >> > On Fri, Nov 28, 2008 at 11:51 AM, Histonet Alias < >> histonetalias@gmail.com>wrote: >> > >> >> They are switching to a more reliable manufacturer. They are being >> >> proactive >> >> and switching them before they fail down the road. My XT has been a >> >> workhorse but we know certain peoples opinions about the company on >> here. >> >> I >> >> will take it any day over the rest. >> >> >> >> On Wed, Nov 26, 2008 at 5:29 PM, Burton, Lynn < >> Lynn.Burton@illinois.gov >> >> >wrote: >> >> >> >> > We have had some trouble but they are planning to replace them. We >> >> received >> >> > a letter that they had changed vendors for parts and found they need >> to >> >> > switch back. They did replace one section earlier when it stopped >> >> working >> >> > completely. >> >> > >> >> > Lynn Burton >> >> > Lab Assoc. I >> >> > Animal Disease Lab >> >> > Galesburg, Il 61401 >> >> > >> >> > ________________________________ >> >> > >> >> > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tracey >> >> Lenek >> >> > Sent: Wed 11/26/2008 10:59 AM >> >> > To: histonet@lists.utsouthwestern.edu >> >> > Cc: Martin Trotter; Joanna Bartczak-McKay >> >> > Subject: [Histonet] Ventana Benchmark XTs >> >> > >> >> > >> >> > >> >> > Hi, >> >> > >> >> > We have had on-going thermal slide pad issues with our XTs since they >> >> were >> >> > installed >> >> > in February. The slide tray assemblies have been replaced not once >> but >> >> > twice and we are still having inconsistent >> >> > results with the temp verifier slides. Has anyone experienced the >> same >> >> > issues with this >> >> > instrumentation? >> >> > >> >> > Thanks >> >> > Tracey Lenek >> >> > Tech III - Anatomic Pathology >> >> > Calgary Laboratory Services >> >> > 403-770-3588 >> >> > >> >> > ________________________________ >> >> > This message and any attached documents are only for the use of the >> >> > intended recipient(s), are confidential and may contain privileged >> >> > information. Any unauthorized review, use, retransmission, or other >> >> > disclosure is strictly prohibited. If you have received this message >> in >> >> > error, please notify the sender immediately, and then delete the >> >> original >> >> > message. Thank you. >> >> > _______________________________________________ >> >> > 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 Unknown HT(ASCP) >> >> _______________________________________________ >> >> Histonet mailing list >> >> Histonet@lists.utsouthwestern.edu >> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > >> > >> >> >> -- >> The Unknown HT(ASCP) >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > The Unknown HT(ASCP) > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From renafail <@t> bellsouth.net Tue Dec 2 15:12:19 2008 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Tue Dec 2 15:12:28 2008 Subject: [Histonet] Ventana Benchmark XTs In-Reply-To: <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> References: <2DFAEEFF192A9141ABACFF88BE613BF30BBC56@EXMBXC1.crha.bewell.ca><4b6c85510811281151w4aec5437ybf85920655ab9def@mail.gmail.com><5b6eb13e0811281210j7fd2f0d5w1c3283bd9bdead11@mail.gmail.com><4b6c85510811290803v7e52528bw2aee194670077f0@mail.gmail.com> <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> Message-ID: <120220082112.22784.4935A4B20009C2280000590022230680329B0A02D2089B9A019C04040A0DBF04070E000E020A9D@att.net> In the past I have seen techs insert a disclaimer stating the opinion is theirs and theirs alone and does not reflect the opinion of their employer nor do they identify their employer. Some managers may feel that it reflects poorly on the Lab if questions are posted. In reality questions allow for the exchange of information and opens a dialogue that may benefit many Rena Fail , retired -------------- Original message from "Histonet Alias" : -------------- > Mr. Della Speranza, I respect what you have done for the histology community > and we have had pleasant conversations in the past but you can not speak for > the Histonet members. You act as if I am hiding my identity to create havoc > and disruption. I have not done this nor will I do this. I have been on > histonet for many years and I will continue to do so. As I explained in my > previous posts my current employer does not want any opinions, help, or > questions associated with the laboratory. That is the only reason that I > have chosen to remain anonymous. Would it make anyone feel better if I made > up a fake name and lab as my signature? I will continue to post and speak > freely without breaking any rules of this listserver. If I respond or ask a > question, those who find my years of experience not "credible" then > I apologize. > > On Mon, Dec 1, 2008 at 11:21 AM, Della Speranza, Vinnie wrote: > > > I'm afraid your opinion will not be credible as long as you insist on > > remaining anonymous. It has nothing to do with whether you like or dislike a > > particular product or instrument. Histonet members appreciate the > > responsibility that comes with posting one's views or knowledge by > > identifying themselves accordingly. In my opinion anyone who insists on > > remaining anonymous should not be permitted to participate on the list. > > > > > > Vinnie Della Speranza > > Manager for Anatomic Pathology Services > > Medical University of South Carolina > > 165 Ashley Avenue Suite 309 > > Charleston, South Carolina 29425 > > Tel: (843) 792-6353 > > Fax: (843) 792-8974 > > > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu [mailto: > > histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias > > Sent: Saturday, November 29, 2008 11:03 AM > > To: Mark Tarango > > Cc: histonet@lists.utsouthwestern.edu; Martin Trotter; Tracey Lenek; > > Burton, Lynn; Joanna Bartczak-McKay > > Subject: Re: [Histonet] Ventana Benchmark XTs > > > > Sorry I don't work for Ventana and I have no beef with them. It > > may disappoint some that I have not had a bad experience. So my opinion > > does > > not count because I have had a different experience with my XT than you? I > > use the name of The Unknown HT so I can express my opinions freely without > > my employer receiving phone calls because certain people don't like my > > opinions. > > > > On Fri, Nov 28, 2008 at 3:10 PM, Mark Tarango > > wrote: > > > > > I don't know how much the opinion of "The Unknown HT(ASCP)" counts. You > > > aren't with Ventana I hope. > > > > > > Mark > > > > > > On Fri, Nov 28, 2008 at 11:51 AM, Histonet Alias < > > histonetalias@gmail.com>wrote: > > > > > >> They are switching to a more reliable manufacturer. They are being > > >> proactive > > >> and switching them before they fail down the road. My XT has been a > > >> workhorse but we know certain peoples opinions about the company on > > here. > > >> I > > >> will take it any day over the rest. > > >> > > >> On Wed, Nov 26, 2008 at 5:29 PM, Burton, Lynn > > >> >wrote: > > >> > > >> > We have had some trouble but they are planning to replace them. We > > >> received > > >> > a letter that they had changed vendors for parts and found they need > > to > > >> > switch back. They did replace one section earlier when it stopped > > >> working > > >> > completely. > > >> > > > >> > Lynn Burton > > >> > Lab Assoc. I > > >> > Animal Disease Lab > > >> > Galesburg, Il 61401 > > >> > > > >> > ________________________________ > > >> > > > >> > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tracey > > >> Lenek > > >> > Sent: Wed 11/26/2008 10:59 AM > > >> > To: histonet@lists.utsouthwestern.edu > > >> > Cc: Martin Trotter; Joanna Bartczak-McKay > > >> > Subject: [Histonet] Ventana Benchmark XTs > > >> > > > >> > > > >> > > > >> > Hi, > > >> > > > >> > We have had on-going thermal slide pad issues with our XTs since they > > >> were > > >> > installed > > >> > in February. The slide tray assemblies have been replaced not once but > > >> > twice and we are still having inconsistent > > >> > results with the temp verifier slides. Has anyone experienced the > > same > > >> > issues with this > > >> > instrumentation? > > >> > > > >> > Thanks > > >> > Tracey Lenek > > >> > Tech III - Anatomic Pathology > > >> > Calgary Laboratory Services > > >> > 403-770-3588 > > >> > > > >> > ________________________________ > > >> > This message and any attached documents are only for the use of the > > >> > intended recipient(s), are confidential and may contain privileged > > >> > information. Any unauthorized review, use, retransmission, or other > > >> > disclosure is strictly prohibited. If you have received this message > > in > > >> > error, please notify the sender immediately, and then delete the > > >> original > > >> > message. Thank you. > > >> > _______________________________________________ > > >> > 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 Unknown HT(ASCP) > > >> _______________________________________________ > > >> Histonet mailing list > > >> Histonet@lists.utsouthwestern.edu > > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >> > > > > > > > > > > > > -- > > The Unknown HT(ASCP) > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > -- > The Unknown HT(ASCP) > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mbisher <@t> Princeton.EDU Tue Dec 2 15:18:21 2008 From: mbisher <@t> Princeton.EDU (Peggy Bisher) Date: Tue Dec 2 15:18:29 2008 Subject: [Histonet] Neurofilament Question Message-ID: I have a student who asked if I could post this question to the Histo-list. I told her how all of you seem to be very helpful and perhaps some of you know the answer to her question: Dear all, I am looking for a good "catch-all" antibody for neuronal intermediate filaments (NF-L, NF-M and NF-H). Since they're generally expressed together in each neurofilament I'm not interested in any one of them specifically, but want to avoid using a cocktail of antibodies, as this could get expensive. Basically, I just want to show that a subset of axons contains neurofilaments with a reliable antibody, but the particular protein or phosphorylation state doesn't matter. Any suggestions you could offer would be incredibly helpful! Sincerely, Eve Schneider Thanks, Margaret E. Bisher Electron Microscopy & Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbisher@princeton.edu From kgrobert <@t> rci.rutgers.edu Tue Dec 2 15:40:47 2008 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Tue Dec 2 15:29:17 2008 Subject: [Histonet] Neurofilament Question In-Reply-To: References: Message-ID: <4935AB5F.70802@rci.rutgers.edu> I have not used this one personally,but I have used antibodies against individual neurofilaments from this same company (before they were taken over by Covance), and they were excellent. If I'm reading this right, it IS a cocktail. Still pricey, but it might be worth a try. Maybe you can get a sample before you buy? I don't know if Covance does that, but it can't hurt to ask. https://store.crpinc.com/datasheet.aspx?UpProd=&catalogno=SMI-312R Good luck, Kathleen Roberts Dept of Pharmacology & Toxicology Rutgers University Peggy Bisher wrote: >I have a student who asked if I could post this question to the Histo-list. >I told her how all of you seem to be very helpful and perhaps some of you >know the answer to her question: > > > >Dear all, > > >I am looking for a good "catch-all" antibody for neuronal intermediate >filaments (NF-L, NF-M and NF-H). Since they're generally expressed >together in each neurofilament I'm not interested in any one of them >specifically, but want to avoid using a cocktail of antibodies, as >this could get expensive. Basically, I just want to show that a subset >of axons contains neurofilaments with a reliable antibody, but the >particular protein or phosphorylation state doesn't matter. Any >suggestions you could offer would be incredibly helpful! > >Sincerely, Eve Schneider > > >Thanks, > > >Margaret E. Bisher >Electron Microscopy & Histology Core Facility Manager >Department of Molecular Biology >Princeton University >Moffett Laboratory, Room 113 >Princeton, New Jersey >Office: (609) 258-7026 >Fax: (609) 258-8468 >mbisher@princeton.edu > > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From mucram11 <@t> comcast.net Tue Dec 2 15:40:41 2008 From: mucram11 <@t> comcast.net (Pamela Marcum) Date: Tue Dec 2 15:40:45 2008 Subject: [Histonet] Large Slide Boxes Message-ID: <517972921.1999491228254041831.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Good Afternoon All, I am looking to purchase some 3 inch by 2 inch (3X2) slide boxes for large slides.? We do some larger specimens and it seems these have become almost impossible to find.? I have tried several sources and no one had them or knew where I could get them at this time.? I have tried myneurolab and several other speciality vendors also to no avail.? HELP Pam Marcum From jim.manavis <@t> imvs.sa.gov.au Tue Dec 2 15:54:55 2008 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Tue Dec 2 15:56:25 2008 Subject: [Histonet] Cleaved Caspase-3 In-Reply-To: <49350B61.90CE.001A.3@umm.edu> References: <49350B61.90CE.001A.3@umm.edu> Message-ID: <003301c954c8$9cdc1070$2c6c140a@41984n> Tumour tissue, I use CNS Lymphoma. Cheers Jim Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases IMVS, Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 / 0401120697 FAX: 61-08-8222 3392 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kimberly Tuttle Sent: Wednesday, 3 December 2008 1:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cleaved Caspase-3 Can anyone recommend control tissue? Thank You Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jim.manavis <@t> imvs.sa.gov.au Tue Dec 2 15:58:00 2008 From: jim.manavis <@t> imvs.sa.gov.au (Jim Manavis) Date: Tue Dec 2 15:58:54 2008 Subject: [Histonet] Neurofilament Question In-Reply-To: References: Message-ID: <003401c954c9$0b3433e0$2c6c140a@41984n> Margaret Get on to the Covance site as they have the Strenberger Neurofilaments. Cheers Jim Jim Manavis Laboratory Manager Hanson Institute Centre for Neurological Diseases IMVS, Adelaide, SA, 5000 Australia Phone: 61-08-8222-3668 / 0401120697 FAX: 61-08-8222 3392 email: jim.manavis@imvs.sa.gov.au Disclaimer: Not this little black duck! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Peggy Bisher Sent: Wednesday, 3 December 2008 7:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Neurofilament Question I have a student who asked if I could post this question to the Histo-list. I told her how all of you seem to be very helpful and perhaps some of you know the answer to her question: Dear all, I am looking for a good "catch-all" antibody for neuronal intermediate filaments (NF-L, NF-M and NF-H). Since they're generally expressed together in each neurofilament I'm not interested in any one of them specifically, but want to avoid using a cocktail of antibodies, as this could get expensive. Basically, I just want to show that a subset of axons contains neurofilaments with a reliable antibody, but the particular protein or phosphorylation state doesn't matter. Any suggestions you could offer would be incredibly helpful! Sincerely, Eve Schneider Thanks, Margaret E. Bisher Electron Microscopy & Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbisher@princeton.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ratliffjack <@t> hotmail.com Tue Dec 2 16:05:12 2008 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Tue Dec 2 16:05:28 2008 Subject: [Histonet] Large Slide Boxes In-Reply-To: <517972921.1999491228254041831.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> References: <517972921.1999491228254041831.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Message-ID: Pamela, I get my boxes (wooden - holds 100 slides) from Brain Research Labs and they cost around $33.00 per box. The website is http://www.brainresearchlab.com/ Jack > Date: Tue, 2 Dec 2008 21:40:41 +0000> From: mucram11@comcast.net> To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] Large Slide Boxes> > > > Good Afternoon All, > > > > I am looking to purchase some 3 inch by 2 inch (3X2) slide boxes for large slides. We do some larger specimens and it seems these have become almost impossible to find. I have tried several sources and no one had them or knew where I could get them at this time. I have tried myneurolab and several other speciality vendors also to no avail. HELP > > > > Pam Marcum> _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail anywhere. No map, no compass. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_anywhere_122008 From cabramley <@t> students.latrobe.edu.au Tue Dec 2 18:12:32 2008 From: cabramley <@t> students.latrobe.edu.au (CLAIRE ANN KENTLER) Date: Tue Dec 2 18:12:49 2008 Subject: [Histonet] Expiration date checks In-Reply-To: References: <16C83872A53F4346AA9C3A18E3A3AAB903F76E9D@VHAV10MSGA1.v10.med.va.gov> Message-ID: <91DC98B386D8614487170C699BE9299F05FB0DF3@stexchange1.students.ltu.edu.au> Hi All, Whilst we're on the subject, we're currently trying to troubleshoot an ELISA assay... This may be a silly question, but does anyone know if (or when) Sodium Bicarbonate loses its buffering capacity? (I.e. does it expire?) Thanks, Claire ************************************************************************ *************************************** Claire Kentler Bachelor of Animal Science (Hons) Postgraduate Student Department of Agricultural Sciences La Trobe University Phone: 9479 1048 E-mail: cabramley@students.latrobe.edu.au -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rande Kline Sent: Wednesday, 3 December 2008 2:18 AM To: Weber, Susan (VHACLE) Cc: histonet Subject: Re: [Histonet] Expiration date checks Patty, Chemicals really don't last forever. Changes take place everytime you open the bottle and some just from age even unopened. Salts used in buffers are hygroscopic. They may appear to look okay but they may not be for your application. How long you can use a chemical is dependent on it's application. I think everyone has experienced a solution not working even though you have followed the correct procedure and a repeat has the same outcome. I have wrote a couple of articles a few years back that appeared in Laboratory Medicine Q&A and Microscopy Today on expiration dating of chemicals. The article in Microscopy Today is a little more indepth. I would not mind faxing you a copy. You may want to dig into more than the dot system. I would also suggest disposing chemicals that you may not know the actual age of, if they exist. On Tue, Dec 2, 2008 at 8:48 AM, Weber, Susan (VHACLE) wrote: > Color coding with sticker "dots" helps a bit. I designate a color for > each year (5 yrs/5 colors max) On the top of that years color I put a > date (usually 6/08 whatever) and make them easier to spot. (Pardon the > pun!)You can find color dots in the office supply catalog, Office Max > (or similar) or the craft stores. Happy Holidays To All! > > Sue Weber > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patti > Loykasek > Sent: Monday, December 01, 2008 6:49 PM > To: histonet > Subject: [Histonet] Expiration date checks > > Hi all. I am hoping for some clever ideas on the best method to check > for expired reagents. We all know that we shouldn't use them, I'm sure > we all actively monitor this (right)! I must admit we occasionally > have expired reagents - a stain or buffer we haven't used in a while > is in a cupboard or fridge & it has expired. What do others do to > monitor & prevent this from happening? We have a weekly safety check > that spot checks a certain # of reagents for expiration, but we > probably have thousands of reagents & don't check every single bottle > every week. Thanks for the input. > > Patti Loykasek > > > > This e-mail message, including any attachments, is for the sole use of > the intended recipients and may contain privileged information. Any > unauthorized review, use, disclosure or distribution is prohibited. If > you are not the intended recipient, please contact the sender by > e-mail and destroy all copies of the original message, or you may call > PhenoPath Laboratories, Seattle, WA U.S.A. > at (206) 374-9000. > > > _______________________________________________ > 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 > -- Rande Kline, HT (ASCP) Histopathology Lab Concepts P-609-744-0803 F-609-939-0270 randek@histopathconcepts.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Tue Dec 2 18:27:14 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Tue Dec 2 18:27:25 2008 Subject: [Histonet] Histo humor :-) References: <4935118102000082000308E8@GWIA1.umsmed.edu> Message-ID: is a casual histo tech like a casual end table? What is it the rest of the time? Oh, yeah, we not only have a great cents of humor, we're real cut ups to. JTT ----- Original Message ----- From: "Dianne Holmes" To: "Histonet" Sent: Tuesday, December 02, 2008 10:44 AM Subject: [Histonet] Histo humor :-) Not only do I get valuable info from this group but histologists have a GREAT sense of humor in common !! I loved the 'casual histo tech' volley!! Individuals who have received this information in error or are not authorized to receive it must promptly return or dispose of the information and notify the sender. Those individuals are hereby notified that they are strictly prohibited from reviewing, forwarding, printing, copying, distributing or using this information in any way. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Tue Dec 2 18:33:07 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Tue Dec 2 18:33:18 2008 Subject: [Histonet] Ventana Benchmark XTs References: <2DFAEEFF192A9141ABACFF88BE613BF30BBC56@EXMBXC1.crha.bewell.ca><4b6c85510811281151w4aec5437ybf85920655ab9def@mail.gmail.com><5b6eb13e0811281210j7fd2f0d5w1c3283bd9bdead11@mail.gmail.com><4b6c85510811290803v7e52528bw2aee194670077f0@mail.gmail.com> <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> Message-ID: I have to agree with Al ( can I call you Al, Alias?) on this one. Remember when I had to sign off of the Histonet a couple of years ago because a certain company contacted my CEO and filed a complaint. I almost lost my job because I posted an honest opinion. An opinion that was greatly shared if I remember correctly. In this era of financial uncertainties, foreclosures and all, I don't blame Al. I'm sure he/she has bills just the rest of us do. Bills that won't go away if he/she loses the job. That's my story and I'm sticking to it. Joe Nocito BS, PA, HT(ASCP)QIHC San Antonio, TX ----- Original Message ----- From: "Histonet Alias" To: "Della Speranza, Vinnie" Cc: ; "Tracey Lenek" ; "Burton,Lynn" ; "Joanna Bartczak-McKay" ; "Martin Trotter" Sent: Tuesday, December 02, 2008 1:19 PM Subject: Re: [Histonet] Ventana Benchmark XTs > Mr. Della Speranza, I respect what you have done for the histology > community > and we have had pleasant conversations in the past but you can not speak > for > the Histonet members. You act as if I am hiding my identity to create > havoc > and disruption. I have not done this nor will I do this. I have been on > histonet for many years and I will continue to do so. As I explained in my > previous posts my current employer does not want any opinions, help, or > questions associated with the laboratory. That is the only reason that I > have chosen to remain anonymous. Would it make anyone feel better if I > made > up a fake name and lab as my signature? I will continue to post and speak > freely without breaking any rules of this listserver. If I respond or ask > a > question, those who find my years of experience not "credible" then > I apologize. > > On Mon, Dec 1, 2008 at 11:21 AM, Della Speranza, Vinnie > wrote: > >> I'm afraid your opinion will not be credible as long as you insist on >> remaining anonymous. It has nothing to do with whether you like or >> dislike a >> particular product or instrument. Histonet members appreciate the >> responsibility that comes with posting one's views or knowledge by >> identifying themselves accordingly. In my opinion anyone who insists on >> remaining anonymous should not be permitted to participate on the list. >> >> >> Vinnie Della Speranza >> Manager for Anatomic Pathology Services >> Medical University of South Carolina >> 165 Ashley Avenue Suite 309 >> Charleston, South Carolina 29425 >> Tel: (843) 792-6353 >> Fax: (843) 792-8974 >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu [mailto: >> histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias >> Sent: Saturday, November 29, 2008 11:03 AM >> To: Mark Tarango >> Cc: histonet@lists.utsouthwestern.edu; Martin Trotter; Tracey Lenek; >> Burton, Lynn; Joanna Bartczak-McKay >> Subject: Re: [Histonet] Ventana Benchmark XTs >> >> Sorry I don't work for Ventana and I have no beef with them. It >> may disappoint some that I have not had a bad experience. So my opinion >> does >> not count because I have had a different experience with my XT than you? >> I >> use the name of The Unknown HT so I can express my opinions freely >> without >> my employer receiving phone calls because certain people don't like my >> opinions. >> >> On Fri, Nov 28, 2008 at 3:10 PM, Mark Tarango >> wrote: >> >> > I don't know how much the opinion of "The Unknown HT(ASCP)" counts. >> > You >> > aren't with Ventana I hope. >> > >> > Mark >> > >> > On Fri, Nov 28, 2008 at 11:51 AM, Histonet Alias < >> histonetalias@gmail.com>wrote: >> > >> >> They are switching to a more reliable manufacturer. They are being >> >> proactive >> >> and switching them before they fail down the road. My XT has been a >> >> workhorse but we know certain peoples opinions about the company on >> here. >> >> I >> >> will take it any day over the rest. >> >> >> >> On Wed, Nov 26, 2008 at 5:29 PM, Burton, Lynn >> >> > >> >wrote: >> >> >> >> > We have had some trouble but they are planning to replace them. We >> >> received >> >> > a letter that they had changed vendors for parts and found they need >> to >> >> > switch back. They did replace one section earlier when it stopped >> >> working >> >> > completely. >> >> > >> >> > Lynn Burton >> >> > Lab Assoc. I >> >> > Animal Disease Lab >> >> > Galesburg, Il 61401 >> >> > >> >> > ________________________________ >> >> > >> >> > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tracey >> >> Lenek >> >> > Sent: Wed 11/26/2008 10:59 AM >> >> > To: histonet@lists.utsouthwestern.edu >> >> > Cc: Martin Trotter; Joanna Bartczak-McKay >> >> > Subject: [Histonet] Ventana Benchmark XTs >> >> > >> >> > >> >> > >> >> > Hi, >> >> > >> >> > We have had on-going thermal slide pad issues with our XTs since >> >> > they >> >> were >> >> > installed >> >> > in February. The slide tray assemblies have been replaced not once >> >> > but >> >> > twice and we are still having inconsistent >> >> > results with the temp verifier slides. Has anyone experienced the >> same >> >> > issues with this >> >> > instrumentation? >> >> > >> >> > Thanks >> >> > Tracey Lenek >> >> > Tech III - Anatomic Pathology >> >> > Calgary Laboratory Services >> >> > 403-770-3588 >> >> > >> >> > ________________________________ >> >> > This message and any attached documents are only for the use of the >> >> > intended recipient(s), are confidential and may contain privileged >> >> > information. Any unauthorized review, use, retransmission, or other >> >> > disclosure is strictly prohibited. If you have received this message >> in >> >> > error, please notify the sender immediately, and then delete the >> >> original >> >> > message. Thank you. >> >> > _______________________________________________ >> >> > 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 Unknown HT(ASCP) >> >> _______________________________________________ >> >> Histonet mailing list >> >> Histonet@lists.utsouthwestern.edu >> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > >> > >> >> >> -- >> The Unknown HT(ASCP) >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > The Unknown HT(ASCP) > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Tue Dec 2 18:36:06 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Tue Dec 2 18:36:16 2008 Subject: [Histonet] Large Slide Boxes References: <517972921.1999491228254041831.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Message-ID: <58CDB4E3235248239CDDF63CCD3D8B9D@yourxhtr8hvc4p> have you tried Lab Storages Systems, New Comer's Supply or Surgipath? JTT ----- Original Message ----- From: "Pamela Marcum" To: Sent: Tuesday, December 02, 2008 3:40 PM Subject: [Histonet] Large Slide Boxes Good Afternoon All, I am looking to purchase some 3 inch by 2 inch (3X2) slide boxes for large slides. We do some larger specimens and it seems these have become almost impossible to find. I have tried several sources and no one had them or knew where I could get them at this time. I have tried myneurolab and several other speciality vendors also to no avail. HELP Pam Marcum _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sheila_adey <@t> hotmail.com Tue Dec 2 18:39:09 2008 From: sheila_adey <@t> hotmail.com (sheila adey) Date: Tue Dec 2 18:39:20 2008 Subject: [Histonet] Casual Histotech In-Reply-To: <1CE1847DFEA0A647B1CCDE4108EA60A70208C5A8@LTA3VS011.ees.hhs.gov> References: <652B88F04HG494005-01@EMF> <1CE1847DFEA0A647B1CCDE4108EA60A70208C5A8@LTA3VS011.ees.hhs.gov> Message-ID: I can't believe so many people are not familiar with the term "Casual" it could also be called contingent. Sheila Adey HT MLT Port Huron Hospital Michigan> Date: Tue, 2 Dec 2008 10:43:35 -0500> From: jqb7@cdc.gov> To: jdhisto@yahoo.com; Histonet@lists.utsouthwestern.edu> Subject: RE: [Histonet] Casual Histotech> CC: > > As opposed to the rest of us...you know, the formal ones. > > > Jeanine Bartlett> Infectious Diseases Pathology Branch> (404) 639-3590 > jeanine.bartlett@cdc.hhs.gov> > > -----Original Message-----> From: histonet-bounces@lists.utsouthwestern.edu> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JD> Sent: Tuesday, December 02, 2008 10:42 AM> To: Histonet@lists.utsouthwestern.edu> Subject: [Histonet] Casual Histotech> > My thoughts exactly. Casual> histotech?_______________________________________________> 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 jkiernan <@t> uwo.ca Tue Dec 2 21:28:20 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Dec 2 21:28:24 2008 Subject: [Histonet] Fontana Masson Message-ID: Probably the best place to start is in Pearse's "Histochemistry" 4 stuff & Fu p.522-527 (19 chemistry of the argentaf Lillie assumed that their readers that had been in use for 100+ years! Unfo chemical information is not in many modern chemist older ones are more helpful.
 
need to follow up the references cited by Pearse and Lillie. If  Unilever's library does not have these books, it's a huge company. Kiernan
Anatomy, UWO
Lond = =
----- Original Message -----< "Kalleberg, Kristopher"
Date: Tuesday, December 2, 2008 15:04< histonet@li All,
> for me to read up on the Fontana Masson staining of melanin hard time finding
> in how FM stain stains the
> me and
> the chemistry and reactions involved. Any information,
> papers, or
> references advance.
> Kalleberg
> Research Scientist
> 40 Merritt Blvd.
> Trumbull, CT 06611
> (203) 381-5765
& ______________________ _______________________ 5F Histonet@lis http://lists.utsouthwester n.edu/mailman/listinfo/histonet

From histonet.nospam <@t> vneubert.com Wed Dec 3 00:30:04 2008 From: histonet.nospam <@t> vneubert.com (V. Neubert) Date: Wed Dec 3 00:30:07 2008 Subject: [Histonet] Staining fungi with Warthin-Starry Message-ID: <616c2a7d372cbbe3888f3ad3955ce98a@vneubert.com> Just a quick question: Did anyone successfully try to stain fungi with Warthin-Starry silver technique? Thank you for our answers, V. Neubert From nbender9 <@t> verizon.net Wed Dec 3 05:18:21 2008 From: nbender9 <@t> verizon.net (Nora Bender) Date: Wed Dec 3 05:18:22 2008 Subject: [Histonet] RE: Large Slide Boxes Message-ID: <0KBA002XISQ8CJA9@vms173005.mailsrvcs.net> Hello Pam, We carry several types of slide boxes for 2"x3" slides as well as boxes to hold slides up to 5"x7". Please feel free to call me toll-free @ 1(888)275-5544. Nora Bender Brain Research Laboratories Tel: 617-965-5544 www.brainresearchlab.com Good Afternoon All, I am looking to purchase some 3 inch by 2 inch (3X2) slide boxes for large slides.B We do some larger specimens and it seems these have become almost impossible to find.B I have tried several sources and no one had them or knew where I could get them at this time.B I have tried myneurolab and several other speciality vendors also to no avail.B HELP Pam Marcum From laurie <@t> conxis.com Wed Dec 3 06:39:41 2008 From: laurie <@t> conxis.com (Laurie Popp) Date: Wed Dec 3 06:40:33 2008 Subject: [Histonet] RE: Casual Histotech Message-ID: <49367E0D.4090504@conxis.com> Casual Histotech is usually considered to either be very part time or a pool position to cover only when needed. Laurie Popp, BA HT (ASCP) From jqb7 <@t> cdc.gov Wed Dec 3 06:50:27 2008 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Wed Dec 3 06:50:39 2008 Subject: [Histonet] RE: Casual Histotech In-Reply-To: <49367E0D.4090504@conxis.com> References: <49367E0D.4090504@conxis.com> Message-ID: <1CE1847DFEA0A647B1CCDE4108EA60A70208C5BB@LTA3VS011.ees.hhs.gov> Who knew? Thanks! Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Laurie Popp Sent: Wednesday, December 03, 2008 7:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Casual Histotech Casual Histotech is usually considered to either be very part time or a pool position to cover only when needed. Laurie Popp, BA HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DixonM <@t> vetmed.ufl.edu Wed Dec 3 07:54:39 2008 From: DixonM <@t> vetmed.ufl.edu (MaryAnn Dixon) Date: Wed Dec 3 07:54:49 2008 Subject: [Histonet] Lysine Message-ID: <530D827EC657DE418C3572ADD63FCDC3249529@EXGVMCNETWORK.vetmed.ufl.edu> Good Morning Histonetters, Can anyone tell me of a supplier and specific grade of L-lysine HCl for the make up of 2% periodate lysine paraformaldehye fixative? Thank you. MaryAnn Dixon BS Biological Scientist Anatomic Pathology UF Veterinary Medical Center (352) 392-2235 Ext. 4517 From histonetalias <@t> gmail.com Wed Dec 3 08:11:21 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Wed Dec 3 08:11:27 2008 Subject: [Histonet] Ventana Benchmark XTs In-Reply-To: References: <2DFAEEFF192A9141ABACFF88BE613BF30BBC56@EXMBXC1.crha.bewell.ca> <4b6c85510811281151w4aec5437ybf85920655ab9def@mail.gmail.com> <5b6eb13e0811281210j7fd2f0d5w1c3283bd9bdead11@mail.gmail.com> <4b6c85510811290803v7e52528bw2aee194670077f0@mail.gmail.com> <4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> Message-ID: <4b6c85510812030611o7516c4e5q26cb6a1a38f5176b@mail.gmail.com> Let's agree to disagree here. I will continue to post when I feel necessary and the people who don't agree with my reasoning can just ignore me. Just for the record I do not work for any vendor and I am a Histology Manager for a private laboratory in the midwest. Will my name make any difference in my posts? Take that for what it is worth. This thread was really ignited by me giving an opinion about a product or company that many people do not care for. If you don't like it don't use it. You can give your opinion about it when asked just like I can. I will use Joe the toes suggestion and change up my signature to avoid any further issue. I work for one of the larger lab companies that do not want their name or employees attached to any opinion other than the companies. If you can not understand that then I can't help you. I have mouths to feed and I do not have the luxury of changing jobs and moving to a new location. Do I like hiding my identity? No. I have been on here for years with an open identity. I have found a way to still contribute and gain knowledge from this listserv and a few of you are all about status and credibility. SO let us just end this thread and save space for real Histology issues. Al Ias HT(ASCP) Histology Manager A MidWest Lab On Tue, Dec 2, 2008 at 7:33 PM, Joe Nocito wrote: > I have to agree with Al ( can I call you Al, Alias?) on this one. Remember > when I had to sign off of the Histonet a couple of years ago because a > certain company contacted my CEO and filed a complaint. I almost lost my job > because I posted an honest opinion. An opinion that was greatly shared if I > remember correctly. In this era of financial uncertainties, foreclosures and > all, I don't blame Al. I'm sure he/she has bills just the rest of us do. > Bills that won't go away if he/she loses the job. > That's my story and I'm sticking to it. > > Joe Nocito BS, PA, HT(ASCP)QIHC > San Antonio, TX > > ----- Original Message ----- From: "Histonet Alias" < > histonetalias@gmail.com> > To: "Della Speranza, Vinnie" > Cc: ; "Tracey Lenek" < > Tracey.Lenek@cls.ab.ca>; "Burton,Lynn" ; "Joanna > Bartczak-McKay" ; "Martin Trotter" < > Martin.Trotter@cls.ab.ca> > Sent: Tuesday, December 02, 2008 1:19 PM > > Subject: Re: [Histonet] Ventana Benchmark XTs > > > Mr. Della Speranza, I respect what you have done for the histology >> community >> and we have had pleasant conversations in the past but you can not speak >> for >> the Histonet members. You act as if I am hiding my identity to create >> havoc >> and disruption. I have not done this nor will I do this. I have been on >> histonet for many years and I will continue to do so. As I explained in my >> previous posts my current employer does not want any opinions, help, or >> questions associated with the laboratory. That is the only reason that I >> have chosen to remain anonymous. Would it make anyone feel better if I >> made >> up a fake name and lab as my signature? I will continue to post and speak >> freely without breaking any rules of this listserver. If I respond or ask >> a >> question, those who find my years of experience not "credible" then >> I apologize. >> >> On Mon, Dec 1, 2008 at 11:21 AM, Della Speranza, Vinnie > >wrote: >> >> I'm afraid your opinion will not be credible as long as you insist on >>> remaining anonymous. It has nothing to do with whether you like or >>> dislike a >>> particular product or instrument. Histonet members appreciate the >>> responsibility that comes with posting one's views or knowledge by >>> identifying themselves accordingly. In my opinion anyone who insists on >>> remaining anonymous should not be permitted to participate on the list. >>> >>> >>> Vinnie Della Speranza >>> Manager for Anatomic Pathology Services >>> Medical University of South Carolina >>> 165 Ashley Avenue Suite 309 >>> Charleston, South Carolina 29425 >>> Tel: (843) 792-6353 >>> Fax: (843) 792-8974 >>> >>> >>> -----Original Message----- >>> From: histonet-bounces@lists.utsouthwestern.edu [mailto: >>> histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias >>> Sent: Saturday, November 29, 2008 11:03 AM >>> To: Mark Tarango >>> Cc: histonet@lists.utsouthwestern.edu; Martin Trotter; Tracey Lenek; >>> Burton, Lynn; Joanna Bartczak-McKay >>> Subject: Re: [Histonet] Ventana Benchmark XTs >>> >>> Sorry I don't work for Ventana and I have no beef with them. It >>> may disappoint some that I have not had a bad experience. So my opinion >>> does >>> not count because I have had a different experience with my XT than you? >>> I >>> use the name of The Unknown HT so I can express my opinions freely >>> without >>> my employer receiving phone calls because certain people don't like my >>> opinions. >>> >>> On Fri, Nov 28, 2008 at 3:10 PM, Mark Tarango >>> wrote: >>> >>> > I don't know how much the opinion of "The Unknown HT(ASCP)" counts. > >>> You >>> > aren't with Ventana I hope. >>> > >>> > Mark >>> > >>> > On Fri, Nov 28, 2008 at 11:51 AM, Histonet Alias < >>> histonetalias@gmail.com>wrote: >>> > >>> >> They are switching to a more reliable manufacturer. They are being >>> >> proactive >>> >> and switching them before they fail down the road. My XT has been a >>> >> workhorse but we know certain peoples opinions about the company on >>> here. >>> >> I >>> >> will take it any day over the rest. >>> >> >>> >> On Wed, Nov 26, 2008 at 5:29 PM, Burton, Lynn >> < >>> Lynn.Burton@illinois.gov >>> >> >wrote: >>> >> >>> >> > We have had some trouble but they are planning to replace them. We >>> >> received >>> >> > a letter that they had changed vendors for parts and found they need >>> to >>> >> > switch back. They did replace one section earlier when it stopped >>> >> working >>> >> > completely. >>> >> > >>> >> > Lynn Burton >>> >> > Lab Assoc. I >>> >> > Animal Disease Lab >>> >> > Galesburg, Il 61401 >>> >> > >>> >> > ________________________________ >>> >> > >>> >> > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Tracey >>> >> Lenek >>> >> > Sent: Wed 11/26/2008 10:59 AM >>> >> > To: histonet@lists.utsouthwestern.edu >>> >> > Cc: Martin Trotter; Joanna Bartczak-McKay >>> >> > Subject: [Histonet] Ventana Benchmark XTs >>> >> > >>> >> > >>> >> > >>> >> > Hi, >>> >> > >>> >> > We have had on-going thermal slide pad issues with our XTs since >> >>> > they >>> >> were >>> >> > installed >>> >> > in February. The slide tray assemblies have been replaced not once >>> >> > but >>> >> > twice and we are still having inconsistent >>> >> > results with the temp verifier slides. Has anyone experienced the >>> same >>> >> > issues with this >>> >> > instrumentation? >>> >> > >>> >> > Thanks >>> >> > Tracey Lenek >>> >> > Tech III - Anatomic Pathology >>> >> > Calgary Laboratory Services >>> >> > 403-770-3588 >>> >> > >>> >> > ________________________________ >>> >> > This message and any attached documents are only for the use of the >>> >> > intended recipient(s), are confidential and may contain privileged >>> >> > information. Any unauthorized review, use, retransmission, or other >>> >> > disclosure is strictly prohibited. If you have received this message >>> in >>> >> > error, please notify the sender immediately, and then delete the >>> >> original >>> >> > message. Thank you. >>> >> > _______________________________________________ >>> >> > 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 Unknown HT(ASCP) >>> >> _______________________________________________ >>> >> Histonet mailing list >>> >> Histonet@lists.utsouthwestern.edu >>> >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >> >>> > >>> > >>> >>> >>> -- >>> The Unknown HT(ASCP) >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>> >> >> >> -- >> The Unknown HT(ASCP) >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > From GDawson <@t> dynacaremilwaukee.com Wed Dec 3 09:09:46 2008 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Wed Dec 3 09:09:54 2008 Subject: [Histonet] Biomeda Corp. In-Reply-To: <530D827EC657DE418C3572ADD63FCDC3249529@EXGVMCNETWORK.vetmed.ufl.edu> Message-ID: All, I vaguely remember a discussion on this, but could someone please refresh me on who bought out Biomeda and perhaps provide contact information for whomever is now dispensing their product? I am looking for their FITC Streptavidin (F72) so if there are any vendors that provide a similar item, please feel free to respond directly to me so I have some options for if it is discontinued all together. On another note, I'm also using Biomeda's Crystal Mount Aqueous Mounting media so alternative product suggestions for this one would be much appreciated as well. Thank-you in Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From fudo <@t> ufl.edu Wed Dec 3 09:16:38 2008 From: fudo <@t> ufl.edu (FU,DONGTAO) Date: Wed Dec 3 09:16:42 2008 Subject: [Histonet] IHC on fresh frozen Message-ID: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> Hi, all Recently I did some IHC(chromagen methods) on mouse fresh frozen tissues, mainly using insulin antibody on pancreas. The image is much fuzzier compare to paraffin embedding tissue. And the staining also smeared to acinar cells which surround the islet. I airdried slide(>30min) and used a general acetone method(-20C 5min) to fix the tissue before I did IHC. How can I get a relatively sharp staining on the fresh frozen tissue?Does anyone here have any experience on it? Any suggestions? Many thanks and have a nice day, Ann From mcauliff <@t> umdnj.edu Wed Dec 3 09:30:14 2008 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Dec 3 09:33:08 2008 Subject: [Histonet] Histo humor :-) In-Reply-To: References: <4935118102000082000308E8@GWIA1.umsmed.edu> Message-ID: <4936A606.8030509@umdnj.edu> Or like an occasional chair? Geoff Joe Nocito wrote: > is a casual histo tech like a casual end table? What is it the rest > of the time? > > Oh, yeah, we not only have a great cents of humor, we're real cut ups to. > > JTT > ----- Original Message ----- From: "Dianne Holmes" > > To: "Histonet" > Sent: Tuesday, December 02, 2008 10:44 AM > Subject: [Histonet] Histo humor :-) > > > Not only do I get valuable info from this group but histologists have > a GREAT sense of humor in common !! > I loved the 'casual histo tech' volley!! > > > Individuals who have received this information in error or are not > authorized to receive it must promptly return or dispose of the > information and notify the sender. Those individuals are hereby > notified that they are strictly prohibited from reviewing, forwarding, > printing, copying, distributing or using this information in any way. > > > > _______________________________________________ > 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 > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583 mcauliff@umdnj.edu ********************************************** From celebrej <@t> HHSC.CA Wed Dec 3 09:38:40 2008 From: celebrej <@t> HHSC.CA (Celebre Julia) Date: Wed Dec 3 09:38:43 2008 Subject: [Histonet] Flame substitute for embedding Message-ID: <4D3667D4B5487546A3139FB918181FEA01F75CBF@ipemail01.hhsc.ca> Hello Histoland!! We have just been told we are no longer allowed to use alcohol burners while embedding and we are to find a safer substitute, something to do with no liking an open flame in the lab. Other than incinerators or 6 pairs of forceps at each centre, we've run out of ideas so we are asking all you experts out there what your current practice is.. oh.. this change needs to be done by this Friday!! Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. From Janet.Bonner <@t> FLHOSP.ORG Wed Dec 3 09:43:46 2008 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Wed Dec 3 09:45:35 2008 Subject: [Histonet] Flame substitute for embedding References: <4D3667D4B5487546A3139FB918181FEA01F75CBF@ipemail01.hhsc.ca> Message-ID: <5F31F38C96781A4FBE3196EBC22D47807F2857@fhosxchmb006.ADVENTISTCORP.NET> We use forceps warmers, find them in any Histology catalog (like Fisher Scientific or Scientific Products/Allegiance). Janet L. Bonner, HTL (ASCP) Pathology Laboratory ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Celebre Julia Sent: Wed 12/3/2008 10:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flame substitute for embedding Hello Histoland!! We have just been told we are no longer allowed to use alcohol burners while embedding and we are to find a safer substitute, something to do with no liking an open flame in the lab. Other than incinerators or 6 pairs of forceps at each centre, we've run out of ideas so we are asking all you experts out there what your current practice is.. oh.. this change needs to be done by this Friday!! Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ======================================================= The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ======================================================= From Jacqueline.Farnsworth <@t> cls.ab.ca Wed Dec 3 09:43:49 2008 From: Jacqueline.Farnsworth <@t> cls.ab.ca (Jacqueline Farnsworth) Date: Wed Dec 3 09:46:15 2008 Subject: [Histonet] RE: Flame substitute for embedding In-Reply-To: <4D3667D4B5487546A3139FB918181FEA01F75CBF@ipemail01.hhsc.ca> References: <4D3667D4B5487546A3139FB918181FEA01F75CBF@ipemail01.hhsc.ca> Message-ID: We also used to have an open flame while embedding! Now, we simply have a policy in place where after EACH cassette is embedded, the Techs MUST thoroughly wipe their forceps with disposable Kleenex (and then dispose of it!). Also, we use toothless forceps as much as possible to reduce the possibility of cross contamination. Jacqueline Farnsworth Anatomic Pathology, Tech III Foothills Medical Centre Calgary Laboratory Services Ph: 403-944-1578 Fax: 403-944-4748 P Please consider the environment before printing this email. ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia [celebrej@HHSC.CA] Sent: December 3, 2008 8:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flame substitute for embedding Hello Histoland!! We have just been told we are no longer allowed to use alcohol burners while embedding and we are to find a safer substitute, something to do with no liking an open flame in the lab. Other than incinerators or 6 pairs of forceps at each centre, we've run out of ideas so we are asking all you experts out there what your current practice is.. oh.. this change needs to be done by this Friday!! Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. From TJJ <@t> Stowers-Institute.org Wed Dec 3 09:49:40 2008 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Dec 3 09:50:16 2008 Subject: [Histonet] Histonet alias Message-ID: It is not uncommon for many/most online forums to have users create screennames that they use for their online moniker. You never really know who they are so everybody, unless they want to divulge their identity, stays anonymous. How can we really know all of the registered names on the histonet are real anyway? And why would it matter? The value of the histonet is information. There are opinions for and against equipment and particular methods. I am not likely to discount good information just because it was given by a company rep, pathologist, med tech, hairdresser, or Mr. Anonymous. I argue that anonymity could be the best opportunity to give the most honest answer to a question posted to the board. It is up to each one of us to evaluate those opinions, take what is important, and leave the rest regardless of who posted it. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From kynemitz <@t> travmax.com Wed Dec 3 10:00:44 2008 From: kynemitz <@t> travmax.com (Kyla Nemitz) Date: Wed Dec 3 10:01:39 2008 Subject: [Histonet] Histology Supervisor position in Frederick, MD Message-ID: <9416D9FA37C1C04FA83D69684E95D2E71F457398@exbk2.maxhealth.com> Hello HistoNet - A facility I work with in Frederick, MD has asked for my assistance in finding a Histology Supervisor. Day shift, rotation of call on evenings and weekends. Salary range is $44,512 - $72,800 and relocation assistance available. If you or anyone you know would be interested in this position, please contact me. I would be more than happy to help you find a new job for 2009! Happy Holidays, hope to hear from you soon! Kyla Nemitz TravelMax Medical Professionals - Maxim Healthcare * 888.800.1855 or 813.371.5175 7 800.294.1248 Search our jobs or apply online at: www.TravelMaxAllied.com From amber.mckenzie <@t> gastrodocs.net Wed Dec 3 10:06:27 2008 From: amber.mckenzie <@t> gastrodocs.net (Amber McKenzie) Date: Wed Dec 3 10:06:30 2008 Subject: [Histonet] Alias identity In-Reply-To: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> References: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> Message-ID: <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> I have to put my 2 cents in about all the statements on having an alias name instead of identifying who each person is after each email...WHAT'S THE BIG DEAL?? We are all supposed to be professionals asking each other for advice/suggestions on the Histonet - who cares who each person is? If I post a question, I don't care if it's Jane Doe answering or John Smith. I assume that if we're on this list that we're all credible, knowledge and professionals. Come on people, we're all in the same boat here. If I'd thought about putting an alias name for myself instead of my real name, I would have! Simply b/c last month I posted a question on the Histonet asking where you all bought your lab chairs, and I ended up having 2 vendors call me at my office trying to sell me some when all I wanted was the advice of other HT's. I love the idea of people not knowing who I am or where I work. Then I wouldn't have to worry about being harassed on the phone. Instead of worrying about the little things on the Histonet - like who each person is, why can't we focus on work related issues? I have to delete so much junk just to get to the material that actually applies to my field b/c of all the multiple emails that don't even matter. From galinadeyneko <@t> yahoo.com Wed Dec 3 10:11:18 2008 From: galinadeyneko <@t> yahoo.com (Galina Deyneko) Date: Wed Dec 3 10:11:21 2008 Subject: [Histonet] Insulin/Glucagon double IHC Message-ID: <182565.81667.qm@web33102.mail.mud.yahoo.com> Dear Colleagues, I would like to ask your advices. I should establish double IHC staining on FFPE?isolated mouse pancreatic islets for alpha and beta cells.?I have very restricted amount of slides and I do not have possibility to test different antibodies and protocols. Could you please give me references what are the best antibodies for insulin and glucagon for mouse and for human (for the future study)?Langerhans islets staining.I think?for double staining it is better?to use the antibodies from?different species.?If you could share with me the protocols, I mean some tips about? antigen retrieval, antibody dilution and incubation time, I will really appreciate it. As a secondary antibody i use the?polymers from Biocare. Thank you in advance. Galina Deyneko. Novartis, Cambridge, MA 617-871-7613 From LSebree <@t> uwhealth.org Wed Dec 3 10:12:51 2008 From: LSebree <@t> uwhealth.org (Sebree Linda A) Date: Wed Dec 3 10:12:56 2008 Subject: [Histonet] IHC on fresh frozen In-Reply-To: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> Message-ID: I was taught to get frozen sections into cold acetone immediately after sectioning, not letting the section air dry. We do C4d staining on frozen renal bxs and immediately fix the sections in cold acetone for 10 minutes. Then we air dry followed by 2 minutes in Ventana Morphosave (not a critical step) then keep the slides wet throughout the immunostain. We get nice clear, crisp staining. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of FU,DONGTAO Sent: Wednesday, December 03, 2008 9:17 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on fresh frozen Hi, all Recently I did some IHC(chromagen methods) on mouse fresh frozen tissues, mainly using insulin antibody on pancreas. The image is much fuzzier compare to paraffin embedding tissue. And the staining also smeared to acinar cells which surround the islet. I airdried slide(>30min) and used a general acetone method(-20C 5min) to fix the tissue before I did IHC. How can I get a relatively sharp staining on the fresh frozen tissue?Does anyone here have any experience on it? Any suggestions? Many thanks and have a nice day, Ann _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From relia1 <@t> earthlink.net Wed Dec 3 10:19:32 2008 From: relia1 <@t> earthlink.net (Pam Barker) Date: Wed Dec 3 10:19:36 2008 Subject: [Histonet] RELIA Histology Management Job Alert 12/03/08 Message-ID: Hi Histonetters!! I hope everybody had a great Thanksgiving. I have several clients that are in need of managers and supervisors so I thought I would put the word out. At this point since it is the end of the year my clients are willing to let you call the shots on how you would like to proceed i.e. interview now or after the holidays. Of course these are all permanent positions offering excellen salaries, benefits and relocation assistance. Here is a list of my current management, supervisory and lead positions: Anatomic Pathology Manager - Los Angeles, CA Histology Supervisor - Los Angeles, CA Histology Supervisor - Frederick MD Histology Supervisor - Cincinnati, OH Histology Manager - Dallas, TX Gross Room Supervisor - Austin, TX Pathology Assistant - Corpus Christi, TX If you or anyone you know is interested call me toll free at 866-607-3542 or e-mail me at relia1@earthlink.net Thanks and Have a Great Day!! 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 <> www.myspace.com/pamatrelia From gayle.callis <@t> bresnan.net Wed Dec 3 10:37:59 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Wed Dec 3 10:38:02 2008 Subject: [Histonet] Detailed Re: Attaching GMA sections to slides Message-ID: <07A847FC24E8492B94ABAA3CE463A648@DHXTS541> Jim, You did not say what you were doing to the sections after mounting them on slides as this may be a factor for section lifting off other than It is important to have perfectly flat sections, so make sure the blades or glass knife is very sharp. GMA is designed for use for thin sections 3 um or thinner, and when we had 4 to 5 um thick sections, lift off was more of a problem. You did not say how thin your sections were? If thick sections are not in flat contact with slide at pickup, this may be a factor. We never had to weigh down GMA sections. We preferred glass knives and used a new knife to section each block. The knife used for a block became the trim knife for the next block. We floated each individual GMA sections onto room temperature distilled water filled to the top of a glass staining dish. It takes a bit of practice to lay the flightly little plastic sections on the water, but it can be done. Don't use warm water baths as this helps to release fumes from the plastic that are sensitizing, avoid becoming allergic to this plastic. Sections were picked up onto a very clean glass slide, with slide held almost vertically. We have used plain uncoated glass slides, washed in hot detergent, rinsed well with tap water then distilled, dipped in acetone or absolute ethanol to dry. The surface of the slide was pristine, without any dirt, dust or oily residue. A labor intensive job to wash slides. We later found Plus charge slides (Erie) worked well. We simply air dried sections, then went to a 56C incubator for 60 minutes. There are tricks to prevent GMA sections from coming off the slide. Our sections did not release from slides unless we overexposed them to 95% alcohol during the H&E staining, and we never bothered to rehydrate a section with distilled water before going into the first staining solution. If the section appeared to be lifting up (swells up) at some point in the staining protocol, a careful rinse then air dry with a stream of compressed air or use a hair dryer on cool setting. GMA is a bit more forgiving with this little trick. . I would not air dry after removal from a stain solution but after a rinse. A small fan or even a cool setting on small hand held hair dryer should work. Forced air was our favorite since it put a stream of air at higher velocity againt the section, drying was instantly done when doing Giemsa's and after alcohol rinse steps. My apologies for a long lecture and good luck on retaining the GMA sections. Gayle M. Callis HTL(ASCP)HT,MT Bozeman MT From ploykasek <@t> phenopath.com Wed Dec 3 10:38:07 2008 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Dec 3 10:38:35 2008 Subject: [Histonet] IHC on fresh frozen In-Reply-To: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> Message-ID: After hearing a presentation by Sharon Lear describing some low temp antigen retrieval that she did, we changed our method for frozen sections. We fix the frozen sections in 10% nuetral buffered formalin for 30'-60', rinse, then do a gentle pretreatment. The gentle pretreatment is usually 10mM citrate buffer pH6 at 70 degrees C for 30'. Slides are cooled, and usual IHC done. This has worked well for us. Our staining with this method is more reliable & intense than with previous methods. Patti Loykasek > Hi, all > > Recently I did some IHC(chromagen methods) on mouse fresh frozen > tissues, mainly using insulin antibody on pancreas. The image is > much fuzzier compare to paraffin embedding tissue. And the > staining also smeared to acinar cells which surround the islet. > > I airdried slide(>30min) and used a general acetone method(-20C > 5min) to fix the tissue before I did IHC. > > How can I get a relatively sharp staining on the fresh frozen > tissue?Does anyone here have any experience on it? Any > suggestions? > > Many thanks and have a nice day, > > Ann > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From gayle.callis <@t> bresnan.net Wed Dec 3 10:43:57 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Wed Dec 3 10:43:57 2008 Subject: [Histonet] Re: Fungus staining with Warthin Starry Message-ID: There is no oxidizing step in Warthin Starry to prepare fungal walls for silver deposition. This stain is for spirochetes. I suggest you use the Grocotts (Gomori's modification) methenamine silver where chromic acid is used to oxidize the fungal cell walls. You can find this on the web, or in the Histonet Archives. It is not difficult to do. Good luck Gayle M. Callis HTL(ASCP)HT,MT Bozeman MT From integrated.histo <@t> gmail.com Wed Dec 3 11:05:36 2008 From: integrated.histo <@t> gmail.com (Cindy DuBois) Date: Wed Dec 3 11:05:41 2008 Subject: [Histonet] Shandon Processor Message-ID: <5d9104a30812030905n6de3c275ncc03aeacf9229bba@mail.gmail.com> Any one in histoland using the Shandon Excelsior? How long have you had it & how many blocks can it process at a time? Cindy DuBois Integrated Pathology From kellerp <@t> ent.wustl.edu Wed Dec 3 11:29:55 2008 From: kellerp <@t> ent.wustl.edu (Pat Keller) Date: Wed Dec 3 11:30:04 2008 Subject: [Histonet] 2x3 slides Message-ID: Fisher has them MILLIPORE CORP MICROSCOPE SLIDES 2"X3" 72/PK Fisher offers many products that do not appear in our catalogs. This may be one of those products, so pictures and detailed descriptions are not available. However, you may be able to order it by adding it to your shopping cart. Item Description Catalog Number Quantity Price MICROSCOPE SLIDES 2"X3" 72/PK XX1007615 Vendor No.:XX1007615 Pack of 72 for $51.26 Patricia Keller, Sr. Res. Tech. Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, MO. 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE 314-747-7166 FAX 314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. From mucram11 <@t> comcast.net Wed Dec 3 11:34:48 2008 From: mucram11 <@t> comcast.net (Pamela Marcum) Date: Wed Dec 3 11:34:52 2008 Subject: [Histonet] 2x3 slides In-Reply-To: Message-ID: <966250373.78611228325688567.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> I got the slides from Thermo Fisher with some problems a long while ago.? The issue was finding a 100 slide box for storage and shipping.? I did order them this morning from Brain Research and they had stock of the boxes.? Thanks to everyone who helped and now I know where to go for the boxes next time. Pam Marcum ----- Original Message ----- From: "Pat Keller" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 3, 2008 12:29:55 PM GMT -05:00 US/Canada Eastern Subject: [Histonet] 2x3 slides Fisher has them MILLIPORE CORP ?MICROSCOPE SLIDES 2"X3" 72/PK Fisher offers many products that do not appear in our catalogs. This may be one of those products, so pictures and detailed descriptions are not available. However, you may be able to order it by adding it to your shopping cart. ? Item Description ? ? Catalog Number ? ? Quantity ? ? Price MICROSCOPE SLIDES 2"X3" 72/PK ?? ?XX1007615 Vendor No.:XX1007615 ? ? ? ? Pack of 72 for $51.26 Patricia Keller, Sr. Res. Tech. ? Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, ? MO. ? 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE ? ?314-747-7166 FAX ? ? ? ? ?314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Margaret.Perry <@t> sdstate.edu Wed Dec 3 11:46:33 2008 From: Margaret.Perry <@t> sdstate.edu (Perry, Margaret) Date: Wed Dec 3 11:46:44 2008 Subject: [Histonet] lids for stainer containers Message-ID: We are looking for a source that sells a lid that looks like wax covered cardboard. We use them on top of the Shandon slide containers holding xylene or formula 83 while waiting to coverslip. They are about 5x7 and very thin. They have been here for years so I have no idea where they came from. Any help in finding them or a substitute will be greatly appreciated. Thanks. Margaret Perry HT (ASCP) IHC Lab Manager Veterinary Science Animal Disease Research and Diagnostic Lab South Dakota State University Box 2175 North Campus Drive Brookings SD 57007 From b-frederick <@t> northwestern.edu Wed Dec 3 11:48:12 2008 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Wed Dec 3 11:48:23 2008 Subject: [Histonet] RE: Flame substitute for embedding In-Reply-To: Message-ID: <000001c9556f$52b14590$d00f7ca5@lurie.northwestern.edu> We have a Shandon Paratrimmer, which melts the wax not only off blocks but does a good job of keeping those forceps clean and hot. We have a bacti-incinerator, but know we can get a tall flame if we leave the paraffin coated forceps in there!! Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jacqueline Farnsworth Sent: Wednesday, December 03, 2008 9:44 AM To: Celebre Julia; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Flame substitute for embedding We also used to have an open flame while embedding! Now, we simply have a policy in place where after EACH cassette is embedded, the Techs MUST thoroughly wipe their forceps with disposable Kleenex (and then dispose of it!). Also, we use toothless forceps as much as possible to reduce the possibility of cross contamination. Jacqueline Farnsworth Anatomic Pathology, Tech III Foothills Medical Centre Calgary Laboratory Services Ph: 403-944-1578 Fax: 403-944-4748 P Please consider the environment before printing this email. ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia [celebrej@HHSC.CA] Sent: December 3, 2008 8:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flame substitute for embedding Hello Histoland!! We have just been told we are no longer allowed to use alcohol burners while embedding and we are to find a safer substitute, something to do with no liking an open flame in the lab. Other than incinerators or 6 pairs of forceps at each centre, we've run out of ideas so we are asking all you experts out there what your current practice is.. oh.. this change needs to be done by this Friday!! Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kimberly.Marshall <@t> ahss.org Wed Dec 3 11:47:34 2008 From: Kimberly.Marshall <@t> ahss.org (Marshall, Kimberly) Date: Wed Dec 3 11:49:10 2008 Subject: [Histonet] Blue haze Message-ID: Howdy all, I have just changed over to Surgi Path's H & E system. With the latest fear of running out of hematoxylin and all my Pathologist wanted us to change over. Its a great stain but we have a bad "haze" in the background. It is not hurting the tissue or DX at all, but to ME it looks awful. Is there anyone else out there using this? We are not using positive charged slides or anything added to my water bath. Any suggestions???? Thanks in advance Kimberly Marshall HT (ASCP) ============================================================================== The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ============================================================================== From gayle.callis <@t> bresnan.net Wed Dec 3 11:56:21 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Wed Dec 3 11:56:29 2008 Subject: [Histonet] Re: PLP fixative lysine Message-ID: For the PLP fixative, the full name for the lysine is L- lysine monohydrochloride. Sigma cat# L5626. Gayle M. Callis HTL(ASCP)HT,MT From histonetalias <@t> gmail.com Wed Dec 3 12:05:03 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Wed Dec 3 12:05:07 2008 Subject: [Histonet] Blue haze In-Reply-To: References: Message-ID: <4b6c85510812031005r447cc8d2v156807ed4921077f@mail.gmail.com> On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < Kimberly.Marshall@ahss.org> wrote: > Howdy all, > > I have just changed over to Surgi Path's H & E system. With the latest > fear of running out of hematoxylin and all my Pathologist wanted us to > change over. Its a great stain but we have a bad "haze" in the background. > It is not hurting the tissue or DX at all, but to ME it looks awful. Is > there anyone else out there using this? We are not using positive charged > slides or anything added to my water bath. Any suggestions???? Thanks in > advance > > Kimberly Marshall HT (ASCP) > > > ============================================================================== > The information contained in this message may be privileged and/or > 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 the sender immediately by replying to > this > message and deleting the material from any computer. > > > ============================================================================== > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States From histonetalias <@t> gmail.com Wed Dec 3 12:08:21 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Wed Dec 3 12:08:25 2008 Subject: [Histonet] Blue haze In-Reply-To: References: Message-ID: <4b6c85510812031008k3902865enbfd57014cba93c4b@mail.gmail.com> Hi Kimberly, Are you using a clarifier step? On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < Kimberly.Marshall@ahss.org> wrote: > Howdy all, > > I have just changed over to Surgi Path's H & E system. With the latest > fear of running out of hematoxylin and all my Pathologist wanted us to > change over. Its a great stain but we have a bad "haze" in the background. > It is not hurting the tissue or DX at all, but to ME it looks awful. Is > there anyone else out there using this? We are not using positive charged > slides or anything added to my water bath. Any suggestions???? Thanks in > advance > > Kimberly Marshall HT (ASCP) > > > ============================================================================== > The information contained in this message may be privileged and/or > 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 the sender immediately by replying to > this > message and deleting the material from any computer. > > > ============================================================================== > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States From Jacqueline.Farnsworth <@t> cls.ab.ca Wed Dec 3 12:08:33 2008 From: Jacqueline.Farnsworth <@t> cls.ab.ca (Jacqueline Farnsworth) Date: Wed Dec 3 12:08:38 2008 Subject: [Histonet] formailin dispensing (large volumes) Message-ID: Hi Histonetters, Can anyone share anything about their formalin preparation and dispensing? We are fortunate enough to be planning for a new morgue. In our existing morgue, we manually make 200L of formalin (from concentrate). It is made in a large tank and then pumped up into the upper tank by a pump that was made by our maintenance dept. (Our morgue staff wear a respirator while making the formalin.) This upper tank, has a gravity fed tap for dispensing. Then, another full tank is made for the bottom tank. Of course, we always have fear of tanks being over-filled or leaking, so we'd like to explore a new system that would be easier, safer and more reliable. We have looked into the formalin dispensing units out there, but the capacity is not large enough for our needs. Anyone have any thoughts or suggestions? Vendors welcome to reply. Thank you, Jacquie Jacqueline Farnsworth Anatomic Pathology, Tech III Foothills Medical Centre Calgary Laboratory Services Ph: 403-944-1578 Fax: 403-944-4748 P Please consider the environment before printing this email. ________________________________ This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. From ploykasek <@t> phenopath.com Wed Dec 3 12:08:30 2008 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Dec 3 12:08:56 2008 Subject: [Histonet] Thanks & new labeler question Message-ID: Thanks to all for the answers on my expired reagents. Now, I have a new question. Is anyone using the Brady label printer or another similar printer for slide labels? I would be interested in hearing your experiences. Thanks, as always. Patti Loykasek This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From RossS <@t> BaylorHealth.edu Wed Dec 3 12:11:32 2008 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Wed Dec 3 12:11:38 2008 Subject: [Histonet] RE: Blue haze In-Reply-To: Message-ID: We tried the Surgipath H/E and had a similar experience. We went back to what we had been using. Since then I've been told that more water rinses or a different Clarifier type reagent may fix this problem, but I haven't had time or a good reason to give it another try. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marshall, Kimberly Sent: Wednesday, December 03, 2008 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Blue haze Howdy all, I have just changed over to Surgi Path's H & E system. With the latest fear of running out of hematoxylin and all my Pathologist wanted us to change over. Its a great stain but we have a bad "haze" in the background. It is not hurting the tissue or DX at all, but to ME it looks awful. Is there anyone else out there using this? We are not using positive charged slides or anything added to my water bath. Any suggestions???? Thanks in advance Kimberly Marshall HT (ASCP) ============================================================================== The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From kellerp <@t> ent.wustl.edu Wed Dec 3 12:28:37 2008 From: kellerp <@t> ent.wustl.edu (Pat Keller) Date: Wed Dec 3 12:29:32 2008 Subject: [Histonet] 2x3 slides boxes Message-ID: Miss read submission but was able to find 2x3 slide boxes at VWR Micro Slide Boxes, Wood Frame Supplier: Jordan Paper Box Find Similar Items in Product CategoryStorage, Boxes, Plates, and Racks Supplier: Jordan Paper Box Micro Slide Boxes, Wood Frame This durable box can hold up to 100 standard slides. It features a clearly numbered plastic insert that provides generous spacing for easy handling of slides. Wood frame is covered with heavy black embossed paper. Hinges and latch are nickel plated. A numbered index lines the cover. 51 x 75 (2 x 3) 24.7 x 19.6 x 5.7 (93/4 x 73/4 x 21/4) 48452-001 Case of 28 $1,172.43 Each $46.69 Call to make sure they are in stock Patricia Keller, Sr. Res. Tech. Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, MO. 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE 314-747-7166 FAX 314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. From Charles.Embrey <@t> carle.com Wed Dec 3 12:30:48 2008 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Wed Dec 3 12:31:04 2008 Subject: [Histonet] Alias identity In-Reply-To: <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> References: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> Message-ID: <44780C571F28624DBB446DE55C4D733A1FE612@EXCHANGEBE1.carle.com> This is one problem you have with an open list. I also belong to the Pathologists' Assistants list server but you must apply for membership that is restricted to AAPA members only. We can be more open with comments and questions without worry of unwanted calls from vendors or junk mail from the growing number of recruiters. Of course anything on the web can be accessed if you search hard enough but we don't have companies monitoring everything we say. I would be careful to "assume that if we're on this list that we're all credible, knowledge and professionals". Over ten years on the list has taught me otherwise. I must admit that I always consider the source. It is sad that some do have to hide their identity but I do understand why they find it necessary and support their choice. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amber McKenzie Sent: Wednesday, December 03, 2008 10:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alias identity I have to put my 2 cents in about all the statements on having an alias name instead of identifying who each person is after each email...WHAT'S THE BIG DEAL?? We are all supposed to be professionals asking each other for advice/suggestions on the Histonet - who cares who each person is? If I post a question, I don't care if it's Jane Doe answering or John Smith. I assume that if we're on this list that we're all credible, knowledge and professionals. Come on people, we're all in the same boat here. If I'd thought about putting an alias name for myself instead of my real name, I would have! Simply b/c last month I posted a question on the Histonet asking where you all bought your lab chairs, and I ended up having 2 vendors call me at my office trying to sell me some when all I wanted was the advice of other HT's. I love the idea of people not knowing who I am or where I work. Then I wouldn't have to worry about being harassed on the phone. Instead of worrying about the little things on the Histonet - like who each person is, why can't we focus on work related issues? I have to delete so much junk just to get to the material that actually applies to my field b/c of all the multiple emails that don't even matter. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Mark.Frei <@t> sial.com Wed Dec 3 12:55:16 2008 From: Mark.Frei <@t> sial.com (Mark Frei) Date: Wed Dec 3 12:55:54 2008 Subject: [Histonet] Flame substitute for embedding Message-ID: This sounds like a similar problem we had in Microbiology decades ago when we had to stop using open flame for sterilizing our wire inoculating loops when streaking Petri plates. Most micro labs now use electric sterilizers which consists of a tightly wound heating coil that the wire loop is placed in. I am not familiar with the amount of paraffin carry-over or the hazards/company warranty issues in using this type of equipment for this purpose, but it might be worth an investigation. Mark Frei MT(ASCP) Sigma-Aldrich Hematology & Histology 3050 Spruce Street / Saint Louis, MO, 63103 / USA P: (800) 771-5765, x4164 / Direct: (314) 286-8080 sigma-aldrich.com This message and any files transmitted with it are the property of Sigma-Aldrich Corporation, are confidential, and are intended solely for the use of the person or entity to whom this e-mail is addressed. If you are not one of the named recipient(s) or otherwise have reason to believe that you have received this message in error, please contact the sender and delete this message immediately from your computer. Any other use, retention, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. From MadaryJ <@t> MedImmune.com Wed Dec 3 13:09:08 2008 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Wed Dec 3 13:09:18 2008 Subject: [Histonet] What is the name of this blue stain? In-Reply-To: Message-ID: Alcian Blue goes into an alkaline alcohol for one hour as part of the old style movat. What is the final name of that blue color? Trying to train someone and could not find it. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Laboratory Mgr One Medimmune Way, Lab 2438-Area 4 Gaithersburg, MD 20878 ph 301.398.4745/6360 fx 301.398.9745 To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. >P Please consider the environment before printing this e-mail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, December 03, 2008 1:04 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 61, Issue 5 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. 2x3 slides (Pat Keller) 2. Re: 2x3 slides (Pamela Marcum) 3. lids for stainer containers (Perry, Margaret) 4. RE: RE: Flame substitute for embedding (Bernice Frederick) 5. Blue haze (Marshall, Kimberly) 6. Re: PLP fixative lysine (Gayle Callis) ---------------------------------------------------------------------- Message: 1 Date: Wed, 03 Dec 2008 11:29:55 -0600 From: Pat Keller Subject: [Histonet] 2x3 slides To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Fisher has them MILLIPORE CORP MICROSCOPE SLIDES 2"X3" 72/PK Fisher offers many products that do not appear in our catalogs. This may be one of those products, so pictures and detailed descriptions are not available. However, you may be able to order it by adding it to your shopping cart. Item Description Catalog Number Quantity Price MICROSCOPE SLIDES 2"X3" 72/PK XX1007615 Vendor No.:XX1007615 Pack of 72 for $51.26 Patricia Keller, Sr. Res. Tech. Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, MO. 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE 314-747-7166 FAX 314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. ------------------------------ Message: 2 Date: Wed, 3 Dec 2008 17:34:48 +0000 (UTC) From: Pamela Marcum Subject: Re: [Histonet] 2x3 slides To: Pat Keller Cc: histonet@lists.utsouthwestern.edu Message-ID: <966250373.78611228325688567.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Content-Type: text/plain; charset=utf-8 I got the slides from Thermo Fisher with some problems a long while ago.?? The issue was finding a 100 slide box for storage and shipping.?? I did order them this morning from Brain Research and they had stock of the boxes.?? Thanks to everyone who helped and now I know where to go for the boxes next time. Pam Marcum ----- Original Message ----- From: "Pat Keller" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 3, 2008 12:29:55 PM GMT -05:00 US/Canada Eastern Subject: [Histonet] 2x3 slides Fisher has them MILLIPORE CORP ??MICROSCOPE SLIDES 2"X3" 72/PK Fisher offers many products that do not appear in our catalogs. This may be one of those products, so pictures and detailed descriptions are not available. However, you may be able to order it by adding it to your shopping cart. ?? Item Description ?? ?? Catalog Number ?? ?? Quantity ?? ?? Price MICROSCOPE SLIDES 2"X3" 72/PK ???? ??XX1007615 Vendor No.:XX1007615 ?? ?? ?? ?? Pack of 72 for $51.26 Patricia Keller, Sr. Res. Tech. ?? Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, ?? MO. ?? 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE ?? ??314-747-7166 FAX ?? ?? ?? ?? ??314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 3 Dec 2008 11:46:33 -0600 From: "Perry, Margaret" Subject: [Histonet] lids for stainer containers To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="us-ascii" We are looking for a source that sells a lid that looks like wax covered cardboard. We use them on top of the Shandon slide containers holding xylene or formula 83 while waiting to coverslip. They are about 5x7 and very thin. They have been here for years so I have no idea where they came from. Any help in finding them or a substitute will be greatly appreciated. Thanks. Margaret Perry HT (ASCP) IHC Lab Manager Veterinary Science Animal Disease Research and Diagnostic Lab South Dakota State University Box 2175 North Campus Drive Brookings SD 57007 ------------------------------ Message: 4 Date: Wed, 3 Dec 2008 11:48:12 -0600 From: "Bernice Frederick" Subject: RE: [Histonet] RE: Flame substitute for embedding To: "'Jacqueline Farnsworth'" , "'Celebre Julia'" , Message-ID: <000001c9556f$52b14590$d00f7ca5@lurie.northwestern.edu> Content-Type: text/plain; charset="us-ascii" We have a Shandon Paratrimmer, which melts the wax not only off blocks but does a good job of keeping those forceps clean and hot. We have a bacti-incinerator, but know we can get a tall flame if we leave the paraffin coated forceps in there!! Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jacqueline Farnsworth Sent: Wednesday, December 03, 2008 9:44 AM To: Celebre Julia; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Flame substitute for embedding We also used to have an open flame while embedding! Now, we simply have a policy in place where after EACH cassette is embedded, the Techs MUST thoroughly wipe their forceps with disposable Kleenex (and then dispose of it!). Also, we use toothless forceps as much as possible to reduce the possibility of cross contamination. Jacqueline Farnsworth Anatomic Pathology, Tech III Foothills Medical Centre Calgary Laboratory Services Ph: 403-944-1578 Fax: 403-944-4748 P Please consider the environment before printing this email. ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia [celebrej@HHSC.CA] Sent: December 3, 2008 8:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flame substitute for embedding Hello Histoland!! We have just been told we are no longer allowed to use alcohol burners while embedding and we are to find a safer substitute, something to do with no liking an open flame in the lab. Other than incinerators or 6 pairs of forceps at each centre, we've run out of ideas so we are asking all you experts out there what your current practice is.. oh.. this change needs to be done by this Friday!! Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 3 Dec 2008 12:47:34 -0500 From: "Marshall, Kimberly" Subject: [Histonet] Blue haze To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Howdy all, I have just changed over to Surgi Path's H & E system. With the latest fear of running out of hematoxylin and all my Pathologist wanted us to change over. Its a great stain but we have a bad "haze" in the background. It is not hurting the tissue or DX at all, but to ME it looks awful. Is there anyone else out there using this? We are not using positive charged slides or anything added to my water bath. Any suggestions???? Thanks in advance Kimberly Marshall HT (ASCP) ============================================================================== The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ============================================================================== ------------------------------ Message: 6 Date: Wed, 3 Dec 2008 10:56:21 -0700 From: "Gayle Callis" Subject: [Histonet] Re: PLP fixative lysine To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" For the PLP fixative, the full name for the lysine is L- lysine monohydrochloride. Sigma cat# L5626. Gayle M. Callis HTL(ASCP)HT,MT ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 61, Issue 5 *************************************** To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary. This communication is expected to be read and/or used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. From renafail <@t> bellsouth.net Wed Dec 3 13:27:47 2008 From: renafail <@t> bellsouth.net (renafail@bellsouth.net) Date: Wed Dec 3 13:27:55 2008 Subject: [Histonet] What is the name of this blue stain? In-Reply-To: References: Message-ID: <120320081927.4026.4936DDB2000270E400000FBA22230703629B0A02D2089B9A019C04040A0DBF04070E000E020A9D@att.net> Monastral fast blue, an insoluble pigment Rena Fail -------------- Original message from "Madary, Joseph" : -------------- Alcian Blue goes into an alkaline alcohol for one hour as part of the old style movat. What is the final name of that blue color? Trying to train someone and > could not find it. > > > Nick Madary, HT/HTL(ASCP)QIHC > Medimmune Histology Laboratory Mgr > One Medimmune Way, Lab 2438-Area 4 > Gaithersburg, MD 20878 > > ph 301.398.4745/6360 > fx 301.398.9745 > To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for > any purpose. Thank you for your cooperation. > >P Please consider the environment before printing this e-mail > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > histonet-request@lists.utsouthwestern.edu > Sent: Wednesday, December 03, 2008 1:04 PM > To: histonet@lists.utsouthwestern.edu > Subject: Histonet Digest, Vol 61, Issue 5 > > 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. 2x3 slides (Pat Keller) > 2. Re: 2x3 slides (Pamela Marcum) > 3. lids for stainer containers (Perry, Margaret) > 4. RE: RE: Flame substitute for embedding (Bernice Frederick) > 5. Blue haze (Marshall, Kimberly) > 6. Re: PLP fixative lysine (Gayle Callis) > > > ---------------------------------------------------------------------- > > Message: 1 > Date: Wed, 03 Dec 2008 11:29:55 -0600 > From: Pat Keller > Subject: [Histonet] 2x3 slides > To: > Message-ID: > Content-Type: text/plain; charset="US-ASCII" > > Fisher has them > > > MILLIPORE CORP MICROSCOPE SLIDES 2"X3" 72/PK > > Fisher offers many products that do not appear in our catalogs. This may be one > of those products, so pictures and detailed descriptions are not available. > However, you may be able to order it by adding it to your shopping cart. > > > Item > Description Catalog Number Quantity Price > MICROSCOPE SLIDES 2"X3" 72/PK > XX1007615 > Vendor No.:XX1007615 Pack of 72 for $51.26 > > > Patricia Keller, > Sr. Res. Tech. Histology Core Manager > Washington University School of Medicine Department of Otolaryngology Campus box > 8115 > 4566 Scott Ave. > St. Louis, MO. 63110 > KellerP@ent.wustl.edu > http://www.otocore.org/ > > OFFICE 314-747-7166 > FAX 314-747-7230 > > > The materials in this message are private and may contain Protected > Healthcare Information. If you are not the intended recipient, be advised that > any unauthorized use, disclosure, copying or the taking of any action in > reliance on the contents of this information is strictly prohibited. If you have > received this email in error, please immediately notify the sender via telephone > or return mail. > > > > ------------------------------ > > Message: 2 > Date: Wed, 3 Dec 2008 17:34:48 +0000 (UTC) > From: Pamela Marcum > Subject: Re: [Histonet] 2x3 slides > To: Pat Keller > Cc: histonet@lists.utsouthwestern.edu > Message-ID: > > <966250373.78611228325688567.JavaMail.root@sz0001a.westchester.pa.mail.comcast.n > et> > > Content-Type: text/plain; charset=utf-8 > > > > > I got the slides from Thermo Fisher with some problems a long while ago. The > issue was finding a 100 slide box for storage and shipping. I did order them > this morning from Brain Research and they had stock of the boxes. Thanks to > everyone who helped and now I know where to go for the boxes next time. > > > > Pam Marcum > > > ----- Original Message ----- > From: "Pat Keller" > To: histonet@lists.utsouthwestern.edu > Sent: Wednesday, December 3, 2008 12:29:55 PM GMT -05:00 US/Canada Eastern > Subject: [Histonet] 2x3 slides > > Fisher has them > > > MILLIPORE CORP MICROSCOPE SLIDES 2"X3" 72/PK > > Fisher offers many products that do not appear in our catalogs. This may be one > of those products, so pictures and detailed descriptions are not available. > However, you may be able to order it by adding it to your shopping cart. > > > Item > Description Catalog Number Quantity Price MICROSCOPE SLIDES > 2"X3" 72/PK XX1007615 Vendor No.:XX1007615 Pack of 72 for > $51.26 > > > Patricia Keller, > Sr. Res. Tech. Histology Core Manager Washington University School of > Medicine Department of Otolaryngology Campus box 8115 > 4566 Scott Ave. > St. Louis, MO. 63110 > KellerP@ent.wustl.edu > http://www.otocore.org/ > > OFFICE 314-747-7166 > FAX 314-747-7230 > > > The materials in this message are private and may contain Protected > Healthcare Information. If you are not the intended recipient, be advised that > any unauthorized use, disclosure, copying or the taking of any action in > reliance on the contents of this information is strictly prohibited. If you have > received this email in error, please immediately notify the sender via telephone > or return mail. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ > > Message: 3 > Date: Wed, 3 Dec 2008 11:46:33 -0600 > From: "Perry, Margaret" > Subject: [Histonet] lids for stainer containers > To: "histonet@lists.utsouthwestern.edu" > > Message-ID: > > Content-Type: text/plain; charset="us-ascii" > > We are looking for a source that sells a lid that looks like wax covered > cardboard. We use them on top of the Shandon slide containers holding xylene or > formula 83 while waiting to coverslip. They are about 5x7 and very thin. They > have been here for years so I have no idea where they came from. Any help in > finding them or a substitute will be greatly appreciated. Thanks. > > Margaret Perry HT (ASCP) > IHC Lab Manager Veterinary Science > Animal Disease Research and Diagnostic Lab > South Dakota State University > Box 2175 North Campus Drive > Brookings SD 57007 > > > > ------------------------------ > > Message: 4 > Date: Wed, 3 Dec 2008 11:48:12 -0600 > From: "Bernice Frederick" > Subject: RE: [Histonet] RE: Flame substitute for embedding > To: "'Jacqueline Farnsworth'" , > "'Celebre Julia'" , > > Message-ID: <000001c9556f$52b14590$d00f7ca5@lurie.northwestern.edu> > Content-Type: text/plain; charset="us-ascii" > > We have a Shandon Paratrimmer, which melts the wax not only off blocks but > does a good job of keeping those forceps clean and hot. > We have a bacti-incinerator, but know we can get a tall flame if we leave > the paraffin coated forceps in there!! > Bernice > > > Bernice Frederick HTL (ASCP) > Northwestern University > Pathology Core Facility > ECOGPCO-RL > 710 N Fairbanks Court > Olson 8-421 > Chicago,IL 60611 > 312-503-3723 > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jacqueline > Farnsworth > Sent: Wednesday, December 03, 2008 9:44 AM > To: Celebre Julia; histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Flame substitute for embedding > > We also used to have an open flame while embedding! Now, we simply have a > policy in place where after EACH cassette is embedded, the Techs MUST > thoroughly wipe their forceps with disposable Kleenex (and then dispose of > it!). Also, we use toothless forceps as much as possible to reduce the > possibility of cross contamination. > > > Jacqueline Farnsworth > Anatomic Pathology, Tech III > Foothills Medical Centre > Calgary Laboratory Services > > Ph: 403-944-1578 > Fax: 403-944-4748 > P Please consider the environment before printing this email. > ________________________________________ > From: histonet-bounces@lists.utsouthwestern.edu > [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia > [celebrej@HHSC.CA] > Sent: December 3, 2008 8:38 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Flame substitute for embedding > > Hello Histoland!! > > We have just been told we are no longer allowed to use alcohol burners > while embedding and we are to find a safer substitute, something to do with > no liking an open flame in the lab. Other than incinerators or 6 pairs of > forceps at each centre, we've run out of ideas so we are asking all you > experts out there what your current practice is.. oh.. this change needs to > be done by this Friday!! > > Julia Celebre MLT > Anatomic Pathology > Hamilton General Hospital > 905-527-0271 ext 46179 > email: celebrej@hhsc.ca > > > > This information is directed in confidence solely to the person > named above and may not otherwise be distributed, copied or > disclosed. Therefore, this information should be considered > strictly confidential. If you have received this email in error, please > notify the sender immediately via a return email for further > direction. Thank you for your assistance. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > This message and any attached documents are only for the use of the intended > recipient(s), are confidential and may contain privileged information. Any > unauthorized review, use, retransmission, or other disclosure is strictly > prohibited. If you have received this message in error, please notify the > sender immediately, and then delete the original message. Thank you. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > ------------------------------ > > Message: 5 > Date: Wed, 3 Dec 2008 12:47:34 -0500 > From: "Marshall, Kimberly" > Subject: [Histonet] Blue haze > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=iso-8859-1 > > Howdy all, > > I have just changed over to Surgi Path's H & E system. With the latest fear > of running out of hematoxylin and all my Pathologist wanted us to change over. > Its a great stain but we have a bad "haze" in the background. It is not > hurting the tissue or DX at all, but to ME it looks awful. Is there anyone else > out there using this? We are not using positive charged slides or anything > added to my water bath. Any suggestions???? Thanks in advance > > Kimberly Marshall HT (ASCP) > > ============================================================================== > The information contained in this message may be privileged and/or 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 the sender immediately by replying to this > message and deleting the material from any computer. > > ============================================================================== > > > ------------------------------ > > Message: 6 > Date: Wed, 3 Dec 2008 10:56:21 -0700 > From: "Gayle Callis" > Subject: [Histonet] Re: PLP fixative lysine > To: "Histonet" > Message-ID: > Content-Type: text/plain; charset="iso-8859-1" > > For the PLP fixative, the full name for the lysine is L- lysine > monohydrochloride. Sigma cat# L5626. > > Gayle M. Callis > HTL(ASCP)HT,MT > > ------------------------------ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > End of Histonet Digest, Vol 61, Issue 5 > *************************************** > > > > To the extent this electronic communication or any of its attachments contain > information that is not in the public domain, such information is considered by > MedImmune to be confidential and proprietary. This communication is expected to > be read and/or used only by the individual(s) for whom it is intended. If you > have received this electronic communication in error, please reply to the sender > advising of the error in transmission and delete the original message and any > accompanying documents from your system immediately, without copying, reviewing > or otherwise using them for any purpose. Thank you for your cooperation. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dellav <@t> musc.edu Wed Dec 3 14:04:59 2008 From: dellav <@t> musc.edu (Della Speranza, Vinnie) Date: Wed Dec 3 14:05:05 2008 Subject: [Histonet] IHC billing with Mohs In-Reply-To: References: Message-ID: If there is anyone on this list knowledgeable about billing of Mohs samples, I'd be grateful if you would contact me back channel to address one or two questions. Thank you, Vinnie Della Speranza Manager for Anatomic Pathology Services Medical University of South Carolina 165 Ashley Avenue Suite 309 Charleston, South Carolina 29425 Tel: (843) 792-6353 Fax: (843) 792-8974 From JMacDonald <@t> mtsac.edu Wed Dec 3 13:55:38 2008 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Wed Dec 3 14:48:15 2008 Subject: [Histonet] What is the name of this blue stain? In-Reply-To: Message-ID: monastral fast blue "Madary, Joseph" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/03/2008 11:15 AM To cc Subject [Histonet] What is the name of this blue stain? Alcian Blue goes into an alkaline alcohol for one hour as part of the old style movat. What is the final name of that blue color? Trying to train someone and could not find it. Nick Madary, HT/HTL(ASCP)QIHC Medimmune Histology Laboratory Mgr One Medimmune Way, Lab 2438-Area 4 Gaithersburg, MD 20878 ph 301.398.4745/6360 fx 301.398.9745 To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. >P Please consider the environment before printing this e-mail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, December 03, 2008 1:04 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 61, Issue 5 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. 2x3 slides (Pat Keller) 2. Re: 2x3 slides (Pamela Marcum) 3. lids for stainer containers (Perry, Margaret) 4. RE: RE: Flame substitute for embedding (Bernice Frederick) 5. Blue haze (Marshall, Kimberly) 6. Re: PLP fixative lysine (Gayle Callis) ---------------------------------------------------------------------- Message: 1 Date: Wed, 03 Dec 2008 11:29:55 -0600 From: Pat Keller Subject: [Histonet] 2x3 slides To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Fisher has them MILLIPORE CORP MICROSCOPE SLIDES 2"X3" 72/PK Fisher offers many products that do not appear in our catalogs. This may be one of those products, so pictures and detailed descriptions are not available. However, you may be able to order it by adding it to your shopping cart. Item Description Catalog Number Quantity Price MICROSCOPE SLIDES 2"X3" 72/PK XX1007615 Vendor No.:XX1007615 Pack of 72 for $51.26 Patricia Keller, Sr. Res. Tech. Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, MO. 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE 314-747-7166 FAX 314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. ------------------------------ Message: 2 Date: Wed, 3 Dec 2008 17:34:48 +0000 (UTC) From: Pamela Marcum Subject: Re: [Histonet] 2x3 slides To: Pat Keller Cc: histonet@lists.utsouthwestern.edu Message-ID: <966250373.78611228325688567.JavaMail.root@sz0001a.westchester.pa.mail.comcast.net> Content-Type: text/plain; charset=utf-8 I got the slides from Thermo Fisher with some problems a long while ago.? The issue was finding a 100 slide box for storage and shipping.? I did order them this morning from Brain Research and they had stock of the boxes.? Thanks to everyone who helped and now I know where to go for the boxes next time. Pam Marcum ----- Original Message ----- From: "Pat Keller" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 3, 2008 12:29:55 PM GMT -05:00 US/Canada Eastern Subject: [Histonet] 2x3 slides Fisher has them MILLIPORE CORP ? MICROSCOPE SLIDES 2"X3" 72/PK Fisher offers many products that do not appear in our catalogs. This may be one of those products, so pictures and detailed descriptions are not available. However, you may be able to order it by adding it to your shopping cart. ? Item Description ? ? Catalog Number ? ? Quantity ? ? Price MICROSCOPE SLIDES 2"X3" 72/PK ? ? ? XX1007615 Vendor No.:XX1007615 ? ? ? ? Pack of 72 for $51.26 Patricia Keller, Sr. Res. Tech. ? Histology Core Manager Washington University School of Medicine Department of Otolaryngology Campus box 8115 4566 Scott Ave. St. Louis, ? MO. ? 63110 KellerP@ent.wustl.edu http://www.otocore.org/ OFFICE ? ? 314-747-7166 FAX ? ? ? ? ? 314-747-7230
The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Wed, 3 Dec 2008 11:46:33 -0600 From: "Perry, Margaret" Subject: [Histonet] lids for stainer containers To: "histonet@lists.utsouthwestern.edu" Message-ID: Content-Type: text/plain; charset="us-ascii" We are looking for a source that sells a lid that looks like wax covered cardboard. We use them on top of the Shandon slide containers holding xylene or formula 83 while waiting to coverslip. They are about 5x7 and very thin. They have been here for years so I have no idea where they came from. Any help in finding them or a substitute will be greatly appreciated. Thanks. Margaret Perry HT (ASCP) IHC Lab Manager Veterinary Science Animal Disease Research and Diagnostic Lab South Dakota State University Box 2175 North Campus Drive Brookings SD 57007 ------------------------------ Message: 4 Date: Wed, 3 Dec 2008 11:48:12 -0600 From: "Bernice Frederick" Subject: RE: [Histonet] RE: Flame substitute for embedding To: "'Jacqueline Farnsworth'" , "'Celebre Julia'" , Message-ID: <000001c9556f$52b14590$d00f7ca5@lurie.northwestern.edu> Content-Type: text/plain; charset="us-ascii" We have a Shandon Paratrimmer, which melts the wax not only off blocks but does a good job of keeping those forceps clean and hot. We have a bacti-incinerator, but know we can get a tall flame if we leave the paraffin coated forceps in there!! Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jacqueline Farnsworth Sent: Wednesday, December 03, 2008 9:44 AM To: Celebre Julia; histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Flame substitute for embedding We also used to have an open flame while embedding! Now, we simply have a policy in place where after EACH cassette is embedded, the Techs MUST thoroughly wipe their forceps with disposable Kleenex (and then dispose of it!). Also, we use toothless forceps as much as possible to reduce the possibility of cross contamination. Jacqueline Farnsworth Anatomic Pathology, Tech III Foothills Medical Centre Calgary Laboratory Services Ph: 403-944-1578 Fax: 403-944-4748 P Please consider the environment before printing this email. ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Celebre Julia [celebrej@HHSC.CA] Sent: December 3, 2008 8:38 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Flame substitute for embedding Hello Histoland!! We have just been told we are no longer allowed to use alcohol burners while embedding and we are to find a safer substitute, something to do with no liking an open flame in the lab. Other than incinerators or 6 pairs of forceps at each centre, we've run out of ideas so we are asking all you experts out there what your current practice is.. oh.. this change needs to be done by this Friday!! Julia Celebre MLT Anatomic Pathology Hamilton General Hospital 905-527-0271 ext 46179 email: celebrej@hhsc.ca This information is directed in confidence solely to the person named above and may not otherwise be distributed, copied or disclosed. Therefore, this information should be considered strictly confidential. If you have received this email in error, please notify the sender immediately via a return email for further direction. Thank you for your assistance. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This message and any attached documents are only for the use of the intended recipient(s), are confidential and may contain privileged information. Any unauthorized review, use, retransmission, or other disclosure is strictly prohibited. If you have received this message in error, please notify the sender immediately, and then delete the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Wed, 3 Dec 2008 12:47:34 -0500 From: "Marshall, Kimberly" Subject: [Histonet] Blue haze To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Howdy all, I have just changed over to Surgi Path's H & E system. With the latest fear of running out of hematoxylin and all my Pathologist wanted us to change over. Its a great stain but we have a bad "haze" in the background. It is not hurting the tissue or DX at all, but to ME it looks awful. Is there anyone else out there using this? We are not using positive charged slides or anything added to my water bath. Any suggestions???? Thanks in advance Kimberly Marshall HT (ASCP) ============================================================================== The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ============================================================================== ------------------------------ Message: 6 Date: Wed, 3 Dec 2008 10:56:21 -0700 From: "Gayle Callis" Subject: [Histonet] Re: PLP fixative lysine To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" For the PLP fixative, the full name for the lysine is L- lysine monohydrochloride. Sigma cat# L5626. Gayle M. Callis HTL(ASCP)HT,MT ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 61, Issue 5 *************************************** To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary. This communication is expected to be read and/or used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dayers1 <@t> jhmi.edu Wed Dec 3 15:16:21 2008 From: dayers1 <@t> jhmi.edu (Deborah Duckworth) Date: Wed Dec 3 15:16:38 2008 Subject: [Histonet] Please remove from mailing list Message-ID: <4936B0D5.008F.0051.0@jhmi.edu> Please remove me from the Histonet mailing list. Thanks for the valuable information I've received through this outreach. From shive003 <@t> umn.edu Wed Dec 3 15:33:57 2008 From: shive003 <@t> umn.edu (Jan Shivers) Date: Wed Dec 3 15:34:01 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed Message-ID: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu> Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu From ryaskovich <@t> dir.nidcr.nih.gov Wed Dec 3 15:36:09 2008 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR) [E]) Date: Wed Dec 3 15:36:16 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu> References: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu> Message-ID: I do it all the time. Yes your antigen-retrieval method will work. Ruth N.I.H. -----Original Message----- From: Jan Shivers [mailto:shive003@umn.edu] Sent: Wednesday, December 03, 2008 4:34 PM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Wed Dec 3 16:16:42 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Dec 3 16:16:55 2008 Subject: [Histonet] Blue haze In-Reply-To: <4b6c85510812031008k3902865enbfd57014cba93c4b@mail.gmail.com> Message-ID: Excuse my ignorance but what is a clarifier step in association with the H&E stain? Or do you mean the differentiation step? Also having been a member of the marvelous Histonet server for many years I think I know who the Histonet Alias is!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias Sent: Thursday, 4 December 2008 5:08 AM To: Marshall, Kimberly Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Blue haze Hi Kimberly, Are you using a clarifier step? On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < Kimberly.Marshall@ahss.org> wrote: > Howdy all, > > I have just changed over to Surgi Path's H & E system. With the > latest fear of running out of hematoxylin and all my Pathologist > wanted us to change over. Its a great stain but we have a bad "haze" > in the background. It is not hurting the tissue or DX at all, but to ME it looks awful. Is > there anyone else out there using this? We are not using positive charged > slides or anything added to my water bath. Any suggestions???? > Thanks in advance > > Kimberly Marshall HT (ASCP) > > > ====================================================================== > ======== > The information contained in this message may be privileged and/or > 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 the sender immediately by replying to > this > message and deleting the material from any computer. > > > ====================================================================== > ======== > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States _______________________________________________ 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 note 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 Wed Dec 3 16:35:09 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Dec 3 16:35:16 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu> Message-ID: Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu _______________________________________________ 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 note 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 sprin119 <@t> umn.edu Wed Dec 3 17:45:55 2008 From: sprin119 <@t> umn.edu (Jennifer Springsteen) Date: Wed Dec 3 17:58:59 2008 Subject: [Histonet] Re: Blue Haze In-Reply-To: Message-ID: I also use the Surgipath SelecTech H&E system and had to solve the blue haze problem in our lab. We've had great success simply using warm tap water to wash the slides after hematoxylin, rather than cold tap water. Hope that helps! -- Jennifer L. Springsteen, B.S. Lab Manager, Assistant Scientist Lillehei Heart Institute Histology & Microscopy Core Facility University of Minnesota School of Medicine, Division of Cardiology 312 Church Street SE 4-266 Nils Hasselmo Hall Minneapolis, MN 55455 Lab: 612-626-3090 Cell: 651-357-7916 Fax: 612-624-8118 > Date: Wed, 3 Dec 2008 12:47:34 -0500 > From: "Marshall, Kimberly" > Subject: [Histonet] Blue haze > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=iso-8859-1 > > Howdy all, > > I have just changed over to Surgi Path's H & E system. With the latest > fear of running out of hematoxylin and all my Pathologist wanted us to change > over. Its a great stain but we have a bad "haze" in the background. It is > not hurting the tissue or DX at all, but to ME it looks awful. Is there > anyone else out there using this? We are not using positive charged slides > or anything added to my water bath. Any suggestions???? Thanks in advance > > Kimberly Marshall HT (ASCP) From CIngles <@t> uwhealth.org Wed Dec 3 18:07:41 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Wed Dec 3 18:09:03 2008 Subject: [Histonet] JCAHO References: Message-ID: <08A0A863637F1349BBFD83A96B27A50A1201C2@uwhis-xchng3.uwhis.hosp.wisc.edu> FYI Everyone: If you are still waiting for your inspection, don't bother putting up Christmas decorations. Claire From ratliffjack <@t> hotmail.com Wed Dec 3 20:12:29 2008 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Wed Dec 3 20:12:33 2008 Subject: [Histonet] GMA embedded section attachment to slides In-Reply-To: <4F820D0A1054E6478FD124A9BE03397271F3B6@MSGEBE35.mfad.mfroot.org> References: <4F820D0A1054E6478FD124A9BE03397271F3B6@MSGEBE35.mfad.mfroot.org> Message-ID: Jim, Is it possible for you to provide more information as to the type of specimen you are working with and how thick you are cutting the sections? Jack> Date: Mon, 1 Dec 2008 16:26:31 -0600> From: Herrick.James@mayo.edu> To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] GMA embedded section attachment to slides> > Hi everyone,> > I have been trying to attach Glycol Methacrylate (GMA) embedded specimens to glass slides and have not been having much luck. So far I have tried gelatin, APES and Histostik. I am using a press and placing them in a 50? C oven for approx. 48 hours. The sections look as if they are attached, until I begin staining. I am pretty sure that the PEG is causing my problem, but have seen an article or two that have attached GMA sections to slides. Does anyone have a protocol that works well with GMA? If so, I would greatly appreciate any help I can get. Thank you much.> > Jim> > > > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail anywhere. No map, no compass. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_anywhere_122008 From jnocito <@t> satx.rr.com Wed Dec 3 20:30:41 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Wed Dec 3 20:30:43 2008 Subject: [Histonet] Blue haze References: <4b6c85510812031008k3902865enbfd57014cba93c4b@mail.gmail.com> Message-ID: <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? Forgive me, we are in the window for CAP and I'm just all in a haze. JTT ----- Original Message ----- From: "Histonet Alias" To: "Marshall, Kimberly" Cc: Sent: Wednesday, December 03, 2008 12:08 PM Subject: Re: [Histonet] Blue haze > Hi Kimberly, > Are you using a clarifier step? > > On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < > Kimberly.Marshall@ahss.org> wrote: > >> Howdy all, >> >> I have just changed over to Surgi Path's H & E system. With the latest >> fear of running out of hematoxylin and all my Pathologist wanted us to >> change over. Its a great stain but we have a bad "haze" in the >> background. >> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >> there anyone else out there using this? We are not using positive >> charged >> slides or anything added to my water bath. Any suggestions???? Thanks >> in >> advance >> >> Kimberly Marshall HT (ASCP) >> >> >> ============================================================================== >> The information contained in this message may be privileged and/or >> 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 the sender immediately by replying >> to >> this >> message and deleting the material from any computer. >> >> >> ============================================================================== >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > Al Ias HT(ASCP) > Histology Manager > Pathology Laboratory > United States > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Wed Dec 3 21:18:47 2008 From: tifei <@t> foxmail.com (tf) Date: Wed Dec 3 21:19:05 2008 Subject: [Histonet] PSA-NCAM IHC on frozen section with BrdU Message-ID: <200812041118418715473@foxmail.com> Hi everyone, it seems that I lost the PSA-NCAM staining when sections are treated with citrate buffer (poring the membrane?) Possibly the membrane was dissociated from membrane then>? or? For co-staining with BrdU, I can only treat sections with HCl - the results sometimes are unstable~~ Anyone worked on this before? 2008-12-04 tf From susanbachus <@t> verizon.net Wed Dec 3 22:17:48 2008 From: susanbachus <@t> verizon.net (Susan Bachus) Date: Wed Dec 3 22:18:15 2008 Subject: [Histonet] Alias identity References: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> <44780C571F28624DBB446DE55C4D733A1FE612@EXCHANGEBE1.carle.com> Message-ID: <195D6FF1BF974E03AA9A1A3428A5D48B@RESLAPTOP> Here's one potential problem with listing real names: a few years ago a very diligent student of mine inadvertently stirred up a lot of controversy when she asked for advice from Histonet on a class assigment and someone was concerned that she was "cheating". I had in fact recommended Histonet as a resource to them (though I didn't intend for them to use it for something so trivial)! One silver lining was that a lot of discussion was aired about what the purpose of Histonet is and several people sprang to the poor student's defense. But later she googled herself when she started worrying about looking for a job and this popped up because she had used her full name, and she was terrified that it would interfere with her finding a job! She wanted to know if it could be taken out of the archives, but of course the problem is that these things are still "cached"! Susan ----- Original Message ----- From: "Charles.Embrey" To: "Amber McKenzie" ; Sent: Wednesday, December 03, 2008 1:30 PM Subject: RE: [Histonet] Alias identity This is one problem you have with an open list. I also belong to the Pathologists' Assistants list server but you must apply for membership that is restricted to AAPA members only. We can be more open with comments and questions without worry of unwanted calls from vendors or junk mail from the growing number of recruiters. Of course anything on the web can be accessed if you search hard enough but we don't have companies monitoring everything we say. I would be careful to "assume that if we're on this list that we're all credible, knowledge and professionals". Over ten years on the list has taught me otherwise. I must admit that I always consider the source. It is sad that some do have to hide their identity but I do understand why they find it necessary and support their choice. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amber McKenzie Sent: Wednesday, December 03, 2008 10:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Alias identity I have to put my 2 cents in about all the statements on having an alias name instead of identifying who each person is after each email...WHAT'S THE BIG DEAL?? We are all supposed to be professionals asking each other for advice/suggestions on the Histonet - who cares who each person is? If I post a question, I don't care if it's Jane Doe answering or John Smith. I assume that if we're on this list that we're all credible, knowledge and professionals. Come on people, we're all in the same boat here. If I'd thought about putting an alias name for myself instead of my real name, I would have! Simply b/c last month I posted a question on the Histonet asking where you all bought your lab chairs, and I ended up having 2 vendors call me at my office trying to sell me some when all I wanted was the advice of other HT's. I love the idea of people not knowing who I am or where I work. Then I wouldn't have to worry about being harassed on the phone. Instead of worrying about the little things on the Histonet - like who each person is, why can't we focus on work related issues? I have to delete so much junk just to get to the material that actually applies to my field b/c of all the multiple emails that don't even matter. _______________________________________________ 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 louise.renton <@t> gmail.com Thu Dec 4 01:51:36 2008 From: louise.renton <@t> gmail.com (louise renton) Date: Thu Dec 4 01:51:46 2008 Subject: [Histonet] lids for stainer containers In-Reply-To: References: Message-ID: These sound like the kind of thing that goes on a foil "take away" or freezer container. Try a home suply depot? We have specialist stores here that supply serious home bakers and chocolatiers and I know that they carry what you describe - bit far for you to come though....... On 12/3/08, Perry, Margaret wrote: > We are looking for a source that sells a lid that looks like wax covered > cardboard. We use them on top of the Shandon slide containers holding > xylene or formula 83 while waiting to coverslip. They are about 5x7 and > very thin. They have been here for years so I have no idea where they came > from. Any help in finding them or a substitute will be greatly > appreciated. Thanks. > > Margaret Perry HT (ASCP) > IHC Lab Manager Veterinary Science > Animal Disease Research and Diagnostic Lab > South Dakota State University > Box 2175 North Campus Drive > Brookings SD 57007 > > _______________________________________________ > 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 "There are nights when the wolves are silent and only the moon howls". George Carlin No trees were killed in the sending of this message. However, many electrons were terribly inconvenienced. From dencrowl <@t> MIT.EDU Thu Dec 4 05:58:05 2008 From: dencrowl <@t> MIT.EDU (Denise Crowley) Date: Thu Dec 4 05:58:16 2008 Subject: [Histonet] blue haze on Surgipath H&E Message-ID: Hi Kimberly, We switched to the Select-Tec system and are very happy with the quality of the hematoxylin and eosin stains. However, the clarifying step and the Blue buffer did not work out for us. So, we kept the stains and went back to using acid alcohol (1% HCl in 70% EtOH) and Scott's tap water substitute. Now the staining is bright, crisp, and the staining solutions last through about 1800 slides. Denise Crowley Facility Manager Histology David H. Koch Institute for Integrative Cancer Research Massachusetts Institute of Technology 40 Ames St. E17-427 Cambridge MA 02139 617-258-8183 dencrowl@mit.edu From kmerriam2003 <@t> yahoo.com Thu Dec 4 06:47:27 2008 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu Dec 4 06:47:32 2008 Subject: [Histonet] IHC on fresh frozen References: Message-ID: <985778.44585.qm@web50308.mail.re2.yahoo.com> We use acetone/ethanol (as recommended by Gayle?Callis in one of her NSH classes).? The slides look great; nice crisp nuclei (which you just won't get with acetone alone).? We air dry the slides and fix for 10 minutes in RT?acetone/ethanol (75% acetone/25% absolute ethanol) and then put the slides into?wash buffer and continue with IHC.? I have heard that this fixative could be problematic with some antigens, but we have not found that to be the case with anything we have used in my lab.? Insulin?should be fine. Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: Patti Loykasek To: "FU,DONGTAO" ; histonet@lists.utsouthwestern.edu Sent: Wednesday, December 3, 2008 11:38:07 AM Subject: Re: [Histonet] IHC on fresh frozen After hearing a presentation by Sharon Lear describing some low temp antigen retrieval that she did, we changed our method for frozen sections. We fix the frozen sections in 10% nuetral buffered formalin for 30'-60', rinse, then do a gentle pretreatment. The gentle pretreatment is usually 10mM citrate buffer pH6 at 70 degrees C for 30'. Slides are cooled, and usual IHC done. This has worked well for us. Our staining with this method is more reliable & intense than with previous methods. Patti Loykasek > Hi, all > > Recently I did some IHC(chromagen methods) on mouse fresh frozen > tissues, mainly using insulin antibody on pancreas. The image is > much fuzzier compare to paraffin embedding tissue. And the > staining also smeared to acinar cells which surround the islet. > > I airdried slide(>30min) and used a general acetone method(-20C > 5min) to fix the tissue before I did IHC. > > How can I get a relatively sharp staining on the fresh frozen > tissue?Does anyone here have any experience on it? Any > suggestions? > > Many thanks and have a nice day, > > Ann > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sbreeden <@t> nmda.nmsu.edu Thu Dec 4 07:00:30 2008 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Thu Dec 4 07:00:34 2008 Subject: [Histonet] Blue Haze Message-ID: <4D14F0FC9316DD41972D5F03C070908B017E65D3@nmdamailsvr.nmda.ad.nmsu.edu> I changed over to the Surgipath H&E "system" and after working out timing and water issues, I resolved the "blue haze" problem by increasing the time in the DEFINE to 45 seconds and I have no blue haze. You might have to tinker with the DEFINE and BLUE BUFFER times to optimize the stain. All in all - an excellent, clear, sharp, pathologist-pleasing stain. And - as we all know - "a happy pathologist is a wondrous thing"! Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 From b-frederick <@t> northwestern.edu Thu Dec 4 08:06:08 2008 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Dec 4 08:06:20 2008 Subject: [Histonet] Blue haze In-Reply-To: <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> Message-ID: <000501c95619$77487c20$d00f7ca5@lurie.northwestern.edu> I'm sure you'll just charm them and they'll never catch on.... Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Wednesday, December 03, 2008 8:31 PM To: Histonet Alias; Marshall, Kimberly Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Blue haze I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? Forgive me, we are in the window for CAP and I'm just all in a haze. JTT ----- Original Message ----- From: "Histonet Alias" To: "Marshall, Kimberly" Cc: Sent: Wednesday, December 03, 2008 12:08 PM Subject: Re: [Histonet] Blue haze > Hi Kimberly, > Are you using a clarifier step? > > On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < > Kimberly.Marshall@ahss.org> wrote: > >> Howdy all, >> >> I have just changed over to Surgi Path's H & E system. With the latest >> fear of running out of hematoxylin and all my Pathologist wanted us to >> change over. Its a great stain but we have a bad "haze" in the >> background. >> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >> there anyone else out there using this? We are not using positive >> charged >> slides or anything added to my water bath. Any suggestions???? Thanks >> in >> advance >> >> Kimberly Marshall HT (ASCP) >> >> >> ============================================================================ == >> The information contained in this message may be privileged and/or >> 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 the sender immediately by replying >> to >> this >> message and deleting the material from any computer. >> >> >> ============================================================================ == >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > Al Ias HT(ASCP) > Histology Manager > Pathology Laboratory > United States > _______________________________________________ > 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 anh2006 <@t> med.cornell.edu Thu Dec 4 08:31:23 2008 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Thu Dec 4 08:33:56 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: References: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu> Message-ID: <1167373032-1228401225-cardhu_decombobulator_blackberry.rim.net-657855511-@bxe173.bisx.prod.on.blackberry> So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ryaskovich <@t> dir.nidcr.nih.gov Thu Dec 4 08:42:07 2008 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR) [E]) Date: Thu Dec 4 08:42:15 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: <1167373032-1228401225-cardhu_decombobulator_blackberry.rim.net-657855511-@bxe173.bisx.prod.on.blackberry> References: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu> <1167373032-1228401225-cardhu_decombobulator_blackberry.rim.net-657855511-@bxe173.bisx.prod.on.blackberry> Message-ID: We buffer our (4% paraformaldehyde). The animals are also perfused with it. I know I tried to get the investigators to just use the 10% formalin and not make up this stuff from scratch but have just given up. Ruth N.I.H. -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] Sent: Thursday, December 04, 2008 9:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Thu Dec 4 08:46:22 2008 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Thu Dec 4 08:48:51 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: References: <6457EB9CA9B846C58BF6720C321FC9E6@auxs.umn.edu><1167373032-1228401225-cardhu_decombobulator_blackberry.rim.net-657855511-@bxe173.bisx.prod.on.blackberry> Message-ID: <2135202035-1228402124-cardhu_decombobulator_blackberry.rim.net-1693561827-@bxe173.bisx.prod.on.blackberry> You can buy 16% PFA ready made. It is one reagent oi refuse to make from scratch myself. -----Original Message----- From: "Yaskovich, Ruth A (NIH/NIDCR) [E]" Date: Thu, 04 Dec 2008 09:42:07 To: ; Tony Henwood; Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed We buffer our (4% paraformaldehyde). The animals are also perfused with it. I know I tried to get the investigators to just use the 10% formalin and not make up this stuff from scratch but have just given up. Ruth N.I.H. -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] Sent: Thursday, December 04, 2008 9:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From leiker <@t> buffalo.edu Thu Dec 4 09:43:07 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Thu Dec 4 09:43:12 2008 Subject: [Histonet] JCAHO In-Reply-To: <08A0A863637F1349BBFD83A96B27A50A1201C2@uwhis-xchng3.uwhis.hosp.wisc.edu> References: <08A0A863637F1349BBFD83A96B27A50A1201C2@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: Sounds like you found out the hard way? --On Wednesday, December 03, 2008 6:07 PM -0600 Ingles Claire wrote: > > FYI Everyone: > If you are still waiting for your inspection, don't bother putting up > Christmas decorations. Claire > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From hej01 <@t> health.state.ny.us Thu Dec 4 09:53:35 2008 From: hej01 <@t> health.state.ny.us (Helen E Johnson) Date: Thu Dec 4 09:53:40 2008 Subject: [Histonet] in utero mouse embryo Message-ID: <18112_1228406017_mB4FrbGV019769_OF5310BD5B.D29FC87B-ON85257515.0056DA19-85257515.00574DF4@notes.health.state.ny.us> How long should in utero mouse embryo be fixed in Bouin's? Helen Johnson (hej01@health.state.ny.us) IMPORTANT NOTICE: This e-mail and any attachments may contain confidential or sensitive information which is, or may be, legally privileged or otherwise protected by law from further disclosure. It is intended only for the addressee. If you received this in error or from someone who was not authorized to send it to you, please do not distribute, copy or use it or any attachments. Please notify the sender immediately by reply e-mail and delete this from your system. Thank you for your cooperation. From anh2006 <@t> med.cornell.edu Thu Dec 4 10:12:06 2008 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Thu Dec 4 10:12:15 2008 Subject: [Histonet] IHC on fresh frozen In-Reply-To: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> References: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> Message-ID: Try fixing for 5-10 min in PFA. At 10:16 AM -0500 12/3/08, FU,DONGTAO wrote: >Hi, all > > Recently I did some IHC(chromagen methods) on mouse fresh frozen >tissues, mainly using insulin antibody on pancreas. The image is >much fuzzier compare to paraffin embedding tissue. And the staining >also smeared to acinar cells which surround the islet. > > I airdried slide(>30min) and used a general acetone method(-20C >5min) to fix the tissue before I did IHC. > > How can I get a relatively sharp staining on the fresh frozen >tissue?Does anyone here have any experience on it? Any suggestions? > > Many thanks and have a nice day, > >Ann > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From ree3 <@t> leicester.ac.uk Thu Dec 4 10:52:37 2008 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Thu Dec 4 10:52:50 2008 Subject: [Histonet] Blue haze In-Reply-To: <000501c95619$77487c20$d00f7ca5@lurie.northwestern.edu> References: <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> <000501c95619$77487c20$d00f7ca5@lurie.northwestern.edu> Message-ID: <7722595275A4DD4FA225B92CDBF174A1744F13BB30@EXC-MBX3.cfs.le.ac.uk> Hey Joe, where you going with that toe in your hand?? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: 04 December 2008 14:06 To: 'Joe Nocito'; 'Histonet Alias'; 'Marshall, Kimberly' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Blue haze I'm sure you'll just charm them and they'll never catch on.... Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Wednesday, December 03, 2008 8:31 PM To: Histonet Alias; Marshall, Kimberly Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Blue haze I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? Forgive me, we are in the window for CAP and I'm just all in a haze. JTT ----- Original Message ----- From: "Histonet Alias" To: "Marshall, Kimberly" Cc: Sent: Wednesday, December 03, 2008 12:08 PM Subject: Re: [Histonet] Blue haze > Hi Kimberly, > Are you using a clarifier step? > > On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < > Kimberly.Marshall@ahss.org> wrote: > >> Howdy all, >> >> I have just changed over to Surgi Path's H & E system. With the latest >> fear of running out of hematoxylin and all my Pathologist wanted us to >> change over. Its a great stain but we have a bad "haze" in the >> background. >> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >> there anyone else out there using this? We are not using positive >> charged >> slides or anything added to my water bath. Any suggestions???? Thanks >> in >> advance >> >> Kimberly Marshall HT (ASCP) >> >> >> ============================================================================ == >> The information contained in this message may be privileged and/or >> 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 the sender immediately by replying >> to >> this >> message and deleting the material from any computer. >> >> >> ============================================================================ == >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > Al Ias HT(ASCP) > Histology Manager > Pathology Laboratory > United States > _______________________________________________ > 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 JCBRITTON <@t> Cheshire-Med.COM Thu Dec 4 10:56:13 2008 From: JCBRITTON <@t> Cheshire-Med.COM (Josie Britton) Date: Thu Dec 4 10:56:22 2008 Subject: [Histonet] Blue haze In-Reply-To: <000501c95619$77487c20$d00f7ca5@lurie.northwestern.edu> References: <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> <000501c95619$77487c20$d00f7ca5@lurie.northwestern.edu> Message-ID: I know this! We have had this problem before. We have now added and extra cap full ( or ? cap full per SurgiPath) to our Define. So instead of 2 we put 3 in. We also have a problem with our water filters clogging. Make sure you have plenty of water pressure also, but this pretty much solved our problem. Josie Britton HT Cheshire Medical Center 580 Court Street Keene, NH 03431 603-354-5454 ext.2454 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: Thursday, December 04, 2008 9:06 AM To: 'Joe Nocito'; 'Histonet Alias'; 'Marshall, Kimberly' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Blue haze I'm sure you'll just charm them and they'll never catch on.... Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Wednesday, December 03, 2008 8:31 PM To: Histonet Alias; Marshall, Kimberly Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Blue haze I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? Forgive me, we are in the window for CAP and I'm just all in a haze. JTT ----- Original Message ----- From: "Histonet Alias" To: "Marshall, Kimberly" Cc: Sent: Wednesday, December 03, 2008 12:08 PM Subject: Re: [Histonet] Blue haze > Hi Kimberly, > Are you using a clarifier step? > > On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < > Kimberly.Marshall@ahss.org> wrote: > >> Howdy all, >> >> I have just changed over to Surgi Path's H & E system. With the latest >> fear of running out of hematoxylin and all my Pathologist wanted us to >> change over. Its a great stain but we have a bad "haze" in the >> background. >> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >> there anyone else out there using this? We are not using positive >> charged >> slides or anything added to my water bath. Any suggestions???? Thanks >> in >> advance >> >> Kimberly Marshall HT (ASCP) >> >> >> ============================================================================ == >> The information contained in this message may be privileged and/or >> 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 the sender immediately by replying >> to >> this >> message and deleting the material from any computer. >> >> >> ============================================================================ == >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > Al Ias HT(ASCP) > Histology Manager > Pathology Laboratory > United States > _______________________________________________ > 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: This electronic message, including any attachments, is for the sole use of the intended recipients 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 electronic mail and destroy all copies of the original message. From b-frederick <@t> northwestern.edu Thu Dec 4 11:21:53 2008 From: b-frederick <@t> northwestern.edu (Bernice Frederick) Date: Thu Dec 4 11:22:06 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: Message-ID: <000001c95634$d024ca90$d00f7ca5@lurie.northwestern.edu> Ruth, I agree with you- As my boss says, they are living in the past. Bernice Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Yaskovich, Ruth A (NIH/NIDCR) [E] Sent: Thursday, December 04, 2008 8:42 AM To: anh2006@med.cornell.edu; Tony Henwood; Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed We buffer our (4% paraformaldehyde). The animals are also perfused with it. I know I tried to get the investigators to just use the 10% formalin and not make up this stuff from scratch but have just given up. Ruth N.I.H. -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] Sent: Thursday, December 04, 2008 9:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 juditw <@t> u.washington.edu Thu Dec 4 11:46:52 2008 From: juditw <@t> u.washington.edu (Judith L. Williams) Date: Thu Dec 4 11:46:58 2008 Subject: [Histonet] Blue haze In-Reply-To: <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> Message-ID: Hi all - I was just waiting for someone to do the "Purple Haze" comment :-) - just too darn good to pass up. almost friday??? yes! Judy in Washington On Wed, 3 Dec 2008, Joe Nocito wrote: > I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? > Forgive me, we are in the window for CAP and I'm just all in a haze. > > JTT > ----- Original Message ----- From: "Histonet Alias" > To: "Marshall, Kimberly" > Cc: > Sent: Wednesday, December 03, 2008 12:08 PM > Subject: Re: [Histonet] Blue haze > > >> Hi Kimberly, >> Are you using a clarifier step? >> >> On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < >> Kimberly.Marshall@ahss.org> wrote: >> >>> Howdy all, >>> >>> I have just changed over to Surgi Path's H & E system. With the latest >>> fear of running out of hematoxylin and all my Pathologist wanted us to >>> change over. Its a great stain but we have a bad "haze" in the >>> background. >>> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >>> there anyone else out there using this? We are not using positive >>> charged >>> slides or anything added to my water bath. Any suggestions???? Thanks in >>> advance >>> >>> Kimberly Marshall HT (ASCP) >>> >>> >>> ============================================================================== >>> The information contained in this message may be privileged and/or >>> 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 the sender immediately by replying >>> to >>> this >>> message and deleting the material from any computer. >>> >>> >>> ============================================================================== >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >> >> >> >> -- >> Al Ias HT(ASCP) >> Histology Manager >> Pathology Laboratory >> United States >> _______________________________________________ >> 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 > Judith Williams, PhD, HT(ASCP) Research Scientist Department of Comparative Medicine University of Washington Seattle, WA 98195 From MadaryJ <@t> MedImmune.com Thu Dec 4 11:49:24 2008 From: MadaryJ <@t> MedImmune.com (Madary, Joseph) Date: Thu Dec 4 11:49:35 2008 Subject: [Histonet] thanks Henwood for the PFA discussion Message-ID: At my previous job,I got so tired of the PFA discussion with a few folks that I would hand them two containers of NBF one labeled PFA and one labeled NBF and tell them to test it and they always went for the "PFA". I mean same stuff! All in their heads man. So those folks always got the special labeled container. Little placebo action never hurt eh? I notice that recycled formalin seems to boil off methanol too which is what I use mainly- recycled. Sometimes I have to add a little full strength, but seems like it reads less alcohol after I run it thru the recycler. I see the billing point of methanol is 64.7 so it would appear to be knocked off. Nick Madary, HT/HTL(ASCP)QIHC Histology Mgr, Medimmune 301.398.6360(lab), 4745(vm),9745(fax) "To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary, and expected to be used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation." To the extent this electronic communication or any of its attachments contain information that is not in the public domain, such information is considered by MedImmune to be confidential and proprietary. This communication is expected to be read and/or used only by the individual(s) for whom it is intended. If you have received this electronic communication in error, please reply to the sender advising of the error in transmission and delete the original message and any accompanying documents from your system immediately, without copying, reviewing or otherwise using them for any purpose. Thank you for your cooperation. From tkngflght <@t> yahoo.com Thu Dec 4 12:06:04 2008 From: tkngflght <@t> yahoo.com (Cheryl) Date: Thu Dec 4 12:06:08 2008 Subject: What's the pH of your tap water? RE: [Histonet] Blue haze In-Reply-To: Message-ID: <15960.97595.qm@web50902.mail.re2.yahoo.com> Hey and Hello- ? I haven't followed this whole string so I apologize if this has been mentioned: ? When I was in a state with really hard water this came up more often than anywhere else...have you checked the pH of your water?? If it isn't close to neutral the haze shows up.? We had to rinse in dH2O to get the extra stain off before rinsing in tap water.? The warm water had a different pH than cold--so that might also help as suggested earlier.? ? This was for a Gill II--not sure of the effect on the new product--just offering up one more variable to try! ? Cheryl ? Cheryl R. Kerry, HT(ASCP), BA Full Staff Inc. Staffing Healthcare Professionals - One GREAT fit at a time! ? 218.913.7285 800.756.3309 resume@fullstaff.org From jwarren23 <@t> cinci.rr.com Thu Dec 4 12:12:44 2008 From: jwarren23 <@t> cinci.rr.com (Jean Warren) Date: Thu Dec 4 12:12:56 2008 Subject: [Histonet] Embedding paraffin blocks Message-ID: <325C5711D4A146F485B98ED4556EFAEA@ownerPC> Is there an average number of blocks that a tech should be able to embed in a defined time period; such as so many blocks per hour? Also, would that number vary if the tech was embedding biopsies, skin or cones versus large tissue blocks, such as uterus? I have not seen this information, but I have heard widely ranging numbers. Thanks Jean in Cincinnati From Dorothy.L.Webb <@t> HealthPartners.Com Thu Dec 4 12:41:06 2008 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Thu Dec 4 12:41:11 2008 Subject: [Histonet] Silanized slides Message-ID: <0E394B648E5284478A6CCB78E5AFDA2705635B63@hpes1.HealthPartners.int> Does anyone have any idea of how long silanized slides made in lab are good for? Would it be the same outdate as is on the solution one is using to prepare the silanized slides? Thanks, as always!! Dorothy Webb, HT Regions Histology Technical Supervisor 651-254-2962 ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. From wdesalvo.cac <@t> hotmail.com Thu Dec 4 12:44:51 2008 From: wdesalvo.cac <@t> hotmail.com (WILLIAM DESALVO) Date: Thu Dec 4 12:44:56 2008 Subject: [Histonet] Embedding paraffin blocks In-Reply-To: <325C5711D4A146F485B98ED4556EFAEA@ownerPC> References: <325C5711D4A146F485B98ED4556EFAEA@ownerPC> Message-ID: There is not a set number or industry guideline. The number of blocks per hour will depend on the tissue type mix, the complexity of the embedding technique and the competency of the persons embedding. If you truley want a number that works for your lab, then perfom a short time study on all the employees assigned to embedding and then look at their performance statistically. The number should be a set as a minimum task expectations, not a maximum. You will want to encourage perfromance excellence. I have found that the number will also fluctuate as there are personnel changes and volume of tissue types change. We have been monitoring performance, coupled w/ quality for 4+ years. William DeSalvo BS HTL(ASCP) > From: jwarren23@cinci.rr.com> To: histonet@lists.utsouthwestern.edu> Date: Thu, 4 Dec 2008 13:12:44 -0500> Subject: [Histonet] Embedding paraffin blocks> > Is there an average number of blocks that a tech should be able to embed in a defined time period; such as so many blocks per hour? Also, would that number vary if the tech was embedding biopsies, skin or cones versus large tissue blocks, such as uterus? I have not seen this information, but I have heard widely ranging numbers. Thanks> > Jean in Cincinnati> _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008 From Valerie.Hannen <@t> parrishmed.com Thu Dec 4 13:22:19 2008 From: Valerie.Hannen <@t> parrishmed.com (Hannen, Valerie) Date: Thu Dec 4 13:22:29 2008 Subject: [Histonet] FW: Leica ST 5020 Message-ID: <5680DA93771F0C48954CC8D38425E72401AB34E6@ISMAIL.parrishmed.local> This message was invertently typed previously using my supervisors email address by me. Please allow this to be sent to the Histonet for me. Thank you. Valerie Hannen,MLT (ASCP),HTL,SU(Fl) Parrish Medical Center Titusville, Florida. ________________________________ From: Shamrock, Rosemary Sent: Thursday, December 04, 2008 1:57 PM To: Hannen, Valerie Subject: FW: Leica ST 5020 ________________________________ From: Shamrock, Rosemary Sent: Tuesday, December 02, 2008 8:45 AM To: 'histonet@lists.utsouthwestern.edu' Subject: Leica ST 5020 Good Morning fellow Histonetters, I am writing to ask if anyone is using the Leica ST 5020 slide stainer with any success? We have had this stainer for approximately 2 months and we can not achieve consistant reproducibility. We have had reps come to our hospital to try to work through the "bugs". Our Pathologists are not happy...we get better staining results when we do our "old school" hand staining than what this instrument has given us. Any insite that you can share would be greatly appreciated. Valerie Hannen,MLT(ASCP),HTL,SU(FL) Parrish Medical Center Titusville, Florida ************************************************************************************************** "This email is intended solely for the use of the individual to whom it is addressed and may contain information that is privileged, confidential or otherwise exempt from disclosure under applicable law. If the reader of this email is not the intended recipient or the employee or agent responsible for delivering the 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 immediately delete this message. Thank you From CIngles <@t> uwhealth.org Thu Dec 4 13:35:04 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Thu Dec 4 13:36:13 2008 Subject: [Histonet] Ventana Benchmark XTs References: <2DFAEEFF192A9141ABACFF88BE613BF30BBC56@EXMBXC1.crha.bewell.ca><4b6c85510811281151w4aec5437ybf85920655ab9def@mail.gmail.com><5b6eb13e0811281210j7fd2f0d5w1c3283bd9bdead11@mail.gmail.com><4b6c85510811290803v7e52528bw2aee194670077f0@mail.gmail.com><4b6c85510812021119h38fce6a3w4cbad348f6cd9c9d@mail.gmail.com> <4b6c85510812030611o7516c4e5q26cb6a1a38f5176b@mail.gmail.com> Message-ID: <08A0A863637F1349BBFD83A96B27A50A1201C3@uwhis-xchng3.uwhis.hosp.wisc.edu> Alright kids, let's all play nice. Santa is watching you know... Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Histonet Alias Sent: Wed 12/3/2008 8:11 AM To: Joe Nocito Cc: histonet@lists.utsouthwestern.edu; Tracey Lenek; Martin Trotter; Joanna Bartczak-McKay; Burton,Lynn Subject: Re: [Histonet] Ventana Benchmark XTs Let's agree to disagree here. I will continue to post when I feel necessary and the people who don't agree with my reasoning can just ignore me. Just for the record I do not work for any vendor and I am a Histology Manager for a private laboratory in the midwest. Will my name make any difference in my posts? Take that for what it is worth. This thread was really ignited by me giving an opinion about a product or company that many people do not care for. If you don't like it don't use it. You can give your opinion about it when asked just like I can. I will use Joe the toes suggestion and change up my signature to avoid any further issue. I work for one of the larger lab companies that do not want their name or employees attached to any opinion other than the companies. If you can not understand that then I can't help you. I have mouths to feed and I do not have the luxury of changing jobs and moving to a new location. Do I like hiding my identity? No. I have been on here for years with an open identity. I have found a way to still contribute and gain knowledge from this listserv and a few of you are all about status and credibility. SO let us just end this thread and save space for real Histology issues. Al Ias HT(ASCP) Histology Manager A MidWest Lab From joelleweaver <@t> hotmail.com Thu Dec 4 13:38:08 2008 From: joelleweaver <@t> hotmail.com (joelle weaver) Date: Thu Dec 4 13:38:14 2008 Subject: [Histonet] Embedding paraffin blocks In-Reply-To: <325C5711D4A146F485B98ED4556EFAEA@ownerPC> References: <325C5711D4A146F485B98ED4556EFAEA@ownerPC> Message-ID: I have recently researched some workload and task analysis in the histo lab related to embedding. I found some good ideas are available to NSH members on their website. True statistics seem to be scarce, but I have seen in a few presentations on increasing standardization and efficiency citing numbers for embedding of 35-50 blocks/hour as an average. I think that I have posted a factor of 0.66-0.69/ block in the past, which correlates to 40/hour. I feel that that time needed for each specimen will vary with tissue type, wrapping methods( papers, sponges etc), and skill level. I think that there is more time involved with embedding skins, biopsies, and "complex" specimens, rather than large, flat pieces. Most labs I have seen, just figure the amount as a miniumum expectation to be an average based on a mix of easy and difficult specimens to embed. I agree that it is good to do some time studies on your own personnel and lab workflow, and then set a minimum with an emphasis of not exchanging speed for quality. I think that the task analysis must be correlated with technical proficiency and quality. However, I know that the numbers above are easy to meet with some experience and practice in any clinical/routine histology lab with paraffin embedding. Joelle Weaver HTL (ASCP) > From: jwarren23@cinci.rr.com> To: > Date: Thu, 4 Dec 2008 13:12:44 -0500> Subject: [Histonet] Embedding paraffin blocks> > Is there an average number of blocks that a tech should be able to embed in a defined time period; such as so many blocks per hour? Also, would that number vary if the tech was embedding biopsies, skin or cones versus large tissue blocks, such as uterus? I have not seen this information, but I have heard widely ranging numbers. Thanks> > Jean in Cincinnati> _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail anywhere. No map, no compass. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_anywhere_122008 From CIngles <@t> uwhealth.org Thu Dec 4 13:47:58 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Thu Dec 4 13:48:50 2008 Subject: [Histonet] Blue haze References: <4b6c85510812031008k3902865enbfd57014cba93c4b@mail.gmail.com> <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609D2@uwhis-xchng3.uwhis.hosp.wisc.edu> No Joe, I think the purple haze is before the tap water wash, then it turns blue... :) Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Joe Nocito Sent: Wed 12/3/2008 8:30 PM To: Histonet Alias; Marshall, Kimberly Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Blue haze I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? Forgive me, we are in the window for CAP and I'm just all in a haze. JTT ----- Original Message ----- From: "Histonet Alias" To: "Marshall, Kimberly" Cc: Sent: Wednesday, December 03, 2008 12:08 PM Subject: Re: [Histonet] Blue haze > Hi Kimberly, > Are you using a clarifier step? > > On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < > Kimberly.Marshall@ahss.org> wrote: > >> Howdy all, >> >> I have just changed over to Surgi Path's H & E system. With the latest >> fear of running out of hematoxylin and all my Pathologist wanted us to >> change over. Its a great stain but we have a bad "haze" in the >> background. >> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >> there anyone else out there using this? We are not using positive >> charged >> slides or anything added to my water bath. Any suggestions???? Thanks >> in >> advance >> >> Kimberly Marshall HT (ASCP) >> >> >> ============================================================================== >> The information contained in this message may be privileged and/or >> 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 the sender immediately by replying >> to >> this >> message and deleting the material from any computer. >> >> >> ============================================================================== >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > Al Ias HT(ASCP) > Histology Manager > Pathology Laboratory > United States > _______________________________________________ > 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 TJJ <@t> Stowers-Institute.org Thu Dec 4 13:54:58 2008 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Dec 4 13:55:33 2008 Subject: [Histonet] Re: in utero mouse embryo Message-ID: Helen, you don't state the age of the embryo or what you plan to do with it afterwards. I would recommend dissecting them out of the uterus and fixing them. Penetration/fixation will be greatly improved by doing this. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From anh2006 <@t> med.cornell.edu Thu Dec 4 13:58:01 2008 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Thu Dec 4 13:58:15 2008 Subject: [Histonet] NG2 Message-ID: Is anyone using NG2 on FFPE sections of mouse tissue? If so, I could use some details of your protocol and antibody. Thanks!! Andrea -- From CIngles <@t> uwhealth.org Thu Dec 4 14:08:12 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Thu Dec 4 14:08:17 2008 Subject: [Histonet] JCAHO References: <08A0A863637F1349BBFD83A96B27A50A1201C2@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609D5@uwhis-xchng3.uwhis.hosp.wisc.edu> Yesterday was the closest I have ever come to going 'Histo'. After 2 pretty much full days of being the only one to decorate the clinic area. Making many of the garlands, not just throwing something on the wall. Buying some of the stuff myself, and being off the clock for some of it (I am proud of all my work, not just slide production). The powers that be have stated that because of fire hazard concerns relayed by the JCAHO inspectors there are not supposed to be any decorations on walls, ceilings, or door frames. It took me an hour just to take everything down. I was so miffed (an understatement) I went home earlier than usual. I also boycotted the 'holiday party' today. ooh, missed out on the cousins subs and sheet cake. Sorry. Still trying to get ahold of the ?logic? involved with this. Not to mention all my wasted time and energy. Thank heavens today is my Friday. I need to get back in the holiday spirit somehow. Claire HTL moonlighting as an unappreciated Interior Decorator ________________________________ Sounds like you found out the hard way? --On Wednesday, December 03, 2008 6:07 PM -0600 Ingles Claire wrote: > > FYI Everyone: > If you are still waiting for your inspection, don't bother putting up > Christmas decorations. Claire > From kdboydhisto <@t> yahoo.com Thu Dec 4 14:40:27 2008 From: kdboydhisto <@t> yahoo.com (KELLY BOYD) Date: Thu Dec 4 14:40:31 2008 Subject: [Histonet] FW: Leica ST 5020 In-Reply-To: <5680DA93771F0C48954CC8D38425E72401AB34E6@ISMAIL.parrishmed.local> Message-ID: <612329.83282.qm@web58604.mail.re3.yahoo.com> I love mine. We have been using it for years now. Contact me directly, I'll see if I can give you some helpful suggestions. Kelly D. Boyd, BS, HTL (ASCP) Lab Manager Harris Histology Services 2025 Eastgate Dr. Ste. F Greenville, NC 27858 www.harrishisto.com? ? Tele (252)-830-6866 Cell? (252)-943-9527 Fax? (252)-830-0032 ? ? --- On Thu, 12/4/08, Hannen, Valerie wrote: From: Hannen, Valerie Subject: [Histonet] FW: Leica ST 5020 To: histonet@lists.utsouthwestern.edu Date: Thursday, December 4, 2008, 2:22 PM This message was invertently typed previously using my supervisors email address by me. Please allow this to be sent to the Histonet for me. Thank you. Valerie Hannen,MLT (ASCP),HTL,SU(Fl) Parrish Medical Center Titusville, Florida. ________________________________ From: Shamrock, Rosemary Sent: Thursday, December 04, 2008 1:57 PM To: Hannen, Valerie Subject: FW: Leica ST 5020 ________________________________ From: Shamrock, Rosemary Sent: Tuesday, December 02, 2008 8:45 AM To: 'histonet@lists.utsouthwestern.edu' Subject: Leica ST 5020 Good Morning fellow Histonetters, I am writing to ask if anyone is using the Leica ST 5020 slide stainer with any success? We have had this stainer for approximately 2 months and we can not achieve consistant reproducibility. We have had reps come to our hospital to try to work through the "bugs". Our Pathologists are not happy...we get better staining results when we do our "old school" hand staining than what this instrument has given us. Any insite that you can share would be greatly appreciated. Valerie Hannen,MLT(ASCP),HTL,SU(FL) Parrish Medical Center Titusville, Florida ************************************************************************************************** "This email is intended solely for the use of the individual to whom it is addressed and may contain information that is privileged, confidential or otherwise exempt from disclosure under applicable law. If the reader of this email is not the intended recipient or the employee or agent responsible for delivering the 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 immediately delete this message. Thank you _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Thu Dec 4 14:41:06 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 4 14:41:11 2008 Subject: [Histonet] Embedding paraffin blocks In-Reply-To: <325C5711D4A146F485B98ED4556EFAEA@ownerPC> Message-ID: <275859.5300.qm@web65711.mail.ac4.yahoo.com> Under separate cover I am forwarding data on the subject and, yes, there are differences between types of specimens and that is defined as "the range" of the averages in my article. Ren? J. --- On Thu, 12/4/08, Jean Warren wrote: From: Jean Warren Subject: [Histonet] Embedding paraffin blocks To: histonet@lists.utsouthwestern.edu Date: Thursday, December 4, 2008, 1:12 PM Is there an average number of blocks that a tech should be able to embed in a defined time period; such as so many blocks per hour? Also, would that number vary if the tech was embedding biopsies, skin or cones versus large tissue blocks, such as uterus? I have not seen this information, but I have heard widely ranging numbers. Thanks Jean in Cincinnati _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Thu Dec 4 15:54:29 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 4 15:54:44 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: <1167373032-1228401225-cardhu_decombobulator_blackberry.rim.net-657855511-@bxe173.bisx.prod.on.blackberry> Message-ID: Interesting point. Since 10% buffered formalin (made from the concentrated 38% formaldehyde) contain about 1% methanol, has it been shown that this has a deleterious effect on ANY antigens or are we expecting this worse case senario as being the norm? I am not aware of any antigens (or antigen-antibody combination) that has been badly effected by 10% formalin that is NOT effected by 10% formaldehyde. Are you aware of any?? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] Sent: Friday, 5 December 2008 1:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 note 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 Dec 4 15:56:04 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 4 15:56:08 2008 Subject: [Histonet] IHC on fresh frozen In-Reply-To: Message-ID: What? Do you roll it in the paraformaldehyde powder? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Hooper Sent: Friday, 5 December 2008 3:12 AM To: FU,DONGTAO; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] IHC on fresh frozen Try fixing for 5-10 min in PFA. At 10:16 AM -0500 12/3/08, FU,DONGTAO wrote: >Hi, all > > Recently I did some IHC(chromagen methods) on mouse fresh frozen >tissues, mainly using insulin antibody on pancreas. The image is >much fuzzier compare to paraffin embedding tissue. And the staining >also smeared to acinar cells which surround the islet. > > I airdried slide(>30min) and used a general acetone method(-20C >5min) to fix the tissue before I did IHC. > > How can I get a relatively sharp staining on the fresh frozen >tissue?Does anyone here have any experience on it? Any suggestions? > > Many thanks and have a nice day, > >Ann > > > > >_______________________________________________ >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 note 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 Vickroy.Jim <@t> mhsil.com Thu Dec 4 16:05:20 2008 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Thu Dec 4 16:06:12 2008 Subject: [Histonet] Don't think I'm nuts. Message-ID: <24A4826E8EF0964D86BC5317306F58A52BA58FA2FD@mmc-mail.ad.mhsil.com> Don't think I'm nuts, but I wouldn't be responding to my employees if I didn't at least ask the question. During the last couple of months several of my employees have noticed that they have been loosing more hair then usual. They are somewhat convinced that it is an environmental issue. I told them I didn't know of anything in the air here that would cause this. The only thing we have noticed is that the humidity levels in the lab have dropped dramatically in the last month or so. We are required to measure our humidity for an inspection and have noticed that the monitors all say "Lo", which indicates that the humidity levels have dropped below 20% relative humidity. Obviously that can cause dry skin, etc. My first thought was it's just our age, but we have a wide range of ages that have complained. Has anybody had these kind of complaints before and does anybody have an idea whether low humidity can cause similar problems? I have also asked our engineering department and human resource department to look into this. Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. From anh2006 <@t> med.cornell.edu Thu Dec 4 17:23:22 2008 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Thu Dec 4 17:23:33 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: References: Message-ID: Good point. I never tested it directly side-by-side to check for the effects of 1% methanol directly. It could be old-fashioned paranoia. However, it was a variable I wanted to omit as I know methanol (we are talking 100%) has deleterious effects to some mouse antigens. Therefore, it wasn't something I wanted to risk ... particularly since 95% of what I do now is bone/hematopoietic tissue work anyway which can be a nightmare to begin with. >Interesting point. >Since 10% buffered formalin (made from the concentrated 38% >formaldehyde) contain about 1% methanol, has it been shown that this has >a deleterious effect on ANY antigens or are we expecting this worse case >senario as being the norm? > >I am not aware of any antigens (or antigen-antibody combination) that >has been badly effected by 10% formalin that is NOT effected by 10% >formaldehyde. Are you aware of any?? > >Regards > >Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) >Laboratory Manager & Senior Scientist >Tel: 612 9845 3306 >Fax: 612 9845 3318 >the children's hospital at westmead >Cnr Hawkesbury Road and Hainsworth Street, Westmead >Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > > >-----Original Message----- >From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] >Sent: Friday, 5 December 2008 1:31 AM >To: Tony Henwood; Jan Shivers; histonet >Subject: Re: [Histonet] IHC on paraformaldehyde-fixed > > >So true. However, be aware that 10% neutral buffered formalin we use has >methanol in it which may affect certain antigens so there may be some >difference in staining (hence why for mouse work we now only use 4% PFA >in pure PBS). It is good to be aware of the other ingredients in your >fixative solutions, whether commercially prepared or a homemaede recipe, >as it isn't only the formaldehyde fixative which can make a difference. > > >-----Original Message----- >From: Tony Henwood > >Date: Thu, 04 Dec 2008 09:35:09 >To: Jan Shivers; >histonet >Subject: RE: [Histonet] IHC on paraformaldehyde-fixed > > >Gee I hate the term paraformaldehyde (as many of you probably know) > >This is an example of how confusion of terms can cause unnecessary work. >Is "4% paraformaldehyde" different from 4 % formaldehyde? > >No > >Should any procedure done to tissues fixed in "4% paraformaldehyde" give >results different to those fixed in 4% formaldehyde or 10% formalin? > >No since they are the same thing. > > >As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state >when paraformaldehyde actually becomes a fixative, it is no longer >paraformaldehyde by chemistry or fixation capacity. Rather, it is >formaldehyde in water without methanol or any other stabiliser. Without >heat and an alkaline environment, paraformaldehyde in water is simply a >paraformaldehyde suspension with little fixation capacity. If the >fixative is prepared from paraformaldehyde then it should be termed 4% >formaldehyde freshly prepared from paraformaldehyde. If a concentrated >formalin solution (40% formaldehyde) is used, then it should be termed >10% formalin. > >If you do a search on Histonet for paraformaldehye, you will find that >this topic has been extensively discussed. > >Regards > >Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory >Manager & Senior Scientist >Tel: 612 9845 3306 >Fax: 612 9845 3318 >the children's hospital at westmead >Cnr Hawkesbury Road and Hainsworth Street, Westmead >Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan >Shivers >Sent: Thursday, 4 December 2008 8:34 AM >To: histonet >Subject: [Histonet] IHC on paraformaldehyde-fixed > > >Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, >how well did it work? Will the same antigen-retrieval methods used with >formalin-fixed tissue be applicable? > >I'm asking for an investigator, who already has his tissues fixed in >paraformaldehyde. > >Jan Shivers >Senior Scientist >Pathology Teaching Program >Histology/IHC/EM Section Head >University of Minnesota >Veterinary Diagnostic Laboratory >1333 Gortner Ave. >St. Paul, MN 55108 >612-624-7297 shive003@umn.edu -- From collette2 <@t> mail.llnl.gov Thu Dec 4 18:40:33 2008 From: collette2 <@t> mail.llnl.gov (Nicole Collette) Date: Thu Dec 4 18:40:05 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: References: Message-ID: Hi, All, I have recently started doing IHC on whole-mount embryos, and one way to fix them at harvest is in a typical formaldehyde/formalin fixative, followed by short- or long-term storage in 100% methanol. The alternative fixation protocol, for antigens that are sensitive to formaldehyde/formalin, is to fix them in 4:1 methanol: DMSO- followed again by short- or long-term storage in methanol. (BTW the protocol works, at least for the antigens I have tested in my limited experience). I guess a counterpoint to other comments... CSH protocols; 2008; doi: 10.1101/pdb.prot 4820 Nicole Collette LLNL postdoc >Good point. I never tested it directly side-by-side to check for the >effects of 1% methanol directly. It could be old-fashioned paranoia. >However, it was a variable I wanted to omit as I know methanol (we >are talking 100%) has deleterious effects to some mouse antigens. >Therefore, it wasn't something I wanted to risk ... particularly >since 95% of what I do now is bone/hematopoietic tissue work anyway >which can be a nightmare to begin with. > > > >>Interesting point. >>Since 10% buffered formalin (made from the concentrated 38% >>formaldehyde) contain about 1% methanol, has it been shown that this has >>a deleterious effect on ANY antigens or are we expecting this worse case >>senario as being the norm? >> >>I am not aware of any antigens (or antigen-antibody combination) that >>has been badly effected by 10% formalin that is NOT effected by 10% >>formaldehyde. Are you aware of any?? >> >>Regards >> >>Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) >>Laboratory Manager & Senior Scientist >>Tel: 612 9845 3306 >>Fax: 612 9845 3318 >>the children's hospital at westmead >>Cnr Hawkesbury Road and Hainsworth Street, Westmead >>Locked Bag 4001, Westmead NSW 2145, AUSTRALIA >> >> >> >> >>-----Original Message----- >>From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] >>Sent: Friday, 5 December 2008 1:31 AM >>To: Tony Henwood; Jan Shivers; histonet >>Subject: Re: [Histonet] IHC on paraformaldehyde-fixed >> >> >>So true. However, be aware that 10% neutral buffered formalin we use has >>methanol in it which may affect certain antigens so there may be some >>difference in staining (hence why for mouse work we now only use 4% PFA >>in pure PBS). It is good to be aware of the other ingredients in your >>fixative solutions, whether commercially prepared or a homemaede recipe, >>as it isn't only the formaldehyde fixative which can make a difference. >> >> >>-----Original Message----- >>From: Tony Henwood >> >>Date: Thu, 04 Dec 2008 09:35:09 >>To: Jan Shivers; >>histonet >>Subject: RE: [Histonet] IHC on paraformaldehyde-fixed >> >> >>Gee I hate the term paraformaldehyde (as many of you probably know) >> >>This is an example of how confusion of terms can cause unnecessary work. >>Is "4% paraformaldehyde" different from 4 % formaldehyde? >> >>No >> >>Should any procedure done to tissues fixed in "4% paraformaldehyde" give >>results different to those fixed in 4% formaldehyde or 10% formalin? >> >>No since they are the same thing. >> >> >>As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state >>when paraformaldehyde actually becomes a fixative, it is no longer >>paraformaldehyde by chemistry or fixation capacity. Rather, it is >>formaldehyde in water without methanol or any other stabiliser. Without >>heat and an alkaline environment, paraformaldehyde in water is simply a >>paraformaldehyde suspension with little fixation capacity. If the >>fixative is prepared from paraformaldehyde then it should be termed 4% >>formaldehyde freshly prepared from paraformaldehyde. If a concentrated >>formalin solution (40% formaldehyde) is used, then it should be termed >>10% formalin. >> >>If you do a search on Histonet for paraformaldehye, you will find that >>this topic has been extensively discussed. >> >>Regards >> >>Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory >>Manager & Senior Scientist >>Tel: 612 9845 3306 >>Fax: 612 9845 3318 >>the children's hospital at westmead >>Cnr Hawkesbury Road and Hainsworth Street, Westmead >>Locked Bag 4001, Westmead NSW 2145, AUSTRALIA >> >> >> >> >>-----Original Message----- >>From: histonet-bounces@lists.utsouthwestern.edu >>[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan >>Shivers >>Sent: Thursday, 4 December 2008 8:34 AM >>To: histonet >>Subject: [Histonet] IHC on paraformaldehyde-fixed >> >> >>Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, >>how well did it work? Will the same antigen-retrieval methods used with >>formalin-fixed tissue be applicable? >> >>I'm asking for an investigator, who already has his tissues fixed in >>paraformaldehyde. >> >>Jan Shivers >>Senior Scientist >>Pathology Teaching Program >>Histology/IHC/EM Section Head >>University of Minnesota >>Veterinary Diagnostic Laboratory >>1333 Gortner Ave. >>St. Paul, MN 55108 >>612-624-7297 >shive003@umn.edu >-- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http:// lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Thu Dec 4 21:11:25 2008 From: tifei <@t> foxmail.com (tf) Date: Thu Dec 4 21:11:47 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed References: Message-ID: <200812051111205608471@foxmail.com> dGhlIGJhc2ljIHByaW5jaXBsZXMgYXJlIHRoZSBzYW1lIGZvciBtb3N0IGNyb3NzLWxpbmtpbmcg Zml4YXRpdmVzIGFuZCBpbmR1Y2Ugc2ltaWxhciBib25kcyANCnRoZSBkaWZmZXJlbmNlIHlvdSBv 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RE: [Histonet] Blue haze References: <15960.97595.qm@web50902.mail.re2.yahoo.com> Message-ID: <200812051112269238582@foxmail.com> Ni44DQoNCg0KMjAwOC0xMi0wNSANCg0KDQoNCnRmIA0KDQoNCg0Kt6K8/sjLo7ogQ2hlcnlsIA0K t6LLzcqxvOSjuiAyMDA4LTEyLTA1ICAwMjoxMDo0OCANCsrVvP7Iy6O6IGhpc3RvbmV0QGxpc3Rz LnV0c291dGh3ZXN0ZXJuLmVkdSANCrOty82juiANCtb3zOKjuiBXaGF0J3MgdGhlIHBIIG9mIHlv dXIgdGFwIHdhdGVyPyBSRTogW0hpc3RvbmV0XSBCbHVlIGhhemUgDQogDQpIZXkgYW5kIEhlbGxv LQ0KPw0KSSBoYXZlbid0IGZvbGxvd2VkIHRoaXMgd2hvbGUgc3RyaW5nIHNvIEkgYXBvbG9naXpl IGlmIHRoaXMgaGFzIGJlZW4gbWVudGlvbmVkOg0KPw0KV2hlbiBJIHdhcyBpbiBhIHN0YXRlIHdp dGggcmVhbGx5IGhhcmQgd2F0ZXIgdGhpcyBjYW1lIHVwIG1vcmUgb2Z0ZW4gdGhhbiBhbnl3aGVy ZSBlbHNlLi4uaGF2ZSB5b3UgY2hlY2tlZCB0aGUgcEggb2YgeW91ciB3YXRlcj8/SWYgaXQgaXNu J3QgY2xvc2UgdG8gbmV1dHJhbCB0aGUgaGF6ZSBzaG93cyB1cC4/V2UgaGFkIHRvIHJpbnNlIGlu IGRIMk8gdG8gZ2V0IHRoZSBleHRyYSBzdGFpbiBvZmYgYmVmb3JlIHJpbnNpbmcgaW4gdGFwIHdh dGVyLj9UaGUgd2FybSB3YXRlciBoYWQgYSBkaWZmZXJlbnQgcEggdGhhbiBjb2xkLS1zbyB0aGF0 IG1pZ2h0IGFsc28gaGVscCBhcyBzdWdnZXN0ZWQgZWFybGllci4/DQo/DQpUaGlzIHdhcyBmb3Ig YSBHaWxsIElJLS1ub3Qgc3VyZSBvZiB0aGUgZWZmZWN0IG9uIHRoZSBuZXcgcHJvZHVjdC0tanVz dCBvZmZlcmluZyB1cCBvbmUgbW9yZSB2YXJpYWJsZSB0byB0cnkhDQo/DQpDaGVyeWwNCj8NCkNo ZXJ5bCBSLiBLZXJyeSwgSFQoQVNDUCksIEJBDQpGdWxsIFN0YWZmIEluYy4NClN0YWZmaW5nIEhl YWx0aGNhcmUgUHJvZmVzc2lvbmFscyAtIE9uZSBHUkVBVCBmaXQgYXQgYSB0aW1lIQ0KPw0KMjE4 LjkxMy43Mjg1DQo4MDAuNzU2LjMzMDkNCnJlc3VtZUBmdWxsc3RhZmYub3JnDQpfX19fX19fX19f X19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fXw0KSGlzdG9uZXQgbWFpbGluZyBs aXN0DQpIaXN0b25ldEBsaXN0cy51dHNvdXRod2VzdGVybi5lZHUNCg== From tifei <@t> foxmail.com Thu Dec 4 21:15:40 2008 From: tifei <@t> foxmail.com (tf) Date: Thu Dec 4 21:15:54 2008 Subject: [Histonet] PSA-NCAM IHC on frozen section with BrdU Message-ID: <200812051115355894806@foxmail.com> Hi everyone, it seems that I lost the PSA-NCAM staining when sections are treated with citrate buffer (poring the membrane?) Possibly the membrane was dissociated from membrane then>? or? For co-staining with BrdU, I can only treat sections with HCl - the results sometimes are unstable~~ Anyone worked on this before? 2008-12-04 tf _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From AnthonyH <@t> chw.edu.au Thu Dec 4 21:29:38 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 4 21:29:55 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: <200812051111205608471@foxmail.com> Message-ID: tf wrote: =20 "I DO believe that one reason some people use 4% PFA rather 10% formalin is= that PFA is a bit more stable, both for storage and transportation~~~." =20 I have not heard this before. Do you have a reference for this? =20 =20 Regards=20 Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20 Laboratory Manager & Senior Scientist=20 Tel: 612 9845 3306=20 Fax: 612 9845 3318=20 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: tf [mailto:tifei@foxmail.com]=20 Sent: Friday, 5 December 2008 2:11 PM To: Tony Henwood; anh2006@med.cornell.edu; Jan Shivers; histonet Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed =09 =09 the basic principles are the same for most cross-linking fixatives and ind= uce similar bonds=20 the difference you observed between may due to any other variability, or t= he co-fixative you used. =20 I DO believe that one reason some people use 4% PFA rather 10% formalin is= that PFA is a bit more stable, both for storage and transportation~~~. =20 =20 =20 =20 2008-12-05=20 =09 ________________________________ tf=20 =09 ________________________________ =B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood=20 =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-05 06:00:03=20 =CA=D5=BC=FE=C8=CB=A3=BA anh2006@med.cornell.edu; Jan Shivers; histonet=20 =B3=AD=CB=CD=A3=BA=20 =D6=F7=CC=E2=A3=BA RE: [Histonet] IHC on paraformaldehyde-fixed=20 =09 =09 Interesting point. Since 10% buffered formalin (made from the concentrated 38% formaldehyde) contain about 1% methanol, has it been shown that this has a deleterious effect on ANY antigens or are we expecting this worse case senario as being the norm? I am not aware of any antigens (or antigen-antibody combination) that has been badly effected by 10% formalin that is NOT effected by 10% formaldehyde. Are you aware of any?? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu]=20 Sent: Friday, 5 December 2008 1:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09=20 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin?=20 No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 sen= der. Views expressed in this message and any attachments are those of the indiv= idual sender, and are not necessarily the views of The Children's Hospital = at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Children= s Hospital at Westmead accepts no liability for any consequential damage re= sulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu ********************************************************************* 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 send= er. Views expressed in this message and any attachments are those of the indivi= dual sender, and are not necessarily the views of The Children's Hospital a= t Westmead This note 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 res= ulting from email containing computer viruses. ********************************************************************** From lenaspencer <@t> insightbb.com Thu Dec 4 21:29:58 2008 From: lenaspencer <@t> insightbb.com (Lena Spencer) Date: Thu Dec 4 21:30:47 2008 Subject: [Histonet] Job Opening Message-ID: <001e01c95689$bfd18f20$3f74ad60$@com> Full time position available - Norton Healthcare, Louisville, KY must be a registered Histotechnician/Histotechnologist. Qualified applicants please contact Mary Beth Knight at 502-629-07884 or Marybeth.Knight@nortonhealthcare.org Lena From carl.hobbs <@t> kcl.ac.uk Fri Dec 5 03:45:42 2008 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Fri Dec 5 03:47:30 2008 Subject: [Histonet] Re: NG2 Message-ID: <11D9615B89C10747B1C985966A63D7CA286ECA98F0@KCL-MAIL04.kclad.ds.kcl.ac.uk> I use Chemicon's AB5320 anti NG2 ( 1/50-100) on mouse and rat pwax brain sections. I need AR and use Citric acid pH6 in a microwaveable pressure cooker.Standard stABCpxDAB detection. Apart from the poor dilution factor, it's a very good reagent, imo. Carl Hobbs Histology Manager Wolfson Centre for Age-Related Diseases King's College London Tel.020 7848 6810 From jnocito <@t> satx.rr.com Fri Dec 5 05:40:14 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Dec 5 05:40:16 2008 Subject: [Histonet] Blue haze References: <777EE70F42D44C11BC99C4C966F939FD@yourxhtr8hvc4p> <000501c95619$77487c20$d00f7ca5@lurie.northwestern.edu> <7722595275A4DD4FA225B92CDBF174A1744F13BB30@EXC-MBX3.cfs.le.ac.uk> Message-ID: <3DDE0EE5E299439EB527CC8C8F692C7C@yourxhtr8hvc4p> lunch!? ----- Original Message ----- From: "Edwards, R.E." To: "'Bernice Frederick'" ; "'Joe Nocito'" ; "'Histonet Alias'" ; "'Marshall, Kimberly'" Cc: Sent: Thursday, December 04, 2008 10:52 AM Subject: RE: [Histonet] Blue haze Hey Joe, where you going with that toe in your hand?? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bernice Frederick Sent: 04 December 2008 14:06 To: 'Joe Nocito'; 'Histonet Alias'; 'Marshall, Kimberly' Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Blue haze I'm sure you'll just charm them and they'll never catch on.... Bernice Frederick HTL (ASCP) Northwestern University Pathology Core Facility ECOGPCO-RL 710 N Fairbanks Court Olson 8-421 Chicago,IL 60611 312-503-3723 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Wednesday, December 03, 2008 8:31 PM To: Histonet Alias; Marshall, Kimberly Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Blue haze I thought that was Purple Haze, by Jimi Hendrix. Oh dear me, is it Friday yet? Forgive me, we are in the window for CAP and I'm just all in a haze. JTT ----- Original Message ----- From: "Histonet Alias" To: "Marshall, Kimberly" Cc: Sent: Wednesday, December 03, 2008 12:08 PM Subject: Re: [Histonet] Blue haze > Hi Kimberly, > Are you using a clarifier step? > > On Wed, Dec 3, 2008 at 12:47 PM, Marshall, Kimberly < > Kimberly.Marshall@ahss.org> wrote: > >> Howdy all, >> >> I have just changed over to Surgi Path's H & E system. With the latest >> fear of running out of hematoxylin and all my Pathologist wanted us to >> change over. Its a great stain but we have a bad "haze" in the >> background. >> It is not hurting the tissue or DX at all, but to ME it looks awful. Is >> there anyone else out there using this? We are not using positive >> charged >> slides or anything added to my water bath. Any suggestions???? Thanks >> in >> advance >> >> Kimberly Marshall HT (ASCP) >> >> >> ============================================================================ == >> The information contained in this message may be privileged and/or >> 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 the sender immediately by replying >> to >> this >> message and deleting the material from any computer. >> >> >> ============================================================================ == >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > -- > Al Ias HT(ASCP) > Histology Manager > Pathology Laboratory > United States > _______________________________________________ > 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 Fri Dec 5 05:43:06 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Fri Dec 5 05:43:05 2008 Subject: [Histonet] JCAHO References: <08A0A863637F1349BBFD83A96B27A50A1201C2@uwhis-xchng3.uwhis.hosp.wisc.edu> <08A0A863637F1349BBFD83A96B27A50A08E609D5@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: can you say "GRINCH"? ----- Original Message ----- From: "Ingles Claire " To: "Merced Leiker" Cc: Sent: Thursday, December 04, 2008 2:08 PM Subject: RE: [Histonet] JCAHO Yesterday was the closest I have ever come to going 'Histo'. After 2 pretty much full days of being the only one to decorate the clinic area. Making many of the garlands, not just throwing something on the wall. Buying some of the stuff myself, and being off the clock for some of it (I am proud of all my work, not just slide production). The powers that be have stated that because of fire hazard concerns relayed by the JCAHO inspectors there are not supposed to be any decorations on walls, ceilings, or door frames. It took me an hour just to take everything down. I was so miffed (an understatement) I went home earlier than usual. I also boycotted the 'holiday party' today. ooh, missed out on the cousins subs and sheet cake. Sorry. Still trying to get ahold of the ?logic? involved with this. Not to mention all my wasted time and energy. Thank heavens today is my Friday. I need to get back in the holiday spirit somehow. Claire HTL moonlighting as an unappreciated Interior Decorator ________________________________ Sounds like you found out the hard way? --On Wednesday, December 03, 2008 6:07 PM -0600 Ingles Claire wrote: > > FYI Everyone: > If you are still waiting for your inspection, don't bother putting up > Christmas decorations. Claire > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MMargiotta <@t> bmhmc.org Fri Dec 5 07:27:25 2008 From: MMargiotta <@t> bmhmc.org (Margiotta, Michele) Date: Fri Dec 5 07:27:30 2008 Subject: [Histonet] thermometer calibration Message-ID: Hi All, We are looking to buy a calibrated thermometer (NIST traceable) that is non-mercury. There's plenty of mercury ones out there but can't find the non-mercury kind. Any one know of a source to check? Thanks! Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. From Kimberly.Marshall <@t> ahss.org Fri Dec 5 07:53:04 2008 From: Kimberly.Marshall <@t> ahss.org (Marshall, Kimberly) Date: Fri Dec 5 07:53:13 2008 Subject: [Histonet] Thanks for all the help Message-ID: Hello again Want to say thanks for all the help with my "haze" I have already tried filtering and it seems to have fixed the problem. Just cuz they say no need to filter dont mean its true. Again thanks for all the help Kim ============================================================================== The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ============================================================================== From rjbuesa <@t> yahoo.com Fri Dec 5 08:02:21 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Dec 5 08:02:24 2008 Subject: [Histonet] thermometer calibration In-Reply-To: Message-ID: <259803.76223.qm@web65716.mail.ac4.yahoo.com> Buy a digital thermometer. They are fast, accurate and all are calibrated. Ren? J. --- On Fri, 12/5/08, Margiotta, Michele wrote: From: Margiotta, Michele Subject: [Histonet] thermometer calibration To: histonet@pathology.swmed.edu Date: Friday, December 5, 2008, 8:27 AM Hi All, We are looking to buy a calibrated thermometer (NIST traceable) that is non-mercury. There's plenty of mercury ones out there but can't find the non-mercury kind. Any one know of a source to check? Thanks! Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Fri Dec 5 08:09:08 2008 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Dec 5 08:09:23 2008 Subject: [Histonet] thermometer calibration In-Reply-To: Message-ID: We can't find non-mercury thermometers that have a serial number on them, so we can't calibrate them because they need an indelible identifier. We are, however, getting away from using thermometers completely by having our digital read-out instruments calibrated instead - i.e., embedding centers, tissue processors, paraffin baths, oven. No more thermometers. Jackie O' "Margiotta, Michele" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/05/2008 07:27 AM To cc Subject [Histonet] thermometer calibration Hi All, We are looking to buy a calibrated thermometer (NIST traceable) that is non-mercury. There's plenty of mercury ones out there but can't find the non-mercury kind. Any one know of a source to check? Thanks! Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Wanda.Smith <@t> HCAhealthcare.com Fri Dec 5 09:06:42 2008 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Dec 5 09:06:48 2008 Subject: [Histonet] Dual Esterase Staining Message-ID: <9E2D36CE2D7CBA4A94D9B22E8328A3BA974609AC@NADCWPMSGCMS03.hca.corpad.net> Happy Friday Everyone! Our HemPath Pathologist is having us do the Dual Esterase stain on blood smears in our histology lab. I have a few questions regarding this stain: 1. Does anyone know what the CPT code is for performing this stain??? 2. Will anyone share what they charge the patient for performing this stain? 3. Does any company other than Sigma sell a kit to perform this stain and how much does the kit cost??? Thanks for any help regarding this matter!!!!! Wanda WANDA G. SMITH, HTL(ASCP)HT Pathology Supervisor TRIDENT MEDICAL CENTER 9330 Medical Plaza Drive Charleston, SC 29406 843-847-4586 843-847-4296 fax From mpence <@t> grhs.net Fri Dec 5 09:27:39 2008 From: mpence <@t> grhs.net (Mike Pence) Date: Fri Dec 5 09:27:44 2008 Subject: [Histonet] thermometer calibration In-Reply-To: Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A39D2@IS-E2K3.grhs.net> Try http://www.ertco.com/non_mercury_ever_safe_thermometers.html This is where I get all my thermometers. Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Friday, December 05, 2008 8:09 AM To: Margiotta, Michele Cc: histonet-bounces@lists.utsouthwestern.edu; histonet@pathology.swmed.edu Subject: Re: [Histonet] thermometer calibration We can't find non-mercury thermometers that have a serial number on them, so we can't calibrate them because they need an indelible identifier. We are, however, getting away from using thermometers completely by having our digital read-out instruments calibrated instead - i.e., embedding centers, tissue processors, paraffin baths, oven. No more thermometers. Jackie O' "Margiotta, Michele" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/05/2008 07:27 AM To cc Subject [Histonet] thermometer calibration Hi All, We are looking to buy a calibrated thermometer (NIST traceable) that is non-mercury. There's plenty of mercury ones out there but can't find the non-mercury kind. Any one know of a source to check? Thanks! Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. _______________________________________________ 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 SEsparza <@t> seton.org Fri Dec 5 09:33:00 2008 From: SEsparza <@t> seton.org (Esparza, Sandra) Date: Fri Dec 5 09:33:08 2008 Subject: [Histonet] thermometer calibration In-Reply-To: <259803.76223.qm@web65716.mail.ac4.yahoo.com> References: <259803.76223.qm@web65716.mail.ac4.yahoo.com> Message-ID: <3D79F47DC92B204F9E5D35C885DFC5CB01003C3C@AUSEX2VS1.seton.org> Our lab uses the Streck thermometers. These meet the CAP and CLIA regulations. The web address is www.streck.com and the phone # 1-800-843-0912. Sandra HT(ASCP)QIHC Dell Children's Hospital Austin Texas -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, December 05, 2008 8:02 AM To: histonet@pathology.swmed.edu; Margiotta, Michele Subject: Re: [Histonet] thermometer calibration Buy a digital thermometer. They are fast, accurate and all are calibrated. Ren? J. --- On Fri, 12/5/08, Margiotta, Michele wrote: From: Margiotta, Michele Subject: [Histonet] thermometer calibration To: histonet@pathology.swmed.edu Date: Friday, December 5, 2008, 8:27 AM Hi All, We are looking to buy a calibrated thermometer (NIST traceable) that is non-mercury. There's plenty of mercury ones out there but can't find the non-mercury kind. Any one know of a source to check? Thanks! Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. _______________________________________________ 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 masaakikitada2007 <@t> gmail.com Fri Dec 5 09:59:36 2008 From: masaakikitada2007 <@t> gmail.com (Masaaki Kitada) Date: Fri Dec 5 09:59:10 2008 Subject: [Histonet] Re: NG2 In-Reply-To: <11D9615B89C10747B1C985966A63D7CA286ECA98F0@KCL-MAIL04.kclad.ds.kcl.ac.uk> References: <11D9615B89C10747B1C985966A63D7CA286ECA98F0@KCL-MAIL04.kclad.ds.kcl.ac.uk> Message-ID: As far as my experience, antigen retrieval with microwave treatment destroyed the antigenicity of NG2, although I used rabbit NG2 antibody given by Dr. Stallcup in immunostaining for cryosections so there should be some differences from your condition. I don't know what's happening in there. Masaaki Kitada _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ Masaaki Kitada, M.D., Ph.D. Associate Professor Department of Stem Cell Biology and Histology Tohoku Univeristy Graduate School of Medicine 2-1, Seiryo-machi, Aoba-ku Sendai, Miyagi 980-8575, Japan Phone: +81-22-717-8029, FAX: +81-22-717-8030 e-mail: Masaaki.Kitada@gmail.com _/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/_/ At 9:45 AM +0000 08.12.5, Hobbs, Carl wrote: >I use Chemicon's AB5320 anti NG2 ( 1/50-100) on mouse and rat pwax brain >sections. I need AR and use Citric acid pH6 in a microwaveable pressure >cooker.Standard stABCpxDAB detection. >Apart from the poor dilution factor, it's a very good reagent, imo. > >Carl Hobbs >Histology Manager >Wolfson Centre for Age-Related Diseases >King's College London >Tel.020 7848 6810 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Reuel.Cornelia <@t> tsrh.org Fri Dec 5 10:08:46 2008 From: Reuel.Cornelia <@t> tsrh.org (Reuel Cornelia) Date: Fri Dec 5 10:09:35 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: References: <200812051111205608471@foxmail.com> Message-ID: <4938FDAE020000C500048F27@nwcl02.tsrh.org> I have been curious about this discussion. we used 4% paraformaldehyde for smaller biopsies only because it has a faster penetration to tissue than 10% formalin. In all my IHC that I have done. I observe that doing an IHC with 4% paraformaldehyde does not necessarily need antigen retrieval in comparison to 10% formalin either it will be human or animal tissue but this depends on how long was it fix, our 4% paraformaldehyde we fix smaller biopsies like nerve,muscle, skin for 6 to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe you can comment on the effect on this to tissue if you say you will use 4% paraformaldehyde for storage and transportation. Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 >>> "Tony Henwood" 12/04/08 9:29 PM >>> tf wrote: "I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~." I have not heard this before. Do you have a reference for this? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: tf [mailto:tifei@foxmail.com] Sent: Friday, 5 December 2008 2:11 PM To: Tony Henwood; anh2006@med.cornell.edu; Jan Shivers; histonet Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed the basic principles are the same for most cross-linking fixatives and induce similar bonds the difference you observed between may due to any other variability, or the co-fixative you used. I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~. 2008-12-05 ________________________________ tf ________________________________ ???? Tony Henwood ???? 2008-12-05 06:00:03 ???? anh2006@med.cornell.edu; Jan Shivers; histonet ??? ?? RE: [Histonet] IHC on paraformaldehyde-fixed Interesting point. Since 10% buffered formalin (made from the concentrated 38% formaldehyde) contain about 1% methanol, has it been shown that this has a deleterious effect on ANY antigens or are we expecting this worse case senario as being the norm? I am not aware of any antigens (or antigen-antibody combination) that has been badly effected by 10% formalin that is NOT effected by 10% formaldehyde. Are you aware of any?? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] Sent: Friday, 5 December 2008 1:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin? No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mlm11 <@t> cornell.edu Fri Dec 5 10:23:32 2008 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Fri Dec 5 10:23:24 2008 Subject: [Histonet] membranes and prices Message-ID: <6.2.1.2.2.20081205111128.04139b58@postoffice9.mail.cornell.edu> Hello All, I have prepared Costar Transwell membrane inserts according to the Corning procedure and I can not keep the little suckers on the slides. Subbed, charged, and celloidin is not working well. With celloidin, they will stay until the last alcohols and then fall off. Any suggestions? I have left the slides in a 60oC oven all night before staining. I seemed to have become a specialty histo lab. I would really appreciate someone else figuring out what to charge but it's left to me. Is there a lab out there that has a great price list I can "borrow"? As a for instance, I have spent 3 days working on these darn membranes with nothing to show yet and have no idea what to charge. I do lots of hand processing also. What's fair for that? Thanks for all the help!! (Corning hasn't gotten back to me yet.) Happy Friday!! Mary Lou From ree3 <@t> leicester.ac.uk Fri Dec 5 10:35:06 2008 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Dec 5 10:35:29 2008 Subject: [Histonet] RE: Don't think I'm nuts. In-Reply-To: <24A4826E8EF0964D86BC5317306F58A52BA58FA2FD@mmc-mail.ad.mhsil.com> References: <24A4826E8EF0964D86BC5317306F58A52BA58FA2FD@mmc-mail.ad.mhsil.com> Message-ID: <7722595275A4DD4FA225B92CDBF174A1744F13BB3A@EXC-MBX3.cfs.le.ac.uk> I believe that low humidity can cause laboratory mice to lose/shed the tips of their tails. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Vickroy, Jim Sent: 04 December 2008 22:05 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Don't think I'm nuts. Don't think I'm nuts, but I wouldn't be responding to my employees if I didn't at least ask the question. During the last couple of months several of my employees have noticed that they have been loosing more hair then usual. They are somewhat convinced that it is an environmental issue. I told them I didn't know of anything in the air here that would cause this. The only thing we have noticed is that the humidity levels in the lab have dropped dramatically in the last month or so. We are required to measure our humidity for an inspection and have noticed that the monitors all say "Lo", which indicates that the humidity levels have dropped below 20% relative humidity. Obviously that can cause dry skin, etc. My first thought was it's just our age, but we have a wide range of ages that have complained. Has anybody had these kind of complaints before and does anybody have an idea whether low humidity can cause similar problems? I have also asked our engineering department and human resource department to look into this. Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ejschmid <@t> ucalgary.ca Fri Dec 5 11:29:33 2008 From: ejschmid <@t> ucalgary.ca (ejschmid@ucalgary.ca) Date: Fri Dec 5 11:29:42 2008 Subject: [Histonet] fixation post-fixation, difference of order used Message-ID: <62915.136.159.150.24.1228498173.squirrel@136.159.150.24> Hi, I was wondering about the effects of using different fixatives in different orders. For example, some of our embryonic material (chick and mouse) is fixed initially with 4% PFA and subsequently is exposed to glutaraldehyde. Chick embryos subject to this are successfully labeled using IHC for anti-HH3. However, our mouse tissues are fixed differently, being first exposed to 4% PFA: 1% Glut solution (Which i think is called a modified Karnovsky's fix). These embryos perform poorly for IHC with anti-HH3. Might it be the glutaraldehyde that is present in our primary fixative used to fix moue embryos? Might it be that the IHC works for chick because the glutaraldehyde is only used as a secondary fix after primary fixation in 4% PFA? Does the order of exposure matter? thanks! eric schmidt University of Calgary Medical Sciences From ejschmid <@t> ucalgary.ca Fri Dec 5 11:30:02 2008 From: ejschmid <@t> ucalgary.ca (ejschmid@ucalgary.ca) Date: Fri Dec 5 11:30:12 2008 Subject: [Histonet] Bouin's fixation and paraffin or vibratome sectioning Message-ID: <62915.136.159.150.24.1228498202.squirrel@136.159.150.24> Hi, Question about use of Bouin's fix on mid-gestational mouse embryos...We fix our ED 10-11 embryos in Bouin's and embed for sectioning, either paraffin or vibratome. What we experience is that the tissues suffer quite a bit of damage, as the sections are often ripped. It sometimes seems to us that the Bouin's fixed tissue is really dry or even brittle. Do others have a similar experience? Eric Schmidt University of Calgary Medical Sciences From Janet.Bonner <@t> FLHOSP.ORG Fri Dec 5 13:43:12 2008 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Dec 5 13:47:04 2008 Subject: [Histonet] Don't think I'm nuts. References: <24A4826E8EF0964D86BC5317306F58A52BA58FA2FD@mmc-mail.ad.mhsil.com> Message-ID: <5F31F38C96781A4FBE3196EBC22D47807F2860@fhosxchmb006.ADVENTISTCORP.NET> The hair loss may be a normal seasonal event, similar to when cats shed their coat in the spring and summer. Did you know that our nails stop growing in the darker months? Instead of having a manicure every two weeks, as in the spring and summer, I get one every three weeks in the winter months. Hair loss will cycle and grow back every three months or so. Perhaps everyone has just now been made aware? Janet Janet L. Bonner, HTL (ASCP) Pathology Laboratory ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Vickroy, Jim Sent: Thu 12/4/2008 5:05 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Don't think I'm nuts. Don't think I'm nuts, but I wouldn't be responding to my employees if I didn't at least ask the question. During the last couple of months several of my employees have noticed that they have been loosing more hair then usual. They are somewhat convinced that it is an environmental issue. I told them I didn't know of anything in the air here that would cause this. The only thing we have noticed is that the humidity levels in the lab have dropped dramatically in the last month or so. We are required to measure our humidity for an inspection and have noticed that the monitors all say "Lo", which indicates that the humidity levels have dropped below 20% relative humidity. Obviously that can cause dry skin, etc. My first thought was it's just our age, but we have a wide range of ages that have complained. Has anybody had these kind of complaints before and does anybody have an idea whether low humidity can cause similar problems? I have also asked our engineering department and human resource department to look into this. Jim Vickroy BS, HT(ASCP) Technical Supervisor - Surgical and Autopsy Pathology Memorial Medical Center 217-788-4046 vickroy.jim@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ======================================================= The information contained in this message may be privileged and/or 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 the sender immediately by replying to this message and deleting the material from any computer. ======================================================= From PMonfils <@t> Lifespan.org Fri Dec 5 14:14:07 2008 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Dec 5 14:14:22 2008 Subject: [Histonet] Polymerizing BIG methyl methacrylate blocks Message-ID: <4EBFF65383B74D49995298C4976D1D5E03835C81@LSRIEXCH1.lsmaster.lifespan.org> Does anyone here have experience with large PMMA blocks, about 150 to 200 ml volume each? Specifically, at what temperature do you recommend polymerizing them, and how long should the polymerization take? Any other helpful hints will be appreciated. Paul M. From leiker <@t> buffalo.edu Fri Dec 5 15:00:14 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Fri Dec 5 15:00:19 2008 Subject: [Histonet] Don't think I'm nuts. In-Reply-To: <5F31F38C96781A4FBE3196EBC22D47807F2860@fhosxchmb006.ADVENTISTCORP.NET> References: <24A4826E8EF0964D86BC5317306F58A52BA58FA2FD@mmc-mail.ad.mhsil.com > <5F31F38C96781A4FBE3196EBC22D47807F2860@fhosxchmb006.ADVENTISTCORP.NET> Message-ID: I've noticed that too, although I think I tend to shed hair more in the summer than in the winter, or when I'm under stress (are your people under more stress these days perhaps??). But I have also noticed also that my nail growth slows down significantly in the winter - I like to keep them as trim as possible and definitely have to cut them less in winter. Merced --On Friday, December 05, 2008 2:43 PM -0500 "Bonner, Janet" wrote: > The hair loss may be a normal seasonal event, similar to when cats shed > their coat in the spring and summer. Did you know that our nails stop > growing in the darker months? Instead of having a manicure every two > weeks, as in the spring and summer, I get one every three weeks in the > winter months. Hair loss will cycle and grow back every three > months or so. Perhaps everyone has just now been made aware? > Janet > > Janet L. Bonner, HTL (ASCP) > Pathology Laboratory > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Vickroy, Jim > Sent: Thu 12/4/2008 5:05 PM > To: 'histonet@lists.utsouthwestern.edu' > Subject: [Histonet] Don't think I'm nuts. > > > > > Don't think I'm nuts, but I wouldn't be responding to my employees if I > didn't at least ask the question. During the last couple of months > several of my employees have noticed that they have been loosing more > hair then usual. They are somewhat convinced that it is an environmental > issue. I told them I didn't know of anything in the air here that would > cause this. The only thing we have noticed is that the humidity levels > in the lab have dropped dramatically in the last month or so. We are > required to measure our humidity for an inspection and have noticed that > the monitors all say "Lo", which indicates that the humidity levels have > dropped below 20% relative humidity. Obviously that can cause dry skin, > etc. My first thought was it's just our age, but we have a wide range of > ages that have complained. Has anybody had these kind of complaints > before and does anybody have an idea whether low humidity can cause > similar problems? I have also asked our engineering department and human > resource department to look into this. > > > Jim Vickroy BS, HT(ASCP) > Technical Supervisor - Surgical and Autopsy Pathology > Memorial Medical Center > 217-788-4046 > vickroy.jim@mhsil.com > > > This message (including any attachments) contains confidential > information intended for a specific individual and purpose, and is > protected by law. If you are not the intended recipient, you should > delete this message. Any disclosure, copying, or distribution of this > message, or the taking of any action based on it, is strictly prohibited. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > ======================================================= > The information contained in this message may be privileged and/or > 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 the sender immediately by replying > to this message and deleting the material from any computer. > ======================================================= > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From barbaraaalbert <@t> yahoo.com Fri Dec 5 15:14:05 2008 From: barbaraaalbert <@t> yahoo.com (Barbara Albert) Date: Fri Dec 5 15:14:08 2008 Subject: [Histonet] solvent recycling Message-ID: <181709.3860.qm@web63707.mail.re1.yahoo.com> We are just now looking into recycling our solvents and alcohol as we're designing a new facility (no room here).? We're particularly interested in those who have?experience with large volume opeations.? We run 4 tissure processors daily, an H&E stainer, 2 glass coverslippers and hand operations for immunos and special stains.? We're also interested in any potential pitfalls.? Feel free to contact me off list. ? Barbara Albert UCSF Medical Center San Francisco From ratliffjack <@t> hotmail.com Fri Dec 5 15:18:39 2008 From: ratliffjack <@t> hotmail.com (Jack Ratliff) Date: Fri Dec 5 15:18:44 2008 Subject: [Histonet] Polymerizing BIG methyl methacrylate blocks In-Reply-To: <4EBFF65383B74D49995298C4976D1D5E03835C81@LSRIEXCH1.lsmaster.lifespan.org> References: <4EBFF65383B74D49995298C4976D1D5E03835C81@LSRIEXCH1.lsmaster.lifespan.org> Message-ID: Paul, I can tell you from many years of experience that you have to be very careful when polymerizing large amounts of MMA. As you probably know, it is an exothermic reaction that can and WILL generate a significant amount of heat depending upon the percentage/amount of catalyst you use, the size and density of the tissue, total volume used, and the temperature at which the solution is left at to polymerize. If you do not control the polymerization, it will reach its v-max too fast which will result in an over-polymerized mess and ruin your tissue. My best advice to you immediately is to take a look at the concentration of catalyst you are using and consider lowering it depending on what you are currently using. I use approximately 0.25% w/w of dry Perkadox-16. A couple of days before I am ready to embed my tissue, I will make up 1000 mL of MMA + DBP and then add 2.5 g of Perkadox. I do this at the end of the working day and let it stir lightly overnight (22.2 - 22.5 C) in the hood at room temperature to 'ripen'. The solution will change from clear to a shade of yellow (lighter or darker) that is somewhat proportional to what room temperature means in your lab. The next piece of advice is how you control the rate of polymerization. The following morning I will put the solution into the fridge overnight to slow the reaction. The very next day, I bring the solution out of the fridge and allow to warm to room temperature. This is to avoid any potential for moisture as you don't want a cooled MMA solution to have the chance to meet with a room temperature tissue and create ANY chance for moisture. It could make your final polymerized block cloudy instead of clear. I then add my embedding solution to the embedding mold w/ pre-polymerized base layer and drop in the tissue. With the lid off, I then place the specimen, within its embedding container, under vacuum (-18 to -20 inHg) until the end of the day (6-8 hours). Don't worry about orientation at this point because you will attend to it before you leave for the evening. At the end of the day I will put the tissue into the desired orientation, put the lid on, and place it in the fridge overnight. The next morning, I simply bring out my embedding container (lid on) and leave it at room temperature (22.2 - 22.5 C) for the rest of the day and then overnight (if you feel that you can trust room temperature to stay consistent) and repeat as necessary. Basically, I babysit my specimens on a daily basis over the next 5-9 days (depending on the tissue size/density and volume of embedding solution) and alternate between room temperature and the fridge to help evenly polymerize my specimen so that I achieve a nice clear block. Once the block is mostly polymerized and definitely above the level of the tissue with only a very thin or tacky layer on top, I place the container in a waterbath (level of water slightly above the block and fluid level) and leave it at 37 C overnight until the block has polymerized. The next day I will then put the container (without the lid) into an oven at 60 C overnight to fully complete polymerization and evaporate most of the residual MMA smell. Believe it or not, but the block still could be polymerizing even though it is firm to the touch after the waterbath step. Also, this oven step helps to drastically reduce the MMA smell when you go to break free from the mold and handle the block. Large tissue polymerization is really something that you cannot predict routinely because of all the variables involved. However, if you can manage catalyst concentration and your room temperature environment, the time estimation of polymerization for specific tissues (size/density and volume of solution needed) becomes easier to calculate and repeat successfully. Try this out and please let me know if you need any clarification. Jack Ratliff > Date: Fri, 5 Dec 2008 15:14:07 -0500> From: PMonfils@Lifespan.org> To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] Polymerizing BIG methyl methacrylate blocks> > Does anyone here have experience with large PMMA blocks, about 150 to 200 ml volume each? Specifically, at what temperature do you recommend polymerizing them, and how long should the polymerization take? Any other helpful hints will be appreciated.> > Paul M.> _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008 From gayle.callis <@t> bresnan.net Fri Dec 5 17:11:42 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Dec 5 17:11:43 2008 Subject: [Histonet] Polymerizing large MMA blocks Message-ID: Paul, I learned this trick from several of my expert bonehead colleagues who worked with some huge bone blocks in MMA. Be prepared to spend time doing this however. It is a called the "layering" technic, but it prevents an explosion of bubbles when the MMA suddenly decides to polymerize. You can use prepolymerized layers, and we used Rubbermaid square plastic containers with snap top lids or Nalgene very wide mouth bottles/jars that have external threads for the white lid. Making prepolymerized layers is a good way to reuse your last MMA infiltration mixture waste. You can make the layers after embedding. It is important at any time to put a lid on top of your containers when making prepolymerized layers or when performing the layering technic. You must maintain the proper mixture balance or the monomer evaporates, leaving behind a seemingly properly polymerized plastic which in fact is not the case. You will end up with an ugly depressed surface that is funky looking, not a good thing to have happen and lousy as flat surface on which to lay your sample. Layering Embedding: Add approximately a half inch a to 3/4 of an inch of embedding mixture to the prepolymerized layer, place bone on top of this, and seal with lid. Stack inside a hood at RT. This embedding mixture should be polymerized before you add another layer of embedding mixture, so it may take several days to layer, polymerize, layer, polymerize, etc until you are finished. Buy a cheap metal cooking spoon or use a wooden tongue depressor to scoop i.e. baste MMA over the top of bone. Replace the lid, and be patient. The bone sometimes turns white due to exposure to the air or whatever but this is prevented by the described "basting" the bone a several times, daily or more - to coat it with the MMA embedding mixture. MMA in large quantities is touchy and really dislikes extra heat. Polymerization will slow if put in refrigerator (not good due to no fume control). Don't worry about adding non-polymerized plastic to the prepolymerized layer, as there is a smooth interface caused by the monomer dissolving a few mm of the pre- polymermized MMA. This is ideal since it forms a very tight bond after polymerization. You will see the differences in the layers as they polymerize, but if you add heat, a horrible mass of polymerized bubbles occurs, which means you have to grind the bubbles away and reembed the infiltrated bone. Been there, done that, and had to close a door due to inappropriate word usage shocking to my fellow workers. Your embedding mixture does not have to be made up daily, and make up no more than 500 mls at a time, but store in a tightly sealed bottle, in the refrigerator, and use it within 7 days. Always warm the MMA to RT before adding to the embedded bone - you must avoid water condensation contamination. We have blocks where some of the bone sticks out of the top of block, and it is coated with MMA, and without sample turning white. The block is perfecty clear, like a museum display piece. The screw top lid was the best method to prevent spills. I stacked containers up in the hood (always left ON) for both bone embedding/polymerization and making prepolymerized layers. After you are finished with embedding, make sure the top layer is above the bone unless you don't need that part of sample, and put the block in 60C oven for an hour or so to cure. You will smell the toxic fumes, so be very careful - the refrig should be vented into a hood or in the hood. Avoid a slightly tacky, sticky top block going into 60C - it will bubble at that part of layer. We did a baby alligator head along with some other large bovine and goat bone, and it took close to a month or longer, but we were ultimately very patient and had perfectly clear embedded sample. There is no STAT work when dealing with large bone in MMA. My favorite MMA mixture came from Diane Sterchi, and I never had problems after I started using her mixture and the layering method. I would be happy to send that procedure to you if you like. Her mixture contained prepolymerized PMMA powder which helped the polymerization get started. I will be off Histonet after this message, so respond to my personal email address Gayle M. Callis HTL(ASCP)HT, MT From tifei <@t> foxmail.com Fri Dec 5 22:00:39 2008 From: tifei <@t> foxmail.com (tf) Date: Fri Dec 5 22:01:03 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed References: <200812051111205608471@foxmail.com>, , <4938FDAE020000C500048F27@nwcl02.tsrh.org> Message-ID: <200812061200339184559@foxmail.com> eW91IHdhbnQgdG8gY2FycnkgYSBib3R0bGUgb2YgdG94aWMgbGlxdWlkIG9uIHlvdXIgY2FyPyBv ciB5b3Ugd2lsbCB0YWtlIGEgYm94IG9mIHBvd2RlciB0aGF0IGNhbiBkaXNzb2x2ZSBpbnRvIHVz ZWZ1bCBzb2x1dGlvbiBlYXNpbHk/DQoNCllvdSBoYXZlIHRvIGFkZCBtZXRoYW5vbCBpbiAxMCUg Zm9ybWFsaW4gJiA0JSBmb3JtYWxkZWh5ZGUsIHJhdGhlciA0JSBwYXJhZm9ybWFsZGh5ZGUuLi4u UEZBIGlzIG1ldGhhbm9sIGZyZWUuLml0J3MgdmVyeSBpbXBvcnRhbnQuDQoNCg0KMjAwOC0xMi0w NiANCg0KDQoNCnRmIA0KDQoNCg0Kt6K8/sjLo7ogUmV1ZWwgQ29ybmVsaWEgDQq3osvNyrG85KO6 IDIwMDgtMTItMDYgIDAwOjA5OjM2IA0KytW8/sjLo7ogVG9ueSBIZW53b29kOyB0aWZlaUBmb3ht YWlsLmNvbTsgaGlzdG9uZXQ7IGFuaDIwMDZAbWVkLmNvcm5lbGwuZWR1OyBKYW4gU2hpdmVycyAN 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KioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKiANCg== From kemlo <@t> f2s.com Sat Dec 6 05:02:40 2008 From: kemlo <@t> f2s.com (kemlo) Date: Sat Dec 6 05:03:04 2008 Subject: [Histonet] fixation post-fixation, difference of order used In-Reply-To: <62915.136.159.150.24.1228498173.squirrel@136.159.150.24> References: <62915.136.159.150.24.1228498173.squirrel@136.159.150.24> Message-ID: <573C34C9A31540919CC0A149A64B96F2@KemloPC> Interesting question..... Post fixation is sometimes carried out to enhance staining. I the old days we fixed in formalin, then in mercury. If you fixed in mercury from the start then the tissue went hard whilst you could leave tissue in formalin for ages; you can therefore control the exposure time in mercury more accurately. Logically the order of fixation and the types of fixative used must make a difference; if you initially fixed in mercury too long then in formalin you'd end up with hard tissue. Fixatives are there to make the tissue ready for processing, so fixatives may also make the tissue ready for being fixed. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of ejschmid@ucalgary.ca Sent: 05 December 2008 17:30 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixation post-fixation, difference of order used Hi, I was wondering about the effects of using different fixatives in different orders. For example, some of our embryonic material (chick and mouse) is fixed initially with 4% PFA and subsequently is exposed to glutaraldehyde. Chick embryos subject to this are successfully labeled using IHC for anti-HH3. However, our mouse tissues are fixed differently, being first exposed to 4% PFA: 1% Glut solution (Which i think is called a modified Karnovsky's fix). These embryos perform poorly for IHC with anti-HH3. Might it be the glutaraldehyde that is present in our primary fixative used to fix moue embryos? Might it be that the IHC works for chick because the glutaraldehyde is only used as a secondary fix after primary fixation in 4% PFA? Does the order of exposure matter? thanks! eric schmidt University of Calgary Medical Sciences _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ejschmid <@t> ucalgary.ca Sat Dec 6 14:30:14 2008 From: ejschmid <@t> ucalgary.ca (ejschmid@ucalgary.ca) Date: Sat Dec 6 14:30:24 2008 Subject: [Histonet] PFA preparation Message-ID: <62735.136.159.150.24.1228595414.squirrel@136.159.150.24> Hi, So then what is the best way to prepare formaldehyde fixative from PFA? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. I've also been taught that too much heat during preparation "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it "degrades." What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences From ejschmid <@t> ucalgary.ca Sat Dec 6 14:32:51 2008 From: ejschmid <@t> ucalgary.ca (ejschmid@ucalgary.ca) Date: Sat Dec 6 14:32:59 2008 Subject: [Histonet] divalent ions in fix Message-ID: <62761.136.159.150.24.1228595571.squirrel@136.159.150.24> Hi, What is the function of divalent ions in fix? It's been suggested to me that by using Dulbeccos PBS to make formaldehyde fix from PFA will better preserve tissues, or at least better preserve proteins. Eric Schmidt University of Calgary Medical Sciences From plucas <@t> biopath.org Sat Dec 6 15:45:31 2008 From: plucas <@t> biopath.org (Paula Lucas) Date: Sat Dec 6 15:45:36 2008 Subject: [Histonet] Cellient Cell Block System Message-ID: <20081206214531.A39FB27C0@courageux.cnchost.com> Hello, Can anyone tell me their experiences with the Hologic's Cellient Cell Block System? I appreciate any thoughts and comments about the system you have. Paula Lucas Lab Manager Bio-Path Medical Group Fountain Valley, CA From rjbuesa <@t> yahoo.com Sat Dec 6 15:49:32 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Dec 6 15:49:35 2008 Subject: [Histonet] PFA preparation In-Reply-To: <62735.136.159.150.24.1228595414.squirrel@136.159.150.24> Message-ID: <276579.63839.qm@web65710.mail.ac4.yahoo.com> You were taught correctly. And the socalled "degradation" is re-polymerization as you wrote. The only thing is that you should add buffer salts bettwen than PBS. Ren? J. --- On Sat, 12/6/08, ejschmid@ucalgary.ca wrote: From: ejschmid@ucalgary.ca Subject: [Histonet] PFA preparation To: histonet@lists.utsouthwestern.edu Date: Saturday, December 6, 2008, 3:30 PM Hi, So then what is the best way to prepare formaldehyde fixative from PFA? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. I've also been taught that too much heat during preparation "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it "degrades." What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Sat Dec 6 15:51:43 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Dec 6 15:51:48 2008 Subject: [Histonet] divalent ions in fix In-Reply-To: <62761.136.159.150.24.1228595571.squirrel@136.159.150.24> Message-ID: <832499.66371.qm@web65714.mail.ac4.yahoo.com> During time, besides the re-polymerization, methanal (= formaldehyde) turns into methanoic (acid) and the buffer salts prevent that acidification. Ren? J. --- On Sat, 12/6/08, ejschmid@ucalgary.ca wrote: From: ejschmid@ucalgary.ca Subject: [Histonet] divalent ions in fix To: histonet@lists.utsouthwestern.edu Date: Saturday, December 6, 2008, 3:32 PM Hi, What is the function of divalent ions in fix? It's been suggested to me that by using Dulbeccos PBS to make formaldehyde fix from PFA will better preserve tissues, or at least better preserve proteins. Eric Schmidt University of Calgary Medical Sciences _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stamptrain <@t> yahoo.com Sat Dec 6 16:48:52 2008 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Sat Dec 6 16:48:56 2008 Subject: [Histonet] PFA preparation References: <276579.63839.qm@web65710.mail.ac4.yahoo.com> Message-ID: <534399.86703.qm@web55808.mail.re3.yahoo.com> I always prepared PFA as at least an 8% solution so that dilution with buffer would give me the final working %.? The protocol was to add the PFA powder to water at 50C and then add 10N NaOH dropwise until the solution cleared.? After cooling to RT, the stock PFA solution was filtered to remove any remaining insoluble particulates.? For a 4% PFA working fixative,?mix the 8% 1:1 with a 2x buffer.? Osmolarity and buffer strength are correct in the final fixative.? Similarly, by adding dH2O one can easily formulate lower conecentrations of formaldehyde in the fixative.? Worked for years in an EM Lab where our standard fixative was 2% PFA and 1% glutaraldehyde. Roger Moretz ----- Original Message ---- From: Rene J Buesa To: histonet@lists.utsouthwestern.edu; ejschmid@ucalgary.ca Sent: Saturday, December 6, 2008 4:49:32 PM Subject: Re: [Histonet] PFA preparation You were taught correctly. And the socalled "degradation" is re-polymerization as you wrote. The only thing is that you should add buffer salts bettwen than PBS. Ren? J. --- On Sat, 12/6/08, ejschmid@ucalgary.ca wrote: From: ejschmid@ucalgary.ca Subject: [Histonet] PFA preparation To: histonet@lists.utsouthwestern.edu Date: Saturday, December 6, 2008, 3:30 PM Hi, ? So then what is the best way to prepare formaldehyde fixative from PFA? ? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. ? I've also been taught that too much heat during preparation "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it "degrades." ? What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences _______________________________________________ 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 AnthonyH <@t> chw.edu.au Sun Dec 7 16:05:41 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Dec 7 16:06:05 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed In-Reply-To: <200812061200339184559@foxmail.com> Message-ID: Tf, In answer to you email: =20 No I do not carry toxic liquid in my car. But does the PFA powder dissolve easily? My experience is that you need to = make the solution alkaline then heat it. I never add methanol to my 10% neutral buffered formalin (it is buffered an= d diluted ie 10%). The risk of polymerisation of the formalin (since it is = diluted) and formic acid formation (since it is buffered) is greatly reduce= d. =20 Regards=20 Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20 Laboratory Manager & Senior Scientist=20 Tel: 612 9845 3306=20 Fax: 612 9845 3318=20 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: tf [mailto:tifei@foxmail.com]=20 Sent: Saturday, 6 December 2008 3:01 PM To: Reuel Cornelia; Tony Henwood; histonet; anh2006@med.cornell.edu; Jan S= hivers Subject: Re: RE: RE: [Histonet] IHC on paraformaldehyde-fixed =09 =09 you want to carry a bottle of toxic liquid on your car? or you will take a= box of powder that can dissolve into useful solution easily? =20 You have to add methanol in 10% formalin & 4% formaldehyde, rather 4% para= formaldhyde....PFA is methanol free..it's very important. =20 =20 2008-12-06=20 =09 ________________________________ tf=20 =09 ________________________________ =B7=A2=BC=FE=C8=CB=A3=BA Reuel Cornelia=20 =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-06 00:09:36=20 =CA=D5=BC=FE=C8=CB=A3=BA Tony Henwood; tifei@foxmail.com; histonet; anh200= 6@med.cornell.edu; Jan Shivers=20 =B3=AD=CB=CD=A3=BA=20 =D6=F7=CC=E2=A3=BA RE: RE: [Histonet] IHC on paraformaldehyde-fixed=20 =09 =09 I have been curious about this discussion. we used 4% paraformaldehyde for smaller biopsies only because it has a faster penetration to tissue than 10% formalin. In all my IHC that I have done. I observe that doing an IHC with 4% paraformaldehyde does not necessarily need antigen retrieval in comparison to 10% formalin either it will be human or animal tissue but this depends on how long was it fix, our 4% paraformaldehyde we fix smaller biopsies like nerve,muscle, skin for 6 to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe you can comment on the effect on this to tissue if you say you will use 4% paraformaldehyde for storage and transportation.=20 Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 >>> "Tony Henwood" 12/04/08 9:29 PM >>> tf wrote: =20 "I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~." =20 I have not heard this before. Do you have a reference for this? =20 =20 Regards=20 Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20 Laboratory Manager & Senior Scientist=20 Tel: 612 9845 3306=20 Fax: 612 9845 3318=20 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: tf [mailto:tifei@foxmail.com]=20 Sent: Friday, 5 December 2008 2:11 PM To: Tony Henwood; anh2006@med.cornell.edu; Jan Shivers; histonet Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed the basic principles are the same for most cross-linking fixatives and induce similar bonds=20 the difference you observed between may due to any other variability, or the co-fixative you used. =20 I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~. =20 =20 =20 =20 2008-12-05=20 ________________________________ tf=20 ________________________________ =B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood=20 =B7=A2=CB=CD=BC=E4=A3=BA 2008-12-05 06:00:03=20 =CA=D5=BC=FE=C8=CB=A3=BA anh2006@med.cornell.edu; Jan Shivers; histonet=20 =B3=AD=CB=CD=A3=BA=20 =D6=F7=A3=BA RE: [Histonet] IHC on paraformaldehyde-fixed=20 Interesting point. Since 10% buffered formalin (made from the concentrated 38% formaldehyde) contain about 1% methanol, has it been shown that this has a deleterious effect on ANY antigens or are we expecting this worse case senario as being the norm? I am not aware of any antigens (or antigen-antibody combination) that has been badly effected by 10% formalin that is NOT effected by 10% formaldehyde. Are you aware of any?? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu]=20 Sent: Friday, 5 December 2008 1:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09=20 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin?=20 No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu=20 [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 **************************************************************************= *****************************************=20 Texas Scottish Rite Hospital for Children is one of the nation's leading p= ediatric centers for the=20 treatment of orthopedic conditions, certain related neurological disorders= and learning disorders,=20 such as dyslexia. This email transmission and/or its attachments may cont= ain confidential health=20 information, intended only for the use of the individual or entity named a= bove.=20 The authorized recipient of this information is prohibited from disclosing= it to any other party unless=20 required to do so by law and is required to delete/destroy the information= after its stated need has been fulfilled. If you are not the intended recipient, any disclosure, co= pying, distribution or action=20 taken in reliance on the contents of this email transmission is prohibited= . If you have received this information in error, please notify the sender immediately and delete this= information.=20 We appreciate your efforts to protect the children's confidential informat= ion.=20 **************************************************************************= *****************************************=20 ********************************************************************* 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 send= er. Views expressed in this message and any attachments are those of the indivi= dual sender, and are not necessarily the views of The Children's Hospital a= t Westmead This note 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 res= ulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Sun Dec 7 16:11:50 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Dec 7 16:11:59 2008 Subject: [Histonet] PFA preparation In-Reply-To: <62735.136.159.150.24.1228595414.squirrel@136.159.150.24> Message-ID: My experience is that when you add paraformaldehyde to water all it forms is a colloidal solution (ie on standing, the paraformaldehyde settles with very little going into solution (personal experience, waited one week, then gave up). Has your experience been different? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of ejschmid@ucalgary.ca Sent: Sunday, 7 December 2008 7:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PFA preparation Hi, So then what is the best way to prepare formaldehyde fixative from PFA? The way I have been taught, which differs from what I have read, is to dissolve 4% into ddH2O at room temperature. After that one could add PBS or buffer. I've also been taught that too much heat during preparation "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored too long will lose freshness because it "degrades." What I read is different. Texts suggest to dissolve the PFA in warm water, and that aging of the fix is due to repolymerization, not through degradation. What should i do? Eric Schmidt University of Calgary Medical Sciences _______________________________________________ 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 note 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 ctanck1 <@t> gmail.com Sun Dec 7 18:11:49 2008 From: ctanck1 <@t> gmail.com (Carol Tanck) Date: Sun Dec 7 18:11:54 2008 Subject: [Histonet] Re:Don't think I'm nuts Message-ID: <2e21cfdc0812071611s4f55fa2dg78044e91a3dbce65@mail.gmail.com> If the technicians involved are female, it could be hormonal and a coincidence they are in sync. From Kemlo.Rogerson <@t> waht.swest.nhs.uk Mon Dec 8 06:15:07 2008 From: Kemlo.Rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Dec 8 06:15:19 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed Message-ID: <86ADE4EB583CE64799A9924684A0FBBF05CD7CA9@wahtntex2.waht.swest.nhs.uk> Isn't petrol toxic?=20 =20 Kemlo Rogerson =20 Pathology Manager DD 01934 647057 or extension 3311 Mob 07749 754194; Pager 07659 597107; Don=A1=AFt be afraid to take a big step when one is indicated. You = can=A1=AFt cross a chasm in two small jumps. --Buckminster Fuller=20 This e-mail is confidential and privileged. If you are not the intended = recipient please accept my 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 me that this message has gone astray before deleting it. = Thank you for your co-operation =20 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu = [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tony = Henwood Sent: 07 December 2008 22:06 To: tifei@foxmail.com; Reuel Cornelia; histonet; = anh2006@med.cornell.edu; Jan Shivers Subject: RE: RE: RE: [Histonet] IHC on paraformaldehyde-fixed Tf, In answer to you email: =20 No I do not carry toxic liquid in my car. But does the PFA powder dissolve easily? My experience is that you need = to make the solution alkaline then heat it. I never add methanol to my 10% neutral buffered formalin (it is buffered = and diluted ie 10%). The risk of polymerisation of the formalin (since = it is diluted) and formic acid formation (since it is buffered) is = greatly reduced. =20 Regards=20 Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory = Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, = Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: tf [mailto:tifei@foxmail.com]=20 Sent: Saturday, 6 December 2008 3:01 PM To: Reuel Cornelia; Tony Henwood; histonet; anh2006@med.cornell.edu; = Jan Shivers Subject: Re: RE: RE: [Histonet] IHC on paraformaldehyde-fixed =09 =09 you want to carry a bottle of toxic liquid on your car? or you will = take a box of powder that can dissolve into useful solution easily? =20 You have to add methanol in 10% formalin & 4% formaldehyde, rather 4% = paraformaldhyde....PFA is methanol free..it's very important. =20 =20 2008-12-06=20 =09 ________________________________ tf=20 =09 ________________________________ =B7=A2=BC=FE=C8=CB=A3=BA Reuel Cornelia=20 =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-06 00:09:36=20 =CA=D5=BC=FE=C8=CB=A3=BA Tony Henwood; tifei@foxmail.com; histonet; = anh2006@med.cornell.edu; Jan Shivers=20 =B3=AD=CB=CD=A3=BA=20 =D6=F7=CC=E2=A3=BA RE: RE: [Histonet] IHC on paraformaldehyde-fixed=20 =09 =09 I have been curious about this discussion. we used 4% paraformaldehyde for smaller biopsies only because it has a faster penetration to tissue than 10% formalin. In all my IHC that I have done. I observe that doing an IHC with 4% paraformaldehyde does not necessarily need antigen retrieval in comparison to 10% formalin either it will be human or animal tissue but this depends on how long was it fix, our 4% paraformaldehyde we fix smaller biopsies like nerve,muscle, skin for 6 to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe you can comment on the effect on this to tissue if you say you will use 4% paraformaldehyde for storage and transportation.=20 Reuel Cornelia, BS MT, AMT Cellular Pathology Texas Scottish Rite Hospital for Children 2222 Welborn Street Dallas, TX 75219 Tel: 214-559-7766 fax: 214-559-7768 >>> "Tony Henwood" 12/04/08 9:29 PM >>> tf wrote: =20 "I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~." =20 I have not heard this before. Do you have a reference for this? =20 =20 Regards=20 Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20 Laboratory Manager & Senior Scientist=20 Tel: 612 9845 3306=20 Fax: 612 9845 3318=20 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: tf [mailto:tifei@foxmail.com]=20 Sent: Friday, 5 December 2008 2:11 PM To: Tony Henwood; anh2006@med.cornell.edu; Jan Shivers; histonet Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed the basic principles are the same for most cross-linking fixatives and induce similar bonds=20 the difference you observed between may due to any other variability, or the co-fixative you used. =20 I DO believe that one reason some people use 4% PFA rather 10% formalin is that PFA is a bit more stable, both for storage and transportation~~~. =20 =20 =20 =20 2008-12-05=20 ________________________________ tf=20 ________________________________ =B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood=20 =B7=A2=CB=CD=BC=E4=A3=BA 2008-12-05 06:00:03=20 =CA=D5=BC=FE=C8=CB=A3=BA anh2006@med.cornell.edu; Jan Shivers; histonet = =B3=AD=CB=CD=A3=BA=20 =D6=F7=A3=BA RE: [Histonet] IHC on paraformaldehyde-fixed=20 Interesting point. Since 10% buffered formalin (made from the concentrated 38% formaldehyde) contain about 1% methanol, has it been shown that this has a deleterious effect on ANY antigens or are we expecting this worse case senario as being the norm? I am not aware of any antigens (or antigen-antibody combination) that has been badly effected by 10% formalin that is NOT effected by 10% formaldehyde. Are you aware of any?? Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu]=20 Sent: Friday, 5 December 2008 1:31 AM To: Tony Henwood; Jan Shivers; histonet Subject: Re: [Histonet] IHC on paraformaldehyde-fixed So true. However, be aware that 10% neutral buffered formalin we use has methanol in it which may affect certain antigens so there may be some difference in staining (hence why for mouse work we now only use 4% PFA in pure PBS). It is good to be aware of the other ingredients in your fixative solutions, whether commercially prepared or a homemaede recipe, as it isn't only the formaldehyde fixative which can make a difference. -----Original Message----- From: Tony Henwood Date: Thu, 04 Dec 2008 09:35:09=20 To: Jan Shivers; histonet Subject: RE: [Histonet] IHC on paraformaldehyde-fixed Gee I hate the term paraformaldehyde (as many of you probably know) This is an example of how confusion of terms can cause unnecessary work. Is "4% paraformaldehyde" different from 4 % formaldehyde? No Should any procedure done to tissues fixed in "4% paraformaldehyde" give results different to those fixed in 4% formaldehyde or 10% formalin?=20 No since they are the same thing. As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) state when paraformaldehyde actually becomes a fixative, it is no longer paraformaldehyde by chemistry or fixation capacity. Rather, it is formaldehyde in water without methanol or any other stabiliser. Without heat and an alkaline environment, paraformaldehyde in water is simply a paraformaldehyde suspension with little fixation capacity. If the fixative is prepared from paraformaldehyde then it should be termed 4% formaldehyde freshly prepared from paraformaldehyde. If a concentrated formalin solution (40% formaldehyde) is used, then it should be termed 10% formalin. If you do a search on Histonet for paraformaldehye, you will find that this topic has been extensively discussed. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu=20 [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jan Shivers Sent: Thursday, 4 December 2008 8:34 AM To: histonet Subject: [Histonet] IHC on paraformaldehyde-fixed Has anyone ever done IHC on parafomaldehyde-fixed tissues, and if so, how well did it work? Will the same antigen-retrieval methods used with formalin-fixed tissue be applicable? I'm asking for an investigator, who already has his tissues fixed in paraformaldehyde. Jan Shivers Senior Scientist Pathology Teaching Program Histology/IHC/EM Section Head University of Minnesota Veterinary Diagnostic Laboratory 1333 Gortner Ave. St. Paul, MN 55108 612-624-7297 shive003@umn.edu_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu=20 http://lists.utsouthwestern.edu/mailman/listinfo/histonet=20 = *************************************************************************= ******************************************=20 Texas Scottish Rite Hospital for Children is one of the nation's = leading pediatric centers for the=20 treatment of orthopedic conditions, certain related neurological = disorders and learning disorders,=20 such as dyslexia. This email transmission and/or its attachments may = contain confidential health=20 information, intended only for the use of the individual or entity = named above.=20 The authorized recipient of this information is prohibited from = disclosing it to any other party unless=20 required to do so by law and is required to delete/destroy the = information after its stated need has been fulfilled. If you are not the intended recipient, any disclosure, = copying, distribution or action=20 taken in reliance on the contents of this email transmission is = prohibited. If you have received this information in error, please notify the sender immediately and delete = this information.=20 We appreciate your efforts to protect the children's confidential = information.=20 = *************************************************************************= ******************************************=20 ********************************************************************* 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From aevans <@t> wellspan.org Mon Dec 8 06:50:36 2008 From: aevans <@t> wellspan.org (Evans, Andria B.) Date: Mon Dec 8 07:04:15 2008 Subject: [Histonet] microwave processor Message-ID: We are currently looking at getting another microwave processor. We currently have the Histos 5 and are looking at getting another one of those or a Pathos (the big guy from milestone). But we have little information on the Pathos. I have a couple questions that maybe someone could answer.... Can you run more then one run at a time on the Pathos? (currently we can run 2 runs at a time on the Histos 5) Does the Pathos do the everything on the machine from fixation, to alcohol rinses, to the end product?? (We have to do all the rinses and changes manually with the Histos 5) If anyone has any other information on the microwave processors that they would like to share that would be great!! Thanks Andria B Evans, HTL(ASCP)CM CONFIDENTIALITY NOTICE: This email may contain confidential health information that is legally privileged. This information is intended for the use of the named recipient(s). The authorized recipient of this information is prohibited from disclosing this information to any party unless required to do so by law or regulation and is required to destroy the information after its stated need has been fulfilled. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or action taken in reliance on the contents of this email is strictly prohibited. If you receive this e-mail message in error, please notify the sender immediately to arrange disposition of the information. ______________________________________________________________________ This e-mail has been scanned by MCI Managed Email Content Service, using Skeptic(tm) technology powered by MessageLabs. For more information on MCI's Managed Email Content Service, visit http://www.mci.com. ______________________________________________________________________ From MMargiotta <@t> bmhmc.org Mon Dec 8 08:27:38 2008 From: MMargiotta <@t> bmhmc.org (Margiotta, Michele) Date: Mon Dec 8 08:27:42 2008 Subject: [Histonet] thermometer calibration Message-ID: Hi All, Just wondering how everyone is doing their thermometer calibrations. Inspection time is right around the corner so I want to be ready! Thanks. Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. From micropathlabs <@t> yahoo.com Mon Dec 8 08:42:16 2008 From: micropathlabs <@t> yahoo.com (Sheila Haas) Date: Mon Dec 8 08:42:19 2008 Subject: [Histonet] Marketing Position Message-ID: <97031.8856.qm@web57804.mail.re3.yahoo.com> We are currently recruiting for a marketing position?in our organization. We are an?Anatomic Pathology?reference lab?located in Lakeland, Florida and are looking for someone with?both lab and marketing experience. If you are interested or?know someone who is, please contact me?toll-free at 866-961-2785. Thank you. ? Sheila Haas Laboratory Supervisor Micro Path Laboratories From kalschev <@t> svm.vetmed.wisc.edu Mon Dec 8 09:30:16 2008 From: kalschev <@t> svm.vetmed.wisc.edu (Vicki Kalscheur) Date: Mon Dec 8 09:30:24 2008 Subject: [Histonet] Paul Monfils re: PMMA blocks Message-ID: <000801c95949$dec44860$c5d76880@vetmed.wisc.edu> Paul, Rather than add to Jack and Gail's experience and expertise, feel free to call me if you need additional hints. I have no technical problems with large PMMA blocks. One hint is to use Pyrex beakers or crystallizing dishes, although expensive, the results are consistent. Best regards, Vicki Kalscheur Department of Surgical Sciences School of Veterinary Medicine University of Wisconsin 2015 Linden Drive Madison, WI 53706-1102 Phone: 608-262-8534 FAX: 608-263-7930 kalschev@svm.vetmed.wisc.edu From leiker <@t> buffalo.edu Mon Dec 8 10:01:23 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Mon Dec 8 10:01:28 2008 Subject: [Histonet] PFA preparation In-Reply-To: References: Message-ID: <681289C0ED95575D3DD453FB@bchwxp2702.ad.med.buffalo.edu> We routinely add paraformaldehyde to alkaline water at room temperature while stirring and wait only about 30-60 mintues for it to dissolve. Then we add a concentrated amount of PBS up to the total required volume (so that the buffer is 1x in the final volume). Then we add acid to bring the pH back down to 7. Then we filter it since not all of the PFA has dissolved (though most of it has). Merced --On Monday, December 08, 2008 9:11 AM +1100 Tony Henwood wrote: > My experience is that when you add paraformaldehyde to water all it > forms is a colloidal solution (ie on standing, the paraformaldehyde > settles with very little going into solution (personal experience, > waited one week, then gave up). > > Has your experience been different? > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > ejschmid@ucalgary.ca > Sent: Sunday, 7 December 2008 7:30 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] PFA preparation > > > Hi, > > So then what is the best way to prepare formaldehyde fixative from > PFA? > > The way I have been taught, which differs from what I have read, is > to dissolve 4% into ddH2O at room temperature. After that one could add > PBS or buffer. > > I've also been taught that too much heat during preparation > "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored > too long will lose freshness because it "degrades." > > What I read is different. Texts suggest to dissolve the PFA in warm > water, and that aging of the fix is due to repolymerization, not through > degradation. > > What should i do? > > Eric Schmidt > > University of Calgary > Medical Sciences > > > > > _______________________________________________ > 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 note 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. > ********************************************************************** > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From ejschmid <@t> ucalgary.ca Mon Dec 8 10:19:35 2008 From: ejschmid <@t> ucalgary.ca (ejschmid@ucalgary.ca) Date: Mon Dec 8 10:19:54 2008 Subject: [Histonet] PFA preparation: Does dissolution also mean that the PFA has depolymerized? In-Reply-To: <681289C0ED95575D3DD453FB@bchwxp2702.ad.med.buffalo.edu> References: <681289C0ED95575D3DD453FB@bchwxp2702.ad.med.buffalo.edu> Message-ID: <63320.136.159.150.24.1228753175.squirrel@136.159.150.24> Hi Merced, Thanks for your reply. So then you manipulate pH rather than temperature to ensure the dissolution of PFA. Does dissolution also mean that the PFA has depolymerized? Are you saying heating to 60 degrees C is not required, or that it is not recommended? I've been told that heating 'degrades' the PFA and one wants to avoid this. But according to other sources, the heating step is required to ensure that the PFA does in indeed degrade, degrade into formaldehyde. Eric University of Calgary Medical Sciences > We routinely add paraformaldehyde to alkaline water at room temperature > while stirring and wait only about 30-60 mintues for it to dissolve. Then > we add a concentrated amount of PBS up to the total required volume (so > that the buffer is 1x in the final volume). Then we add acid to bring the > pH back down to 7. Then we filter it since not all of the PFA has > dissolved (though most of it has). > > Merced > > --On Monday, December 08, 2008 9:11 AM +1100 Tony Henwood > wrote: > >> My experience is that when you add paraformaldehyde to water all it >> forms is a colloidal solution (ie on standing, the paraformaldehyde >> settles with very little going into solution (personal experience, >> waited one week, then gave up). >> >> Has your experience been different? >> >> Regards >> >> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) >> Laboratory Manager & Senior Scientist >> Tel: 612 9845 3306 >> Fax: 612 9845 3318 >> the children's hospital at westmead >> Cnr Hawkesbury Road and Hainsworth Street, Westmead >> Locked Bag 4001, Westmead NSW 2145, AUSTRALIA >> >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> ejschmid@ucalgary.ca >> Sent: Sunday, 7 December 2008 7:30 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] PFA preparation >> >> >> Hi, >> >> So then what is the best way to prepare formaldehyde fixative from >> PFA? >> >> The way I have been taught, which differs from what I have read, is >> to dissolve 4% into ddH2O at room temperature. After that one could add >> PBS or buffer. >> >> I've also been taught that too much heat during preparation >> "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored >> too long will lose freshness because it "degrades." >> >> What I read is different. Texts suggest to dissolve the PFA in warm >> water, and that aging of the fix is due to repolymerization, not >> through >> degradation. >> >> What should i do? >> >> Eric Schmidt >> >> University of Calgary >> Medical Sciences >> >> >> >> >> _______________________________________________ >> 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 note 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. >> ********************************************************************** >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > From Kathleen.Caleri <@t> RoswellPark.org Mon Dec 8 10:27:12 2008 From: Kathleen.Caleri <@t> RoswellPark.org (Caleri, Kathleen) Date: Mon Dec 8 10:27:17 2008 Subject: [Histonet] processing times Message-ID: <97101976F8A044468CA74FE11883B90E29B9EAEF@VISTA.roswellpark.org> Hello-I have just started at Roswell Park Cancer Institute and we are hoping to decrease our TAT by decreasing our processing times-having come from a facility where we hadn't changed processor times in 20 years I would appreciate it if someone could supply their schedules so I could do a comparison...we currently do a short run for bx's (5 hr), a standard surgical run (13 hr) , and a longer fatty/breast run (16 hr) Regards, Kate Kathleen Caleri, BS, MLT, HT (ASCP) Histology Lab Supervisor Pathology Roswell Park Cancer Institute (716) 845-1329 "the future of mankind lies waiting for those who come to understand their lives and take up their responsibilities to help others" This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. From histonetalias <@t> gmail.com Mon Dec 8 10:48:44 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Mon Dec 8 10:50:02 2008 Subject: [Histonet] microwave processor In-Reply-To: References: Message-ID: <4b6c85510812080848i7a4ce45eq22e78776bd88a24b@mail.gmail.com> The Pathos can only run one run at a time. It does everything a conventional processor does. It is fully automated. On Mon, Dec 8, 2008 at 7:50 AM, Evans, Andria B. wrote: > We are currently looking at getting another microwave processor. We > currently have the Histos 5 and are looking at getting another one of those > or a Pathos (the big guy from milestone). But we have little information on > the Pathos. I have a couple questions that maybe someone could answer.... > > Can you run more then one run at a time on the Pathos? (currently we can > run 2 runs at a time on the Histos 5) > Does the Pathos do the everything on the machine from fixation, to alcohol > rinses, to the end product?? (We have to do all the rinses and changes > manually with the Histos 5) > > If anyone has any other information on the microwave processors that they > would like to share that would be great!! > > Thanks > > Andria B Evans, HTL(ASCP)CM > > > CONFIDENTIALITY NOTICE: > > This email may contain confidential health information that is legally > privileged. This information is intended for the use of the named > recipient(s). The authorized recipient of this information is prohibited > from disclosing this information to any party unless required to do so by > law or regulation and is required to destroy the information after its > stated need has been fulfilled. If you are not the intended recipient, you > are hereby notified that any disclosure, copying, distribution, or action > taken in reliance on the contents of this email is strictly prohibited. If > you receive this e-mail message in error, please notify the sender > immediately to arrange disposition of the information. > > > ______________________________________________________________________ > This e-mail has been scanned by MCI Managed Email Content Service, using > Skeptic(tm) technology powered by MessageLabs. For more information on MCI's > Managed Email Content Service, visit http://www.mci.com. > ______________________________________________________________________ > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States From leiker <@t> buffalo.edu Mon Dec 8 11:14:42 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Mon Dec 8 11:14:48 2008 Subject: [Histonet] PFA preparation: Does dissolution also mean that the PFA has depolymerized? In-Reply-To: <63320.136.159.150.24.1228753175.squirrel@136.159.150.24> References: <681289C0ED95575D3DD453FB@bchwxp2702.ad.med.buffalo.edu> <63320.136.159.150.24.1228753175.squirrel@136.159.150.24> Message-ID: Yes, we would say that heating is not necessary to depolymerize PFA. Only the high pH is. After 30-60 minutes of stirring we obtain a crystal clear solution with only a few particles of undissolved PFA swimming around at the bottom (which is why we filter). --On Monday, December 08, 2008 9:19 AM -0700 ejschmid@ucalgary.ca wrote: > Hi Merced, > > Thanks for your reply. So then you manipulate pH rather than temperature > to ensure the dissolution of PFA. Does dissolution also mean that the PFA > has depolymerized? Are you saying heating to 60 degrees C is not required, > or that it is not recommended? I've been told that heating 'degrades' the > PFA and one wants to avoid this. But according to other sources, the > heating step is required to ensure that the PFA does in indeed degrade, > degrade into formaldehyde. > > Eric > > University of Calgary > Medical Sciences > > >> > We routinely add paraformaldehyde to alkaline water at room temperature >> while stirring and wait only about 30-60 mintues for it to dissolve. >> Then we add a concentrated amount of PBS up to the total required volume >> (so that the buffer is 1x in the final volume). Then we add acid to >> bring the pH back down to 7. Then we filter it since not all of the PFA >> has dissolved (though most of it has). >> >> Merced >> >> --On Monday, December 08, 2008 9:11 AM +1100 Tony Henwood >> wrote: >> >>> My experience is that when you add paraformaldehyde to water all it >>> forms is a colloidal solution (ie on standing, the paraformaldehyde >>> settles with very little going into solution (personal experience, >>> waited one week, then gave up). >>> >>> Has your experience been different? >>> >>> Regards >>> >>> Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) >>> Laboratory Manager & Senior Scientist >>> Tel: 612 9845 3306 >>> Fax: 612 9845 3318 >>> the children's hospital at westmead >>> Cnr Hawkesbury Road and Hainsworth Street, Westmead >>> Locked Bag 4001, Westmead NSW 2145, AUSTRALIA >>> >>> >>> >>> >>> -----Original Message----- >>> From: histonet-bounces@lists.utsouthwestern.edu >>> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >>> ejschmid@ucalgary.ca >>> Sent: Sunday, 7 December 2008 7:30 AM >>> To: histonet@lists.utsouthwestern.edu >>> Subject: [Histonet] PFA preparation >>> >>> >>> Hi, >>> >>> So then what is the best way to prepare formaldehyde fixative from >>> PFA? >>> >>> The way I have been taught, which differs from what I have read, is >>> to dissolve 4% into ddH2O at room temperature. After that one could add >>> PBS or buffer. >>> >>> I've also been taught that too much heat during preparation >>> "degrades" PFA, and that PFA (or formaldehyde solution, rather) stored >>> too long will lose freshness because it "degrades." >>> >>> What I read is different. Texts suggest to dissolve the PFA in warm >>> water, and that aging of the fix is due to repolymerization, not >>> through >>> degradation. >>> >>> What should i do? >>> >>> Eric Schmidt >>> >>> University of Calgary >>> Medical Sciences >>> >>> >>> >>> >>> _______________________________________________ >>> 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 note 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. >>> ********************************************************************** >>> >>> >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >> >> >> >> Merced M Leiker >> Research Technician II >> 354 BRB (pkgs) / 140 Farber Hall (letters) >> School of Medicine and Biomedical Sciences >> State University of New York at Buffalo >> 3435 Main St, Buffalo, NY 14214 >> Ph: (716) 829-6033 >> Fx: (716) 829-2725 >> >> "Without my flaws I'm really very boring." >> - random internet blog commentator >> >> >> > > > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From rjbuesa <@t> yahoo.com Mon Dec 8 11:16:06 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 8 11:16:10 2008 Subject: [Histonet] processing times In-Reply-To: <97101976F8A044468CA74FE11883B90E29B9EAEF@VISTA.roswellpark.org> Message-ID: <376994.56747.qm@web65715.mail.ac4.yahoo.com> Kathleen: Call me "dinosaur", but before starting to change your known procedures (that in time seem to be OK), answer yourself the following questions: ? 1- why do you want to reduce your TAT? 2- how short your protocols have to get in order to reduce your lab total?TAT significantly? 3- what changes are going to be done in the?REST of the work flow to assure a TAT reduction? 4- is tissue processing the only aspect to experience time reductions? ? If you are satisfied with your answers, then go ahead with your protocol reductions (when you get answers to your initial question), but remember taht?tissue processing is usually LESS than20% of the whole processing time, the rest are the PRE and POST processing tasks, how are you going to reduce them also? Ren? J. --- On Mon, 12/8/08, Caleri, Kathleen wrote: From: Caleri, Kathleen Subject: [Histonet] processing times To: histonet@lists.utsouthwestern.edu Date: Monday, December 8, 2008, 11:27 AM Hello-I have just started at Roswell Park Cancer Institute and we are hoping to decrease our TAT by decreasing our processing times-having come from a facility where we hadn't changed processor times in 20 years I would appreciate it if someone could supply their schedules so I could do a comparison...we currently do a short run for bx's (5 hr), a standard surgical run (13 hr) , and a longer fatty/breast run (16 hr) Regards, Kate Kathleen Caleri, BS, MLT, HT (ASCP) Histology Lab Supervisor Pathology Roswell Park Cancer Institute (716) 845-1329 "the future of mankind lies waiting for those who come to understand their lives and take up their responsibilities to help others" This email message may contain legally privileged and/or confidential information. If you are not the intended recipient(s), or the employee or agent responsible for the delivery of this message to the intended recipient(s), you are hereby notified that any disclosure, copying, distribution, or use of this email message is prohibited. If you have received this message in error, please notify the sender immediately by e-mail and delete this email message from your computer. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Dec 8 12:43:40 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Dec 8 12:44:09 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed Message-ID: It is impossible for "4% paraformaldehyde" to penetrate tissues faster than "10% formalin" because both solutions contain the same concentrations of methylene glycol and formaldehyde if they are made up in a solution within several orders of magnitude of neutrality (pH 4 to 9). The usual phosphate buffers fix the pH of a formaldehyde fixative at 7.2 to 7.6, and the composition of the solution is almost exactly the same whether the solution is derived from high polymers (paraformaldehyde) or low polymers (formalin). Solutions of formaldehyde diluted from formalin contain a little methanol and sodium formate, which are not present if paraformaldehyde is used, but these extra ingredients do not influence penetration and have little or no effect on structural preservation, even at EM level, if the osmotic pressure of the fixative is right. See Freida Carson's textbook (Histotechnology, 2nd ed.) and also her important paper in Am. J. Clin. Path. 59: 365-373 (1973). John Kiernan Anatomy, UWO London, Canada = = = histonet , anh2006@med.cornell.edu, Jan Shivers > I have been curious about this discussion. we used 4% paraformaldehyde > for smaller biopsies only because it has a faster penetration to > tissuethan 10% formalin. In all my IHC that I have done. I > observe that doing > an IHC with 4% paraformaldehyde does not necessarily need > antigenretrieval in comparison to 10% formalin either it > will be human or > animal tissue but this depends on how long was it fix, our 4% > paraformaldehyde we fix smaller biopsies like nerve,muscle, skin > for 6 > to 12 hrs. and for formalin it is 12 to 48 hours or more. Maybe > you can > comment on the effect on this to tissue if you say you will use 4% > paraformaldehyde for storage and transportation. > > > > > Reuel Cornelia, BS MT, AMT > Cellular Pathology > Texas Scottish Rite Hospital for Children > 2222 Welborn Street > Dallas, TX 75219 > Tel: 214-559-7766 > fax: 214-559-7768 > > >>> "Tony Henwood" 12/04/08 9:29 PM >>> > tf wrote: > > "I DO believe that one reason some people use 4% PFA rather 10% > formalin is that PFA is a bit more stable, both for storage and > transportation~~~." > > I have not heard this before. > Do you have a reference for this? > > > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > -----Original Message----- > From: tf [mailto:tifei@foxmail.com] > Sent: Friday, 5 December 2008 2:11 PM > To: Tony Henwood; anh2006@med.cornell.edu; Jan Shivers; > histonet > Subject: Re: RE: [Histonet] IHC on paraformaldehyde-fixed > > > the basic principles are the same for most cross-linking > fixatives and induce similar bonds > the difference you observed between may due to any other > variability, or the co-fixative you used. > > I DO believe that one reason some people use 4% PFA rather 10% > formalin is that PFA is a bit more stable, both for storage and > transportation~~~. > > > > > 2008-12-05 > > ________________________________ > > tf > > ________________________________ > > ???? Tony Henwood > ???? 2008-12-05 06:00:03 > ???? anh2006@med.cornell.edu; Jan Shivers; histonet > ??? > ?? RE: [Histonet] IHC on paraformaldehyde-fixed > > > Interesting point. > Since 10% buffered formalin (made from the concentrated 38% > formaldehyde) contain about 1% methanol, has it been shown that > this has > a deleterious effect on ANY antigens or are we expecting this > worse case > senario as being the norm? > I am not aware of any antigens (or antigen-antibody combination) > that > has been badly effected by 10% formalin that is NOT effected by > 10% > formaldehyde. Are you aware of any?? > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > -----Original Message----- > From: anh2006@med.cornell.edu [mailto:anh2006@med.cornell.edu] > Sent: Friday, 5 December 2008 1:31 AM > To: Tony Henwood; Jan Shivers; histonet > Subject: Re: [Histonet] IHC on paraformaldehyde-fixed > So true. However, be aware that 10% neutral buffered formalin we > use has > methanol in it which may affect certain antigens so there may be > some > difference in staining (hence why for mouse work we now only use > 4% PFA > in pure PBS). It is good to be aware of the other ingredients in > your > fixative solutions, whether commercially prepared or a homemaede > recipe, > as it isn't only the formaldehyde fixative which can make a > difference. > -----Original Message----- > From: Tony Henwood > Date: Thu, 04 Dec 2008 09:35:09 > To: Jan Shivers; > histonet > Subject: RE: [Histonet] IHC on paraformaldehyde-fixed > Gee I hate the term paraformaldehyde (as many of you probably > know) > This is an example of how confusion of terms can cause > unnecessary work. > Is "4% paraformaldehyde" different from 4 % formaldehyde? > No > Should any procedure done to tissues fixed in "4% > paraformaldehyde" give > results different to those fixed in 4% formaldehyde or 10% > formalin? > No since they are the same thing. > As Manoonkitiwongsa and Schultz (Histochem J 34: 365-367, 2002) > state > when paraformaldehyde actually becomes a fixative, it is no > longer > paraformaldehyde by chemistry or fixation capacity. Rather, it > is > formaldehyde in water without methanol or any other stabiliser. > Without > heat and an alkaline environment, paraformaldehyde in water is > simply a > paraformaldehyde suspension with little fixation capacity. If > the > fixative is prepared from paraformaldehyde then it should be > termed 4% > formaldehyde freshly prepared from paraformaldehyde. If a > concentrated > formalin solution (40% formaldehyde) is used, then it should be > termed > 10% formalin. > If you do a search on Histonet for paraformaldehye, you will > find that > this topic has been extensively discussed. > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory > Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Jan > Shivers > Sent: Thursday, 4 December 2008 8:34 AM > To: histonet > Subject: [Histonet] IHC on paraformaldehyde-fixed > Has anyone ever done IHC on parafomaldehyde-fixed tissues, and > if so, > how well did it work? Will the same antigen-retrieval methods > used with > formalin-fixed tissue be applicable? > I'm asking for an investigator, who already has his tissues > fixed in > paraformaldehyde. > Jan Shivers > Senior Scientist > Pathology Teaching Program > Histology/IHC/EM Section Head > University of Minnesota > Veterinary Diagnostic Laboratory > 1333 Gortner Ave. > St. Paul, MN 55108 > 612-624-7297 > shive003@umn.edu_______________________________________________ > > 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 note 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. > ********************************************************************** > _______________________________________________ > 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 note 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. > ********************************************************************** > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > > > ********************************************************************* > 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 note 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. > ********************************************************************** > > _______________________________________________ > 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 jkiernan <@t> uwo.ca Mon Dec 8 13:43:42 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Dec 8 13:43:47 2008 Subject: [Histonet] Silanized slides Message-ID: The solution for making silanized slides (APES in acetone; also needs a trace of water, which does not need to be added specially) has to be used right away: it's stable for perhaps a few hours. The resulting slides, washed and dried, should be stable indefinitely if kept clean. See Microscopy Today (June 1999) pp.22-24; also available at http://publish.uwo.ca/~jkiernan/adhesivs.htm John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: "Webb, Dorothy L" Date: Thursday, December 4, 2008 13:42 Subject: [Histonet] Silanized slides To: histonet@lists.utsouthwestern.edu > Does anyone have any idea of how long silanized slides made in > lab are > good for? Would it be the same outdate as is on the > solution one is > using to prepare the silanized slides? Thanks, as always!! > > Dorothy Webb, HT > Regions Histology Technical Supervisor > 651-254-2962 > ________________________________________ > This e-mail and any files transmitted with it are confidential > and are intended solely for the use of the individual or entity > to whom they are addressed. If you are not the intended > recipient or the individual responsible for delivering the e- > mail to the intended recipient, please be advised that you have > received this e-mail in error and that any use, dissemination, > forwarding, printing, or copying of this e-mail is strictly > prohibited. > If you have received this e-mail in error, please immediately > notify the HealthPartners Support Center by telephone at (952) > 967-6600. You will be reimbursed for reasonable costs incurred > in notifying us. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CHRISH <@t> HEALTHCARESCOUTS.COM Mon Dec 8 14:02:49 2008 From: CHRISH <@t> HEALTHCARESCOUTS.COM (Chris Handrahan) Date: Mon Dec 8 14:11:30 2008 Subject: [Histonet] Immediate Need for Histotech in Pittsburgh, PA Message-ID: The #1 Recruiter for Laboratory/Biotech Specialists Healthcare Scouts is a medical laboratory/biotech recruitment firm that places professionals in full-time, permanent positions throughout the United States. As the leading nationally recognized permanent placement solution for medical laboratory/biotech professionals, Healthcare Scouts has the following histology opening in Pittsburgh Day shift - 5am start time ASCP required Full time permanent position Reference Lab Referrals With employment opportunities all over the US, we are always looking for qualified and motivated professionals. Recommend a friend or family member, and you could be eligible for up to $1,000 through Healthcare Scouts' referral program. Call us today to learn more! For immediate consideration please contact Chris Handrahan Managing Director of Allied Health Healthcare Scouts 800-708-0605 office 321-231-5427 cell chrish@healthcarescouts.com www.healthcarescouts.com From jkiernan <@t> uwo.ca Mon Dec 8 14:18:12 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Dec 8 14:18:28 2008 Subject: [Histonet] Alias identity In-Reply-To: <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> References: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> Message-ID: Amber McKenzie wrote: > . . .We are all supposed to be professionals asking each > other for advice/suggestions on the Histonet - who cares who > each person is? If I post a question, I don't care if it's Jane Doe > answering or John Smith ... That's true, and it certainly makes no difference if the person asking a question hides behind an alias. But what about the person answering? Can you act upon advice from an unknown source? Jane Doe might be experienced and wise, whereas John Smith might be some pimply youth who only thinks he knows all the answers. John Kiernan Anatomy, UWO London, Canada = = = From ksecrest <@t> hsc.wvu.edu Mon Dec 8 14:32:37 2008 From: ksecrest <@t> hsc.wvu.edu (Kimberly Secrest) Date: Mon Dec 8 14:33:37 2008 Subject: [Histonet] HTL lab donations Message-ID: <493D3E150200007800005734@newgwia.hsc.wvu.edu> Hello All, I am calling out to those of you willing and capable to make a tax deductible donation to a good cause, the start up of an HTL student laboratory. I am currently in the need of embedding centers, water baths, Coplin jars, staining racks, slides, cassettes, basically all basic histology supplies. I?m also interested in staining reagents, special stain powders, etc. Any donation would be greatly appreciated. Thank you Kimberly Secrest, HTL, QIHC Instructor Department of Pathology West Virginia University 304-293-7628 ksecrest@hsc.wvu.edu From sccrshlly <@t> yahoo.com Mon Dec 8 17:10:27 2008 From: sccrshlly <@t> yahoo.com (Shelly Coker) Date: Mon Dec 8 17:10:30 2008 Subject: [Histonet] Histology Technician position Message-ID: <665489.54369.qm@web90305.mail.mud.yahoo.com> We are currently seeking an ASCP registered histology technician.?We are a small GI pathology lab.?We offer a competetive salary and compensation package.? Our lab is located in Austin, Texas, an area still experiencing economic growth.? Send resumes or inquiries to mcoker@austingastro.com. ? Thanks, Shelly Coker Laboratory Supervisor Austin Gastroenterology Histology Lab From asmith <@t> mail.barry.edu Mon Dec 8 17:43:51 2008 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Mon Dec 8 17:46:57 2008 Subject: [Histonet] Alias identity In-Reply-To: References: <1859408067.197601228317398502.JavaMail.osg@osgjas02.cns.ufl.edu> <03C921A1EAF7F541B16543F6EC6A4B37020E673F@giamail2.Gia.com> Message-ID: While I prefer to go by my own name, I recognize that some people rightly fear to do so. An anonymous source may still have something worthwhile to say. Experience does not always teach well. The wise and experienced Linus Pauling and Erwin Chargaff were wrong about the structure of DNA; the brash young James Watson was right. -Allen A. Smith Professor of Anatomy School of Podiatric Medicine Barry University Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Monday, December 08, 2008 3:18 PM To: Amber McKenzie Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Alias identity Amber McKenzie wrote: > . . .We are all supposed to be professionals asking each other for > advice/suggestions on the Histonet - who cares who each person is? If > I post a question, I don't care if it's Jane Doe answering or John > Smith ... That's true, and it certainly makes no difference if the person asking a question hides behind an alias. But what about the person answering? Can you act upon advice from an unknown source? Jane Doe might be experienced and wise, whereas John Smith might be some pimply youth who only thinks he knows all the answers. John Kiernan Anatomy, UWO London, Canada = = = From CIngles <@t> uwhealth.org Mon Dec 8 18:41:33 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Mon Dec 8 18:42:44 2008 Subject: [Histonet] RE: Don't think I'm nuts. References: <24A4826E8EF0964D86BC5317306F58A52BA58FA2FD@mmc-mail.ad.mhsil.com> <7722595275A4DD4FA225B92CDBF174A1744F13BB3A@EXC-MBX3.cfs.le.ac.uk> Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609DA@uwhis-xchng3.uwhis.hosp.wisc.edu> And since we're all lab rats anyway... Hey, I've already lost my tail! ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Edwards, R.E. Sent: Fri 12/5/2008 10:35 AM To: 'Vickroy, Jim'; 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] RE: Don't think I'm nuts. I believe that low humidity can cause laboratory mice to lose/shed the tips of their tails. From tifei <@t> foxmail.com Tue Dec 9 00:31:26 2008 From: tifei <@t> foxmail.com (tf) Date: Tue Dec 9 00:32:12 2008 Subject: [Histonet] IHC on paraformaldehyde-fixed References: Message-ID: <200812091431213276324@foxmail.com> VGhhdCBtYWtlcyBzZW5zZS4gQnV0IHdoeSB3ZSBhcmUgbm90IHVzaW5nIDEwJSBmb3JtYWxpbiBu b3c/DQoNCg0KMjAwOC0xMi0wOSANCg0KDQoNCnRmIA0KDQoNCg0Kt6K8/sjLo7ogSm9obiBLaWVy bmFuIA0Kt6LLzcqxvOSjuiAyMDA4LTEyLTA5ICAwMjo0NDowNyANCsrVvP7Iy6O6IFJldWVsIENv cm5lbGlhIA0Ks63LzaO6IFRvbnkgSGVud29vZDsgdGlmZWlAZm94bWFpbC5jb207IGhpc3RvbmV0 OyBhbmgyMDA2QG1lZC5jb3JuZWxsLmVkdTsgSmFuIFNoaXZlcnMgDQrW98zio7ogUmU6IFJFOiBS RTogW0hpc3RvbmV0XSBJSEMgb24gcGFyYWZvcm1hbGRlaHlkZS1maXhlZCANCiANCkl0IGlzIGlt cG9zc2libGUgZm9yICI0JSBwYXJhZm9ybWFsZGVoeWRlIiB0byBwZW5ldHJhdGUgdGlzc3VlcyBm YXN0ZXIgdGhhbiAiMTAlIGZvcm1hbGluIiBiZWNhdXNlIGJvdGggc29sdXRpb25zIGNvbnRhaW4g dGhlIHNhbWUgY29uY2VudHJhdGlvbnMgb2YgbWV0aHlsZW5lIGdseWNvbCBhbmQgZm9ybWFsZGVo eWRlIGlmIHRoZXkgYXJlIG1hZGUgdXAgaW4gYSBzb2x1dGlvbiB3aXRoaW4gc2V2ZXJhbCBvcmRl 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References: , <681289C0ED95575D3DD453FB@bchwxp2702.ad.med.buffalo.edu>, <63320.136.159.150.24.1228753175.squirrel@136.159.150.24>, Message-ID: <200812091432567529818@foxmail.com> SSBrbm93IHNvbWUgd29ya2VycyBzdGlycmluZyB0aGUgUEZBIHNvbHV0aW9uIG92ZXJuaWdodCBh bmQgdGhlbiBmaWx0ZXIuDQoNCg0KMjAwOC0xMi0wOSANCg0KDQoNCnRmIA0KDQoNCg0Kt6K8/sjL o7ogTWVyY2VkIExlaWtlciANCreiy83Ksbzko7ogMjAwOC0xMi0wOSAgMDE6MTg6NTUgDQrK1bz+ yMujuiBlanNjaG1pZEB1Y2FsZ2FyeS5jYSANCrOty82juiBoaXN0b25ldEBsaXN0cy51dHNvdXRo d2VzdGVybi5lZHU7IFRvbnkgSGVud29vZCANCtb3zOKjuiBSRTogW0hpc3RvbmV0XSBQRkEgcHJl cGFyYXRpb246IERvZXMgZGlzc29sdXRpb24gYWxzbyBtZWFuIHRoYXR0aGUgUEZBIGhhcyBkZXBv bHltZXJpemVkPyANCiANClllcywgd2Ugd291bGQgc2F5IHRoYXQgaGVhdGluZyBpcyBub3QgbmVj ZXNzYXJ5IHRvIGRlcG9seW1lcml6ZSBQRkEuIE9ubHkgDQp0aGUgaGlnaCBwSCBpcy4NCkFmdGVy IDMwLTYwIG1pbnV0ZXMgb2Ygc3RpcnJpbmcgd2Ugb2J0YWluIGEgY3J5c3RhbCBjbGVhciBzb2x1 dGlvbiB3aXRoIA0Kb25seSBhIGZldyBwYXJ0aWNsZXMgb2YgdW5kaXNzb2x2ZWQgUEZBIHN3aW1t 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Are you not forgetting about Rosalind Franklin? Ren? J. --- On Mon, 12/8/08, Smith, Allen wrote: From: Smith, Allen Subject: RE: [Histonet] Alias identity To: "'John Kiernan'" Cc: "'Histonet@lists.utsouthwestern.edu'" Date: Monday, December 8, 2008, 6:43 PM While I prefer to go by my own name, I recognize that some people rightly fear to do so. An anonymous source may still have something worthwhile to say. Experience does not always teach well. The wise and experienced Linus Pauling and Erwin Chargaff were wrong about the structure of DNA; the brash young James Watson was right. -Allen A. Smith Professor of Anatomy School of Podiatric Medicine Barry University Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Monday, December 08, 2008 3:18 PM To: Amber McKenzie Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Alias identity Amber McKenzie wrote: > . . .We are all supposed to be professionals asking each other for > advice/suggestions on the Histonet - who cares who each person is? If > I post a question, I don't care if it's Jane Doe answering or John > Smith ... That's true, and it certainly makes no difference if the person asking a question hides behind an alias. But what about the person answering? Can you act upon advice from an unknown source? Jane Doe might be experienced and wise, whereas John Smith might be some pimply youth who only thinks he knows all the answers. John Kiernan Anatomy, UWO London, Canada = = = _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Tue Dec 9 08:06:39 2008 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Tue Dec 9 08:09:45 2008 Subject: [Histonet] Alias identity In-Reply-To: <511796.20395.qm@web65706.mail.ac4.yahoo.com> References: <511796.20395.qm@web65706.mail.ac4.yahoo.com> Message-ID: The wise and experienced Rosalind Franklin also had the structure wrong. From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Tuesday, December 09, 2008 8:19 AM To: 'John Kiernan'; Smith, Allen Cc: 'Histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Alias identity Barry: Are you not forgetting about Rosalind Franklin? Ren? J. --- On Mon, 12/8/08, Smith, Allen wrote: From: Smith, Allen Subject: RE: [Histonet] Alias identity To: "'John Kiernan'" Cc: "'Histonet@lists.utsouthwestern.edu'" Date: Monday, December 8, 2008, 6:43 PM While I prefer to go by my own name, I recognize that some people rightly fear to do so. An anonymous source may still have something worthwhile to say. Experience does not always teach well. The wise and experienced Linus Pauling and Erwin Chargaff were wrong about the structure of DNA; the brash young James Watson was right. -Allen A. Smith Professor of Anatomy School of Podiatric Medicine Barry University Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Monday, December 08, 2008 3:18 PM To: Amber McKenzie Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Alias identity Amber McKenzie wrote: > . . .We are all supposed to be professionals asking each other for > advice/suggestions on the Histonet - who cares who each person is? If > I post a question, I don't care if it's Jane Doe answering or John > Smith ... That's true, and it certainly makes no difference if the person asking a question hides behind an alias. But what about the person answering? Can you act upon advice from an unknown source? Jane Doe might be experienced and wise, whereas John Smith might be some pimply youth who only thinks he knows all the answers. John Kiernan Anatomy, UWO London, Canada = = = _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rachelr <@t> mail.nih.gov Tue Dec 9 08:23:30 2008 From: rachelr <@t> mail.nih.gov (Rivka Rachel) Date: Tue Dec 9 08:24:40 2008 Subject: [Histonet] Alias identity In-Reply-To: Message-ID: Rosalind Franklin was wise and experienced because she was a genius who started her scientific career at the age of 21. By the time she passed away at the age of 37, she had made significant contributions in many more fields than Watson ever did. For more information, see http://www.pbs.org/wgbh/aso/databank/entries/bofran.html. On 12/9/08 9:06 AM, "Smith, Allen" wrote: > The wise and experienced Rosalind Franklin also had the structure wrong. > > From: Rene J Buesa [mailto:rjbuesa@yahoo.com] > Sent: Tuesday, December 09, 2008 8:19 AM > To: 'John Kiernan'; Smith, Allen > Cc: 'Histonet@lists.utsouthwestern.edu' > Subject: RE: [Histonet] Alias identity > > Barry: > Are you not forgetting about Rosalind Franklin? > Ren? J. > > --- On Mon, 12/8/08, Smith, Allen wrote: > From: Smith, Allen > Subject: RE: [Histonet] Alias identity > To: "'John Kiernan'" > Cc: "'Histonet@lists.utsouthwestern.edu'" > Date: Monday, December 8, 2008, 6:43 PM > > While I prefer to go by my own name, I recognize that some people rightly fear > > to do so. An anonymous source may still have something worthwhile to say. > > Experience does not always teach well. The wise and experienced Linus Pauling > > and Erwin Chargaff were wrong about the structure of DNA; the brash young > James > > Watson was right. > > -Allen A. Smith > > Professor of Anatomy > > School of Podiatric Medicine > > Barry University > > Miami Shores, FL > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan > > Sent: Monday, December 08, 2008 3:18 PM > > To: Amber McKenzie > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] Alias identity > > > > Amber McKenzie wrote: > >> . . .We are all supposed to be professionals asking each other for > >> advice/suggestions on the Histonet - who cares who each person is? If > >> I post a question, I don't care if it's Jane Doe answering or John > >> Smith ... > > > > That's true, and it certainly makes no difference if the person asking a > > question hides behind an alias. But what about the person answering? Can you > act > > upon advice from an unknown source? Jane Doe might be experienced and wise, > > whereas John Smith might be some pimply youth who only thinks he knows all the > > answers. > > > > John Kiernan > > Anatomy, UWO > > London, Canada > > = = = > > > > > > > > _______________________________________________ > > 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 -- Rivka A. Rachel, MD, PhD Staff Scientist, National Eye Institute Neurobiology, Neurodegeneration & Repair Laboratory Bldg 10 Rm 10D43 Tel: 301 443-4906 From je2 <@t> sanger.ac.uk Tue Dec 9 08:33:52 2008 From: je2 <@t> sanger.ac.uk (Jeanne Estabel) Date: Tue Dec 9 08:34:03 2008 Subject: [Histonet] Dmetrix scanner Message-ID: <8DAC27FBBF33764B9F3DCF0A3E6B0962028F22AE@exchsrv2.internal.sanger.ac.uk> Hi, I would like any feedback on Dmetrix scanner. We have one here and we experienced problems as no reading barcodes. Regards Jeanne Estabel, PhD MGP Histologist, Team 109 Wellcome Trust/Sanger Institute Hinxton CB10 1SA Tel: 01223 495338 -- The Wellcome Trust Sanger Institute is operated by Genome Research Limited, a charity registered in England with number 1021457 and a compa ny registered in England with number 2742969, whose registered office is 2 15 Euston Road, London, NW1 2BE. From rjbuesa <@t> yahoo.com Tue Dec 9 09:03:26 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 9 09:03:31 2008 Subject: [Histonet] Alias identity In-Reply-To: Message-ID: <965265.3424.qm@web65706.mail.ac4.yahoo.com> And neither Watson nor Crick even mentioned her in their Nobel acceptance speech when she was the one who showed them the route and "enlightened" their minds. Ren? J. --- On Tue, 12/9/08, Rivka Rachel wrote: From: Rivka Rachel Subject: Re: [Histonet] Alias identity To: "Smith, Allen" , "'rjbuesa@yahoo.com'" Cc: "'Histonet@lists.utsouthwestern.edu'" Date: Tuesday, December 9, 2008, 9:23 AM Rosalind Franklin was wise and experienced because she was a genius who started her scientific career at the age of 21. By the time she passed away at the age of 37, she had made significant contributions in many more fields than Watson ever did. For more information, see http://www.pbs.org/wgbh/aso/databank/entries/bofran.html. On 12/9/08 9:06 AM, "Smith, Allen" wrote: > The wise and experienced Rosalind Franklin also had the structure wrong. > > From: Rene J Buesa [mailto:rjbuesa@yahoo.com] > Sent: Tuesday, December 09, 2008 8:19 AM > To: 'John Kiernan'; Smith, Allen > Cc: 'Histonet@lists.utsouthwestern.edu' > Subject: RE: [Histonet] Alias identity > > Barry: > Are you not forgetting about Rosalind Franklin? > Ren? J. > > --- On Mon, 12/8/08, Smith, Allen wrote: > From: Smith, Allen > Subject: RE: [Histonet] Alias identity > To: "'John Kiernan'" > Cc: "'Histonet@lists.utsouthwestern.edu'" > Date: Monday, December 8, 2008, 6:43 PM > > While I prefer to go by my own name, I recognize that some people rightly fear > > to do so. An anonymous source may still have something worthwhile to say. > > Experience does not always teach well. The wise and experienced Linus Pauling > > and Erwin Chargaff were wrong about the structure of DNA; the brash young > James > > Watson was right. > > -Allen A. Smith > > Professor of Anatomy > > School of Podiatric Medicine > > Barry University > > Miami Shores, FL > > > > -----Original Message----- > > From: histonet-bounces@lists.utsouthwestern.edu > > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan > > Sent: Monday, December 08, 2008 3:18 PM > > To: Amber McKenzie > > Cc: histonet@lists.utsouthwestern.edu > > Subject: Re: [Histonet] Alias identity > > > > Amber McKenzie wrote: > >> . . .We are all supposed to be professionals asking each other for > >> advice/suggestions on the Histonet - who cares who each person is? If > >> I post a question, I don't care if it's Jane Doe answering or John > >> Smith ... > > > > That's true, and it certainly makes no difference if the person asking a > > question hides behind an alias. But what about the person answering? Can you > act > > upon advice from an unknown source? Jane Doe might be experienced and wise, > > whereas John Smith might be some pimply youth who only thinks he knows all the > > answers. > > > > John Kiernan > > Anatomy, UWO > > London, Canada > > = = = > > > > > > > > _______________________________________________ > > 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 -- Rivka A. Rachel, MD, PhD Staff Scientist, National Eye Institute Neurobiology, Neurodegeneration & Repair Laboratory Bldg 10 Rm 10D43 Tel: 301 443-4906 From relia1 <@t> earthlink.net Tue Dec 9 09:37:54 2008 From: relia1 <@t> earthlink.net (Pam Barker) Date: Tue Dec 9 09:38:00 2008 Subject: [Histonet] RELIA Solutions Special Job Alert for Pathologist's Assistants and Histology Supervisors Message-ID: Hi Histonetters!! I have several exciting opportunities for experienced Pathologist?s Assistants and Histology Supervisors and Managers in hospital and private lab environments in several locations nationwide. The compensation packages are excellent, the work is challenging and the sky is the limit. The positions are of course full time and permanent. Here are the locations for my PA positions: Seattle, WA Cincinnati, OH Corpus Christi, TX Tampa, FL Here are the locations for my Management Positions: Los Angeles, CA - Histology Supervisor and AP Manager Cincinnati, OH Frederick, MD Austin, TX If you would like more information or know of someone else who might be interested, please contact me at relia1@earthlink.net or 866-607-3542. I am available to discuss the opportunity at your convenience including after hours. Thanks-Pam 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 < mailto:relia1@earthlink.net> << http://home.earthlink.net/~relia1>> www.myspace.com/pamatrelia < http://www.myspace.com/pamatrelia> From gu.lang <@t> gmx.at Tue Dec 9 10:03:03 2008 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Dec 9 10:10:10 2008 Subject: [Histonet] commercial decalcifier ingredients? Message-ID: <8AFBCAFD23C142FA8A6D831DAC860F5C@dielangs.at> Hi listmembers! Does anyone know the ingredients of this decalcifier? DC 1 decalcifier slow from Labonord. We have used it two years ago and I can't find the MSDS any more. The internet didn't help either. I know it consists of formic acid and formaldehyd, but I don't remember, if it also contents hydrochloric acid. Please help Gudrun Lang From rjbuesa <@t> yahoo.com Tue Dec 9 11:13:07 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 9 11:13:11 2008 Subject: [Histonet] commercial decalcifier ingredients? In-Reply-To: <8AFBCAFD23C142FA8A6D831DAC860F5C@dielangs.at> Message-ID: <946937.94476.qm@web65703.mail.ac4.yahoo.com> Gudrun: According with the manufacturer, it contains a combination (melage) of acids and folmaldehyde. They do not post a MSDS Ren? J. --- On Tue, 12/9/08, Gudrun Lang wrote: From: Gudrun Lang Subject: [Histonet] commercial decalcifier ingredients? To: histonet@lists.utsouthwestern.edu Date: Tuesday, December 9, 2008, 11:03 AM Hi listmembers! Does anyone know the ingredients of this decalcifier? DC 1 decalcifier slow from Labonord. We have used it two years ago and I can't find the MSDS any more. The internet didn't help either. I know it consists of formic acid and formaldehyd, but I don't remember, if it also contents hydrochloric acid. Please help Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kdboydhisto <@t> yahoo.com Tue Dec 9 11:15:30 2008 From: kdboydhisto <@t> yahoo.com (KELLY BOYD) Date: Tue Dec 9 11:15:34 2008 Subject: [Histonet] Billing question Message-ID: <776883.63385.qm@web58604.mail.re3.yahoo.com> Hi all! ? Question for those familiar with all the billing regulations: If you use an antibody that is for RUO (research use only)?or ASR (analyte specific reagent), can you bill the patient for these immunos? Kelly D. Boyd, BS, HTL (ASCP) Lab Manager Harris Histology Services 2025 Eastgate Dr. Ste. F Greenville, NC 27858 www.harrishisto.com? ? Tele (252)-830-6866 Cell? (252)-943-9527 Fax? (252)-830-0032 ? ? From JWeems <@t> sjha.org Tue Dec 9 11:23:07 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Tue Dec 9 11:23:17 2008 Subject: [Histonet] Billing question In-Reply-To: <776883.63385.qm@web58604.mail.re3.yahoo.com> References: <776883.63385.qm@web58604.mail.re3.yahoo.com> Message-ID: <5D64396A0D4A5346BEBC759022AAEAA51BA391@ITSSSXM01V6.one.ads.che.org> If I understand correctly... RUO - No ASR - Yes with a disclaimer Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of KELLY BOYD Sent: Tuesday, December 09, 2008 12:16 PM To: histonet Subject: [Histonet] Billing question Hi all! ? Question for those familiar with all the billing regulations: If you use an antibody that is for RUO (research use only)?or ASR (analyte specific reagent), can you bill the patient for these immunos? Kelly D. Boyd, BS, HTL (ASCP) Lab Manager Harris Histology Services 2025 Eastgate Dr. Ste. F Greenville, NC 27858 www.harrishisto.com? ? Tele (252)-830-6866 Cell? (252)-943-9527 Fax? (252)-830-0032 ? ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From rjbuesa <@t> yahoo.com Tue Dec 9 11:53:16 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 9 11:53:20 2008 Subject: [Histonet] Billing question In-Reply-To: <776883.63385.qm@web58604.mail.re3.yahoo.com> Message-ID: <701488.65144.qm@web65707.mail.ac4.yahoo.com> As your pathologists first because even when we charged for IHC to the patients, there was always a disclaimer saying that the Ab was for research, in spite of which, the charges were done (they are essentially for the pathologist interpretation).Hope this will help you. Ren? J. --- On Tue, 12/9/08, KELLY BOYD wrote: From: KELLY BOYD Subject: [Histonet] Billing question To: "histonet" Date: Tuesday, December 9, 2008, 12:15 PM Hi all! ? Question for those familiar with all the billing regulations: If you use an antibody that is for RUO (research use only)?or ASR (analyte specific reagent), can you bill the patient for these immunos? Kelly D. Boyd, BS, HTL (ASCP) Lab Manager Harris Histology Services 2025 Eastgate Dr. Ste. F Greenville, NC 27858 www.harrishisto.com? ? Tele (252)-830-6866 Cell? (252)-943-9527 Fax? (252)-830-0032 ? ? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Tue Dec 9 12:15:09 2008 From: jcline <@t> wchsys.org (Joyce Cline) Date: Tue Dec 9 12:15:15 2008 Subject: [Histonet] thermometer calibration In-Reply-To: Message-ID: I have purchased a calibrated digital thermometer. I take the temps using the calibrated thermometer and the regular thermometer. Compare the difference and list it at the top of my Quality Control sheets for the instruments. When the temps are taken everyday with the regular thermometer they are listed on the daily record sheet. These temps should fall within the designated temps. This worked for my last inspection. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Margiotta, Michele Sent: Monday, December 08, 2008 9:28 AM To: histonet@pathology.swmed.edu Subject: [Histonet] thermometer calibration Hi All, Just wondering how everyone is doing their thermometer calibrations. Inspection time is right around the corner so I want to be ready! Thanks. Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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 dencrowl <@t> MIT.EDU Tue Dec 9 13:12:31 2008 From: dencrowl <@t> MIT.EDU (Denise Crowley) Date: Tue Dec 9 13:16:58 2008 Subject: [Histonet] coverslip issues Message-ID: Hi all, I'm looking for some help in choosing a new vendor for coverslips for the Thermo Consul coverslipper. We have been buying coverslips from Thermo since we bought the instrument, and the last shipment (of 10 cases) is from a new supplier and is not working in the instrument. The coverslip cannot be detected and the mountant is not dispensing. I spoke with the technician (Diane is wonderful) and she walked me through a diagnostic on the Consul and the instrument is not the problem. Does anyone out there have a suggestion as to a coverslip that they know works in the Consul? I have already tried the Vista from VWR (which is okay, but not perfect) and am eagerly awaiting requested samples. Thanks for your help. Denise Crowley Facility Manager Histology David H. Koch Institute for Integrative Cancer Research Massachusetts Institute of Technology 40 Ames St. E17-427 Cambridge MA 02139 617-258-8183 dencrowl@mit.edu From stillmanlouise <@t> yahoo.com Tue Dec 9 15:17:30 2008 From: stillmanlouise <@t> yahoo.com (Louise Stillman) Date: Tue Dec 9 15:17:35 2008 Subject: [Histonet] (no subject) Message-ID: <29294.10789.qm@web46305.mail.sp1.yahoo.com> unsubscribe? E-Mail address:? stillmanlouise@yahoo.com From carrolpb <@t> umdnj.edu Tue Dec 9 15:30:21 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Tue Dec 9 15:31:33 2008 Subject: [Histonet] (no subject) In-Reply-To: <29294.10789.qm@web46305.mail.sp1.yahoo.com> References: <29294.10789.qm@web46305.mail.sp1.yahoo.com> Message-ID: <493EE36D.8050005@umdnj.edu> Hey Louise, See that link below, at the bottom of your email? The one that connects you to the page where you can unsubscribe yourself? Click it! Just another friendly reminder that theres no Unsubscribe Fairy lurking about this list making your unsubscription dreams come true... you can actually do it yourself with merely a few clicks and a little typing! Louise Stillman wrote: > unsubscribe E-Mail address: stillmanlouise@yahoo.com > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From pattennj <@t> mail.nih.gov Wed Dec 10 09:09:45 2008 From: pattennj <@t> mail.nih.gov (Patten, Nicole (NIH/NIAAA) [F]) Date: Wed Dec 10 09:09:49 2008 Subject: [Histonet] Primary works with ABC but not IF In-Reply-To: References: Message-ID: Hello- I am in a situation where my primary antibody works using the ABC method, but I do not see staining with immunofluorescence. My IF protocol works well with other antibodies so I do not know why it's not working with this particular antibody. Does anybody have any suggestions? I have considered using an avidin-Alexa Fluor conjugate but I am not sure if this is even possible (I am new to IHC) or how I would go about it. Would I need to first biotinylate my primary? Does anyone have any advice/protocols? Thanks! Any help would be greatly appreciated. Nicole Patten Post-Baccalaureate Fellow/IRTA NIAAA/National Institutes of Health From gagnone <@t> KGH.KARI.NET Wed Dec 10 09:10:34 2008 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Wed Dec 10 09:10:39 2008 Subject: [Histonet] Histology Workload Guidelines Message-ID: To add to the recent discussion about how many blocks can be embedded per hour, the College of Medical Laboratory Technologists of Ontario recently published MLT practice guidelines which may be of use. The practice guidelines are "intended to support, not replace, the exercise of professional judgment by medical laboratory technologists practising in histology...and are maintained...in consultation with CMLTO members and stakeholders." Check out quality assurance at: http://cmlto.com or more specifically: http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_hsto_gdlne.pdf Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada From mpence <@t> grhs.net Wed Dec 10 09:33:04 2008 From: mpence <@t> grhs.net (Mike Pence) Date: Wed Dec 10 09:33:20 2008 Subject: [Histonet] Histology Workload Guidelines In-Reply-To: Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A39DB@IS-E2K3.grhs.net> Why are times bases on 37.5 hour works per week? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, Eric Sent: Wednesday, December 10, 2008 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Workload Guidelines To add to the recent discussion about how many blocks can be embedded per hour, the College of Medical Laboratory Technologists of Ontario recently published MLT practice guidelines which may be of use. The practice guidelines are "intended to support, not replace, the exercise of professional judgment by medical laboratory technologists practising in histology...and are maintained...in consultation with CMLTO members and stakeholders." Check out quality assurance at: http://cmlto.com or more specifically: http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_h sto_gdlne.pdf Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Wed Dec 10 10:03:04 2008 From: bhewlett <@t> cogeco.ca (bhewlett@cogeco.ca) Date: Wed Dec 10 10:03:11 2008 Subject: [Histonet] Histology Workload Guidelines Message-ID: <493fe838.182.25af.14976@cogeco.ca> Because that is the length of time for a standard work week in this province. Bryan > Why are times bases on 37.5 hour works per week? > > Mike > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, > Eric > Sent: Wednesday, December 10, 2008 9:11 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histology Workload Guidelines > > > To add to the recent discussion about how many blocks can be embedded > per hour, the College of Medical Laboratory Technologists of Ontario > recently published MLT practice guidelines which may be of use. The > practice guidelines are "intended to support, not replace, the exercise > of professional judgment by medical laboratory technologists practising > in histology...and are maintained...in consultation with CMLTO members > and stakeholders." Check out quality assurance at: > > http://cmlto.com > > or more specifically: > > http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_h > sto_gdlne.pdf > > Eric Gagnon MLT > > Histology Laboratory > > Kingston General Hospital, > > Kingston, Ontario, Canada > > > > > > _______________________________________________ > 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 kmerriam2003 <@t> yahoo.com Wed Dec 10 10:05:40 2008 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Wed Dec 10 10:05:44 2008 Subject: [Histonet] Primary works with ABC but not IF References: Message-ID: <858018.38695.qm@web50301.mail.re2.yahoo.com> Hi Nicole, I don't know what kind of?secondary antibody you are using, but we have had very good luck with the Alexa-Fluor secondaries.??Often, when you go from an amplified?chromagenic method (such as ABC) to?a less-amplified staining method (such as with a?fluorescently-labeled secondary), the primary antibody needs to be retitrated.? Perhaps there is not?enough amplification with a directly labeled seconday; you may need to add another lay or do additional amplification, such as with tyramide - we are having very good luck with Invitrogen's?TSA?kits that contain AlexaFluor dyes.? The Perkin Elmer FITC-TSA kits also work well, but cost about 3X as much as Invitrogens. If you send more information about your staining protocols, we could probably give you more specific advice. Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: "Patten, Nicole (NIH/NIAAA) [F]" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 10, 2008 10:09:45 AM Subject: [Histonet] Primary works with ABC but not IF Hello- I am in a situation where my primary antibody works using the ABC method, but I do not see staining with immunofluorescence. My IF protocol works well with other antibodies so I do not know why it's not working with this particular antibody. Does anybody have any suggestions? I have considered using an avidin-Alexa Fluor conjugate but I am not sure if this is even possible (I am new to IHC) or how I would go about it. Would I need to first biotinylate my primary? Does anyone have any advice/protocols? Thanks! Any help would be greatly appreciated. Nicole Patten Post-Baccalaureate Fellow/IRTA NIAAA/National Institutes of Health _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kgrobert <@t> rci.rutgers.edu Wed Dec 10 10:54:32 2008 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Wed Dec 10 10:42:45 2008 Subject: [Histonet] Processing mouse seminal vesicles Message-ID: <493FF448.7020001@rci.rutgers.edu> Does anybody have a protocol for this? My last batch of these came out VERY dry and crunchy when run with other tissues on my standard protocol, which is as follows: (They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before processing.) 70%: 30 min 80%: 30 min 95%: 45 min 95%: 45 min 100%: 45 min 100%: 45 min xylene: 45 min xylene: 45 min Paraffin: 30 min Paraffin: 30min Paraffin: 30 min My other thought is that something is up with our VIP 5 processor, though no error messages are showing up. Any and all suggestions are most welcome. Thanks in advance, Kathleen Roberts Rutgers University From mbisher <@t> Princeton.EDU Wed Dec 10 11:12:57 2008 From: mbisher <@t> Princeton.EDU (Peggy Bisher) Date: Wed Dec 10 11:13:12 2008 Subject: [Histonet] Histology Workload Guidelines In-Reply-To: <493fe838.182.25af.14976@cogeco.ca> Message-ID: We (Princeton University) are also on a 37.5 hour week. That takes into account 30 minutes for a lunch break. Peggy On 12/10/08 11:03 AM, "bhewlett@cogeco.ca" wrote: > Because that is the length of time for a standard work week in this province. > > Bryan > > > >> Why are times bases on 37.5 hour works per week? >> >> Mike >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, >> Eric >> Sent: Wednesday, December 10, 2008 9:11 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Histology Workload Guidelines >> >> >> To add to the recent discussion about how many blocks can be embedded >> per hour, the College of Medical Laboratory Technologists of Ontario >> recently published MLT practice guidelines which may be of use. The >> practice guidelines are "intended to support, not replace, the exercise >> of professional judgment by medical laboratory technologists practising >> in histology...and are maintained...in consultation with CMLTO members >> and stakeholders." Check out quality assurance at: >> >> http://cmlto.com >> >> or more specifically: >> >> http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_h >> sto_gdlne.pdf >> >> Eric Gagnon MLT >> >> Histology Laboratory >> >> Kingston General Hospital, >> >> Kingston, Ontario, Canada >> >> >> >> >> >> _______________________________________________ >> 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 rjbuesa <@t> yahoo.com Wed Dec 10 11:32:08 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 10 11:32:11 2008 Subject: [Histonet] Processing mouse seminal vesicles In-Reply-To: <493FF448.7020001@rci.rutgers.edu> Message-ID: <213720.77503.qm@web65712.mail.ac4.yahoo.com> Mouse tissues, any one, will dry out too much with ethanol and xylene. I advise you to stop using ethanol and xylene and substitute BOTH with iso-propanol at the concentrations and times you are using now. Instead of the first paraffin, use a mixture 1:1 of iso-propanol and paraffin, followed by the 2 paraffins. Try it and you will notice the difference (and the saings). Ren? J. --- On Wed, 12/10/08, Kathleen Roberts wrote: From: Kathleen Roberts Subject: [Histonet] Processing mouse seminal vesicles To: "'histonet@lists.utsouthwestern.edu'" Date: Wednesday, December 10, 2008, 11:54 AM Does anybody have a protocol for this? My last batch of these came out VERY dry and crunchy when run with other tissues on my standard protocol, which is as follows: (They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before processing.) 70%: 30 min 80%: 30 min 95%: 45 min 95%: 45 min 100%: 45 min 100%: 45 min xylene: 45 min xylene: 45 min Paraffin: 30 min Paraffin: 30min Paraffin: 30 min My other thought is that something is up with our VIP 5 processor, though no error messages are showing up. Any and all suggestions are most welcome. Thanks in advance, Kathleen Roberts Rutgers University _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Stacy_McLaughlin <@t> cooley-dickinson.org Wed Dec 10 12:28:55 2008 From: Stacy_McLaughlin <@t> cooley-dickinson.org (Stacy McLaughlin) Date: Wed Dec 10 12:29:00 2008 Subject: [Histonet] old slide disposal Message-ID: We are going to dispose of very old histology slides. I was told by our environmental services manager that they were OK to box up and put them in a trash compactor to go to the landfill. (as long as any names were blacked out to avoid HIPAA violations). 2 years ago when we did this, I was told I had to put them in a sharps container, inside of a biohazard box for incineration. Quite a contrast of instructions! Makes us feel quite uncomfortable. Does any one know of any info on the regulations covering this topic? How do your institutions handle this? Thanks for your help!!! Stacy McLaughlin HT(ASCP) Lead Histology Tech./Laboratory Safety From kgrobert <@t> rci.rutgers.edu Wed Dec 10 12:42:09 2008 From: kgrobert <@t> rci.rutgers.edu (Kathleen Roberts) Date: Wed Dec 10 12:30:26 2008 Subject: [Histonet] Processing mouse seminal vesicles-thanks! In-Reply-To: <213720.77503.qm@web65712.mail.ac4.yahoo.com> References: <213720.77503.qm@web65712.mail.ac4.yahoo.com> Message-ID: <49400D81.7000606@rci.rutgers.edu> Well, I have a number of suggestions now; once I get past the holidays I'll start trying them out when the next batch of samples comes in. We provide histology and pathology services for our dept and university, so it's all animal tissue. Thanks again to everybody, and I'll let you know how it turns out. -Kathleen Rutgers University Rene J Buesa wrote: > Mouse tissues, any one, will dry out too much with ethanol and xylene. > I advise you to stop using ethanol and xylene and substitute BOTH with > iso-propanol at the concentrations and times you are using now. > Instead of the first paraffin, use a mixture 1:1 of iso-propanol and > paraffin, followed by the 2 paraffins. > Try it and you will notice the difference (and the saings). > Ren? J. > > --- On Wed, 12/10/08, Kathleen Roberts wrote: > > From: Kathleen Roberts > Subject: [Histonet] Processing mouse seminal vesicles > To: "'histonet@lists.utsouthwestern.edu'" > > Date: Wednesday, December 10, 2008, 11:54 AM > >Does anybody have a protocol for this? My last batch of these came out VERY dry >and crunchy when run with other tissues on my standard protocol, which is as >follows: >(They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before >processing.) > >70%: 30 min >80%: 30 min >95%: 45 min >95%: 45 min >100%: 45 min >100%: 45 min >xylene: 45 min >xylene: 45 min >Paraffin: 30 min >Paraffin: 30min >Paraffin: 30 min > >My other thought is that something is up with our VIP 5 processor, though no >error messages are showing up. Any and all suggestions are most welcome. > >Thanks in advance, >Kathleen Roberts >Rutgers University > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From TJJ <@t> stowers-institute.org Wed Dec 10 12:40:07 2008 From: TJJ <@t> stowers-institute.org (Johnson, Teri) Date: Wed Dec 10 12:40:34 2008 Subject: [Histonet] Re: Processing mouse seminal vesicles Message-ID: Kathleen, Before changing your entire processing set-up, try decreasing the time in your alcohols and xylene to 10-15 minutes per station. Mouse seminal vesicles are very small and you don't need such rigorous dehydration and clearing to get proper processing. Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110 From leiker <@t> buffalo.edu Wed Dec 10 13:07:34 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Wed Dec 10 13:07:41 2008 Subject: [Histonet] PCNA staining on paraffin In-Reply-To: <661949901A768E4F9CC16D8AF8F2838C017A39DB@IS-E2K3.grhs.net> References: <661949901A768E4F9CC16D8AF8F2838C017A39DB@IS-E2K3.grhs.net> Message-ID: <47184CF1787BB2007276F94C@bchwxp2702.ad.med.buffalo.edu> Does anyone have any suggestions for staining PCNA on paraffin? We are using Santa Cruz's PCNA (FL261) SC-7907. We have already stained with it successfully on frozen sections, but it does not appear to be working on paraffin. Thank you. Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From CIngles <@t> uwhealth.org Wed Dec 10 13:09:23 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Wed Dec 10 13:09:58 2008 Subject: [Histonet] old slide disposal References: Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609DD@uwhis-xchng3.uwhis.hosp.wisc.edu> We usually put them in the sharps container. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Stacy McLaughlin Sent: Wed 12/10/2008 12:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] old slide disposal We are going to dispose of very old histology slides. I was told by our environmental services manager that they were OK to box up and put them in a trash compactor to go to the landfill. (as long as any names were blacked out to avoid HIPAA violations). 2 years ago when we did this, I was told I had to put them in a sharps container, inside of a biohazard box for incineration. Quite a contrast of instructions! Makes us feel quite uncomfortable. Does any one know of any info on the regulations covering this topic? How do your institutions handle this? Thanks for your help!!! From CIngles <@t> uwhealth.org Wed Dec 10 13:17:58 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Wed Dec 10 13:20:27 2008 Subject: [Histonet] For Vinnie Della Speranza Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609E0@uwhis-xchng3.uwhis.hosp.wisc.edu> Vinnie: I have talked with my coders and they have said that because of the way things are listed on path reports, etc. The powers that be regard billing for more than one block per specimen essentially double billing. Claire From nfournier <@t> sasktel.net Wed Dec 10 13:41:31 2008 From: nfournier <@t> sasktel.net (Neil Fournier) Date: Wed Dec 10 13:41:37 2008 Subject: [Histonet] perfusion question Message-ID: <000d01c95aff$4d8a2830$9f2de143@NEIL> Is there a rationale for using normal saline (0.9% (w/v) NaCl dissolved in dH2O) over 0.1 M PBS (pH 7.4) as a rinsing solution during intraventricular perfusion of a rat. Would one yield better results over the other? Also is there a raionale for why some people perfuse using PBS made only from monobasic and dibasic sodium phosphate (with 0.9% NaCl) vs. using PBS that also include KCl, sodium phosphate dibasic, NaCl, and potassium phosphate monobasic in the recipe. Thanks for the help Neil From as3323 <@t> columbia.edu Wed Dec 10 14:12:31 2008 From: as3323 <@t> columbia.edu (Anna K. Schultz) Date: Wed Dec 10 14:12:36 2008 Subject: [Histonet] de-waxing / staining station question Message-ID: <494022AF.8040905@columbia.edu> Hi all, In an effort to reduce space usage, it has been suggested that I eliminate one of two xylene steps (and lengthen the time in the remaining xylene) when de-waxing paraffin sections. I do not see any protocols that only use one step of xylene. Are two strictly necessary? I would like to know the reasoning. Also, do gill hematoxylins and alcoholic eosin need to be used and stored under a hood? I've read the MSDS but it wasn't entirely clear if they should just be used with caution or strictly under the hood. They are in staining buckets with lids (the lids are sometimes leaky). I thought I would seek some advice here. Thanks, Anna basic science research core tech From pruegg <@t> ihctech.net Wed Dec 10 14:25:28 2008 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Wed Dec 10 14:25:32 2008 Subject: [Histonet] de-waxing / staining station question Message-ID: <20081210132528.f86bd30e73b823f57b516b5451216a98.de51b6b659.wbe@email.secureserver.net> i would definately always use at least 2 xylenes and change the f irst often where most of the paraffin is removed, or at least rotate it and -------- Original Message -------- Subject: [Histonet] de-waxing From: "Anna K. Schultz" bu.edu Wed Dec 10 14:48:38 2008 From: mburton1 <@t> bu.edu (Mark A Burton) Date: Wed Dec 10 14:49:05 2008 Subject: [Histonet] Double IHC Staining Message-ID: <000001c95b08$ad7e4790$087ad6b0$@edu> Hello, Can someone recommend reagents/kits and or favorite protocols for double IHC staining? We might use double immunofluorescence as well but I'd like to try IHC. Thank you! Mark Mark A Burton HTL ASCP Molecular Aging & Development Lab Boston University Medical Campus From jmartin <@t> lrgh.org Wed Dec 10 14:45:29 2008 From: jmartin <@t> lrgh.org (Martin, Jessica) Date: Wed Dec 10 14:50:15 2008 Subject: [Histonet] RE: Histonet Digest, Vol 61, Issue 16 In-Reply-To: <56bb908c-5c1f-4ed1-aa40-688f24622e50@LRGHEXEDGE1.lrgh.org> References: <56bb908c-5c1f-4ed1-aa40-688f24622e50@LRGHEXEDGE1.lrgh.org> Message-ID: <53ADC5744236FC43B956FBCA32DEBBE30138DB7848@LRGHEXVS2.practice.lrgh.org> Hi all, Diane, We also have the Thermo Consul coverslipper and we switched to Mercedes Medical coverslips some time ago. We are actually having much better luck with them and they are way less expensive. Contact Jake Smith at 1-800-359-8807. The order number that we use is r2450. He sent us samples and we liked them. I hope this helps. (No, I don't get any kick-backs for this!) Jessica Martin HT, ASCP x-3231 jmartin@lrgh.org ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu [histonet-request@lists.utsouthwestern.edu] Sent: Wednesday, December 10, 2008 1:04 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 61, Issue 16 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: thermometer calibration (Joyce Cline) 2. coverslip issues (Denise Crowley) 3. (no subject) (Louise Stillman) 4. Re: (no subject) (Peter Carroll) 5. Primary works with ABC but not IF (Patten, Nicole (NIH/NIAAA) [F]) 6. Histology Workload Guidelines (Gagnon, Eric) 7. RE: Histology Workload Guidelines (Mike Pence) 8. RE: Histology Workload Guidelines (bhewlett@cogeco.ca) 9. Re: Primary works with ABC but not IF (Kim Merriam) 10. Processing mouse seminal vesicles (Kathleen Roberts) 11. Re: Histology Workload Guidelines (Peggy Bisher) 12. Re: Processing mouse seminal vesicles (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Tue, 9 Dec 2008 13:15:09 -0500 From: "Joyce Cline" Subject: RE: [Histonet] thermometer calibration To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" I have purchased a calibrated digital thermometer. I take the temps using the calibrated thermometer and the regular thermometer. Compare the difference and list it at the top of my Quality Control sheets for the instruments. When the temps are taken everyday with the regular thermometer they are listed on the daily record sheet. These temps should fall within the designated temps. This worked for my last inspection. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Margiotta, Michele Sent: Monday, December 08, 2008 9:28 AM To: histonet@pathology.swmed.edu Subject: [Histonet] thermometer calibration Hi All, Just wondering how everyone is doing their thermometer calibrations. Inspection time is right around the corner so I want to be ready! Thanks. Michele Margiotta BMHMC Histology Supervisor 631-654-7192 DISCLAIMER: This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to which they are addressed. This communication may contain material protected by the attorney-client privilege. If you are not the intended recipient or the person responsible for delivering the e-mail to the intended recipient, be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the sender via return e-mail or call Brookhaven Memorial Hospital Medical Center at (631) 654-7282. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** 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. ------------------------------ Message: 2 Date: Tue, 9 Dec 2008 14:12:31 -0500 From: Denise Crowley Subject: [Histonet] coverslip issues To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Hi all, I'm looking for some help in choosing a new vendor for coverslips for the Thermo Consul coverslipper. We have been buying coverslips from Thermo since we bought the instrument, and the last shipment (of 10 cases) is from a new supplier and is not working in the instrument. The coverslip cannot be detected and the mountant is not dispensing. I spoke with the technician (Diane is wonderful) and she walked me through a diagnostic on the Consul and the instrument is not the problem. Does anyone out there have a suggestion as to a coverslip that they know works in the Consul? I have already tried the Vista from VWR (which is okay, but not perfect) and am eagerly awaiting requested samples. Thanks for your help. Denise Crowley Facility Manager Histology David H. Koch Institute for Integrative Cancer Research Massachusetts Institute of Technology 40 Ames St. E17-427 Cambridge MA 02139 617-258-8183 dencrowl@mit.edu ------------------------------ Message: 3 Date: Tue, 9 Dec 2008 13:17:30 -0800 (PST) From: Louise Stillman Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu Message-ID: <29294.10789.qm@web46305.mail.sp1.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 unsubscribe? E-Mail address:? stillmanlouise@yahoo.com ------------------------------ Message: 4 Date: Tue, 09 Dec 2008 16:30:21 -0500 From: Peter Carroll Subject: Re: [Histonet] (no subject) To: Louise Stillman Cc: histonet@lists.utsouthwestern.edu Message-ID: <493EE36D.8050005@umdnj.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hey Louise, See that link below, at the bottom of your email? The one that connects you to the page where you can unsubscribe yourself? Click it! Just another friendly reminder that theres no Unsubscribe Fairy lurking about this list making your unsubscription dreams come true... you can actually do it yourself with merely a few clicks and a little typing! Louise Stillman wrote: > unsubscribe E-Mail address: stillmanlouise@yahoo.com > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > ------------------------------ Message: 5 Date: Wed, 10 Dec 2008 10:09:45 -0500 From: "Patten, Nicole (NIH/NIAAA) [F]" Subject: [Histonet] Primary works with ABC but not IF To: Message-ID: Content-Type: text/plain; charset="US-ASCII" Hello- I am in a situation where my primary antibody works using the ABC method, but I do not see staining with immunofluorescence. My IF protocol works well with other antibodies so I do not know why it's not working with this particular antibody. Does anybody have any suggestions? I have considered using an avidin-Alexa Fluor conjugate but I am not sure if this is even possible (I am new to IHC) or how I would go about it. Would I need to first biotinylate my primary? Does anyone have any advice/protocols? Thanks! Any help would be greatly appreciated. Nicole Patten Post-Baccalaureate Fellow/IRTA NIAAA/National Institutes of Health ------------------------------ Message: 6 Date: Wed, 10 Dec 2008 10:10:34 -0500 From: "Gagnon, Eric" Subject: [Histonet] Histology Workload Guidelines To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" To add to the recent discussion about how many blocks can be embedded per hour, the College of Medical Laboratory Technologists of Ontario recently published MLT practice guidelines which may be of use. The practice guidelines are "intended to support, not replace, the exercise of professional judgment by medical laboratory technologists practising in histology...and are maintained...in consultation with CMLTO members and stakeholders." Check out quality assurance at: http://cmlto.com or more specifically: http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_hsto_gdlne.pdf Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada ------------------------------ Message: 7 Date: Wed, 10 Dec 2008 09:33:04 -0600 From: "Mike Pence" Subject: RE: [Histonet] Histology Workload Guidelines To: "Gagnon, Eric" , Message-ID: <661949901A768E4F9CC16D8AF8F2838C017A39DB@IS-E2K3.grhs.net> Content-Type: text/plain; charset="us-ascii" Why are times bases on 37.5 hour works per week? Mike -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, Eric Sent: Wednesday, December 10, 2008 9:11 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology Workload Guidelines To add to the recent discussion about how many blocks can be embedded per hour, the College of Medical Laboratory Technologists of Ontario recently published MLT practice guidelines which may be of use. The practice guidelines are "intended to support, not replace, the exercise of professional judgment by medical laboratory technologists practising in histology...and are maintained...in consultation with CMLTO members and stakeholders." Check out quality assurance at: http://cmlto.com or more specifically: http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_h sto_gdlne.pdf Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Wed, 10 Dec 2008 11:03:04 -0500 From: bhewlett@cogeco.ca Subject: RE: [Histonet] Histology Workload Guidelines To: "Mike Pence" , "Gagnon, Eric" , Message-ID: <493fe838.182.25af.14976@cogeco.ca> Because that is the length of time for a standard work week in this province. Bryan > Why are times bases on 37.5 hour works per week? > > Mike > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, > Eric > Sent: Wednesday, December 10, 2008 9:11 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Histology Workload Guidelines > > > To add to the recent discussion about how many blocks can be embedded > per hour, the College of Medical Laboratory Technologists of Ontario > recently published MLT practice guidelines which may be of use. The > practice guidelines are "intended to support, not replace, the exercise > of professional judgment by medical laboratory technologists practising > in histology...and are maintained...in consultation with CMLTO members > and stakeholders." Check out quality assurance at: > > http://cmlto.com > > or more specifically: > > http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_h > sto_gdlne.pdf > > Eric Gagnon MLT > > Histology Laboratory > > Kingston General Hospital, > > Kingston, Ontario, Canada > > > > > > _______________________________________________ > 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: 9 Date: Wed, 10 Dec 2008 08:05:40 -0800 (PST) From: Kim Merriam Subject: Re: [Histonet] Primary works with ABC but not IF To: "Patten, Nicole \(NIH/NIAAA\) \[F\]" , histonet@lists.utsouthwestern.edu Message-ID: <858018.38695.qm@web50301.mail.re2.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Nicole, I don't know what kind of?secondary antibody you are using, but we have had very good luck with the Alexa-Fluor secondaries.??Often, when you go from an amplified?chromagenic method (such as ABC) to?a less-amplified staining method (such as with a?fluorescently-labeled secondary), the primary antibody needs to be retitrated.? Perhaps there is not?enough amplification with a directly labeled seconday; you may need to add another lay or do additional amplification, such as with tyramide - we are having very good luck with Invitrogen's?TSA?kits that contain AlexaFluor dyes.? The Perkin Elmer FITC-TSA kits also work well, but cost about 3X as much as Invitrogens. If you send more information about your staining protocols, we could probably give you more specific advice. Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: "Patten, Nicole (NIH/NIAAA) [F]" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 10, 2008 10:09:45 AM Subject: [Histonet] Primary works with ABC but not IF Hello- I am in a situation where my primary antibody works using the ABC method, but I do not see staining with immunofluorescence. My IF protocol works well with other antibodies so I do not know why it's not working with this particular antibody. Does anybody have any suggestions? I have considered using an avidin-Alexa Fluor conjugate but I am not sure if this is even possible (I am new to IHC) or how I would go about it. Would I need to first biotinylate my primary? Does anyone have any advice/protocols? Thanks! Any help would be greatly appreciated. Nicole Patten Post-Baccalaureate Fellow/IRTA NIAAA/National Institutes of Health _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Wed, 10 Dec 2008 11:54:32 -0500 From: Kathleen Roberts Subject: [Histonet] Processing mouse seminal vesicles To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <493FF448.7020001@rci.rutgers.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Does anybody have a protocol for this? My last batch of these came out VERY dry and crunchy when run with other tissues on my standard protocol, which is as follows: (They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before processing.) 70%: 30 min 80%: 30 min 95%: 45 min 95%: 45 min 100%: 45 min 100%: 45 min xylene: 45 min xylene: 45 min Paraffin: 30 min Paraffin: 30min Paraffin: 30 min My other thought is that something is up with our VIP 5 processor, though no error messages are showing up. Any and all suggestions are most welcome. Thanks in advance, Kathleen Roberts Rutgers University ------------------------------ Message: 11 Date: Wed, 10 Dec 2008 12:12:57 -0500 From: Peggy Bisher Subject: Re: [Histonet] Histology Workload Guidelines To: , Mike Pence , "Gagnon, Eric" , Message-ID: Content-Type: text/plain; charset="US-ASCII" We (Princeton University) are also on a 37.5 hour week. That takes into account 30 minutes for a lunch break. Peggy On 12/10/08 11:03 AM, "bhewlett@cogeco.ca" wrote: > Because that is the length of time for a standard work week in this province. > > Bryan > > > >> Why are times bases on 37.5 hour works per week? >> >> Mike >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gagnon, >> Eric >> Sent: Wednesday, December 10, 2008 9:11 AM >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Histology Workload Guidelines >> >> >> To add to the recent discussion about how many blocks can be embedded >> per hour, the College of Medical Laboratory Technologists of Ontario >> recently published MLT practice guidelines which may be of use. The >> practice guidelines are "intended to support, not replace, the exercise >> of professional judgment by medical laboratory technologists practising >> in histology...and are maintained...in consultation with CMLTO members >> and stakeholders." Check out quality assurance at: >> >> http://cmlto.com >> >> or more specifically: >> >> http://cmlto.com/quality_assurance/MLT_practice_guidelines/learning/10_h >> sto_gdlne.pdf >> >> Eric Gagnon MLT >> >> Histology Laboratory >> >> Kingston General Hospital, >> >> Kingston, Ontario, Canada >> >> >> >> >> >> _______________________________________________ >> 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 ------------------------------ Message: 12 Date: Wed, 10 Dec 2008 09:32:08 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Processing mouse seminal vesicles To: "'histonet@lists.utsouthwestern.edu'" , Kathleen Roberts Message-ID: <213720.77503.qm@web65712.mail.ac4.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Mouse tissues, any one, will dry out too much with ethanol and xylene. I advise you to stop using ethanol and xylene and substitute BOTH with iso-propanol at the concentrations and times you are using now. Instead of the first paraffin, use a mixture 1:1 of iso-propanol and paraffin, followed by the 2 paraffins. Try it and you will notice the difference (and the saings). Ren? J. --- On Wed, 12/10/08, Kathleen Roberts wrote: From: Kathleen Roberts Subject: [Histonet] Processing mouse seminal vesicles To: "'histonet@lists.utsouthwestern.edu'" Date: Wednesday, December 10, 2008, 11:54 AM Does anybody have a protocol for this? My last batch of these came out VERY dry and crunchy when run with other tissues on my standard protocol, which is as follows: (They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before processing.) 70%: 30 min 80%: 30 min 95%: 45 min 95%: 45 min 100%: 45 min 100%: 45 min xylene: 45 min xylene: 45 min Paraffin: 30 min Paraffin: 30min Paraffin: 30 min My other thought is that something is up with our VIP 5 processor, though no error messages are showing up. Any and all suggestions are most welcome. Thanks in advance, Kathleen Roberts Rutgers University _______________________________________________ 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 61, Issue 16 **************************************** THIS MESSAGE IS CONFIDENTIAL. This e-mail message and any attachments are proprietary and confidential information intended only for the use of the recipient(s) named above. If you are not the intended recipient, you may not print,distribute, or copy this message or any attachments. If you have received this communication in error, please notify the sender by return e-mail and delete this message and any attachments from your computer. Any views or opinions expressed are solely those of the author and do not necessarily represent those of LRGHealthcare. From mcauliff <@t> umdnj.edu Wed Dec 10 15:12:48 2008 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Dec 10 15:14:23 2008 Subject: [Histonet] de-waxing / staining station question In-Reply-To: <494022AF.8040905@columbia.edu> References: <494022AF.8040905@columbia.edu> Message-ID: <494030D0.3050400@umdnj.edu> Two changes of xylene is a minimum. Also, almost every time someone asks about uneven staining it is due to inadequate removal of wax. Geoff Anna K. Schultz wrote: > Hi all, > In an effort to reduce space usage, it has been suggested that I > eliminate one of two xylene steps (and lengthen the time in the > remaining xylene) when de-waxing paraffin sections. I do not see any > protocols that only use one step of xylene. Are two strictly > necessary? I would like to know the reasoning. > Also, do gill hematoxylins and alcoholic eosin need to be used and > stored under a hood? I've read the MSDS but it wasn't entirely clear > if they should just be used with caution or strictly under the hood. > They are in staining buckets with lids (the lids are sometimes leaky). > I thought I would seek some advice here. > Thanks, > Anna > basic science research core tech > > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From mcauliff <@t> umdnj.edu Wed Dec 10 15:16:04 2008 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Dec 10 15:17:40 2008 Subject: [Histonet] perfusion question In-Reply-To: <000d01c95aff$4d8a2830$9f2de143@NEIL> References: <000d01c95aff$4d8a2830$9f2de143@NEIL> Message-ID: <49403194.1070205@umdnj.edu> Does not matter. The only error I see people making is spending a lot of time on the washout step. You will never get all of the blood out so there is no reason to try. Given a 250 g rat, I would use 25-30 ml of washout instilled in 15-20 seconds, then switch over to fix. Geoff Neil Fournier wrote: > Is there a rationale for using normal saline (0.9% (w/v) NaCl dissolved in dH2O) over 0.1 M PBS (pH 7.4) as a rinsing solution during intraventricular perfusion of a rat. Would one yield better results over the other? > > Also is there a raionale for why some people perfuse using PBS made only from monobasic and dibasic sodium phosphate (with 0.9% NaCl) vs. using PBS that also include KCl, sodium phosphate dibasic, NaCl, and potassium phosphate monobasic in the recipe. > > Thanks for the help > > Neil > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From rjbuesa <@t> yahoo.com Wed Dec 10 15:21:54 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 10 15:21:59 2008 Subject: [Histonet] de-waxing / staining station question In-Reply-To: <494022AF.8040905@columbia.edu> Message-ID: <239286.89575.qm@web65707.mail.ac4.yahoo.com> Anna: Neither Gill's hematoxylin not alcoholic eosin need to be used or?stored in a fume hood. As for the elimination of 2 xylene stations for dewaxing, IF you are going to dewax with xylene, that is not a good idea. A better idea though would be to eliminate xylene altogether; use instead a 2% aqueous diswasher soap solution at 90?C (2 stations), followed by 2 tap water stations at 90?C and one tap water station at 45?C and a final wash with distilled water. Stain as usual. wrote: From: Anna K. Schultz Subject: [Histonet] de-waxing / staining station question To: histonet@lists.utsouthwestern.edu Date: Wednesday, December 10, 2008, 3:12 PM Hi all, In an effort to reduce space usage, it has been suggested that I eliminate one of two xylene steps (and lengthen the time in the remaining xylene) when de-waxing paraffin sections. I do not see any protocols that only use one step of xylene. Are two strictly necessary? I would like to know the reasoning. Also, do gill hematoxylins and alcoholic eosin need to be used and stored under a hood? I've read the MSDS but it wasn't entirely clear if they should just be used with caution or strictly under the hood. They are in staining buckets with lids (the lids are sometimes leaky). I thought I would seek some advice here. Thanks, Anna basic science research core tech _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Dec 10 15:25:28 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 10 15:25:31 2008 Subject: [Histonet] old slide disposal In-Reply-To: Message-ID: <882952.67380.qm@web65712.mail.ac4.yahoo.com> Those regulations vary so widely that what I suggest you is to consult with your safety officer and follow his/her instructions. Ren? J. --- On Wed, 12/10/08, Stacy McLaughlin wrote: From: Stacy McLaughlin Subject: [Histonet] old slide disposal To: histonet@lists.utsouthwestern.edu Date: Wednesday, December 10, 2008, 1:28 PM We are going to dispose of very old histology slides. I was told by our environmental services manager that they were OK to box up and put them in a trash compactor to go to the landfill. (as long as any names were blacked out to avoid HIPAA violations). 2 years ago when we did this, I was told I had to put them in a sharps container, inside of a biohazard box for incineration. Quite a contrast of instructions! Makes us feel quite uncomfortable. Does any one know of any info on the regulations covering this topic? How do your institutions handle this? Thanks for your help!!! Stacy McLaughlin HT(ASCP) Lead Histology Tech./Laboratory Safety _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From steely0511 <@t> yahoo.com Wed Dec 10 17:37:06 2008 From: steely0511 <@t> yahoo.com (John Steel) Date: Wed Dec 10 17:37:12 2008 Subject: [Histonet] Primary works with ABC but not IF References: <858018.38695.qm@web50301.mail.re2.yahoo.com> Message-ID: <491229.89394.qm@web59709.mail.ac4.yahoo.com> Hi All, I do agree with titration - try this first.? You may consider pre-blocking?before use of the primary antibody.? In my experience, tryamide amplification is expensive, and may not be necessary - put this on the back burner.? Not sure on your specific applications, but am always interested in saving $$$$ while focusing on quality outcomes. Bests regards, John Steele ________________________________ From: Kim Merriam To: "Patten, Nicole (NIH/NIAAA) [F]" ; histonet@lists.utsouthwestern.edu Sent: Wednesday, December 10, 2008 11:05:40 AM Subject: Re: [Histonet] Primary works with ABC but not IF Hi Nicole, I don't know what kind of?secondary antibody you are using, but we have had very good luck with the Alexa-Fluor secondaries.??Often, when you go from an amplified?chromagenic method (such as ABC) to?a less-amplified staining method (such as with a?fluorescently-labeled secondary), the primary antibody needs to be retitrated.? Perhaps there is not?enough amplification with a directly labeled seconday; you may need to add another lay or do additional amplification, such as with tyramide - we are having very good luck with Invitrogen's?TSA?kits that contain AlexaFluor dyes.? The Perkin Elmer FITC-TSA kits also work well, but cost about 3X as much as Invitrogens. If you send more information about your staining protocols, we could probably give you more specific advice. Kim ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: "Patten, Nicole (NIH/NIAAA) [F]" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 10, 2008 10:09:45 AM Subject: [Histonet] Primary works with ABC but not IF Hello- I am in a situation where my primary antibody works using the ABC method, but I do not see staining with immunofluorescence. My IF protocol works well with other antibodies so I do not know why it's not working with this particular antibody.. Does anybody have any suggestions? I have considered using an avidin-Alexa Fluor conjugate but I am not sure if this is even possible (I am new to IHC) or how I would go about it. Would I need to first biotinylate my primary? Does anyone have any advice/protocols? Thanks! Any help would be greatly appreciated. Nicole Patten Post-Baccalaureate Fellow/IRTA NIAAA/National Institutes of Health _______________________________________________ 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 tifei <@t> foxmail.com Wed Dec 10 20:14:09 2008 From: tifei <@t> foxmail.com (tf) Date: Wed Dec 10 20:14:21 2008 Subject: [Histonet] perfusion question References: <000d01c95aff$4d8a2830$9f2de143@NEIL>, <49403194.1070205@umdnj.edu> Message-ID: <200812111014040355383@foxmail.com> SSBwZXJzb25hbGx5IHByZWZlciB0byB3YXNoIG91dCBsb25nZXIgLSB1bnRpbCB0aGUgZmxvdyBv dXQgdHVybmVkIHRvIGJlIGNsZWFyLg0KDQpPciB5b3Ugd2lsbCBoYXZlIHNvbWUgYmxvb2QgY2Vs bHMgaW5zaWRlIHRoZSBsaXZlci9icmFpbiwgZGVwZW5kaW5nIG9uIHlvdXIgdGlzc3VlcyBvZiBp bnRlcmVzdC4gT3IgZm9yIElIQyBzdGFpbmluZyB0aGlzIG1pZ2h0IGFyaXNlIHNvbWUgcHJvYmxl bXMuDQoNCg0KMjAwOC0xMi0xMSANCg0KDQoNCnRmIA0KDQoNCg0Kt6K8/sjLo7ogR2VvZmYgTWNB dWxpZmZlIA0Kt6LLzcqxvOSjuiAyMDA4LTEyLTExICAwNToyMDoyNCANCsrVvP7Iy6O6IE5laWwg Rm91cm5pZXIgDQqzrcvNo7ogaGlzdG9uZXRAbGlzdHMudXRzb3V0aHdlc3Rlcm4uZWR1IA0K1vfM 4qO6IFJlOiBbSGlzdG9uZXRdIHBlcmZ1c2lvbiBxdWVzdGlvbiANCiANCkRvZXMgbm90IG1hdHRl ci4gVGhlIG9ubHkgZXJyb3IgSSBzZWUgcGVvcGxlIG1ha2luZyBpcyBzcGVuZGluZyBhIGxvdCBv ZiANCnRpbWUgb24gdGhlIHdhc2hvdXQgc3RlcC4gWW91IHdpbGwgbmV2ZXIgZ2V0IGFsbCBvZiB0 aGUgYmxvb2Qgb3V0IHNvIA0KdGhlcmUgaXMgbm8gcmVhc29uIHRvIHRyeS4gR2l2ZW4gYSAyNTAg ZyByYXQsIEkgd291bGQgdXNlIDI1LTMwIG1sIG9mIA0Kd2FzaG91dCBpbnN0aWxsZWQgaW4gMTUt MjAgc2Vjb25kcywgdGhlbiBzd2l0Y2ggb3ZlciB0byBmaXguDQpHZW9mZg0KTmVpbCBGb3Vybmll ciB3cm90ZToNCj4gSXMgdGhlcmUgYSByYXRpb25hbGUgZm9yIHVzaW5nIG5vcm1hbCBzYWxpbmUg KDAuOSUgKHcvdikgTmFDbCBkaXNzb2x2ZWQgaW4gZEgyTykgb3ZlciAwLjEgTSBQQlMgKHBIIDcu NCkgYXMgYSByaW5zaW5nIHNvbHV0aW9uIGR1cmluZyBpbnRyYXZlbnRyaWN1bGFyIHBlcmZ1c2lv biBvZiBhIHJhdC4gV291bGQgb25lIHlpZWxkIGJldHRlciByZXN1bHRzIG92ZXIgdGhlIG90aGVy Pw0KPg0KPiBBbHNvIGlzIHRoZXJlIGEgcmFpb25hbGUgZm9yIHdoeSBzb21lIHBlb3BsZSBwZXJm dXNlIHVzaW5nIFBCUyBtYWRlIG9ubHkgZnJvbSBtb25vYmFzaWMgYW5kIGRpYmFzaWMgc29kaXVt IHBob3NwaGF0ZSAod2l0aCAwLjklIE5hQ2wpIHZzLiB1c2luZyBQQlMgdGhhdCBhbHNvIGluY2x1 ZGUgS0NsLCBzb2RpdW0gcGhvc3BoYXRlIGRpYmFzaWMsIE5hQ2wsIGFuZCBwb3Rhc3NpdW0gcGhv c3BoYXRlIG1vbm9iYXNpYyBpbiB0aGUgcmVjaXBlLg0KPg0KPiBUaGFua3MgZm9yIHRoZSBoZWxw DQo+DQo+IE5laWwgDQo+IF9fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19f X19fX19fDQo+IEhpc3RvbmV0IG1haWxpbmcgbGlzdA0KPiBIaXN0b25ldEBsaXN0cy51dHNvdXRo d2VzdGVybi5lZHUNCj4gaHR0cDovL2xpc3RzLnV0c291dGh3ZXN0ZXJuLmVkdS9tYWlsbWFuL2xp c3RpbmZvL2hpc3RvbmV0DQo+DQo+DQo+ICAgDQotLSANCi0tDQoqKioqKioqKioqKioqKioqKioq KioqKioqKioqKioqKioqKioqKioqKioqKioqDQpHZW9mZiBNY0F1bGlmZmUsIFBoLkQuDQpOZXVy b3NjaWVuY2UgYW5kIENlbGwgQmlvbG9neQ0KUm9iZXJ0IFdvb2QgSm9obnNvbiBNZWRpY2FsIFNj aG9vbA0KNjc1IEhvZXMgTGFuZSwgUGlzY2F0YXdheSwgTkogMDg4NTQNCnZvaWNlOiAoNzMyKS0y MzUtNDU4MyANCm1jYXVsaWZmQHVtZG5qLmVkdQ0KKioqKioqKioqKioqKioqKioqKioqKioqKioq KioqKioqKioqKioqKioqKioqKg0KX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19f X19fX19fX19fX18NCkhpc3RvbmV0IG1haWxpbmcgbGlzdA0KSGlzdG9uZXRAbGlzdHMudXRzb3V0 aHdlc3Rlcm4uZWR1DQo= From Robert.Schoonhoven <@t> mpiresearch.com Thu Dec 11 06:49:33 2008 From: Robert.Schoonhoven <@t> mpiresearch.com (Robert Schoonhoven) Date: Thu Dec 11 06:49:42 2008 Subject: [Histonet] PCNA staining on paraffin In-Reply-To: <47184CF1787BB2007276F94C@bchwxp2702.ad.med.buffalo.edu> Message-ID: Merced, I haven't seen any replies to your question. Under separate E-mail I'll send you the procedure I developed at UNC-Ch which will work on paraffin processed tissue from all mammals, fish, yeast and some plants. Please note that "drying" the slides at room temp. is important. Do not use a slide dryer. Bob Robert Schoonhoven BS, HT, HTL (ASCP) Scientist, Pathology MPI Research 54943 North Main Street Mattawan, MI 49071-9399 E-Mail: robert.schoonhoven@mpiresearch.com Office 269.668.3336 X-1768 Lab. 269.668.3336 X-2009 Cell 269.615.0576 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Merced Leiker Sent: Wednesday, December 10, 2008 2:08 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PCNA staining on paraffin Does anyone have any suggestions for staining PCNA on paraffin? We are using Santa Cruz's PCNA (FL261) SC-7907. We have already stained with it successfully on frozen sections, but it does not appear to be working on paraffin. Thank you. Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This communication, including attachments, is for the exclusive use of addressee and may contain proprietary, confidential and/or privileged information. If you are not the intended recipient, any use, copying, disclosure, dissemination or distribution is strictly prohibited. If you are not the intended recipient, please notify the sender immediately by return e-mail, delete this communication and destroy all copies. From rcharles <@t> state.pa.us Thu Dec 11 09:00:06 2008 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Thu Dec 11 09:00:10 2008 Subject: [Histonet] Paraffin wax Message-ID: <1B2F365C5F88A946B7D602E64ACD9CDD0447FD1EF0@ENHBGMBX04.PA.LCL> Hello, Could someone share their knowledge of the Paraplast line of paraffin with me? We are being forced to change from the TissuePrep line. Thanks Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 From histonetalias <@t> gmail.com Thu Dec 11 09:06:40 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Thu Dec 11 09:06:45 2008 Subject: [Histonet] Paraffin wax In-Reply-To: <1B2F365C5F88A946B7D602E64ACD9CDD0447FD1EF0@ENHBGMBX04.PA.LCL> References: <1B2F365C5F88A946B7D602E64ACD9CDD0447FD1EF0@ENHBGMBX04.PA.LCL> Message-ID: <4b6c85510812110706h7864d3a3pf4c0da66ac68442@mail.gmail.com> I have had very good luck with Paraplast Plus for infiltration and using the Paraplast Xtra for embedding. On Thu, Dec 11, 2008 at 10:00 AM, Charles, Roger wrote: > Hello, > Could someone share their knowledge of the Paraplast line of paraffin with > me? We are being forced to change from the TissuePrep line. > Thanks > > Roger Charles > Microbiologist > Pennsylvania Veterinary Laboratory > 2305 N Cameron St > Harrisburg, PA 17110 > 717-787-8808 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States From rjbuesa <@t> yahoo.com Thu Dec 11 09:08:24 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 11 09:08:29 2008 Subject: [Histonet] Paraffin wax In-Reply-To: <1B2F365C5F88A946B7D602E64ACD9CDD0447FD1EF0@ENHBGMBX04.PA.LCL> Message-ID: <384412.49738.qm@web65701.mail.ac4.yahoo.com> I always used Paraplast and I consider it as the best paraffin wax there is. Ren? J. --- On Thu, 12/11/08, Charles, Roger wrote: From: Charles, Roger Subject: [Histonet] Paraffin wax To: "Histonet (histonet@lists.utsouthwestern.edu)" Date: Thursday, December 11, 2008, 10:00 AM Hello, Could someone share their knowledge of the Paraplast line of paraffin with me? We are being forced to change from the TissuePrep line. Thanks Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MLafrini <@t> csmlab.com Thu Dec 11 09:27:57 2008 From: MLafrini <@t> csmlab.com (Michael LaFriniere) Date: Thu Dec 11 09:28:38 2008 Subject: [Histonet] Billing question In-Reply-To: <701488.65144.qm@web65707.mail.ac4.yahoo.com> References: <776883.63385.qm@web58604.mail.re3.yahoo.com> <701488.65144.qm@web65707.mail.ac4.yahoo.com> Message-ID: <4940EAED.588C.00AF.0@csmlab.com> It is my understand that , Yes you can bill as long as the ASR disclaimer is on the report and you heave validation of such antibodies that do did not have FDA approvals yet Michael LaFriniere >>> On 12/9/2008 at 12:53:16 PM, Rene J Buesa wrote: As your pathologists first because even when we charged for IHC to the patients, there was always a disclaimer saying that the Ab was for research, in spite of which, the charges were done (they are essentially for the pathologist interpretation).Hope this will help you. Ren? J. --- On Tue, 12/9/08, KELLY BOYD wrote: From: KELLY BOYD Subject: [Histonet] Billing question To: "histonet" Date: Tuesday, December 9, 2008, 12:15 PM Hi all! Question for those familiar with all the billing regulations: If you use an antibody that is for RUO (research use only) or ASR (analyte specific reagent), can you bill the patient for these immunos? Kelly D. Boyd, BS, HTL (ASCP) Lab Manager Harris Histology Services 2025 Eastgate Dr. Ste. F Greenville, NC 27858 www.harrishisto.com Tele (252)-830-6866 Cell (252)-943-9527 Fax (252)-830-0032 _______________________________________________ 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 MLafrini <@t> csmlab.com Thu Dec 11 09:37:42 2008 From: MLafrini <@t> csmlab.com (Michael LaFriniere) Date: Thu Dec 11 09:38:15 2008 Subject: [Histonet] Offended Message-ID: <4940ED37.588C.00AF.0@csmlab.com> I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia From shive003 <@t> umn.edu Thu Dec 11 10:01:38 2008 From: shive003 <@t> umn.edu (Jan Shivers) Date: Thu Dec 11 10:01:45 2008 Subject: [Histonet] PCNA staining on paraffin References: <661949901A768E4F9CC16D8AF8F2838C017A39DB@IS-E2K3.grhs.net> <47184CF1787BB2007276F94C@bchwxp2702.ad.med.buffalo.edu> Message-ID: <31F43506AA01410C940AD893DEE15376@auxs.umn.edu> I use Dako's PCNA (cat. # M0789), mouse monoclonal, clone PC10, HIER with pressure cooker (Decloaker, 30" at 121C, 21psi) in Target Retrieval buffer, 45 min in primary antibody (current dilution is 1:200, but that varies with each new lot), 30 min in goat anti-Ms EnVision+/HRP, 15 min in AEC. I use an autostainer, but it can be done manually with the same incubation times at RT. This works for me on many mammalian species. Jan Shivers UMN VetDiagLab St. Paul, MN ----- Original Message ----- From: "Merced Leiker" To: Sent: Wednesday, December 10, 2008 1:07 PM Subject: [Histonet] PCNA staining on paraffin > Does anyone have any suggestions for staining PCNA on paraffin? We are > using Santa Cruz's PCNA (FL261) SC-7907. > > We have already stained with it successfully on frozen sections, but it > does not appear to be working on paraffin. Thank you. > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From anh2006 <@t> med.cornell.edu Thu Dec 11 10:19:26 2008 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Thu Dec 11 10:21:55 2008 Subject: [Histonet] PCNA staining on paraffin In-Reply-To: <31F43506AA01410C940AD893DEE15376@auxs.umn.edu> References: <661949901A768E4F9CC16D8AF8F2838C017A39DB@IS-E2K3.grhs.net><47184CF1787BB2007276F94C@bchwxp2702.ad.med.buffalo.edu><31F43506AA01410C940AD893DEE15376@auxs.umn.edu> Message-ID: <218117745-1229012509-cardhu_decombobulator_blackberry.rim.net-1231399437-@bxe173.bisx.prod.on.blackberry> Second this, the DAKO clone works great. -----Original Message----- From: Jan Shivers Date: Thu, 11 Dec 2008 10:01:38 To: Merced Leiker Cc: histonet Subject: Re: [Histonet] PCNA staining on paraffin I use Dako's PCNA (cat. # M0789), mouse monoclonal, clone PC10, HIER with pressure cooker (Decloaker, 30" at 121C, 21psi) in Target Retrieval buffer, 45 min in primary antibody (current dilution is 1:200, but that varies with each new lot), 30 min in goat anti-Ms EnVision+/HRP, 15 min in AEC. I use an autostainer, but it can be done manually with the same incubation times at RT. This works for me on many mammalian species. Jan Shivers UMN VetDiagLab St. Paul, MN ----- Original Message ----- From: "Merced Leiker" To: Sent: Wednesday, December 10, 2008 1:07 PM Subject: [Histonet] PCNA staining on paraffin > Does anyone have any suggestions for staining PCNA on paraffin? We are > using Santa Cruz's PCNA (FL261) SC-7907. > > We have already stained with it successfully on frozen sections, but it > does not appear to be working on paraffin. Thank you. > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > _______________________________________________ > 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 pjfnefro <@t> duke.edu Thu Dec 11 10:58:49 2008 From: pjfnefro <@t> duke.edu (Pat Flannery) Date: Thu Dec 11 10:58:53 2008 Subject: [Histonet] Silly Question? Message-ID: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) From jmjohnson34 <@t> hotmail.com Thu Dec 11 10:59:17 2008 From: jmjohnson34 <@t> hotmail.com (Jennifer Johnson) Date: Thu Dec 11 10:59:23 2008 Subject: [Histonet] training materials Message-ID: Can anyone suggest a really good book, atlas, etc. for embedding? The girl that took my place at my last job is having a really hard time (especially with skin) and I told her I would ask the experts. Thanks, Jennifer Johnson, HTL (ASCP) _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008 From rosenfeldtek <@t> hotmail.com Thu Dec 11 11:09:50 2008 From: rosenfeldtek <@t> hotmail.com (JR R) Date: Thu Dec 11 11:09:55 2008 Subject: [Histonet] RE: Offended In-Reply-To: <4940ED37.588C.00AF.0@csmlab.com> References: <4940ED37.588C.00AF.0@csmlab.com> Message-ID: Hi Michael, Yeah, I can see how that would be annoying to you. If you get a solicitation call at work that bugs you--tell them to buzz off. I don't know that you ought to let it hurt your feelings or something, though. In the world of business, it is reasonable to expect that head hunters are at all times seeking to hire your best and brightest out from under you--the way to stop them is by making it worth people's while to stick around. To my knowledge, histotechnologists are, as a rule of thumb, over-worked and under-appreciated. And yet they are darn handy to have around. I suggest that you make it your personal mission to ensure that your histotechs ARE NOT looking for another job. That way, when a recruiter calls, your techs say "Leave my current position? What are you crazy??!!!" Jerry Ricks Research Scientist University of Washington Department of Pathology. > Date: Thu, 11 Dec 2008 10:37:42 -0500 > From: MLafrini@csmlab.com > To: Histonet@lists.utsouthwestern.edu > CC: > Subject: [Histonet] Offended > > I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! > > Michael LaFriniere > Executive Director > CSM > Washington DC/Maryland/Virginia > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008 From Linda.Watson <@t> bms.com Thu Dec 11 11:10:35 2008 From: Linda.Watson <@t> bms.com (Linda M Watson) Date: Thu Dec 11 11:10:57 2008 Subject: [Histonet] Silly Question? In-Reply-To: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> References: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Message-ID: <4941498B.4020307@bms.com> Hi Pat, Paraformaldehyde does not contain any additives and is considered more "pure" than formaldehyde which often contains methanol which in some cases is undesirable depending on the type of assay being conducted. Linda Pat Flannery wrote: > Please humor me on this if it's obvious (to everyone but me): why do > we use paraformaldehyde (which is so inconvenient to make up) rather > than buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde (either > 2% or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where > you were trained (i.e. force of habit) or is there a valid reason to > use each solution (basically the same chemical once in solution, > merely buffered or not)? The only answer I've gotten when I've asked > is, "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWeems <@t> sjha.org Thu Dec 11 11:12:51 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Thu Dec 11 11:13:01 2008 Subject: [Histonet] Silly Question? - Need help quickly! In-Reply-To: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> References: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Message-ID: <5D64396A0D4A5346BEBC759022AAEAA51BA6C4@ITSSSXM01V6.one.ads.che.org> I was just going to post a question regarding paraformaldhyde myself! Just last week I believe I remember someone saying that paraformaldehyde and formalin are the same and they had put the same solution in two different containers for one of their researchers because they were so insistent to have two different solutions. Are they the same? Well, today I have a request to put tissue for a researcher in formalin and paraformaldehyde. So.... Without percentage required, do I use 10% NBF? Do I call somewhere and get paraformaldehyde and make 4% paraformaldehyde? I have asked the surgeon twice for the number for the lab so I can find out - don't have it yet. I have two fresh adrenals in the fridge. Help!! Thanks in advance... Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Thursday, December 11, 2008 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From dlschneider <@t> gmail.com Thu Dec 11 11:20:40 2008 From: dlschneider <@t> gmail.com (Daniel Schneider) Date: Thu Dec 11 11:20:44 2008 Subject: [Histonet] training materials In-Reply-To: References: Message-ID: <1085e7000812110920s5a810ed6i56dc005fc858a26c@mail.gmail.com> I would be very interested in these suggestions as well, as we would like to improve the quality of skin embedding. Thanks! On Thu, Dec 11, 2008 at 10:59 AM, Jennifer Johnson wrote: > > Can anyone suggest a really good book, atlas, etc. for embedding? The girl > that took my place at my last job is having a really hard time (especially > with skin) and I told her I would ask the experts. > > Thanks, > Jennifer Johnson, HTL (ASCP) > _________________________________________________________________ > Send e-mail faster without improving your typing skills. > > http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008_______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pjfnefro <@t> duke.edu Thu Dec 11 11:22:17 2008 From: pjfnefro <@t> duke.edu (Pat Flannery) Date: Thu Dec 11 11:22:20 2008 Subject: [Histonet] Silly Question? Message-ID: <886EE175-ABA6-4325-BE60-369E64F324CE@duke.edu> Thank you, Jeanine, Joyce, and Linda. The tissues I'm cutting are just for H&E staining and light microscopy; they won't be used for Immunostaining, antigen retrieval, or anything too fancy. I don't see what difference it would make, but of course I'll put them in whatever people ask for. I know that commercial formaldehyde is "stabilized" for storage so I can understand some folks wouldn't want the MeOH contamination, no matter how slight. -Pat From rjbuesa <@t> yahoo.com Thu Dec 11 11:58:23 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 11 11:58:27 2008 Subject: [Histonet] Silly Question? - Need help quickly! In-Reply-To: <5D64396A0D4A5346BEBC759022AAEAA51BA6C4@ITSSSXM01V6.one.ads.che.org> Message-ID: <520038.88648.qm@web65711.mail.ac4.yahoo.com> Joyce: Methanal, which is the chemical name of formaldehyde, polymerizes. If it forms a polymer of at least 50 molecules or more, it gets solid = para-formaldehyde. Formalin (a trade name as formol is also another trade name)is the 37-50% aqueous solution of formaldehyde (with some additiveses to prevent polymerization). You can prepare BNF using the formalin solution or dissolving the amount of solid para-formaldehydede to get to the concentrationon you desire. The chemical in both solutions is the same = methanal or formaldehyde.Ren? J. --- On Thu, 12/11/08, Weems, Joyce wrote: From: Weems, Joyce Subject: RE: [Histonet] Silly Question? - Need help quickly! To: "Pat Flannery" , histonet@lists.utsouthwestern.edu Date: Thursday, December 11, 2008, 12:12 PM I was just going to post a question regarding paraformaldhyde myself! Just last week I believe I remember someone saying that paraformaldehyde and formalin are the same and they had put the same solution in two different containers for one of their researchers because they were so insistent to have two different solutions. Are they the same? Well, today I have a request to put tissue for a researcher in formalin and paraformaldehyde. So.... Without percentage required, do I use 10% NBF? Do I call somewhere and get paraformaldehyde and make 4% paraformaldehyde? I have asked the surgeon twice for the number for the lab so I can find out - don't have it yet. I have two fresh adrenals in the fridge. Help!! Thanks in advance... Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Thursday, December 11, 2008 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Thu Dec 11 12:06:23 2008 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Thu Dec 11 12:06:26 2008 Subject: [Histonet] training materials References: <1085e7000812110920s5a810ed6i56dc005fc858a26c@mail.gmail.com> Message-ID: <896775.88566.qm@web50311.mail.re2.yahoo.com> The AFIP book (Laboratory Methods in Histotechnology, edited by Edna Prophet, et al)?has?nice chapters on specimen orientation and embedding? It is?not really a training manual, but it?has some nice pictures as to how certain tissues should be placed into the molds. ?Kim Merriam, MA, HT(ASCP)QIHC Cambridge, MA ________________________________ From: Daniel Schneider To: histonet@lists.utsouthwestern.edu Sent: Thursday, December 11, 2008 12:20:40 PM Subject: Re: [Histonet] training materials I would be very interested in these suggestions as well, as we would like to improve the quality of skin embedding. Thanks! On Thu, Dec 11, 2008 at 10:59 AM, Jennifer Johnson wrote: > > Can anyone suggest a really good book, atlas, etc. for embedding?? The girl > that took my place at my last job is having a really hard time (especially > with skin) and I told her I would ask the experts. > > Thanks, > Jennifer Johnson, HTL (ASCP) > _________________________________________________________________ > Send e-mail faster without improving your typing skills. > > http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008_______________________________________________ > 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 rjbuesa <@t> yahoo.com Thu Dec 11 12:06:51 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 11 12:06:54 2008 Subject: [Histonet] Silly Question? In-Reply-To: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Message-ID: <433726.40630.qm@web65716.mail.ac4.yahoo.com> The "advantage" of para-formaldehyde is that you can prepare more easily any type of formaldehyde solution, at any concentration and also it does not have any additives (methanol) as "pure" formalin (37-50%formaldehydede) does. In reality the fixation mechanism is the same and?sometimes the whole issue boils down to personal preferences. Ren? J. --- On Thu, 12/11/08, Pat Flannery wrote: From: Pat Flannery Subject: [Histonet] Silly Question? To: histonet@lists.utsouthwestern.edu Date: Thursday, December 11, 2008, 11:58 AM Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Cathy.Crumpton <@t> tuality.org Thu Dec 11 12:10:31 2008 From: Cathy.Crumpton <@t> tuality.org (Cathy.Crumpton@tuality.org) Date: Thu Dec 11 12:10:36 2008 Subject: [Histonet] Non-gyn cytology prep - amount processed? In-Reply-To: <20081211180042.2A7CC132CB20@tumbleweed.corp.tuality.net> References: <20081211180042.2A7CC132CB20@tumbleweed.corp.tuality.net> Message-ID: Hi all, for those of you who also work with cytology - what is procedure for fluids that have a large volume? How much of the fluid do you process? I am wanting to compare our polic instance this week where we received processed 100 mL of that after making sure it first cell blocks only contained fibrin, but the malignant cells. When we went back to the contai settled after 24 hours so we poured off the top part of t and only processed the gunk at the bottom. The second time t were malignant cells in the cell block. My pathologist is pushin for us to process 100% of the fluids, 100% of the time, saying what we cu get those From leiker <@t> buffalo.edu Thu Dec 11 12:18:43 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Thu Dec 11 12:18:49 2008 Subject: [Histonet] Silly Question? - Need help quickly! In-Reply-To: <520038.88648.qm@web65711.mail.ac4.yahoo.com> References: <520038.88648.qm@web65711.mail.ac4.yahoo.com> Message-ID: <8332CB03EAE51284E4AE67A6@bchwxp2702.ad.med.buffalo.edu> So...is a polymer of paraformaldehyde considered "depolymerized" if it remains somewhere between 1-50 molecules long once it's been dissolved in solution, however it's dissolved? (the dissolving being a separate topic of debate on Histonet). Does it matter for tissue fixation purposes if there are formaldehyde chain lengths of 50 molecules present in solution - not long enough to precipitate out, but perhaps long enough to affect its penetration and fixing of tissues? Any ideas? Merced --On Thursday, December 11, 2008 9:58 AM -0800 Rene J Buesa wrote: > Joyce: > Methanal, which is the chemical name of formaldehyde, polymerizes. If it > forms a polymer of at least 50 molecules or more, it gets solid = > para-formaldehyde. Formalin (a trade name as formol is also another trade > name)is the 37-50% aqueous solution of formaldehyde (with some > additiveses to prevent polymerization). You can prepare BNF using the > formalin solution or dissolving the amount of solid para-formaldehydede > to get to the concentrationon you desire. The chemical in both solutions > is the same = methanal or formaldehyde.Ren? J. > > --- On Thu, 12/11/08, Weems, Joyce wrote: > > > From: Weems, Joyce > Subject: RE: [Histonet] Silly Question? - Need help quickly! > To: "Pat Flannery" , histonet@lists.utsouthwestern.edu > Date: Thursday, December 11, 2008, 12:12 PM > > I was just going to post a question regarding paraformaldhyde myself! > Just last week I believe I remember someone saying that paraformaldehyde > and formalin are the same and they had put the same solution in two > different containers for one of their researchers because they were so > insistent to have two different solutions. Are they the same? > > Well, today I have a request to put tissue for a researcher in formalin > and paraformaldehyde. So.... Without percentage required, do I use 10% > NBF? Do I call somewhere and get paraformaldehyde and make 4% > paraformaldehyde? > > I have asked the surgeon twice for the number for the lab so I can find > out - don't have it yet. I have two fresh adrenals in the fridge. Help!! > > > Thanks in advance... > Joyce > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 678-843-7376 - Phone > 678-843-7831 - Fax > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Thursday, December 11, 2008 11:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > > Please humor me on this if it's obvious (to everyone but me): why do we > use paraformaldehyde (which is so inconvenient to make up) rather than > buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde (either 2% > or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Confidentiality Notice: > This email, including any attachments is the > property of Catholic Health East and is intended > for the sole use of the intended recipient(s). > It may contain information that is privileged and > confidential. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are > not the intended recipient, please reply to the > sender that you have received the message in > error, then delete this 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 > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From jkiernan <@t> uwo.ca Thu Dec 11 13:11:51 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Thu Dec 11 13:11:54 2008 Subject: [Histonet] perfusion question In-Reply-To: <000d01c95aff$4d8a2830$9f2de143@NEIL> References: <000d01c95aff$4d8a2830$9f2de143@NEIL> Message-ID: The wash-out solution should have pH and osmotic pressure close to those of the animal's extracellular fluid, to avoid shrinkage or swelling of cells, collagen fibres etc. This can be achieved with simple saline (0.9% NaCl). A buffer prevents acidification of the extracellular fluid by products released from dying cells. Calcium ions (not compatible with phosphate buffers) enhance the preservation of phospholipids of cell membranes, myelin etc. Potassium ions are included in "physiological" saline solutions such as Ringer-Locke in which tissues and small organs can be kept alive, sometimes for several hours. I don't know of any study of effects of potassium on fixation, but probably someone has looked into it. The formaldehyde should also be dissolved in an isosmotic buffer because the chemical events of fixation occur slowly (several hours). Brain tissue still responds to changes in ambient osmotic pressure after several hours in neutral buffered formaldehyde. In glutaraldehyde, however, the cells are stabilized in 20 minutes. See: Paljarvi L, Garcia JH, Kalimo H (1979) The efficiency of aldehyde fixation for electron microscopy: stabilization of rat brain tissue to withstand osmotic stress. Histochem. J. 11: 267-276. This paper has also has references to several other studies. Traditional fixative mixtures are mostly acidic and rapidly acting, stabilizing the structure of the tissue (for light microscopy) before the development of adverse effects of low pH or osmotic pressure. The subject was also reviewed by J.R.Baker in his book Principles of Biological Microtechnique (1958), pp.75-86. John Kiernan Anatomy, UWO = = = ----- Original Message ----- From: Neil Fournier Date: Wednesday, December 10, 2008 14:42 Subject: [Histonet] perfusion question To: histonet@lists.utsouthwestern.edu > Is there a rationale for using normal saline (0.9% (w/v) NaCl > dissolved in dH2O) over 0.1 M PBS (pH 7.4) as a rinsing solution > during intraventricular perfusion of a rat. Would one yield > better results over the other? > > Also is there a raionale for why some people perfuse using PBS > made only from monobasic and dibasic sodium phosphate (with 0.9% > NaCl) vs. using PBS that also include KCl, sodium phosphate > dibasic, NaCl, and potassium phosphate monobasic in the recipe. > > Thanks for the help > > Neil > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----- Original Message ----- From: Neil Fournier Date: Wednesday, December 10, 2008 14:42 Subject: [Histonet] perfusion question To: histonet@lists.utsouthwestern.edu > Is there a rationale for using normal saline (0.9% (w/v) NaCl > dissolved in dH2O) over 0.1 M PBS (pH 7.4) as a rinsing solution > during intraventricular perfusion of a rat. Would one yield > better results over the other? > > Also is there a raionale for why some people perfuse using PBS > made only from monobasic and dibasic sodium phosphate (with 0.9% > NaCl) vs. using PBS that also include KCl, sodium phosphate > dibasic, NaCl, and potassium phosphate monobasic in the recipe. > > Thanks for the help > > Neil > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dcrippen <@t> buckinstitute.org Thu Dec 11 13:30:21 2008 From: dcrippen <@t> buckinstitute.org (Danielle Crippen) Date: Thu Dec 11 13:30:28 2008 Subject: [Histonet] paraffin processing rat tissue Message-ID: <69142A827D247F45877B305E991DB61F18DCB4@inverness.buckcenter.org> Dear Histo-Experts, I'm interested in paraffin processing protocols for rat kidneys, spleens and livers. Specifically, I'm interested in fixation time (immersion fixation in formalin) and dehydration and paraffin infiltration times. I've done some using my best guess based on the thickness of the tissue and resulting sections (5-7um) have "spaces" in them (after H and E) which may be due to inadequate fixation or dehydration times (I'm guessing). The spaces don't look like tears from sectioning...they look more like tissue separation... If you wouldn't mind sharing your protocols with me that would be WONDERFUL!!! A thousand thanks in advance! danielle Danielle Crippen Morphology and Imaging Core From carl.hobbs <@t> kcl.ac.uk Thu Dec 11 14:00:00 2008 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Thu Dec 11 14:00:40 2008 Subject: [Histonet] Re: PCNA Message-ID: <11D9615B89C10747B1C985966A63D7CA293069EC5D@KCL-MAIL04.kclad.ds.kcl.ac.uk> Use a PC10 mouse clone for pwax sections: works very well on many species that have been fixed in routine Formalin. Imho, no special procedures reqd. Sure, AR is best. If you wish to check out this site: http://www.immunoportal.com/index.php you might find further help? Also , be very careful if you use PCNA as a proliferation marker......it has a very long half-life compared with Ki67. So, use an anti Ki67 Ab instead for general proliferation studies that cannot include use of BrdU. Sure, if you want to assess a Mitotic index...use pH3 Ab on pwax sections. carl From CIngles <@t> uwhealth.org Thu Dec 11 14:07:30 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Thu Dec 11 14:10:49 2008 Subject: [Histonet] RE: Offended References: <4940ED37.588C.00AF.0@csmlab.com> Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> A certain company has recently started sending histo related fliers TO MY HOUSE. I want to know where they got my home address, especially since I have never actually done business with this company. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of JR R Sent: Thu 12/11/2008 11:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Offended Hi Michael, Yeah, I can see how that would be annoying to you. If you get a solicitation call at work that bugs you--tell them to buzz off. I don't know that you ought to let it hurt your feelings or something, though. In the world of business, it is reasonable to expect that head hunters are at all times seeking to hire your best and brightest out from under you--the way to stop them is by making it worth people's while to stick around. To my knowledge, histotechnologists are, as a rule of thumb, over-worked and under-appreciated. And yet they are darn handy to have around. I suggest that you make it your personal mission to ensure that your histotechs ARE NOT looking for another job. That way, when a recruiter calls, your techs say "Leave my current position? What are you crazy??!!!" Jerry Ricks Research Scientist University of Washington Department of Pathology. > Date: Thu, 11 Dec 2008 10:37:42 -0500 > From: MLafrini@csmlab.com > To: Histonet@lists.utsouthwestern.edu > CC: > Subject: [Histonet] Offended > > I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! > > Michael LaFriniere > Executive Director > CSM > Washington DC/Maryland/Virginia > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sraibley <@t> yahoo.com Thu Dec 11 14:13:54 2008 From: sraibley <@t> yahoo.com (Susan Raibley) Date: Thu Dec 11 14:13:57 2008 Subject: [Histonet] Re: processing mouse seminal vesicles Message-ID: <448976.49759.qm@web56005.mail.re3.yahoo.com> Try this processing schudule for mouse tissue: ? 70% 15 min 95% 15 min 95% 15 min 100% 15 min 100% 15 min 100% 15 min 100% 15 min Xylene 15 min Xylene 15 min Paraffin 15 min Paraffin 15 min Paraffin 15 min Paraffin 15 min ? Good luck! Susan Bincsik ? ? ? ? Does anybody have a protocol for this?? My last batch of these came out VERY dry and crunchy when run with other tissues on my standard protocol, which is as follows: (They are fixed on the benchtop in 10% NBF for 4-5 days, then rinsed out before processing.) 70%: 30 min 80%: 30 min 95%: 45 min 95%: 45 min 100%: 45 min 100%: 45 min xylene: 45 min xylene: 45 min Paraffin: 30 min Paraffin: 30min Paraffin: 30 min My other thought is that something is up with our VIP 5 processor, though no error messages are showing up.? Any and all suggestions are most welcome. Thanks in advance, Kathleen Roberts Rutgers University From mcauliff <@t> umdnj.edu Thu Dec 11 14:53:12 2008 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Thu Dec 11 14:55:22 2008 Subject: [Histonet] Silly Question? In-Reply-To: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> References: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Message-ID: <49417DB8.1040704@umdnj.edu> Hi Pat: The differences are largely in the minds of the investigators. Confusion comes from inexact nomenclature. One part of formaldehyde (37-40%) plus 9 parts of buffer makes formalin or 10% formalin which is about 4% formaldehyde. Yes, the 37-40% formaldehyde you buy has some methanol added to keep it from polymerizing but it makes no difference in the quality of fixation, especially given the (too) short times used to fix clinical specimens. Sure, you can go to the trouble to make 4% formaldehyde solution fresh from paraformaldehyde but for the vast majority of applications it just does not matter. Geoff Pat Flannery wrote: > Please humor me on this if it's obvious (to everyone but me): why do > we use paraformaldehyde (which is so inconvenient to make up) rather > than buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde (either > 2% or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > 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 mcauliff@umdnj.edu ********************************************** From bakevictoria <@t> gmail.com Thu Dec 11 15:03:01 2008 From: bakevictoria <@t> gmail.com (Victoria Baker) Date: Thu Dec 11 15:08:34 2008 Subject: [Histonet] RE: Offended In-Reply-To: <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> References: <4940ED37.588C.00AF.0@csmlab.com> <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: <4f016b690812111303v14c3297eq9ebf1991458ee04@mail.gmail.com> Hi Between symposiums, memberships or subscriptions it is possible for vendors and recruiters to get your home address. Sometimes you will be asked if you can refer someone to them as well if you are not interested. My dealings with recruiters/headhunters/vendors has always been pretty good and if or when I'm asked if I can refer someone - I always check with that person first as they may not want to be contacted. Everything in today's work or even private environment is not always so kept confidential or given the right to privacy that many of us are accustomed to and at times can be both intrusive as well as personally damaging. In business or work environments getting contacts is currently the catch-phrase 'networking' and do they ever work the 'net! Vikki On Thu, Dec 11, 2008 at 3:07 PM, Ingles Claire wrote: > A certain company has recently started sending histo related fliers TO MY HOUSE. I want to know where they got my home address, especially since I have never actually done business with this company. > Claire > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of JR R > Sent: Thu 12/11/2008 11:09 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Offended > > > > > Hi Michael, > > Yeah, I can see how that would be annoying to you. If you get a solicitation call at work that bugs you--tell them to buzz off. > > > I don't know that you ought to let it hurt your feelings or something, though. > > In the world of business, it is reasonable to expect that head hunters are at all times seeking to hire your best and brightest out from under you--the way to stop them is by making it worth people's while to stick around. > > To my knowledge, histotechnologists are, as a rule of thumb, over-worked and under-appreciated. > > And yet they are darn handy to have around. > > > I suggest that you make it your personal mission to ensure that your histotechs ARE NOT looking for another job. That way, when a recruiter calls, your techs say "Leave my current position? What are you crazy??!!!" > > Jerry Ricks > Research Scientist > University of Washington > Department of Pathology. > > >> Date: Thu, 11 Dec 2008 10:37:42 -0500 >> From: MLafrini@csmlab.com >> To: Histonet@lists.utsouthwestern.edu >> CC: >> Subject: [Histonet] Offended >> >> I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! >> >> Michael LaFriniere >> Executive Director >> CSM >> Washington DC/Maryland/Virginia >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ > Send e-mail faster without improving your typing skills. > http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008_______________________________________________ > 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 Thu Dec 11 15:14:57 2008 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Thu Dec 11 15:15:01 2008 Subject: [Histonet] posting for Tim Message-ID: <20081211141457.f86bd30e73b823f57b516b5451216a98.33a2126612.wbe@email.secureserver.net> Subject: respond directly to Tim on this at&nbs HELP ! HISTONET ADDRESS From: "Timothy Malloy" <[2]tmalloy@cox.net> ([3]Add as Preferred Sender) [4] 3D? < Date: Thu, Dec 11, 2008 2:57 am To: <[5]PRUEGG@IHCTECH.NET>, <[6]tmalloy77@hotm <[7]tmalloy@cox.net> Hi Patsy, I am a fan of yours on histonet. Just recently I moved my mail fr hotmail to outlook and lost histonets address. Could you supply me with likely to answ and we are in need o of a microwave. We have iss age of the building. Would you kno pneumocystisis on a hot plate?? Thank You Tim Malloy [8]tmalloy@cox.net or [9]tmalloy77@hotmail. Timothy Malloy HT (ASCP) A.A.S. References 1. 3D"http://em=/ 2. 3D"http://email.secureserver.net/addressBookQuickAdd.php?contact 3. 3D"http://email.secureserver.net/w 4. 3D"http://email.secureserver.net/webm 5. 3D"http://email.secureserver.net/addressBookQuickAdd.php?cont 6. 3D"http://email.secureserver.net/addr 7. 3D"http://emai=/ 8. file://localhost/tmp/3D"m 9. 3D"mailto:tmalloy77@hotmail.com" From Shakun.Aswani <@t> acologix.com Thu Dec 11 15:16:49 2008 From: Shakun.Aswani <@t> acologix.com (Shakun Aswani) Date: Thu Dec 11 15:16:52 2008 Subject: [Histonet] AxioVision Message-ID: <777AB0DE519C8E46A6220E2287C5BAD3019BB350@EXCHANGE.acologix.com> Hi, Is any one using Zeiss AxioVision software to analyze images for histomorphometery? Shakun P. Aswani Scientist I, Preclinical Development Acologix, Inc. 3960 Point Eden Way Hayward CA 94545 (510) 512-7231 phone (510) 786-1116 facsimile shakun.aswani@acologix.com From liz <@t> premierlab.com Thu Dec 11 15:33:11 2008 From: liz <@t> premierlab.com (Liz Chlipala) Date: Thu Dec 11 15:33:19 2008 Subject: [Histonet] AxioVision In-Reply-To: <777AB0DE519C8E46A6220E2287C5BAD3019BB350@EXCHANGE.acologix.com> References: <777AB0DE519C8E46A6220E2287C5BAD3019BB350@EXCHANGE.acologix.com> Message-ID: We have used Automeasure from Zeiss in the past for image analysis. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, CO 80308 phone (303) 682-3949 fax (303) 682-9060 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Shakun Aswani Sent: Thursday, December 11, 2008 2:17 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] AxioVision Hi, Is any one using Zeiss AxioVision software to analyze images for histomorphometery? Shakun P. Aswani Scientist I, Preclinical Development Acologix, Inc. 3960 Point Eden Way Hayward CA 94545 (510) 512-7231 phone (510) 786-1116 facsimile shakun.aswani@acologix.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carrolpb <@t> umdnj.edu Thu Dec 11 15:46:29 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Thu Dec 11 15:50:14 2008 Subject: [Histonet] RE: Offended In-Reply-To: <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> References: <4940ED37.588C.00AF.0@csmlab.com> <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: <49418A35.5050106@umdnj.edu> do you get any trade publications, by any chance? those people are the *worst* about selling your info... I signed up for one or two, now I get calls to my office almost every day for other publications, seminars and workshops that aren't even remotely relevant to my field... Guess its true there is in fact no free lunch these days... Ingles Claire wrote: > A certain company has recently started sending histo related fliers TO MY HOUSE. I want to know where they got my home address, especially since I have never actually done business with this company. > Claire > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of JR R > Sent: Thu 12/11/2008 11:09 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Offended > > > > > Hi Michael, > > Yeah, I can see how that would be annoying to you. If you get a solicitation call at work that bugs you--tell them to buzz off. > > > I don't know that you ought to let it hurt your feelings or something, though. > > In the world of business, it is reasonable to expect that head hunters are at all times seeking to hire your best and brightest out from under you--the way to stop them is by making it worth people's while to stick around. > > To my knowledge, histotechnologists are, as a rule of thumb, over-worked and under-appreciated. > > And yet they are darn handy to have around. > > > I suggest that you make it your personal mission to ensure that your histotechs ARE NOT looking for another job. That way, when a recruiter calls, your techs say "Leave my current position? What are you crazy??!!!" > > Jerry Ricks > Research Scientist > University of Washington > Department of Pathology. > > > >> Date: Thu, 11 Dec 2008 10:37:42 -0500 >> From: MLafrini@csmlab.com >> To: Histonet@lists.utsouthwestern.edu >> CC: >> Subject: [Histonet] Offended >> >> I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! >> >> Michael LaFriniere >> Executive Director >> CSM >> Washington DC/Maryland/Virginia >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > _________________________________________________________________ > Send e-mail faster without improving your typing skills. > http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008_______________________________________________ > 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 ejschmid <@t> ucalgary.ca Thu Dec 11 15:50:13 2008 From: ejschmid <@t> ucalgary.ca (ejschmid@ucalgary.ca) Date: Thu Dec 11 15:50:35 2008 Subject: [Histonet] glutaraldehyde autofluoresence Message-ID: <60668.136.159.150.24.1229032213.squirrel@136.159.150.24> Hi, My glutaraldehyde fixed mouse embryos fluoresce slightly reddish orange under rhodamine. I have read on this website's archives that using a solution of "bland amino acids" can help get rid of this. Does anyone have a protocol to fix this problem or can someone explain to me how and why this works? Eric Schmidt From leiker <@t> buffalo.edu Thu Dec 11 16:10:43 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Thu Dec 11 16:10:49 2008 Subject: [Histonet] RE: Offended In-Reply-To: <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> References: <4940ED37.588C.00AF.0@csmlab.com> <08A0A863637F1349BBFD83A96B27A50A08E609E3@uwhis-xchng3.uwhis.hosp.wisc.edu> Message-ID: <864B65D8E171539B216AB058@bchwxp2702.ad.med.buffalo.edu> 1.) Bio companies may sell their mailing lists to other companies... 2.) Do you have any online subscriptions to anything? They may sell your info. 3.) I've heard of companies scanning Facebook and MySpace - is your home address is on them and set to public? Just some guesses.... --On Thursday, December 11, 2008 2:07 PM -0600 Ingles Claire wrote: > A certain company has recently started sending histo related fliers TO MY > HOUSE. I want to know where they got my home address, especially since I > have never actually done business with this company. Claire > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of JR R > Sent: Thu 12/11/2008 11:09 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] RE: Offended > > > > > Hi Michael, > > Yeah, I can see how that would be annoying to you. If you get a > solicitation call at work that bugs you--tell them to buzz off. > > > I don't know that you ought to let it hurt your feelings or something, > though. > > In the world of business, it is reasonable to expect that head hunters > are at all times seeking to hire your best and brightest out from under > you--the way to stop them is by making it worth people's while to stick > around. > > To my knowledge, histotechnologists are, as a rule of thumb, over-worked > and under-appreciated. > > And yet they are darn handy to have around. > > > I suggest that you make it your personal mission to ensure that your > histotechs ARE NOT looking for another job. That way, when a recruiter > calls, your techs say "Leave my current position? What are you > crazy??!!!" > > Jerry Ricks > Research Scientist > University of Washington > Department of Pathology. > > >> Date: Thu, 11 Dec 2008 10:37:42 -0500 >> From: MLafrini@csmlab.com >> To: Histonet@lists.utsouthwestern.edu >> CC: >> Subject: [Histonet] Offended >> >> I Wanted to obtained opinions of Managers/Supervisors in the histo world >> and how you handle situations regarding recruiters calling directly into >> Pathology Labs to specific tech's and stating they are seeking referrals >> for job placements. I recently had an AP staffing solution company call >> one of my labs to talk to any "histologist" I felt was to recruit >> staff. I feel this is an unprofessional practice to call directly into >> the laboratory and to hide behind the wording "referrals" when actually >> it appears they are trying to get information from the tech on the phone >> seeing if they are looking for a "new job". I feel that this is an >> intrusion as well as inappropriate in our line of business and we are >> not in the business to give referrals to companies in the staffing >> solution arena for their monetary gain as well as trying to entice our >> staff. What the staff does with these types of companies on personal >> time is their business, however, I think it is offending for companies >> to demonstrate this behavior. If anyone would like to know which >> company and person called into my lab I will personally inform you on >> private email! >> >> Michael LaFriniere >> Executive Director >> CSM >> Washington DC/Maryland/Virginia >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ > Send e-mail faster without improving your typing skills. > http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_spee > d_122008_______________________________________________ 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 > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From AnthonyH <@t> chw.edu.au Thu Dec 11 16:16:02 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 11 16:16:13 2008 Subject: [Histonet] Silly Question? In-Reply-To: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Message-ID: Pat, I agree with you. In a routine diagnostic histopathology laboratory, it makes little difference what you use. Around the world for over 100 years most labs use 10% neutral buffered formalin made from concentrated 38%(or there abouts) formalin (or formaldehyde). Researchers, though, are a different kettle of fish. They will tend to hang on to misinformed, "mystical" methods believing they are being scientific. Funny, you would think that they, as a group, would be the ones pushing the boundaries and critically assessing each step of their research, ensuring that they understand what and why they are doing it. (Disclaimer - not all researchers are like this, thank heavens!!) Using a formaldehyde solution made from polyformaldehyde can cause problems. One researcher used it and wondered why their morphology was sub-optimal and their p53 immunohistochemistry was negative. He assured me that he had fixed small samples of tissue for 6 hours in 4% formaldehyde and then processed them using ethanol, xylene and wax. I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!! This also explains the loss of p53 staining. I gave him some of our routine 10% phosphate buffered fomalin, asked him to fix overnight, and try agin. Low and behold problem solved. How's that for a Friday Flamming!!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Friday, 12 December 2008 3:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ 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 note 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 Dec 11 16:18:05 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 11 16:18:10 2008 Subject: [Histonet] Silly Question? In-Reply-To: <4941498B.4020307@bms.com> Message-ID: Where is the evidence - or is this another mythical beast some of us believe? I don't believe it. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda M Watson Sent: Friday, 12 December 2008 4:11 AM To: Pat Flannery Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Silly Question? Hi Pat, Paraformaldehyde does not contain any additives and is considered more "pure" than formaldehyde which often contains methanol which in some cases is undesirable depending on the type of assay being conducted. Linda Pat Flannery wrote: > Please humor me on this if it's obvious (to everyone but me): why do > we use paraformaldehyde (which is so inconvenient to make up) rather > than buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde (either > 2% or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where > you were trained (i.e. force of habit) or is there a valid reason to > use each solution (basically the same chemical once in solution, > merely buffered or not)? The only answer I've gotten when I've asked > is, "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > 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 note 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 Dec 11 16:23:42 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 11 16:23:50 2008 Subject: [Histonet] Silly Question? In-Reply-To: <886EE175-ABA6-4325-BE60-369E64F324CE@duke.edu> Message-ID: And I won't stop!! I would prefer a slight methanol rather than no formaldehyde (see previous post) or formic acid being present. Also if some one sent me tissues fixed in a mercury containing fixative, I would report them and send it back. If researches or surgeons want quality results from my lab, they follow my rules or take a different road. But having said this I will negotiate, hoping to get the best quality that we can. Communication (and sometimes education) is the key Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Friday, 12 December 2008 4:22 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Silly Question? Thank you, Jeanine, Joyce, and Linda. The tissues I'm cutting are just for H&E staining and light microscopy; they won't be used for Immunostaining, antigen retrieval, or anything too fancy. I don't see what difference it would make, but of course I'll put them in whatever people ask for. I know that commercial formaldehyde is "stabilized" for storage so I can understand some folks wouldn't want the MeOH contamination, no matter how slight. -Pat _______________________________________________ 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 note 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 Dec 11 16:29:40 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Dec 11 16:29:48 2008 Subject: [Histonet] Silly Question? - Need help quickly! In-Reply-To: <8332CB03EAE51284E4AE67A6@bchwxp2702.ad.med.buffalo.edu> Message-ID: Good point Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Merced Leiker Sent: Friday, 12 December 2008 5:19 AM To: rjbuesa@yahoo.com; Pat Flannery; histonet@lists.utsouthwestern.edu; Weems, Joyce Subject: RE: [Histonet] Silly Question? - Need help quickly! So...is a polymer of paraformaldehyde considered "depolymerized" if it remains somewhere between 1-50 molecules long once it's been dissolved in solution, however it's dissolved? (the dissolving being a separate topic of debate on Histonet). Does it matter for tissue fixation purposes if there are formaldehyde chain lengths of 50 molecules present in solution - not long enough to precipitate out, but perhaps long enough to affect its penetration and fixing of tissues? Any ideas? Merced --On Thursday, December 11, 2008 9:58 AM -0800 Rene J Buesa wrote: > Joyce: > Methanal, which is the chemical name of formaldehyde, polymerizes. If > it forms a polymer of at least 50 molecules or more, it gets solid = > para-formaldehyde. Formalin (a trade name as formol is also another > trade name)is the 37-50% aqueous solution of formaldehyde (with some > additiveses to prevent polymerization). You can prepare BNF using the > formalin solution or dissolving the amount of solid > para-formaldehydede to get to the concentrationon you desire. The > chemical in both solutions is the same = methanal or formaldehyde.Ren? > J. > > --- On Thu, 12/11/08, Weems, Joyce wrote: > > > From: Weems, Joyce > Subject: RE: [Histonet] Silly Question? - Need help quickly! > To: "Pat Flannery" , > histonet@lists.utsouthwestern.edu > Date: Thursday, December 11, 2008, 12:12 PM > > I was just going to post a question regarding paraformaldhyde myself! > Just last week I believe I remember someone saying that > paraformaldehyde and formalin are the same and they had put the same > solution in two different containers for one of their researchers > because they were so insistent to have two different solutions. Are > they the same? > > Well, today I have a request to put tissue for a researcher in > formalin and paraformaldehyde. So.... Without percentage required, do > I use 10% NBF? Do I call somewhere and get paraformaldehyde and make > 4% paraformaldehyde? > > I have asked the surgeon twice for the number for the lab so I can > find out - don't have it yet. I have two fresh adrenals in the fridge. > Help!! > > > Thanks in advance... > Joyce > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 678-843-7376 - Phone > 678-843-7831 - Fax > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Thursday, December 11, 2008 11:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > > Please humor me on this if it's obvious (to everyone but me): why do > we use paraformaldehyde (which is so inconvenient to make up) rather > than buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde (either > 2% or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Confidentiality Notice: > This email, including any attachments is the > property of Catholic Health East and is intended > for the sole use of the intended recipient(s). > It may contain information that is privileged and confidential. Any > unauthorized review, use, disclosure, or distribution is prohibited. > If you are not the intended recipient, please reply to the > sender that you have received the message in > error, then delete this 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 > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator _______________________________________________ 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 note 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 laurie.colbert <@t> huntingtonhospital.com Thu Dec 11 17:51:01 2008 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Thu Dec 11 17:51:10 2008 Subject: [Histonet] Coverslipping Fume Hoods Message-ID: <57BE698966D5C54EAE8612E8941D76830469774D@EXCHANGE3.huntingtonhospital.com> Can anyone recommend a heavy-duty fume hood that we can use for coverslipping? We are currently using a Fume Guard hood that is probably 20 years old. We use HistoClear to coverslip, and we would like to cut down on the vapors/smell. Laurie Colbert From bethcoxx <@t> gmail.com Thu Dec 11 17:58:46 2008 From: bethcoxx <@t> gmail.com (Beth Cox) Date: Thu Dec 11 17:58:43 2008 Subject: [Histonet] Re: Non-gyn cytology prep - amount processed? Message-ID: <4941A936.4090508@gmail.com> Hi Cathy, Your message came through a little garbled, but I think I get the jist of it. Standard procedure for large volume cytology fluid specimens is to mix the specimen well, and then take off a 50 ml aliquot to centrifuge down and make the slides and cell block. If the specimen is not mixed, you are likely to miss the settled malignant cells. It is not prudent (or recommended!) to centrifuge down the entire specimen. If a specimen were extremely sparsely cellular, I might occasionally spin down two 50 ml aliquots. I agree that if a specimen has set for a while and settled naturally, pouring off the supernatent and using the concentrated precipitate for a 50 ml aliquat can be a nice bonus. BUT, there should have been malignant cells in the WELL-MIXED original aliquot. What prep procedures are you using for the slides?? ThinPrep processing can be subject to problems with these kind of specimens because sometimes the filter becomes 'clogged' with fibrin and protein, and the processor thinks it has cells, but not much is there. ThinPrep may not be the best choice for body fluid specimens. A well done cell block should also have shown cells in it. If you are simply scraping loose cells into a lens paper to process and try to embed for a cell block, I'm not surprised if you lost the cells and only recovered the more prominent fibrin. It's always a good idea to use some method to actually hold (block!) the cells together - such as agar, thrombin-prothrombin, or albumin clot. Beth Cox, SCT/HT(ASCP) _____________________________________ Message: 3 Date: Thu, 11 Dec 2008 10:10:31 -0800 From: Cathy.Crumpton@tuality.org Subject: [Histonet] Non-gyn cytology prep - amount processed? To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="ISO-8859-1" Hi all, for those of you who also work with cytology - what is procedure for fluids that have a large volume? How much of the fluid do you process? I am wanting to compare our polic instance this week where we received processed 100 mL of that after making sure it first cell blocks only contained fibrin, but the malignant cells. When we went back to the contai settled after 24 hours so we poured off the top part of t and only processed the gunk at the bottom. The second time t were malignant cells in the cell block. My pathologist is pushin for us to process 100% of the fluids, 100% of the time, saying what we cu get those From jnocito <@t> satx.rr.com Thu Dec 11 19:19:15 2008 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Thu Dec 11 19:19:18 2008 Subject: [Histonet] Offended References: <4940ED37.588C.00AF.0@csmlab.com> Message-ID: <1AA7429A27114D9DBCFD8C4D56BF4F9A@yourxhtr8hvc4p> here we go and it's almost Friday. I received a call just like that at my last job. Me being me, I gave her a piece of my mind ( not too much, I don't have that much to go around). Told her she had some big testicles (not the word I used, but you get the idea) and hung up. I then called in the tech and had a talk with him. Asked him if he was looking for a job. He told me that he wasn't and didn't know how she got his name. I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door. There are other ways to go about this. Calling a lab is not one of them, but I'm sure some people give their work number for different reasons, trying to use it as leverage for a pay raise, to get the supervisors nervous, because the techs are that brazen, or belong to the id10t club. Let's face it, we are becoming a scarce breed and people have quotas to meet. Some will stop at nothing. That's my story and I'm sticking to it. JTT ----- Original Message ----- From: "Michael LaFriniere" To: Sent: Thursday, December 11, 2008 9:37 AM Subject: [Histonet] Offended I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Thu Dec 11 19:29:59 2008 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Thu Dec 11 19:32:31 2008 Subject: [Histonet] Offended Message-ID: <1628650306-1229045542-cardhu_decombobulator_blackberry.rim.net-627675921-@bxe173.bisx.prod.on.blackberry> Joe, I think firing a tech because a head hunter calls is unfair and maybe a case for a lawsuit of improper termination. Often the heah hunters get your name through the grapevine, from former employees or from publications. --------------------------------- Joe said: "I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door." From RSRICHMOND <@t> aol.com Thu Dec 11 20:20:33 2008 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Thu Dec 11 20:20:39 2008 Subject: [Histonet] Re: Non-gyn cytology prep - amount processed? Message-ID: Cathy Crumpton asks (the message seems to have gotten garbled in transmission): >> Hi all, for those of you who also work with cytology - what is procedure for fluids that have a large volume? How much of the fluid do you process? - I am wanting to compare our polic instance this week where we received processed 100 mL of that after making sure it first cell blocks only contained fibrin, but the malignant cells. When we went back to the contai settled after 24 hours so we poured off the top part of t and only processed the gunk at the bottom. The second time t were malignant cells in the cell block. My pathologist is pushin for us to process 100% of the fluids, 100% of the time, saying what we cu get those<< You certainly cannot process an entire large liquid specimen for cytologic examination - many pleural fluid specimens are well over a litre. There are a number of ways to prepare a cell block from such a specimen - an important adjunct to the cytologic examination examination of such fluids. A lot of laboratories have trouble preparing cell blocks consistently. I'd suggested getting a method from a cytopathology service that has it working - and probably visiting the service. If your pathologist thinks you can process 100% of a one litre specimen, he isn't going to be of much help to you in getting a procedure working. Bob Richmond Samurai Pathologist Knoxville TN From geoweigel <@t> hotmail.com Thu Dec 11 22:50:49 2008 From: geoweigel <@t> hotmail.com (Georgia Weigel) Date: Thu Dec 11 22:50:52 2008 Subject: [Histonet] Re: Non-gyn cytology prep Message-ID: When doing cell blocks you should not process the fibrin tissue or what is floating on top. At least that has always been the preference of the pathologist's that I've worked with. They don't want to see the fibrin or the junk that's floating on top. You will not get the cells that they want to see from the fibrin. Your cells are going to be in the button, or the so called "gunk" on the bottom of the conical tube. You should centrifuge at least a good 50ml for a cell block. Now, if your button after centrifugation is still rather small and you have additional fluid then pour off the top and spin down more fluid until your button is a good size. G Weigel, HT (ASCP) _________________________________________________________________ Suspicious message? There?s an alert for that. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_broad2_122008 From tifei <@t> foxmail.com Fri Dec 12 02:21:50 2008 From: tifei <@t> foxmail.com (tf) Date: Fri Dec 12 02:22:18 2008 Subject: [Histonet] perfusion question References: <000d01c95aff$4d8a2830$9f2de143@NEIL>, Message-ID: <200812121621456839103@foxmail.com> SGksIEJlY2F1c2Ugd2FzaCBvdXQgc29sdXRpb24gY2FuIGJlIDAuOSUgc2FsaW5lIG9yIDAuMDEg TSBQQlMuLi4ubmVhciB0byBwaHlzaW9sb2dpY2FsIG9zbW90aWMgcHJlc3N1cmUuLi4NCg0KSG93 ZXZlciB0aGUgcGVyZnVzaW9uIHNvbHV0aW9uICg0JSBQRkEgaW4gMC4xTSBQQikgaXMgbW9yZSBj b25jZW50cmF0ZWQsIGluIHRoZSBzZW5zZSBvZiBvc21vdGljIHByZXNzdXJlLi4uLkNhbiB5b3Ug Y29tbWVudCBvbiB0aGlzPw0KDQpZb3UgY2FuIG5vdGUgdGhlIHNocmlua2FnZSBvZiBicmFpbiBh ZnRlciBQRkEgcGVyZnVzaW9uLn4NCg0KDQoyMDA4LTEyLTEyIA0KDQoNCg0KdGYgDQoNCg0KDQq3 orz+yMujuiBKb2huIEtpZXJuYW4gDQq3osvNyrG85KO6IDIwMDgtMTItMTIgIDAzOjE2OjMyIA0K ytW8/sjLo7ogTmVpbCBGb3VybmllciANCrOty82juiBoaXN0b25ldEBsaXN0cy51dHNvdXRod2Vz 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DQo= From tifei <@t> foxmail.com Fri Dec 12 02:35:28 2008 From: tifei <@t> foxmail.com (tf) Date: Fri Dec 12 02:35:51 2008 Subject: [Histonet] (reply) silly questions.---PFA References: Message-ID: <200812121635232792765@foxmail.com> IkkgbG9va2VkIGF0IHRoZSBzZWN0aW9ucyBhbmQgdGhlIGNlbGwgc2hyaW5rYWdlIChhbmQgcHJv bWluZW50IHNwYWNlcw0KYmV0d2VlbiBjZWxscyBhbmQgY29ubmVjdGl2ZSB0aXNzdWUpIGluZGlj YXRlZCB0aGF0IG1vc3Qgb2YgdGhlDQoiZml4YXRpb24iIHNlZW1lZCB0byBoYXZlIG9jY3VyZWQg aW4gdGhlIHByb2Nlc3NpbmcgZXRoYW5vbHMuIEkgYXNrZWQNCmhpbSBmb3Igc29tZSBvZiB0aGUg Zml4YXRpdmUgaGUgdXNlZCwgdGVzdGVkIHRoZSBmb3JtYWxkZWh5ZGUNCmNvbmNlbnRyYXRpb24g YW5kIGZvdW5kIGl0IHRvIGJlIGxlc3MgdGhhbiAwLjUlISEiDQoNClRvbnk6IERvIHlvdSB0aGlu ayB0aGlzIGlzIGJlY2F1c2Ugb2YgaW5wcm9wZXIgcHJlcGFyYXRpb24gb2YgUEZBIGluIGhpcyBs YWIsIG9yIHRoZSBjb21tb24gcHJvYmxlbSBpbiBhbGwgcmVzZWFyY2hlcnMgdXNpbmcgUEZBPw0K ICAgICAgICAgSSBkbyB0aGluayBtb3N0IGJpb21lZGljYWwgbGFicyBjdXJyZW50bHkgYXJlIHVz 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aGFua3MuDQotUGF0IEZsYW5uZXJ5IChub3QgYSAicmVhbCIgaGlzdG9sb2dpc3QgLSBJIGp1c3Qg cGxheSBvbmUgaW4gdGhlIGxhYikNCl9fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19f X19fX19fX19fX19fDQpIaXN0b25ldCBtYWlsaW5nIGxpc3QNCkhpc3RvbmV0QGxpc3RzLnV0c291 dGh3ZXN0ZXJuLmVkdQ0KaHR0cDovL2xpc3RzLnV0c291dGh3ZXN0ZXJuLmVkdS9tYWlsbWFuL2xp c3RpbmZvL2hpc3RvbmV0DQoqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioq KioqKioqKioqKioqKioqKioqKioqKioqKioqKioNClRoaXMgZW1haWwgYW5kIGFueSBmaWxlcyB0 cmFuc21pdHRlZCB3aXRoIGl0IGFyZSBjb25maWRlbnRpYWwgYW5kIGludGVuZGVkIHNvbGVseSBm b3IgdGhlIHVzZSBvZiB0aGUgaW5kaXZpZHVhbCBvciBlbnRpdHkgdG8gd2hvbSB0aGV5IGFyZSBh ZGRyZXNzZWQuIElmIHlvdSBhcmUgbm90IHRoZSBpbnRlbmRlZCByZWNpcGllbnQsIHBsZWFzZSBk ZWxldGUgaXQgYW5kIG5vdGlmeSB0aGUgc2VuZGVyLg0KVmlld3MgZXhwcmVzc2VkIGluIHRoaXMg bWVzc2FnZSBhbmQgYW55IGF0dGFjaG1lbnRzIGFyZSB0aG9zZSBvZiB0aGUgaW5kaXZpZHVhbCBz ZW5kZXIsIGFuZCBhcmUgbm90IG5lY2Vzc2FyaWx5IHRoZSB2aWV3cyBvZiBUaGUgQ2hpbGRyZW4n cyBIb3NwaXRhbCBhdCBXZXN0bWVhZA0KVGhpcyBub3RlIGFsc28gY29uZmlybXMgdGhhdCB0aGlz IGVtYWlsIG1lc3NhZ2UgaGFzIGJlZW4NCnZpcnVzIHNjYW5uZWQgYW5kIGFsdGhvdWdoIG5vIGNv bXB1dGVyIHZpcnVzZXMgd2VyZSBkZXRlY3RlZCwgVGhlIENoaWxkcmVucyBIb3NwaXRhbCBhdCBX ZXN0bWVhZCBhY2NlcHRzIG5vIGxpYWJpbGl0eSBmb3IgYW55IGNvbnNlcXVlbnRpYWwgZGFtYWdl IHJlc3VsdGluZyBmcm9tIGVtYWlsIGNvbnRhaW5pbmcgY29tcHV0ZXIgdmlydXNlcy4NCioqKioq KioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioq KioqKioqKioNCl9fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19fX19f DQpIaXN0b25ldCBtYWlsaW5nIGxpc3QNCkhpc3RvbmV0QGxpc3RzLnV0c291dGh3ZXN0ZXJuLmVk dQ0K From tmalloy77 <@t> hotmail.com Fri Dec 12 03:52:19 2008 From: tmalloy77 <@t> hotmail.com (TIMOTHY MALLOY) Date: Fri Dec 12 03:52:27 2008 Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS Message-ID: We currently use a stain called grocotts silver methenimine solution for staining pnuemocystis. We would like to use a hot plate for the procedure instead of a unvented microwave. Does anybody have any formula's the would like to share with the hot plate? Timothy G. Malloy, HT ( ASCP ) A.A.S. tmalloy77@hotmail.com This email communication may contain CONFIDENTIAL INFORMATION WHICH ALSO MAY BE LEGALLY PRIVILEGED and is intended only for the use of the intended recipients identified above. If you are not the intended recipient of this communication, you are hereby notified that any unauthorized review, use, dissemination, distribution, downloading, or copying of this communication is strictly prohibited. If you have received this communication in error, please immediately notify us by reply email, delete the communication and destroy all copies. From lpwenk <@t> sbcglobal.net Fri Dec 12 04:03:03 2008 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Dec 12 04:03:13 2008 Subject: [Histonet] training materials In-Reply-To: Message-ID: <001001c95c40$d3019b50$0202a8c0@HPPav2> Not an immediate help, but there will be a NSH teleconference May 27, 2009 on embedding. PARAFFIN EMBEDDING AND PROCESS IMPROVEMENT Date: May 27, 2009 Time: 1:00PM EST Presented by Joelle Weaver, HTL(ASCP), Blanchard Valley Hospital, Findlay, Ohio; Riverside Methodist Hospital, Columbus, Ohio; Histology Education Coordinator, Columbus State Community College, Columbus, Ohio This presentation is a good review of fundamentals for everyone who routinely embeds a variety of tissue types. The embedding step is often overlooked in process improvement initiatives. At many labs, the embedding step has remained relatively static. What is being done at your facility during embedding to eliminate contamination as well as orientate tissue correctly and optimally for sectioning quality? This teleconference will begin with a review of the fundamentals of tissue embedding and orientation, covering basic principles of tissue sampling and specimen inking that may assist the embedding histologist. The impact of well infiltrated, correctly spaced, properly oriented specimens is strongly correlated to both ease of sectioning as well as section quality. Therefore, it is worth taking the time to consider this topic with "fresh eyes" and to re-evaluate embedding methodology when other changes are made in the overall process. For info on this and other NSH Teleconferences, go to: http://www.e-guana.net/organizations.php3?orgid=111&typeID=1184&action=print ContentTypeHome&User_Session=41327fea4e36d34fede5e76bf55697d2 If having problems with the link, go to: www.nsh.org Click on Continuing Education on the left Click on Earn Continuing Education Click on Partipate in Teleconferences, about half way down the page The 2009 Schedule is available. All are on Wednesdays, from 1-2 pm Eastern time. Cost is $125 per teleconference, or if order all 11 by Jan. 27, 2009, it's only $1100, which is $100 each. You can have as many people listen as you want, and they all earn CE certificates. (If you have 10 people working in your lab, that bring the cost down to $12.50 per person for each to earn 1 hour CE. The Jan 2009 is "Working with Difficult People", so invite everyone from all the labs and the pathology office and the pathologists, and everyone can learn to get along with earning 1 hour CE!) About 1 week before the teleconference, your lab will be sent a link to the PowerPoint and any additional handouts. The day of the talk, call up a phone number, listen to the speaker through speaker phone and be able to ask questions, and have everyone sign in, which is then faxed to NSH. Everyone who attended and signed in earns 1 hour CE. About 1-2 months after the teleconference, the lab will receive a CD with the PowerPoint, speaker's talks, any additional handouts, and a 4 question test. So if anyone couldn't attend the original teleconference, they can sit down when there is some free time in the lab, listen to the conference, view the PowerPoint, read the additional handouts, and now take a 4 question test, which they take and fax/mail into NSH, and they can get 1 hour CE, up to 2 years later. Also, your lab now has good training modules for anyone at any time. Peggy A. Wenk, HTL(ASCP)SLS Representing NSH as the NSH Teleconference Coordinator -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jennifer Johnson Sent: Thursday, December 11, 2008 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] training materials Can anyone suggest a really good book, atlas, etc. for embedding? The girl that took my place at my last job is having a really hard time (especially with skin) and I told her I would ask the experts. Thanks, Jennifer Johnson, HTL (ASCP) _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_1 22008_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Fri Dec 12 05:38:00 2008 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Fri Dec 12 05:37:41 2008 Subject: [Histonet] Paraffin wax In-Reply-To: <4b6c85510812110706h7864d3a3pf4c0da66ac68442@mail.gmail.com> References: <1B2F365C5F88A946B7D602E64ACD9CDD0447FD1EF0@ENHBGMBX04.PA.LCL> <4b6c85510812110706h7864d3a3pf4c0da66ac68442@mail.gmail.com> Message-ID: We have used Paraplast Plus for years. For both processing and embedding. Betsy Molinari HT(ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave MC 1-283 Houston, TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Histonet Alias Sent: Thursday, December 11, 2008 9:07 AM To: Charles, Roger Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Paraffin wax I have had very good luck with Paraplast Plus for infiltration and using the Paraplast Xtra for embedding. On Thu, Dec 11, 2008 at 10:00 AM, Charles, Roger wrote: > Hello, > Could someone share their knowledge of the Paraplast line of paraffin with > me? We are being forced to change from the TissuePrep line. > Thanks > > Roger Charles > Microbiologist > Pennsylvania Veterinary Laboratory > 2305 N Cameron St > Harrisburg, PA 17110 > 717-787-8808 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katherine-walters <@t> uiowa.edu Fri Dec 12 07:48:18 2008 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Fri Dec 12 07:49:29 2008 Subject: [Histonet] (reply) silly questions.---PFA In-Reply-To: <200812121635232792765@foxmail.com> References: <200812121635232792765@foxmail.com> Message-ID: This may be another silly question, but how does one test the concentration= of formaldehyde in solution? Thanks, Kathy Notice: This UI Health Care e-mail (including attachments) is covered by = the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confiden= tial and may be legally privileged. If you are not the intended recipient,= you are hereby notified that any retention, dissemination, distribution, o= r copying of this communication is strictly prohibited. Please reply to th= e sender that you have received the message in error, then delete it. Than= k you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@= lists.utsouthwestern.edu] On Behalf Of tf Sent: Friday, December 12, 2008 2:35 AM To: Tony Henwood; Pat Flannery; histonet@lists.utsouthwestern.ed Subject: [Histonet] (reply) silly questions.---PFA "I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!!" Tony: Do you think this is because of inproper preparation of PFA in his = lab, or the common problem in all researchers using PFA? I do think most biomedical labs currently are using PFA to prepa= re the fixatives! So, anyone has the idea on a correction preparation procedure of 4% PFA? I noticed some of you dissolve PFA powder in NaOH-conditioned alkaline wa= ter, then add concentrated PB solution. We here dissolve PFA in concentrated PB solution directly (heat & stir fo= r 2-3 hours), then adjust pH to 7.4. We dont have big problem in tissue quaility....except when one want to cu= t the brain in a cryostat rather sliding microtome. Many times the brain sections from the cryostat have "cheese" like holes/= cavities, which almost never appear on sliding microtome-prepared sections. 2008-12-12 tf =B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-12 06:18:47 =CA=D5=BC=FE=C8=CB=A3=BA Pat Flannery; histonet@lists.utsouthwestern.edu =B3=AD=CB=CD=A3=BA =D6=F7=CC=E2=A3=BA RE: [Histonet] Silly Question? Pat, I agree with you. In a routine diagnostic histopathology laboratory, it makes little difference what you use. Around the world for over 100 years most labs use 10% neutral buffered formalin made from concentrated 38%(or there abouts) formalin (or formaldehyde). Researchers, though, are a different kettle of fish. They will tend to hang on to misinformed, "mystical" methods believing they are being scientific. Funny, you would think that they, as a group, would be the ones pushing the boundaries and critically assessing each step of their research, ensuring that they understand what and why they are doing it. (Disclaimer - not all researchers are like this, thank heavens!!) Using a formaldehyde solution made from polyformaldehyde can cause problems. One researcher used it and wondered why their morphology was sub-optimal and their p53 immunohistochemistry was negative. He assured me that he had fixed small samples of tissue for 6 hours in 4% formaldehyde and then processed them using ethanol, xylene and wax. I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!! This also explains the loss of p53 staining. I gave him some of our routine 10% phosphate buffered fomalin, asked him to fix overnight, and try agin. Low and behold problem solved. How's that for a Friday Flamming!!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Friday, 12 December 2008 3:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ 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 intende= d 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 se= nder. Views expressed in this message and any attachments are those of the indi= vidual sender, and are not necessarily the views of The Children's Hospital= at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Childre= ns Hospital at Westmead accepts no liability for any consequential damage r= esulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From ree3 <@t> leicester.ac.uk Fri Dec 12 08:04:29 2008 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Dec 12 08:04:35 2008 Subject: [Histonet] Silly Question? In-Reply-To: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> References: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> Message-ID: <7722595275A4DD4FA225B92CDBF174A174501D423F@EXC-MBX3.cfs.le.ac.uk> You hit the nail on the head "That's what we always use", fear of change is a common human condition. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: 11 December 2008 16:59 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Fri Dec 12 08:14:18 2008 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Dec 12 08:14:25 2008 Subject: [Histonet] Offended In-Reply-To: <1AA7429A27114D9DBCFD8C4D56BF4F9A@yourxhtr8hvc4p> References: <4940ED37.588C.00AF.0@csmlab.com> <1AA7429A27114D9DBCFD8C4D56BF4F9A@yourxhtr8hvc4p> Message-ID: <44780C571F28624DBB446DE55C4D733A1FE619@EXCHANGEBE1.carle.com> Joe, I can hardly believe you could be that grouchy :). When I was looking hard for a tech I called my buddy Matt Chase at Children's to try and coax him over to my lab. When he said no, I asked if he would let me chat with his techs and see how they were doing. Alas, he also replied negatively to that. The truth is that techs are becoming harder to find and are going at a premium price. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, December 11, 2008 7:19 PM To: Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Offended here we go and it's almost Friday. I received a call just like that at my last job. Me being me, I gave her a piece of my mind ( not too much, I don't have that much to go around). Told her she had some big testicles (not the word I used, but you get the idea) and hung up. I then called in the tech and had a talk with him. Asked him if he was looking for a job. He told me that he wasn't and didn't know how she got his name. I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door. There are other ways to go about this. Calling a lab is not one of them, but I'm sure some people give their work number for different reasons, trying to use it as leverage for a pay raise, to get the supervisors nervous, because the techs are that brazen, or belong to the id10t club. Let's face it, we are becoming a scarce breed and people have quotas to meet. Some will stop at nothing. That's my story and I'm sticking to it. JTT ----- Original Message ----- From: "Michael LaFriniere" To: Sent: Thursday, December 11, 2008 9:37 AM Subject: [Histonet] Offended I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia _______________________________________________ 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 lblazek <@t> digestivespecialists.com Fri Dec 12 08:22:11 2008 From: lblazek <@t> digestivespecialists.com (Blazek, Linda) Date: Fri Dec 12 08:20:44 2008 Subject: [Histonet] Offended In-Reply-To: <44780C571F28624DBB446DE55C4D733A1FE619@EXCHANGEBE1.carle.com> References: <4940ED37.588C.00AF.0@csmlab.com> <1AA7429A27114D9DBCFD8C4D56BF4F9A@yourxhtr8hvc4p> <44780C571F28624DBB446DE55C4D733A1FE619@EXCHANGEBE1.carle.com> Message-ID: <5A2BD13465E061429D6455C8D6B40E3907C253440D@IBMB7Exchange.digestivespecialists.com> Matt may have said no but we still found out! Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lblazek@digestivespecialists.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Friday, December 12, 2008 9:14 AM To: Joe Nocito; Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Offended Joe, I can hardly believe you could be that grouchy :). When I was looking hard for a tech I called my buddy Matt Chase at Children's to try and coax him over to my lab. When he said no, I asked if he would let me chat with his techs and see how they were doing. Alas, he also replied negatively to that. The truth is that techs are becoming harder to find and are going at a premium price. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, December 11, 2008 7:19 PM To: Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Offended here we go and it's almost Friday. I received a call just like that at my last job. Me being me, I gave her a piece of my mind ( not too much, I don't have that much to go around). Told her she had some big testicles (not the word I used, but you get the idea) and hung up. I then called in the tech and had a talk with him. Asked him if he was looking for a job. He told me that he wasn't and didn't know how she got his name. I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door. There are other ways to go about this. Calling a lab is not one of them, but I'm sure some people give their work number for different reasons, trying to use it as leverage for a pay raise, to get the supervisors nervous, because the techs are that brazen, or belong to the id10t club. Let's face it, we are becoming a scarce breed and people have quotas to meet. Some will stop at nothing. That's my story and I'm sticking to it. JTT ----- Original Message ----- From: "Michael LaFriniere" To: Sent: Thursday, December 11, 2008 9:37 AM Subject: [Histonet] Offended I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia _______________________________________________ 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 tifei <@t> foxmail.com Fri Dec 12 09:12:07 2008 From: tifei <@t> foxmail.com (tf) Date: Fri Dec 12 09:12:27 2008 Subject: [Histonet] re: concentration of formaldehyde in soultion: ASTM D2194 - 02(2007) Standard Test Method for Concentration of Formaldehyde Solution References: , <200812121635232792765@foxmail.com>, Message-ID: <200812122312023456365@foxmail.com> 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(Merced Leiker) Date: Fri Dec 12 09:12:37 2008 Subject: [Histonet] Silly Question? In-Reply-To: <7722595275A4DD4FA225B92CDBF174A174501D423F@EXC-MBX3.cfs.le.ac.uk> References: <5CB1BA86-6596-436A-8897-B10BBAB0B8F7@duke.edu> <7722595275A4DD4FA225B92CDBF174A174501D423F@EXC-MBX3.cfs.le.ac.uk> Message-ID: In research lab situations particularly, one does not have the time or technique for nailing down the ways of making each of the buffers, reagents, and procedures work the "right" way or the most optimum way...a lot of times it's students or postdocs just focused on getting their project done and not caring how their fixative is made as long as it "works" to some degree and, alas, it's up to us already over-booked technicians to figure out the best way to make the PFA....and we usually don't have a whole day (week, or year) to spend researching the back-and-forth arguments, either! ;-) Merced --On Friday, December 12, 2008 2:04 PM +0000 "Edwards, R.E." wrote: > You hit the nail on the head "That's what we always use", fear of > change is a common human condition. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery Sent: 11 December 2008 16:59 > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > > Please humor me on this if it's obvious (to everyone but me): why do > we use paraformaldehyde (which is so inconvenient to make up) rather > than buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde (either > 2% or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > 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 > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From leiker <@t> buffalo.edu Fri Dec 12 09:16:20 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Fri Dec 12 09:16:28 2008 Subject: [Histonet] (reply) silly questions.---PFA In-Reply-To: References: <200812121635232792765@foxmail.com> Message-ID: <8B5E25FBBBBB9B13F09E4237@bchwxp2702.ad.med.buffalo.edu> Oh, I'd like to know that, too, please! --On Friday, December 12, 2008 7:48 AM -0600 "Walters, Katherine S" wrote: > This may be another silly question, but how does one test the > concentration of formaldehyde in solution? > > Thanks, > Kathy > > > > Notice: This UI Health Care e-mail (including attachments) is covered by > the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is > confidential and may be legally privileged. If you are not the intended > recipient, you are hereby notified that any retention, dissemination, > distribution, or copying of this communication is strictly prohibited. > Please reply to the sender that you have received the message in error, > then delete it. Thank you. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of tf Sent: > Friday, December 12, 2008 2:35 AM > To: Tony Henwood; Pat Flannery; histonet@lists.utsouthwestern.ed > Subject: [Histonet] (reply) silly questions.---PFA > > "I looked at the sections and the cell shrinkage (and prominent spaces > between cells and connective tissue) indicated that most of the > "fixation" seemed to have occured in the processing ethanols. I asked > him for some of the fixative he used, tested the formaldehyde > concentration and found it to be less than 0.5%!!" > > Tony: Do you think this is because of inproper preparation of PFA in his > lab, or the common problem in all researchers using PFA? I do > think most biomedical labs currently are using PFA to prepare the > fixatives! > So, anyone has the idea on a correction preparation procedure of 4% PFA? > I noticed some of you dissolve PFA powder in NaOH-conditioned alkaline > water, then add concentrated PB solution. We here dissolve PFA in > concentrated PB solution directly (heat & stir for 2-3 hours), then > adjust pH to 7.4. > > We dont have big problem in tissue quaility....except when one want to > cut the brain in a cryostat rather sliding microtome. Many times the > brain sections from the cryostat have "cheese" like holes/cavities, which > almost never appear on sliding microtome-prepared sections. > > 2008-12-12 > > > > tf > > > > ???????? Tony Henwood > ?????????? 2008-12-12 06:18:47 > ???????? Pat Flannery; histonet@lists.utsouthwestern.edu > ?????? > ?????? RE: [Histonet] Silly Question? > > Pat, > I agree with you. > In a routine diagnostic histopathology laboratory, it makes little > difference what you use. Around the world for over 100 years most labs > use 10% neutral buffered formalin made from concentrated 38%(or there > abouts) formalin (or formaldehyde). > Researchers, though, are a different kettle of fish. They will tend to > hang on to misinformed, "mystical" methods believing they are being > scientific. Funny, you would think that they, as a group, would be the > ones pushing the boundaries and critically assessing each step of their > research, ensuring that they understand what and why they are doing it. > (Disclaimer - not all researchers are like this, thank heavens!!) > Using a formaldehyde solution made from polyformaldehyde can cause > problems. One researcher used it and wondered why their morphology was > sub-optimal and their p53 immunohistochemistry was negative. He assured > me that he had fixed small samples of tissue for 6 hours in 4% > formaldehyde and then processed them using ethanol, xylene and wax. > I looked at the sections and the cell shrinkage (and prominent spaces > between cells and connective tissue) indicated that most of the > "fixation" seemed to have occured in the processing ethanols. I asked > him for some of the fixative he used, tested the formaldehyde > concentration and found it to be less than 0.5%!! > This also explains the loss of p53 staining. I gave him some of our > routine 10% phosphate buffered fomalin, asked him to fix overnight, and > try agin. Low and behold problem solved. > How's that for a Friday Flamming!!! > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Friday, 12 December 2008 3:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > Please humor me on this if it's obvious (to everyone but me): why do > we use paraformaldehyde (which is so inconvenient to make up) rather > than buffered formalin or just diluted formaldehyde itself? > It seems that around here, some folks prefer paraformaldehyde (either > 2% or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've asked is, > "That's what we always use." > Thanks. > -Pat Flannery (not a "real" histologist - I just play one in the lab) > _______________________________________________ > 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 note 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. > ********************************************************************** > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From michelle.steinkrauss <@t> novartis.com Fri Dec 12 09:34:40 2008 From: michelle.steinkrauss <@t> novartis.com (michelle.steinkrauss@novartis.com) Date: Fri Dec 12 09:34:48 2008 Subject: [Histonet] Michelle Steinkrauss is out of the office. Message-ID: I will be out of the office starting 12/12/2008 and will not return until 12/15/2008. If you require immediate assistance, please contact Michelle Broome at x 47477. From PVerden <@t> UROPARTNERS.COM Fri Dec 12 09:35:13 2008 From: PVerden <@t> UROPARTNERS.COM (Paul Verden) Date: Fri Dec 12 09:35:18 2008 Subject: [Histonet] formalin concentration Message-ID: If you are interested in knowing the concentration of formaldehyde, i.e., making sure you have 10% formalin, you can contact CBG Biotech at 800-941-9484. They sell and service one of the best solvent recyclers I have ever used. When I was at Rockford Memorial Hospital, we used one of their recyclers. The calculation of formaldehyde/formalin is a very easy test. I used it whenever we recycled formalin, which was all the time! It can be used to test both buffered and non-buffered solutions. The test procedures and reagents were provided by CBG. Please contact them with your request. They may be able to help you. R. Paul Verden Supervisor - Cytology and Molecular UroPartners, LLC Westchester, IL 708-486-0076 From smit_d3 <@t> yahoo.com Fri Dec 12 10:52:18 2008 From: smit_d3 <@t> yahoo.com (smit dangaria) Date: Fri Dec 12 10:52:25 2008 Subject: [Histonet] regarding problems with decalcification of rat maxilla's Message-ID: <775887.25260.qm@web8507.mail.in.yahoo.com> Hi, I am a graduate student at University of Illinois, chicago.? I have been having tremendous problems with sectioning of paraffin embedded rat maxilla with molars and I searched the Histonet archives and found that people here have invaluable experience, insight and suggestions for various problems. Heres my problem. : As I section the block the section would crumble and tear and has a dusty dry look to it and if I run my fingers over the block I can clean off the dusty apperance of the block.?(does that mean it wasnt decalcified well, or?paraffin infiltration wasnt enough??or is it possible there is some?EDTA salts or left over paraformaldehyde?but I did wash the samples with water after being in?paraformaldehyde and EDTA?) ?After decalcification the samples felt very soft and I could bend the maxilla and were flexible and could be cut with an ordinary blade. Here is the?protocol I used for decalcifying the samples and the processing details. I would really appreciate if you guys can give me some suggestions as to what I can do about these samples and what must have went worng. Here's the method that I used for rat maxilla's with teeth in them. 1. Fixed in 4% paraformaldehyde for 4 days and then washed with distilled water. 2. Decalcified using 8% EDTA with change to fresh solution everyday for 3 weeks using the microwave technique. (eg: the maxilla from one rat were decalcified in 40 mL EDTA) 3. After decalcification the samples were washed in distilled water for couple of hours with fresh changes every 30 mins. 4. The samples were then transferred to 50% ethanol for an hour and then 70% ethanol for an hour and then overnight in 70% ethanol. 5. Then 95% ethanol for 1.5 hours with fresh changes every 30 mins 6. Then 100% ethanol for 1.5 hours with fresh changes every 30 mins 7. Then half 100% ethanol half 100% xylene for 20 mins 8. Then Xylene for 3 hours with fresh change every 45 mins (samples could very clear after this step). 9. Half Xylene half paraffin for 30 mins. 10. 100% paraffin (paraplast plus) for 3 hours with fresh change every 45 mins and then embedded in paraffin. I would really appreciate if you guys would guide me as to what I can do so that I can use these samples to cut sections. Thankyou for your time and help. Sincerely, Smit. Add more friends to your messenger and enjoy! Go to http://messenger.yahoo.com/invite/ From tbraud <@t> holyredeemer.com Fri Dec 12 10:53:12 2008 From: tbraud <@t> holyredeemer.com (Terri Braud) Date: Fri Dec 12 10:53:17 2008 Subject: [Histonet] RE: Histonet Digest, Vol 61, Issue 20 In-Reply-To: <72a5b1aa0002ee54@HolyRedeemer.com> Message-ID: Hi - Here we perform the silver step with a hot plate under the hood. As the slides are going into the sodium metabisulfite step, we turn on a hot plate on high. Then we mix the silver solution according to the procedure, put it in a beaker on the hot plate, rinse our slides with microwaved hot DI water a few times, leaving them in hot DI water. We gently swirl the silver solution as it heats and just when it starts to turn color (right before boiling), we dump the hot water off the slides and pour on the silver. The tissue usually stains within 45-60 seconds, a little longer if your silver is not really hot. Rinse with hot DI water afterwards. I was a little dubious when I first heard about it, but we too, do not have a vented microwave, using it only to heat water. This works great, though no one here could tell us who came up with the idea. You do have to use hot water before and after to keep from breaking coplin jars or slides from drastic temp changes. Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Laboratory Holy Redeemer Hospital and Medical Center 1648 Huntingdon Pike Meadowbrook, PA 19046 (215) 938-3676 phone (215) 938-3689 fax -----Original Message----- Message: 9 Date: Fri, 12 Dec 2008 04:52:19 -0500 From: TIMOTHY MALLOY Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS We currently use a stain called grocotts silver methenimine solution for staining pnuemocystis. We would like to use a hot plate for the procedure instead of a unvented microwave. Does anybody have any formula's the would like to share with the hot plate? Timothy G. Malloy, HT ( ASCP ) A.A.S. --------------------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. From jseaton <@t> wlgore.com Fri Dec 12 11:01:08 2008 From: jseaton <@t> wlgore.com (Janella Seaton) Date: Fri Dec 12 11:06:09 2008 Subject: [Histonet] Janella Seaton/WLGORE is out of the office. Message-ID: I will be out of the office starting 12/12/2008 and will not return until 12/15/2008. I will respond to your message when I return. Any histology-related requests or questions can be sent to: histology@wlgore.com. From pruegg <@t> ihctech.net Fri Dec 12 11:36:45 2008 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Dec 12 11:36:50 2008 Subject: [Histonet] Silly =?UTF-8?Q?Question=3F?= Message-ID: <20081212103645.f86bd30e73b823f57b516b5451216a98.b14b18c9f2.wbe@email.secureserver.net> i have had this argument with the researchers at the University f 30 years, somewhere back in the day they were told that commercially mad not hurt their tissue for and make it fresh themsel it often does not get issue is miss placed as they for consistancy and paying more attent fixation in reg formalin. Another anoying myth that is difficult to combat with them is that "we will cross always getting ti 24 hrs. to protect it fr proteins are not cross linked they c washed away forever), the people in research kno linking fo aldehydes but do not know that cross linking o is a good thing and they also do not know that we have advanced methods HIER or EIER for unmasking the proteins, but we have no way of gett sample was there i will get off my Friday soap box.................. Happy Holidays to all! Patsy -------- Original Message -------- Subject: RE: [Histonet] Silly From: Merced Leiker Date: Fri, To: "Edwards, R.E." Cc: "'histonet@lists.uts In re or techn reagents, way...a lot of project "works" to technicians t usually don't have back-and-forth arg Merced --On Friday, December 12, 2008 2:0 wrote: & of &g > > -----Original Messag > From: histonet-bounces@lists.utsouthwestern.edu > [[1]mailto:histonet-bounces@lists.utsouthwestern.edu] Pat > Flannery Sent: 11 December 2008 16:59 > To: > Subject: [Histonet] Silly Questi > > Please humor me on this if it's obvious (to everyone bu do > we use paraformaldehyde (which is so inconvenient to rather > than buffered formalin or just diluted formaldehyde > > It seems that around here, some folks prefer paraf (either > 2% or 4%) and others use formalin, while some o diluted > formaldehyde (I see all 4 on labels for spec > histology). Is it mostly a matter of personal p you > were trained (i.e. force of habit) or is the use > each solution (basically the same chemical > buffered or not)? The only answer I've go > "That's what we always use." > &g > > -Pat Flannery (not a "real" histologist - I just lab) > > > _____________________________ > Histonet mailing list > Histonet@lists.uts > [2]http://lists.utsouthwestern.edu/mailman/list > > _________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern. > [3]http://lists.utsouthwestern.edu/mailman/listinfo/histone > Merced M Leiker Research Technician II 3 School of Medicine and Biomedi State University of New York at Buffalo 3435 Main St, Bu Ph: (716) 829-6033 Fx: (716) 829-2725 "Without - random internet blog commentator _______________________________________________ Histonet mailin Histonet@lists.utsouthwestern.edu [4]http://lists.utsouthw References 1. 3D"http://email.secureserver.=/ 2. 3D"http://lists.utsouthwestern.edu/mailman/ 3. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/his 4. 3D"http://lists.utso=/ From LSebree <@t> uwhealth.org Fri Dec 12 12:45:53 2008 From: LSebree <@t> uwhealth.org (Sebree Linda A) Date: Fri Dec 12 12:45:58 2008 Subject: [Histonet] 7B11 clone of Ki67 Message-ID: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDEA7@UWHC-MAIL01.uwhis.hosp.wisc.edu> Good afternoon, 'netters, I am in desperate need of Ki67 antibody, clone 7B11 preferably, to use on Ventana instruments. I didn't realize we were running low and our usual vendor has no stock. I'd prefer a predilute but only if it is known to work on our instruments. I don't have the time to search this out myself so I am turning to all of you. Thanks, Linda Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Laboratory DB1-223 VAH 600 Highland Ave. Madison, WI 53792 (608)265-6596 From histonetalias <@t> gmail.com Fri Dec 12 12:52:49 2008 From: histonetalias <@t> gmail.com (Histonet Alias) Date: Fri Dec 12 12:52:57 2008 Subject: [Histonet] 7B11 clone of Ki67 In-Reply-To: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDEA7@UWHC-MAIL01.uwhis.hosp.wisc.edu> References: <5998F3BDFF7AAC4091C7AE93A7A1A5892CDEA7@UWHC-MAIL01.uwhis.hosp.wisc.edu> Message-ID: <4b6c85510812121052r52cb2a1q67cc6af4b24d6832@mail.gmail.com> Why not use the (30-9) clone? It is much easier to come by. I sue it on my Benchmark, cat# 790-4286. On Fri, Dec 12, 2008 at 1:45 PM, Sebree Linda A wrote: > Good afternoon, 'netters, > > I am in desperate need of Ki67 antibody, clone 7B11 preferably, to use > on Ventana instruments. I didn't realize we were running low and our > usual vendor has no stock. I'd prefer a predilute but only if it is > known to work on our instruments. I don't have the time to search this > out myself so I am turning to all of you. > > Thanks, > Linda > > Linda A. Sebree > University of Wisconsin Hospital & Clinics > IHC/ISH Laboratory > DB1-223 VAH > 600 Highland Ave. > Madison, WI 53792 > (608)265-6596 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Al Ias HT(ASCP) Histology Manager Pathology Laboratory United States From leiker <@t> buffalo.edu Fri Dec 12 13:03:58 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Fri Dec 12 13:04:03 2008 Subject: [Histonet] Silly Question? In-Reply-To: <20081212103645.f86bd30e73b823f57b516b5451216a98.b14b18c9f2.wbe@email.secureserver.net> References: <20081212103645.f86bd30e73b823f57b516b5451216a98.b14b18c9f2.wbe@ email.secureserver.net> Message-ID: So I have a question about the cross-linking aspect of PFA...while I agree I need it to keep my epitope in place, is there such a thing as OVER-crosslinking (i.e., tissue spending TOO much time in formalin - weeks? months?) that would make my epitope difficult to near-impossible to retrieve? Loving the formaldehyde soap-boxes histonetters get onto... --On Friday, December 12, 2008 10:36 AM -0700 pruegg@ihctech.net wrote: > > i have had this argument with the researchers at the University for 30 > years, somewhere back in the day they were told that commercially made > formalin had methanol in it (it does but just a little and does not hurt > anything in my experience) and that methanol would damage their tissue > for IHC, so they think they must use paraformaldehyde and make it fresh > themselves. Since new people make it all the time it often does not get > made up correctly and their stress over this issue is miss placed as they > should be using something commercial for consistancy and paying more > attention to adequate time for fixation in reg formalin. > Another anoying myth that is difficult to combat with them is that "we > should limit the fixation time in aldehyde fixatives because it will > cross link the proteins masking them for IHC", there fore i am always > getting tissue that has not been fixed long enough (at least 24 hrs. to > protect it from paraffin processing, because if the proteins are not > cross linked they can be alcohol fixed and/or washed away forever), the > people in research know about the cross linking fo aldehydes but do not > know that cross linking of proteins is a good thing and they also do not > know that we have advanced methods HIER or EIER for unmasking the > proteins, but we have no way of getting a protein back that has been lost > in processing because the sample was not adequately fixed. > > there i will get off my Friday soap box.................. > > Happy Holidays to all! > > Patsy > > > > -------- Original Message -------- > Subject: RE: [Histonet] Silly Question? > From: Merced Leiker > Date: Fri, December 12, 2008 8:12 am > To: "Edwards, R.E." , 'Pat Flannery' > > Cc: "'histonet@lists.utsouthwestern.edu'" > > > In research lab situations particularly, one does not have the time or > technique for nailing down the ways of making each of the buffers, > reagents, and procedures work the "right" way or the most optimum way...a > lot of times it's students or postdocs just focused on getting their > project done and not caring how their fixative is made as long as it > "works" to some degree and, alas, it's up to us already over-booked > technicians to figure out the best way to make the PFA....and we usually > don't have a whole day (week, or year) to spend researching the > back-and-forth arguments, either! ;-) > > Merced > > --On Friday, December 12, 2008 2:04 PM +0000 "Edwards, R.E." > wrote: > >> You hit the nail on the head "That's what we always use", fear of >> change is a common human condition. >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat >> Flannery Sent: 11 December 2008 16:59 >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Silly Question? >> >> Please humor me on this if it's obvious (to everyone but me): why do >> we use paraformaldehyde (which is so inconvenient to make up) rather >> than buffered formalin or just diluted formaldehyde itself? >> >> It seems that around here, some folks prefer paraformaldehyde (either >> 2% or 4%) and others use formalin, while some others stick to diluted >> formaldehyde (I see all 4 on labels for specimens submitted for >> histology). Is it mostly a matter of personal preference or where you >> were trained (i.e. force of habit) or is there a valid reason to use >> each solution (basically the same chemical once in solution, merely >> buffered or not)? The only answer I've gotten when I've asked is, >> "That's what we always use." >> >> Thanks. >> >> -Pat Flannery (not a "real" histologist - I just play one in the lab) >> >> >> _______________________________________________ >> 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 >> > > > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From rjbuesa <@t> yahoo.com Fri Dec 12 15:09:19 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Dec 12 15:09:24 2008 Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS In-Reply-To: Message-ID: <746229.92433.qm@web65711.mail.ac4.yahoo.com> I used the MW oven to heat-boost the slutions but the actual staining was done in a water bath. Why don't you try a water bath at 60?C? ren? J. --- On Fri, 12/12/08, TIMOTHY MALLOY wrote: From: TIMOTHY MALLOY Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS To: "HISTONET ..." , "TIM MALLOY" Date: Friday, December 12, 2008, 4:52 AM We currently use a stain called grocotts silver methenimine solution for staining pnuemocystis. We would like to use a hot plate for the procedure instead of a unvented microwave. Does anybody have any formula's the would like to share with the hot plate? Timothy G. Malloy, HT ( ASCP ) A.A.S. tmalloy77@hotmail.com This email communication may contain CONFIDENTIAL INFORMATION WHICH ALSO MAY BE LEGALLY PRIVILEGED and is intended only for the use of the intended recipients identified above. If you are not the intended recipient of this communication, you are hereby notified that any unauthorized review, use, dissemination, distribution, downloading, or copying of this communication is strictly prohibited. If you have received this communication in error, please immediately notify us by reply email, delete the communication and destroy all copies._______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Fri Dec 12 15:19:37 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Dec 12 15:19:42 2008 Subject: [Histonet] Silly Question? In-Reply-To: Message-ID: <871437.41100.qm@web65714.mail.ac4.yahoo.com> Just two things about crosslinking: 1- there is not such a thing as over crosslinkage, because when all the reaction sites have reacted, there are no reaction sites left to "overreact"; and 2- formaldehyde fixation is a reversible reaction and the tissue will lose the croslinkage even by placing them in distilled water instead of HIER (although it will take much more time). Ren? J. --- On Fri, 12/12/08, Merced Leiker wrote: From: Merced Leiker Subject: RE: [Histonet] Silly Question? To: pruegg@ihctech.net Cc: "'histonet@lists.utsouthwestern.edu'" , "'Pat Flannery'" Date: Friday, December 12, 2008, 2:03 PM So I have a question about the cross-linking aspect of PFA...while I agree I need it to keep my epitope in place, is there such a thing as OVER-crosslinking (i.e., tissue spending TOO much time in formalin - weeks? months?) that would make my epitope difficult to near-impossible to retrieve? Loving the formaldehyde soap-boxes histonetters get onto... --On Friday, December 12, 2008 10:36 AM -0700 pruegg@ihctech.net wrote: > > i have had this argument with the researchers at the University for 30 > years, somewhere back in the day they were told that commercially made > formalin had methanol in it (it does but just a little and does not hurt > anything in my experience) and that methanol would damage their tissue > for IHC, so they think they must use paraformaldehyde and make it fresh > themselves. Since new people make it all the time it often does not get > made up correctly and their stress over this issue is miss placed as they > should be using something commercial for consistancy and paying more > attention to adequate time for fixation in reg formalin. > Another anoying myth that is difficult to combat with them is that "we > should limit the fixation time in aldehyde fixatives because it will > cross link the proteins masking them for IHC", there fore i am always > getting tissue that has not been fixed long enough (at least 24 hrs. to > protect it from paraffin processing, because if the proteins are not > cross linked they can be alcohol fixed and/or washed away forever), the > people in research know about the cross linking fo aldehydes but do not > know that cross linking of proteins is a good thing and they also do not > know that we have advanced methods HIER or EIER for unmasking the > proteins, but we have no way of getting a protein back that has been lost > in processing because the sample was not adequately fixed. > > there i will get off my Friday soap box.................. > > Happy Holidays to all! > > Patsy > > > > -------- Original Message -------- > Subject: RE: [Histonet] Silly Question? > From: Merced Leiker > Date: Fri, December 12, 2008 8:12 am > To: "Edwards, R.E." , 'Pat Flannery' > > Cc: "'histonet@lists.utsouthwestern.edu'" > > > In research lab situations particularly, one does not have the time or > technique for nailing down the ways of making each of the buffers, > reagents, and procedures work the "right" way or the most optimum way...a > lot of times it's students or postdocs just focused on getting their > project done and not caring how their fixative is made as long as it > "works" to some degree and, alas, it's up to us already over-booked > technicians to figure out the best way to make the PFA....and we usually > don't have a whole day (week, or year) to spend researching the > back-and-forth arguments, either! ;-) > > Merced > > --On Friday, December 12, 2008 2:04 PM +0000 "Edwards, R.E." > wrote: > >> You hit the nail on the head "That's what we always use", fear of >> change is a common human condition. >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat >> Flannery Sent: 11 December 2008 16:59 >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Silly Question? >> >> Please humor me on this if it's obvious (to everyone but me): why do >> we use paraformaldehyde (which is so inconvenient to make up) rather >> than buffered formalin or just diluted formaldehyde itself? >> >> It seems that around here, some folks prefer paraformaldehyde (either >> 2% or 4%) and others use formalin, while some others stick to diluted >> formaldehyde (I see all 4 on labels for specimens submitted for >> histology). Is it mostly a matter of personal preference or where you >> were trained (i.e. force of habit) or is there a valid reason to use >> each solution (basically the same chemical once in solution, merely >> buffered or not)? The only answer I've gotten when I've asked is, >> "That's what we always use." >> >> Thanks. >> >> -Pat Flannery (not a "real" histologist - I just play one in the lab) >> >> >> _______________________________________________ >> 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 >> > > > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Fri Dec 12 15:20:08 2008 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Dec 12 15:20:24 2008 Subject: [Histonet] CD123 and TCL-1 Message-ID: <49428F380200007700007833@gwmail4.harthosp.org> Anyone doing IHC for CD123 and TCL-1 on formalin-fixed, paraffin-embedded tissue? Richard Richard W. Cartun, Ph.D. Director, Histology & Immunopathology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From PMonfils <@t> Lifespan.org Fri Dec 12 15:53:43 2008 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Dec 12 15:53:50 2008 Subject: [Histonet] de-waxing / staining station question In-Reply-To: <494022AF.8040905@columbia.edu> Message-ID: <4EBFF65383B74D49995298C4976D1D5E03835C82@LSRIEXCH1.lsmaster.lifespan.org> Dewaxing in one change of solvent, regardless of time in the solvent, is not a good idea. After a rack of slides has passed through the first xylene, that dish no longer contains xylene. It contains a solution of paraffin in xylene, and with each additional rack, that paraffin solution becomes more concentrated. Therefore you will not be going from xylene into absolute alcohol, but from a paraffin solution into absolute alcohol. Since the paraffin in that solution is not soluble in alcohol, it will precipitate on the slides and in the tissue. The purpose of having additional alcohols is not that one xylene can't dissolve all the paraffin out of the tissue. It is to get rid of the dissolved paraffin so that you are not carrying paraffin into the first alcohol. From JWeems <@t> sjha.org Fri Dec 12 15:55:48 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Fri Dec 12 15:55:52 2008 Subject: [Histonet] Biogenex Message-ID: <5D64396A0D4A5346BEBC759022AAEAA51BA910@ITSSSXM01V6.one.ads.che.org> Would anyone using the Biogenix Xmatrix be willing to speak with me about your experience? I'd appreciate your feedback! Thanks, j Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From susanbachus <@t> verizon.net Fri Dec 12 15:31:49 2008 From: susanbachus <@t> verizon.net (Susan Bachus) Date: Fri Dec 12 16:02:02 2008 Subject: [Histonet] re: concentration of formaldehyde in soultion: ASTM D2194 - 02(2007) Standard Test Method for Concentration of Formaldehyde Solution References: , <200812121635232792765@foxmail.com>, <200812122312023456365@foxmail.com> Message-ID: I believe that the "swiss cheese" holes are due to ice crystal format= ion=20 during freezing, at least that's the rationale we were always taught = for=20 using additional fixation in sucrose-formalin after the initial fixat= ion in=20 formalin, i.e. the sucrose would prevent ice crystal formation. Sus= an ----- Original Message -----=20 =46rom: "tf" To: "Walters, Katherine S" ; "Tony Henwo= od"=20 ; "Pat Flannery" ;=20 "histonet@lists.utsouthwestern.ed" Sent: Friday, December 12, 2008 10:12 AM Subject: [Histonet] re: concentration of formaldehyde in soultion: AS= TM=20 D2194 - 02(2007) Standard Test Method for Concentration of Formaldehy= de=20 Solution > http://www.astm.org/Standards/D2194.htm > > ASTM D2194 - 02(2007) > > > ASTM D2194 - 02(2007) Standard Test Method for Concentration of= =20 > Formaldehyde Solutions > > > 2008-12-12 > > > > tf > > > > =B7=A2=BC=FE=C8=CB=A3=BA Walters, Katherine S > =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-12 21:49:26 > =CA=D5=BC=FE=C8=CB=A3=BA tifei@foxmail.com; Tony Henwood; Pat Flann= ery;=20 > histonet@lists.utsouthwestern.ed > =B3=AD=CB=CD=A3=BA > =D6=F7=CC=E2=A3=BA RE: [Histonet] (reply) silly questions.---PFA > > This may be another silly question, but how does one test the=20 > concentration of formaldehyde in solution? > Thanks, > Kathy > Notice: This UI Health Care e-mail (including attachments) is cover= ed by=20 > the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is= =20 > confidential and may be legally privileged. If you are not the int= ended=20 > recipient, you are hereby notified that any retention, disseminatio= n,=20 > distribution, or copying of this communication is strictly prohibit= ed.=20 > Please reply to the sender that you have received the message in er= ror,=20 > then delete it. Thank you. > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu=20 > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of tf > Sent: Friday, December 12, 2008 2:35 AM > To: Tony Henwood; Pat Flannery; histonet@lists.utsouthwestern.ed > Subject: [Histonet] (reply) silly questions.---PFA > "I looked at the sections and the cell shrinkage (and prominent spa= ces > between cells and connective tissue) indicated that most of the > "fixation" seemed to have occured in the processing ethanols. I ask= ed > him for some of the fixative he used, tested the formaldehyde > concentration and found it to be less than 0.5%!!" > Tony: Do you think this is because of inproper preparation of PFA i= n his=20 > lab, or the common problem in all researchers using PFA? > I do think most biomedical labs currently are using PFA to = prepare=20 > the fixatives! > > So, anyone has the idea on a correction preparation procedure of 4%= PFA? > I noticed some of you dissolve PFA powder in NaOH-conditioned alkal= ine=20 > water, then add concentrated PB solution. > We here dissolve PFA in concentrated PB solution directly (heat & s= tir for=20 > 2-3 hours), then adjust pH to 7.4. > We dont have big problem in tissue quaility....except when one want= to cut=20 > the brain in a cryostat rather sliding microtome. > Many times the brain sections from the cryostat have "cheese" like= =20 > holes/cavities, which almost never appear on sliding microtome-prep= ared=20 > sections. > 2008-12-12 > tf > =B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood > =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-12 06:18:47 > =CA=D5=BC=FE=C8=CB=A3=BA Pat Flannery; histonet@lists.utsouthwester= n.edu > =B3=AD=CB=CD=A3=BA > =D6=F7=CC=E2=A3=BA RE: [Histonet] Silly Question? > > Pat, > I agree with you. > In a routine diagnostic histopathology laboratory, it makes little > difference what you use. Around the world for over 100 years most l= abs > use 10% neutral buffered formalin made from concentrated 38%(or the= re > abouts) formalin (or formaldehyde). > Researchers, though, are a different kettle of fish. They will tend= to > hang on to misinformed, "mystical" methods believing they are being > scientific. Funny, you would think that they, as a group, would be = the > ones pushing the boundaries and critically assessing each step of t= heir > research, ensuring that they understand what and why they are doing= it. > (Disclaimer - not all researchers are like this, thank heavens!!) > Using a formaldehyde solution made from polyformaldehyde can cause > problems. One researcher used it and wondered why their morphology = was > sub-optimal and their p53 immunohistochemistry was negative. He ass= ured > me that he had fixed small samples of tissue for 6 hours in 4% > formaldehyde and then processed them using ethanol, xylene and wax. > I looked at the sections and the cell shrinkage (and prominent spac= es > between cells and connective tissue) indicated that most of the > "fixation" seemed to have occured in the processing ethanols. I ask= ed > him for some of the fixative he used, tested the formaldehyde > concentration and found it to be less than 0.5%!! > This also explains the loss of p53 staining. I gave him some of our > routine 10% phosphate buffered fomalin, asked him to fix overnight,= and > try agin. Low and behold problem solved. > How's that for a Friday Flamming!!! > Regards > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Friday, 12 December 2008 3:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > Please humor me on this if it's obvious (to everyone but me): why = do > we use paraformaldehyde (which is so inconvenient to make up) rathe= r > than buffered formalin or just diluted formaldehyde itself? > It seems that around here, some folks prefer paraformaldehyde (eith= er > 2% or 4%) and others use formalin, while some others stick to dilut= ed > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference or where = you > were trained (i.e. force of habit) or is there a valid reason to us= e > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've asked is, > "That's what we always use." > Thanks. > -Pat Flannery (not a "real" histologist - I just play one in the la= b) > _______________________________________________ > 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 i= ntended=20 > solely for the use of the individual or entity to whom they are add= ressed.=20 > If you are not the intended recipient, please delete it and notify = the=20 > sender. > Views expressed in this message and any attachments are those of th= e=20 > individual sender, and are not necessarily the views of The Childre= n's=20 > Hospital at Westmead > This note also confirms that this email message has been > virus scanned and although no computer viruses were detected, The= =20 > Childrens Hospital at Westmead accepts no liability for any consequ= ential=20 > damage resulting from email containing computer viruses. > *******************************************************************= *** > _______________________________________________ > Histonet mailing list > ---------------------------------------------------------------------= ----------- > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >=20 From gayle.callis <@t> bresnan.net Fri Dec 12 16:08:45 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Dec 12 16:08:47 2008 Subject: [Histonet] Re: Reduction of autofluorescence using glycine Message-ID: To reduce aldehyde induced autofluorescence, you can use 100 - 300 mM glycine in pH 7.4 buffer. TRIS buffer or even Dulbeccos PBS will work. You rehydrate the section and then immerse into the glycine solution for 20 minutes, maybe even longer. Glycine works by getting rid (binding?) of free aldehyde groups. You can either treat the tissue prior to processing (after fixation) by immersing for an hour or so, but we simply did the glycine treatment on individual sections. It worked best for us when we did a short length fixation in NBF. This has been discussed at length on Histonet in the past, so do an archive search. One person put a summary together on various methods and what worked best for him. There are other methods for getting rid of autofluorescence although some are less successful than others and one is made from a chemical that is explosive. Try IHCworld website, fluorescence topics or Google access this discussion written by Wright Cell Imaging Faculty, Toronto Western Research Institute, titled: Autofluorescence, Causes and Cures, a must read on the subject. Another trick is to use fluorophores in the near infrared range, the camera sees the fluorescence but no autofluorescence and you cannot see this red fluorophore with the naked eye. Alexa 750 will work if you have the filters and excitation wavelength available. Good luck Gayle M. Callis HTL(ASCP)HT,MT Bozeman MT From anh2006 <@t> med.cornell.edu Fri Dec 12 16:11:13 2008 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Fri Dec 12 16:11:20 2008 Subject: [Histonet] mouse dendritic cells Message-ID: Is anyone staining for mouse dendritic cells either in paraffin or frozen sections? If so, what markers/antibodies are you using? Thanks, ANDREA -- From awatanabe <@t> tgen.org Fri Dec 12 16:26:10 2008 From: awatanabe <@t> tgen.org (Aprill Watanabe) Date: Fri Dec 12 16:26:17 2008 Subject: [Histonet] Mucin blocking reagent for IHC Message-ID: I am staining CA19-9 on pancreas and have a good amount of mucin trapping the DAB. Is there a product that either blocks mucin or something else that will limit the mucin effect? Aprill Watanabe, B.S. Research Associate Integrated Cancer Genomics Division Tissue Microarray Center (TMA) Translational Genomics Research Institute (TGen) main: 602-343-8822 Fax: 602-343-8840 awatanabe@tgen.org www.tgen.org From gayle.callis <@t> bresnan.net Fri Dec 12 16:41:40 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Dec 12 16:41:43 2008 Subject: [Histonet] Re: rat maxilla processing Message-ID: <55F643C92AF6475EB23741A86C7F9CBE@DHXTS541> Dear Smit You wrote: I am a graduate student at University of Illinois, chicago. I have been having tremendous problems with sectioning of paraffin embedded rat maxilla with molars and I searched the Histonet archives and found that people here have invaluable experience, insight and suggestions for various problems. Heres my problem. : As I section the block the section would crumble and tear and has a dusty dry look to it and if I run my fingers over the block I can clean off the dusty apperance of the block. (does that mean it wasnt decalcified well, or paraffin infiltration wasnt enough? or is it possible there is some EDTA salts or left over paraformaldehyde but I did wash the samples with water after being in paraformaldehyde and EDTA ) After decalcification the samples felt very soft and I could bend the maxilla and were flexible and could be cut with an ordinary blade. Here is the protocol I used for decalcifying the samples and the processing details. I would really appreciate if you guys can give me some suggestions as to what I can do about these samples and what must have went worng. Here's the method that I used for rat maxilla's with teeth in them. 1. Fixed in 4% paraformaldehyde for 4 days and then washed with distilled water. 2. Decalcified using 8% EDTA with change to fresh solution everyday for 3 weeks using the microwave technique. (eg: the maxilla from one rat were decalcified in 40 mL EDTA) 3. After decalcification the samples were washed in distilled water for couple of hours with fresh changes every 30 mins. 4. The samples were then transferred to 50% ethanol for an hour and then 70% ethanol for an hour and then overnight in 70% ethanol. 5. Then 95% ethanol for 1.5 hours with fresh changes every 30 mins 6. Then 100% ethanol for 1.5 hours with fresh changes every 30 mins 7. Then half 100% ethanol half 100% xylene for 20 mins 8. Then Xylene for 3 hours with fresh change every 45 mins (samples could very clear after this step). 9. Half Xylene half paraffin for 30 mins. 10. 100% paraffin (paraplast plus) for 3 hours with fresh change every 45 mins and then embedded in paraffin. I would really appreciate if you guys would guide me as to what I can do so that I can use these samples to cut sections. Thankyou for your time and help. Sincerely, Smit.Reply: You bone sample is underprocessed, poorly infiltrated with paraffin and may not be totally decalcified. When you do the fixation, fix longer than 4 days, as rat maxilla is not a small sample and complicated by teeth in situ. It would be better to perfuse the animal with your PFA solution, then dissect off the maxilla, and immerse into fresh PFA for several days. If perfused then 4 days may be adequate. Rinse with running tap water, and immerse into your EDTA (what is pH?) Also, use 20 times the volume of EDTA to size of tissue, and change the EDTA after a couple of days to replenish the EDTA. EDTA is safe to use over a weekend so you do not have to change it so often. You need to do endpoint determination to know when the bone is decalcified and I will be happy to send a simple weight loss/weight gain method that works for EDTA or use a FAXITRON xray determination. After EDTA, rinse with running tap water for several hours, not distilled water . Please contact me for the endpoint determination, I will send as a separate attachment. To process, start the bone in 70% alcohol. The time changes in each of the alcohols, clearing agent and paraffin are the same. 2 hours per change (30 minutes per change for a total of 1.5 hours is not doing the job!) If you only have a small portion of the maxilla, you can reduce the time to 1.5 hours per change. I hope you have a tissue processor to do this for you, as you need to use vacuum and pressure for the best results. 70% ethanol80% ethanol95% ethanol X 2 changes100% ethanol X 2 changesXylene X 2 changes and to get rid of brittleness, use one change of Richard Allan Clearite 3, then 1 change of xylene. You can reverse the order of these two clearing agents without problems. Paraffin X 3 changes (minimum) at times suggested at NO more than 60C. Heat is an enemy that contributes to hardness of decalcified bone. To achieve vacuum pressure with hand processing, use a vacuum dessicator, and hopefully a heated vacuum oven for the paraffin steps. Use a harder paraffin - there are several on the marked. Tissue Prep 3 is excellent for bone as you want to try and match the hardness of your embedding media to hardness of decalcified bone as much as possible. What is meant by an ordinary blade? A low profile or high profile disposable blade? or are you using a c profile steel knife? High profile blades give much more stability when sectioning bone, and after a very careful trim of block face, soak the block on ice water for a short time, return to holder, and use a brand new sharp edge to section. Do NOT cut away what you have soaked, but over soaking will cause the decalcified bone to swell out of the block face, not a good thing. You should be able to get good sections mounted on plus charge slides, but dry them flat at 37C to 40C overnight but several days is even better. Do NOT go to a hot 60C oven. Over drying can cause precious section lift off. Good luckGayle Callis HTL(ASCP)HT,MT, Bozeman MT From gayle.callis <@t> bresnan.net Fri Dec 12 17:22:17 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Fri Dec 12 17:22:21 2008 Subject: [Histonet] mouse dendritic cells References: Message-ID: <4775D39ED66D45F999FB39BC5819427E@DHXTS541> Andrea, We stain for murine dendritic cells very frequently. CD11c (HL3 clone) is excellent from BD Biosciences using fresh snap frozen tissue with spleen as the positive control. Fixation is with our beloved 25% ethanol/75% acetone for 5 min at RT then going directly to buffer from fixative. The sections are air dried overnight before fixation. Dec 205, NLDC 145 from Serotec works but we have superior results with the BD Pharm rat antiMouse CD11c HL3 clone. There are a whole series of new DC antibodies out there, used by our FACS tech, but I have not tried them on frozen sections yet. I would still used the same tissue preparation and fixation if we did try them but also toss in cold acetone just for posteriety sake. The CD11c we work with is biotinylated, which simplifies everything since we come back with Streptavidin Alexa dyes for single and double immunofluorescence work these days. It is an Armenian hamster host IgG1. Happy Holidays to you Gayle Callis HTL(ASCP)HT,MT Bozeman MT ----- Original Message ----- From: "Andrea Hooper" To: "Histonet" Sent: Friday, December 12, 2008 3:11 PM Subject: [Histonet] mouse dendritic cells > Is anyone staining for mouse dendritic cells either in paraffin or frozen > sections? If so, what markers/antibodies are you using? > > Thanks, ANDREA > -- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bob.nienhuis <@t> gmail.com Fri Dec 12 19:16:27 2008 From: bob.nienhuis <@t> gmail.com (Bob Nienhuis) Date: Fri Dec 12 19:16:32 2008 Subject: [Histonet] Re: Reduction of autofluorescence using glycine In-Reply-To: References: Message-ID: <45109da50812121716j14d93b03m595f239a442caaa8@mail.gmail.com> If this works by binding free aldehyde groups that attach to antibodies/ or fluorochromes, or biotinylated whatever. shouldn't it also work for DAB or ABC immunolabeling and reduce background labeling? Bob UCLA / VA Medical Center On Fri, Dec 12, 2008 at 2:08 PM, Gayle Callis wrote: > To reduce aldehyde induced autofluorescence, you can use 100 - 300 mM > glycine in pH 7.4 buffer. TRIS buffer or even Dulbeccos PBS will work. You > rehydrate the section and then immerse into the glycine solution for 20 > minutes, maybe even longer. Glycine works by getting rid (binding?) of free > aldehyde groups. You can either treat the tissue prior to processing (after > fixation) by immersing for an hour or so, but we simply did the glycine > treatment on individual sections. It worked best for us when we did a short > length fixation in NBF. > > This has been discussed at length on Histonet in the past, so do an archive > search. One person put a summary together on various methods and what > worked best for him. > > There are other methods for getting rid of autofluorescence although some > are less successful than others and one is made from a chemical that is > explosive. Try IHCworld website, fluorescence topics or Google access > this discussion written by Wright Cell Imaging Faculty, Toronto Western > Research Institute, titled: Autofluorescence, Causes and Cures, a must read > on the subject. > > Another trick is to use fluorophores in the near infrared range, the camera > sees the fluorescence but no autofluorescence and you cannot see this red > fluorophore with the naked eye. Alexa 750 will work if you have the filters > and excitation wavelength available. > > Good luck > > Gayle M. Callis > HTL(ASCP)HT,MT > Bozeman MT > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From susanbachus <@t> verizon.net Fri Dec 12 19:32:03 2008 From: susanbachus <@t> verizon.net (Susan Bachus) Date: Fri Dec 12 19:32:33 2008 Subject: [Histonet] Silly Question? - Need help quickly! References: <520038.88648.qm@web65711.mail.ac4.yahoo.com> Message-ID: <0E6551E75A13431FBFA4B343E63FC968@RESLAPTOP> I tried earlier to send, for this thread, a wonderful paper by Nauta, chronicaling the history his discovery of his tract tracing method, in which serendipity and degradation of formalin played critical roles, not realizing that the size of the attachment would prevent it from going through, so I am trying again with a URL for this paper: http://www.jneurosci.org/cgi/reprint/13/4/1337 Susan ----- Original Message ----- From: "Rene J Buesa" To: "Pat Flannery" ; ; "Weems, Joyce" Sent: Thursday, December 11, 2008 12:58 PM Subject: RE: [Histonet] Silly Question? - Need help quickly! Joyce: Methanal, which is the chemical name of formaldehyde, polymerizes. If it forms a polymer of at least 50 molecules or more, it gets solid = para-formaldehyde. Formalin (a trade name as formol is also another trade name)is the 37-50% aqueous solution of formaldehyde (with some additiveses to prevent polymerization). You can prepare BNF using the formalin solution or dissolving the amount of solid para-formaldehydede to get to the concentrationon you desire. The chemical in both solutions is the same = methanal or formaldehyde.Ren? J. --- On Thu, 12/11/08, Weems, Joyce wrote: From: Weems, Joyce Subject: RE: [Histonet] Silly Question? - Need help quickly! To: "Pat Flannery" , histonet@lists.utsouthwestern.edu Date: Thursday, December 11, 2008, 12:12 PM I was just going to post a question regarding paraformaldhyde myself! Just last week I believe I remember someone saying that paraformaldehyde and formalin are the same and they had put the same solution in two different containers for one of their researchers because they were so insistent to have two different solutions. Are they the same? Well, today I have a request to put tissue for a researcher in formalin and paraformaldehyde. So.... Without percentage required, do I use 10% NBF? Do I call somewhere and get paraformaldehyde and make 4% paraformaldehyde? I have asked the surgeon twice for the number for the lab so I can find out - don't have it yet. I have two fresh adrenals in the fridge. Help!! Thanks in advance... Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital 5665 Peachtree Dunwoody Rd NE Atlanta, GA 30342 678-843-7376 - Phone 678-843-7831 - Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Thursday, December 11, 2008 11:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this 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 anh2006 <@t> med.cornell.edu Fri Dec 12 20:19:56 2008 From: anh2006 <@t> med.cornell.edu (anh2006@med.cornell.edu) Date: Fri Dec 12 20:22:31 2008 Subject: [Histonet] Re: Reduction of autofluorescence using glycine In-Reply-To: <45109da50812121716j14d93b03m595f239a442caaa8@mail.gmail.com> References: <45109da50812121716j14d93b03m595f239a442caaa8@mail.gmail.com> Message-ID: <1550118692-1229134945-cardhu_decombobulator_blackberry.rim.net-360559008-@bxe173.bisx.prod.on.blackberry> No, because the glycine acts by reducing the autofluorescene of the free aldehydes (maybe Dr. Kiernan or another knowledgeable person in the chemistry can tell us precisely how) rather than reducing the binding of other staining components to the aldehydes. -----Original Message----- From: Bob Nienhuis Date: Fri, 12 Dec 2008 17:16:27 To: Gayle Callis Cc: Subject: Re: [Histonet] Re: Reduction of autofluorescence using glycine If this works by binding free aldehyde groups that attach to antibodies/ or fluorochromes, or biotinylated whatever. shouldn't it also work for DAB or ABC immunolabeling and reduce background labeling? Bob UCLA / VA Medical Center On Fri, Dec 12, 2008 at 2:08 PM, Gayle Callis wrote: > To reduce aldehyde induced autofluorescence, you can use 100 - 300 mM > glycine in pH 7.4 buffer. TRIS buffer or even Dulbeccos PBS will work. You > rehydrate the section and then immerse into the glycine solution for 20 > minutes, maybe even longer. Glycine works by getting rid (binding?) of free > aldehyde groups. You can either treat the tissue prior to processing (after > fixation) by immersing for an hour or so, but we simply did the glycine > treatment on individual sections. It worked best for us when we did a short > length fixation in NBF. > > This has been discussed at length on Histonet in the past, so do an archive > search. One person put a summary together on various methods and what > worked best for him. > > There are other methods for getting rid of autofluorescence although some > are less successful than others and one is made from a chemical that is > explosive. Try IHCworld website, fluorescence topics or Google access > this discussion written by Wright Cell Imaging Faculty, Toronto Western > Research Institute, titled: Autofluorescence, Causes and Cures, a must read > on the subject. > > Another trick is to use fluorophores in the near infrared range, the camera > sees the fluorescence but no autofluorescence and you cannot see this red > fluorophore with the naked eye. Alexa 750 will work if you have the filters > and excitation wavelength available. > > Good luck > > Gayle M. Callis > HTL(ASCP)HT,MT > Bozeman MT > > > _______________________________________________ > 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 lee2323 <@t> comcast.net Sat Dec 13 07:46:57 2008 From: lee2323 <@t> comcast.net (lee2323@comcast.net) Date: Sat Dec 13 07:47:01 2008 Subject: [Histonet] Offended In-Reply-To: <5A2BD13465E061429D6455C8D6B40E3907C253440D@IBMB7Exchange.digestivespecialists.com> Message-ID: <579789926.409031229176017940.JavaMail.root@sz0101a.westchester.pa.mail.comcast.net> I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. ----- Original Message ----- From: "Linda Blazek" To: "Charles.Embrey" , "Joe Nocito" , Histonet@lists.utsouthwestern.edu Sent: Friday, December 12, 2008 9:22:11 AM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] Offended Matt may have said no but we still found out! Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lblazek@digestivespecialists.com ? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Friday, December 12, 2008 9:14 AM To: Joe Nocito; Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Offended Joe, I can hardly believe you could be that grouchy :). ?When I was looking hard for a tech I called my buddy Matt Chase at Children's to try and coax him over to my lab. ?When he said no, I asked if he would let me chat with his techs and see how they were doing. ?Alas, he also replied negatively to that. ?The truth is that techs are becoming harder to find and are going at a premium price. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, December 11, 2008 7:19 PM To: Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Offended here we go and it's almost Friday. I received a call just like that at my last job. Me being me, I gave her a piece of my mind ( not too much, I don't have that much to go around). Told her she had some big testicles (not the word I used, but you get the idea) and hung up. I then called in the tech and had a talk with him. Asked him if he was looking for a job. He told me that he wasn't and didn't know how she got his name. I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door. There are other ways to go about this. Calling a lab is not one of them, but I'm sure some people give their work number for different reasons, trying to use it as leverage for a pay raise, to get the supervisors nervous, because the techs are that brazen, or belong to the id10t ?club. Let's face it, we are becoming a scarce breed and people have quotas to meet. Some will stop at nothing. That's my story and I'm sticking to it. JTT ----- Original Message ----- From: "Michael LaFriniere" To: Sent: Thursday, December 11, 2008 9:37 AM Subject: [Histonet] Offended I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" ?I felt was to recruit staff. ?I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". ?I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice ?our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. ?If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia _______________________________________________ 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 tifei <@t> foxmail.com Sat Dec 13 07:48:03 2008 From: tifei <@t> foxmail.com (=?utf-8?B?dGY=?=) Date: Sat Dec 13 07:48:29 2008 Subject: =?utf-8?B?UmU6IFJFOiBbSGlzdG9uZXRdIFNpbGx5IFF1ZXN0aW9uPw==?= References: <871437.41100.qm@web65714.mail.ac4.yahoo.com> Message-ID: <200812132147578756444@foxmail.com> No. Once the tisse was fixed, u can never get it fresh again. 2008-12-13 tf ???? Rene J Buesa ????? 2008-12-13 05:23:06 ???? pruegg; Merced Leiker ??? 'histonet@lists.utsouthwestern.edu'; 'Pat Flannery' ??? RE: [Histonet] Silly Question? Just two things about crosslinking: 1- there is not such a thing as over crosslinkage, because when all the reaction sites have reacted, there are no reaction sites left to "overreact"; and 2- formaldehyde fixation is a reversible reaction and the tissue will lose the croslinkage even by placing them in distilled water instead of HIER (although it will take much more time). Ren?J. --- On Fri, 12/12/08, Merced Leiker wrote: From: Merced Leiker Subject: RE: [Histonet] Silly Question? To: pruegg@ihctech.net Cc: "'histonet@lists.utsouthwestern.edu'" , "'Pat Flannery'" Date: Friday, December 12, 2008, 2:03 PM So I have a question about the cross-linking aspect of PFA...while I agree I need it to keep my epitope in place, is there such a thing as OVER-crosslinking (i.e., tissue spending TOO much time in formalin - weeks? months?) that would make my epitope difficult to near-impossible to retrieve? Loving the formaldehyde soap-boxes histonetters get onto... --On Friday, December 12, 2008 10:36 AM -0700 pruegg@ihctech.net wrote: > > i have had this argument with the researchers at the University for 30 > years, somewhere back in the day they were told that commercially made > formalin had methanol in it (it does but just a little and does not hurt > anything in my experience) and that methanol would damage their tissue > for IHC, so they think they must use paraformaldehyde and make it fresh > themselves. Since new people make it all the time it often does not get > made up correctly and their stress over this issue is miss placed as they > should be using something commercial for consistancy and paying more > attention to adequate time for fixation in reg formalin. > Another anoying myth that is difficult to combat with them is that "we > should limit the fixation time in aldehyde fixatives because it will > cross link the proteins masking them for IHC", there fore i am always > getting tissue that has not been fixed long enough (at least 24 hrs. to > protect it from paraffin processing, because if the proteins are not > cross linked they can be alcohol fixed and/or washed away forever), the > people in research know about the cross linking fo aldehydes but do not > know that cross linking of proteins is a good thing and they also do not > know that we have advanced methods HIER or EIER for unmasking the > proteins, but we have no way of getting a protein back that has been lost > in processing because the sample was not adequately fixed. > > there i will get off my Friday soap box.................. > > Happy Holidays to all! > > Patsy > > > > -------- Original Message -------- > Subject: RE: [Histonet] Silly Question? > From: Merced Leiker > Date: Fri, December 12, 2008 8:12 am > To: "Edwards, R.E." , 'Pat Flannery' > > Cc: "'histonet@lists.utsouthwestern.edu'" > > > In research lab situations particularly, one does not have the time or > technique for nailing down the ways of making each of the buffers, > reagents, and procedures work the "right" way or the most optimum way...a > lot of times it's students or postdocs just focused on getting their > project done and not caring how their fixative is made as long as it > "works" to some degree and, alas, it's up to us already over-booked > technicians to figure out the best way to make the PFA....and we usually > don't have a whole day (week, or year) to spend researching the > back-and-forth arguments, either! ;-) > > Merced > > --On Friday, December 12, 2008 2:04 PM +0000 "Edwards, R.E." > wrote: > >> You hit the nail on the head "That's what we always use", fear of >> change is a common human condition. >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat >> Flannery Sent: 11 December 2008 16:59 >> To: histonet@lists.utsouthwestern.edu >> Subject: [Histonet] Silly Question? >> >> Please humor me on this if it's obvious (to everyone but me): why do >> we use paraformaldehyde (which is so inconvenient to make up) rather >> than buffered formalin or just diluted formaldehyde itself? >> >> It seems that around here, some folks prefer paraformaldehyde (either >> 2% or 4%) and others use formalin, while some others stick to diluted >> formaldehyde (I see all 4 on labels for specimens submitted for >> histology). Is it mostly a matter of personal preference or where you >> were trained (i.e. force of habit) or is there a valid reason to use >> each solution (basically the same chemical once in solution, merely >> buffered or not)? The only answer I've gotten when I've asked is, >> "That's what we always use." >> >> Thanks. >> >> -Pat Flannery (not a "real" histologist - I just play one in the lab) >> >> >> _______________________________________________ >> 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 >> > > > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From thisisann <@t> aol.com Sat Dec 13 08:44:39 2008 From: thisisann <@t> aol.com (thisisann@aol.com) Date: Sat Dec 13 08:44:51 2008 Subject: [Histonet] Grosser Requirements Message-ID: <8CB2B487641A762-5A8-116C@mblk-d22.sysops.aol.com> I was recently informed the CAP's grossing requirements changed....I can not find anything about a change on their website. Does anyone have the current regs in reference to educational requirements for Grossing? Ann From rjbuesa <@t> yahoo.com Sat Dec 13 09:11:24 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Dec 13 09:11:29 2008 Subject: [Histonet] Silly Question? In-Reply-To: <200812132147578756444@foxmail.com> Message-ID: <776754.14752.qm@web65712.mail.ac4.yahoo.com> Check it out, YES formaldehyde fixation is reversible and that is the basis for HIER. Ren? J. --- On Sat, 12/13/08, tf wrote: From: tf Subject: Re: RE: [Histonet] Silly Question? To: "rjbuesa" , "pruegg" , "Merced Leiker" Cc: "'histonet@lists.utsouthwestern.edu'" , "'Pat Flannery'" Date: Saturday, December 13, 2008, 8:48 AM ? _filtered #yiv1373040317 { font-family:??; } _filtered #yiv1373040317 { font-family:Verdana; } _filtered #yiv1373040317 { } _filtered #yiv1373040317 {margin:72.0pt 90.0pt 72.0pt 90.0pt;} #yiv1373040317 P.MsoNormal { TEXT-JUSTIFY:inter-ideograph;FONT-SIZE:10.5pt;MARGIN:0cm 0cm 0pt;FONT-FAMILY:"Times New Roman";TEXT-ALIGN:justify;} #yiv1373040317 LI.MsoNormal { TEXT-JUSTIFY:inter-ideograph;FONT-SIZE:10.5pt;MARGIN:0cm 0cm 0pt;FONT-FAMILY:"Times New Roman";TEXT-ALIGN:justify;} #yiv1373040317 DIV.MsoNormal { TEXT-JUSTIFY:inter-ideograph;FONT-SIZE:10.5pt;MARGIN:0cm 0cm 0pt;FONT-FAMILY:"Times New Roman";TEXT-ALIGN:justify;} #yiv1373040317 A:link { COLOR:blue;TEXT-DECORATION:underline;} #yiv1373040317 SPAN.MsoHyperlink { COLOR:blue;TEXT-DECORATION:underline;} #yiv1373040317 A:visited { COLOR:purple;TEXT-DECORATION:underline;} #yiv1373040317 SPAN.MsoHyperlinkFollowed { COLOR:purple;TEXT-DECORATION:underline;} #yiv1373040317 SPAN.EmailStyle17 { FONT-WEIGHT:normal;COLOR:windowtext;FONT-STYLE:normal;FONT-FAMILY:Verdana;TEXT-DECORATION:none;} #yiv1373040317 DIV.Section1 { } #yiv1373040317 UNKNOWN { FONT-SIZE:10pt;} #yiv1373040317 BLOCKQUOTE { MARGIN-TOP:0px;MARGIN-BOTTOM:0px;MARGIN-LEFT:2em;} #yiv1373040317 OL { MARGIN-TOP:0px;MARGIN-BOTTOM:0px;} #yiv1373040317 UL { MARGIN-TOP:0px;MARGIN-BOTTOM:0px;} No. Once the tisse was fixed, u can never get it fresh again. ? ? 2008-12-13 tf ???? Rene J Buesa ????? 2008-12-13? 05:23:06 ???? pruegg; Merced Leiker ??? 'histonet@lists.utsouthwestern.edu'; 'Pat Flannery' ??? RE: [Histonet] Silly Question? Just?two?things?about?crosslinking: 1-?there?is?not?such?a?thing?as?over?crosslinkage,?because?when?all?the?reaction?sites?have?reacted,?there?are?no?reaction?sites?left?to?"overreact";?and 2-?formaldehyde?fixation?is?a?reversible?reaction?and?the?tissue?will?lose?the?croslinkage?even?by?placing?them?in?distilled?water?instead?of?HIER?(although?it?will?take?much?more?time). Ren?J. ---?On?Fri,?12/12/08,?Merced?Leiker??wrote: From:?Merced?Leiker? Subject:?RE:?[Histonet]?Silly?Question? To:?pruegg@ihctech.net Cc:?"'histonet@lists.utsouthwestern.edu'"?,?"'Pat?Flannery'"? Date:?Friday,?December?12,?2008,?2:03?PM So?I?have?a?question?about?the?cross-linking?aspect?of?PFA...while?I?agree? I?need?it?to?keep?my?epitope?in?place,?is?there?such?a?thing?as? OVER-crosslinking?(i.e.,?tissue?spending?TOO?much?time?in?formalin?-?weeks?? months?)?that?would?make?my?epitope?difficult?to?near-impossible?to? retrieve? Loving?the?formaldehyde?soap-boxes?histonetters?get?onto... --On?Friday,?December?12,?2008?10:36?AM?-0700?pruegg@ihctech.net?wrote: > >?i?have?had?this?argument?with?the?researchers?at?the?University?for?30 >?years,?somewhere?back?in?the?day?they?were?told?that?commercially?made >?formalin?had?methanol?in?it?(it?does?but?just?a?little?and?does?not?hurt >?anything?in?my?experience)?and?that?methanol?would?damage?their?tissue >?for?IHC,?so?they?think?they?must?use?paraformaldehyde?and?make?it?fresh >?themselves.??Since?new?people?make?it?all?the?time?it?often?does?not?get >?made?up?correctly?and?their?stress?over?this?issue?is?miss?placed?as?they >?should?be?using?something?commercial?for?consistancy?and?paying?more >?attention?to?adequate?time?for?fixation?in?reg?formalin. >?Another?anoying?myth?that?is?difficult?to?combat?with?them?is?that "we >?should?limit?the?fixation?time?in?aldehyde?fixatives?because?it?will >?cross?link?the?proteins?masking?them?for?IHC",?there?fore?i?am?always >?getting?tissue?that?has?not?been?fixed?long?enough?(at?least?24?hrs.?to >?protect?it?from?paraffin?processing,?because?if?the?proteins?are?not >?cross?linked?they?can?be?alcohol?fixed?and/or?washed?away?forever),?the >?people?in?research?know?about?the?cross?linking?fo?aldehydes?but?do?not >?know?that?cross?linking?of?proteins?is?a?good?thing?and?they?also?do?not >?know?that?we?have?advanced?methods?HIER?or?EIER?for?unmasking?the >?proteins,?but?we?have?no?way?of?getting?a?protein?back?that?has?been?lost >?in?processing?because?the?sample?was?not?adequately?fixed. > >?there?i?will?get?off?my?Friday?soap?box.................. > >?Happy?Holidays?to?all! > >?Patsy > > > >?--------?Original?Message?-------- >?Subject:?RE:?[Histonet]?Silly?Question? >?From:?Merced?Leiker? >?Date:?Fri,?December?12,?2008?8:12?am >?To:?"Edwards,?R.E."?,?'Pat Flannery' >? >?Cc:?"'histonet@lists.utsouthwestern.edu'" >? > >?In?research?lab?situations?particularly,?one?does?not?have?the?time?or >?technique?for?nailing?down?the?ways?of?making?each?of?the?buffers, >?reagents,?and?procedures?work?the?"right"?way?or?the?most optimum?way...a >?lot?of?times?it's?students?or?postdocs?just?focused?on?getting?their >?project?done?and?not?caring?how?their?fixative?is?made?as?long?as?it >?"works"?to?some?degree?and,?alas,?it's?up?to?us?already over-booked >?technicians?to?figure?out?the?best?way?to?make?the?PFA....and?we?usually >?don't?have?a?whole?day?(week,?or?year)?to?spend?researching?the >?back-and-forth?arguments,?either!?;-) > >?Merced > >?--On?Friday,?December?12,?2008?2:04?PM?+0000?"Edwards,?R.E." >??wrote: > >>?You?hit?the?nail?on?the?head?"That's?what?we?always use",?fear?of >>?change?is?a?common?human?condition. >> >>?-----Original?Message----- >>?From:?histonet-bounces@lists.utsouthwestern.edu >>?[mailto:histonet-bounces@lists.utsouthwestern.edu]?On?Behalf?Of?Pat >>?Flannery?Sent:?11?December?2008?16:59 >>?To:?histonet@lists.utsouthwestern.edu >>?Subject:?[Histonet]?Silly?Question? >> >>?Please?humor?me?on?this?if?it's?obvious?(to?everyone?but?me):?why do >>?we?use?paraformaldehyde?(which?is?so?inconvenient?to?make?up)?rather >>?than?buffered?formalin?or?just?diluted?formaldehyde?itself? >> >>?It?seems?that?around?here,?some?folks?prefer?paraformaldehyde?(either >>?2%?or?4%)?and?others?use?formalin,?while?some?others?stick?to?diluted >>?formaldehyde?(I?see?all?4?on?labels?for?specimens?submitted?for >>?histology).?Is?it?mostly?a?matter?of?personal?preference?or?where?you >>?were?trained?(i.e.?force?of?habit)?or?is?there?a?valid?reason?to?use >>?each?solution?(basically?the?same?chemical?once?in?solution,?merely >>?buffered?or?not)??The?only?answer?I've?gotten?when?I've?asked is, >>?"That's?what?we?always?use." >> >>?Thanks. >> >>?-Pat?Flannery?(not?a?"real"?histologist?-?I?just?play?one?in the?lab) >> >> >>?_______________________________________________ >>?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 >> > > > >?Merced?M?Leiker >?Research?Technician?II >?354?BRB?(pkgs)?/?140?Farber?Hall?(letters) >?School?of?Medicine?and?Biomedical?Sciences >?State?University?of?New?York?at?Buffalo >?3435?Main?St,?Buffalo,?NY?14214 >?Ph:?(716)?829-6033 >?Fx:?(716)?829-2725 > >?"Without?my?flaws?I'm?really?very?boring." >?-?random?internet?blog?commentator > > >?_______________________________________________ >?Histonet?mailing?list >?Histonet@lists.utsouthwestern.edu >?http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced?M?Leiker Research?Technician?II 354?BRB?(pkgs)?/?140?Farber?Hall?(letters) School?of?Medicine?and?Biomedical?Sciences State?University?of?New?York?at?Buffalo 3435?Main?St,?Buffalo,?NY?14214 Ph:?(716)?829-6033 Fx:?(716)?829-2725 "Without?my?flaws?I'm?really?very?boring." -?random?internet?blog?commentator _______________________________________________ Histonet?mailing?list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ?????? _______________________________________________ Histonet?mailing?list Histonet@lists.utsouthwestern.edu From gayle.callis <@t> bresnan.net Sat Dec 13 17:45:47 2008 From: gayle.callis <@t> bresnan.net (Gayle Callis) Date: Sat Dec 13 17:45:50 2008 Subject: More on Re: [Histonet] Reduction of autofluorescence using glycine References: <45109da50812121716j14d93b03m595f239a442caaa8@mail.gmail.com> <1550118692-1229134945-cardhu_decombobulator_blackberry.rim.net-360559008-@bxe173.bisx.prod.on.blackberry> Message-ID: <2625636BF77C49689A7CB4A2342AECCD@DHXTS541> For information on how (chemically) aldehyde fixatives cause autofluorescence and also a link to JAK e.g. John A Kiernan in Histonet archives, access the Wright Cell Imaging Facility, Toronto Western Resedarch Institute. This website is linked via www.IHCworld and simply found by Googling aldehyde induced autofluorescence. You will find protocols for reducing fixative induced fluorescence, and a tutorial on natural to aldehyde induced autofluorescence, plus a great deal more on the subject. There is also an excellent reference list. As for a quick chemical explanatioon on how the aldehyde groups are gotten rid of is "by reducing the -CHO groups to -OH with sodium borohydride or as they stated, "feeding [the aldehyde groups] bland amino groups" e.g. glycine or lysine. This has been discussed at length on Histonet in the past, and by searching with key words, you will probably pick up John Kiernan's comments, plus a reference on a Review of Autofluorescence. If anyone needs the latter publication, I will be happy to send via personal email. As for increased background with ABC methods or DAB, Jules Elias's book says antibodies can bind to free aldehyde groups, and the tissue can be pretreated in the same way as labs do when working with immunofluorescence. One would need to determine if the background is caused by another source (endogenous biotin, nonspecific binding of antibodies to tissue immunoglobulins, or overdevelopment of DAB) so if the pretreatment to get rid of free aldhehydes doesn't work and background still exists - then the sleuthing continues. Gluteraldehyde is the worst offender, along with long fixation in NBF, or warm temperatures during NBF fixation. Gayle M. Callis HTL(ASCP)HT,MT Bozeman MT ----- Original Message ----- From: To: "Bob Nienhuis" ; "Gayle Callis" Cc: Sent: Friday, December 12, 2008 7:19 PM Subject: Re: [Histonet] Re: Reduction of autofluorescence using glycine > No, because the glycine acts by reducing the autofluorescene of the free > aldehydes (maybe Dr. Kiernan or another knowledgeable person in the > chemistry can tell us precisely how) rather than reducing the binding of > other staining components to the aldehydes. > > -----Original Message----- > From: Bob Nienhuis > > Date: Fri, 12 Dec 2008 17:16:27 > To: Gayle Callis > Cc: > Subject: Re: [Histonet] Re: Reduction of autofluorescence using glycine > > > If this works by binding free aldehyde groups that attach to antibodies/ > or > fluorochromes, or biotinylated whatever. shouldn't it also work for DAB > or > ABC immunolabeling and > reduce background labeling? > > Bob > UCLA / VA Medical Center > > On Fri, Dec 12, 2008 at 2:08 PM, Gayle Callis > wrote: > >> To reduce aldehyde induced autofluorescence, you can use 100 - 300 mM >> glycine in pH 7.4 buffer. TRIS buffer or even Dulbeccos PBS will work. >> You >> rehydrate the section and then immerse into the glycine solution for 20 >> minutes, maybe even longer. Glycine works by getting rid (binding?) of >> free >> aldehyde groups. You can either treat the tissue prior to processing >> (after >> fixation) by immersing for an hour or so, but we simply did the glycine >> treatment on individual sections. It worked best for us when we did a >> short >> length fixation in NBF. >> >> This has been discussed at length on Histonet in the past, so do an >> archive >> search. One person put a summary together on various methods and what >> worked best for him. >> >> There are other methods for getting rid of autofluorescence although some >> are less successful than others and one is made from a chemical that is >> explosive. Try IHCworld website, fluorescence topics or Google access >> this discussion written by Wright Cell Imaging Faculty, Toronto Western >> Research Institute, titled: Autofluorescence, Causes and Cures, a must >> read >> on the subject. >> >> Another trick is to use fluorophores in the near infrared range, the >> camera >> sees the fluorescence but no autofluorescence and you cannot see this red >> fluorophore with the naked eye. Alexa 750 will work if you have the >> filters >> and excitation wavelength available. >> >> Good luck >> >> Gayle M. Callis >> HTL(ASCP)HT,MT >> Bozeman MT >> >> >> _______________________________________________ >> 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 jkiernan <@t> uwo.ca Sun Dec 14 23:37:28 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sun Dec 14 23:37:34 2008 Subject: Silver & ammonia (Was Re: [Histonet] Silly Question? , , ,) Message-ID: Dear Susan, Thank you for bringing Walle Nauta's 1993 review article to Histonetters' attention. I hadn't seen it before, and I agree that it was well worth downloading, reading and filing for future reference. More than 50 years ago Nauta & Gygax published a very easily understood account of ammoniacal silver solutions and the effects of changing their pH. This is fundamental to all techniques involving silver nitrate and ammonia, and the exact information is not in chemistry textbooks. Here's the reference. Nauta WJH, Gygax PA (1951) Silver impregnation of degenerating axon terminals in the central nervous system: (1) Technic. (2) Chemical notes. Stain Technology 26: 5-11. Part 2 was an appendix to their paper on the early (non-suppressive) Nauta-Gygax method, which stains both normal and degenerating axons. Many of the principles explained in Part 2 of this paper apply also to other silver methods. If your library subscribes to journals published by Informa, you can download this Nauta-Gygax paper (and anything else in Stain Technology) as a PDF file from the InformaWorld web site: http://www.informaworld.com/smpp/title~content=t713692932~bb=all The journal's name shows on the present publisher's web site in its present form (Biotechnic & Histochemistry) even though the name was really Stain Technology before about 1990. This is a problem with quite a few journals that have changed their names. The Springer web site, for example, doesn't seem to recognize the name changes of Histochemie --> Histochemistry --> Histochemistry and Cell Biology. Library catalogues usually record name changes faithfully. For journals, changing the name probably isn't always a good move! John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: Susan Bachus Date: Friday, December 12, 2008 20:33 Subject: Re: [Histonet] Silly Question? - Need help quickly! To: rjbuesa@yahoo.com, Pat Flannery , histonet@lists.utsouthwestern.edu, "Weems, Joyce" > I tried earlier to send, for this thread, a wonderful paper by > Nauta, > chronicaling the history his discovery of his tract tracing > method, in which > serendipity and degradation of formalin played critical roles, > not realizing > that the size of the attachment would prevent it from going > through, so I am > trying again with a URL for this paper: > http://www.jneurosci.org/cgi/reprint/13/4/1337 > > Susan > > ----- Original Message ----- > From: "Rene J Buesa" > To: "Pat Flannery" ; > ; > "Weems, Joyce" > Sent: Thursday, December 11, 2008 12:58 PM > Subject: RE: [Histonet] Silly Question? - Need help quickly! > > > Joyce: > Methanal, which is the chemical name of formaldehyde, > polymerizes. If it > forms a polymer of at least 50 molecules or more, it gets solid > = > para-formaldehyde. > Formalin (a trade name as formol is also another trade name)is > the 37-50% > aqueous solution of formaldehyde (with some additiveses to > prevent > polymerization). > You can prepare BNF using the formalin solution or dissolving > the amount of > solid para-formaldehydede to get to the concentrationon you desire. > The chemical in both solutions is the same = methanal or > formaldehyde.Ren? > J. > > --- On Thu, 12/11/08, Weems, Joyce wrote: > > > From: Weems, Joyce > Subject: RE: [Histonet] Silly Question? - Need help quickly! > To: "Pat Flannery" , > histonet@lists.utsouthwestern.eduDate: Thursday, December 11, > 2008, 12:12 PM > > I was just going to post a question regarding paraformaldhyde myself! > Just last week I believe I remember someone saying that > paraformaldehydeand formalin are the same and they had put the > same solution in two > different containers for one of their researchers because they > were so > insistent to have two different solutions. Are they the same? > > Well, today I have a request to put tissue for a researcher in > formalinand paraformaldehyde. So.... Without percentage > required, do I use 10% > NBF? Do I call somewhere and get paraformaldehyde and make 4% > paraformaldehyde? > > I have asked the surgeon twice for the number for the lab so I > can find > out - don't have it yet. I have two fresh adrenals in the > fridge. Help!! > > > Thanks in advance... > Joyce > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 678-843-7376 - Phone > 678-843-7831 - Fax > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Thursday, December 11, 2008 11:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > > Please humor me on this if it's obvious (to everyone but > me): why do we > use paraformaldehyde (which is so inconvenient to make up) > rather than > buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde > (either 2% > or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference > or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've > asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Confidentiality Notice: > This email, including any attachments is the > property of Catholic Health East and is intended > for the sole use of the intended recipient(s). > It may contain information that is privileged and > confidential. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are > not the intended recipient, please reply to the > sender that you have received the message in > error, then delete this 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Dec 15 00:15:02 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Dec 15 00:15:05 2008 Subject: [Histonet] Mucin blocking reagent for IHC In-Reply-To: References: Message-ID: Please explain "mucin trapping the DAB". Are you sure that this is not genuine immunohistochemical localization of the antigen? At neutral pH DAB cations are attracted to tissue polyanions (in mucus, cartilage etc). The attracted DAB should not be oxidized to a brown insoluble polymer in the absence of peroxidase. The only working peroxidase molecules in an immunohistochemical preparation should be those specifically bound at the site recognized by the primary antibody. Brown staining of mucus suggests the presence of peroxidase in the DAB-H2O2 final incbation solution. This would catalyze the oxidation of tissue-bound DAB. More washing after the last HRP-containing reagent might help. John Kiernan Anatomy, UWO London, Canada - - - ----- Original Message ----- From: Aprill Watanabe Date: Friday, December 12, 2008 17:26 Subject: [Histonet] Mucin blocking reagent for IHC To: histonet@lists.utsouthwestern.edu > I am staining CA19-9 on pancreas and have a good amount of mucin > trappingthe DAB. Is there a product that either blocks > mucin or something else that > will limit the mucin effect? > > Aprill Watanabe, B.S. > Research Associate > Integrated Cancer Genomics Division > Tissue Microarray Center (TMA) > Translational Genomics Research Institute (TGen) > main: 602-343-8822 > Fax: 602-343-8840 > awatanabe@tgen.org > www.tgen.org > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micropathlabs <@t> yahoo.com Mon Dec 15 04:00:05 2008 From: micropathlabs <@t> yahoo.com (Sheila Haas) Date: Mon Dec 15 04:00:14 2008 Subject: [Histonet] Grosser Requirements References: <8CB2B487641A762-5A8-116C@mblk-d22.sysops.aol.com> Message-ID: <204088.26635.qm@web57807.mail.re3.yahoo.com> I'd appreciate information on changes?also. This is the first I've heard of any. Thanks! ? Sheila Haas Laboratory Supervisor Micro Path Laboratories ? ________________________________ From: "thisisann@aol.com" To: histonet@lists.utsouthwestern.edu Sent: Saturday, December 13, 2008 9:44:39 AM Subject: [Histonet] Grosser Requirements I was recently informed the CAP's grossing requirements changed....I can not find anything about a change on their website. Does anyone have the current regs in reference to educational requirements for Grossing? Ann _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Mon Dec 15 04:56:28 2008 From: tifei <@t> foxmail.com (=?utf-8?B?dGY=?=) Date: Mon Dec 15 04:57:10 2008 Subject: [Histonet] Silver staing for degenerating axons Message-ID: <200812151856234435185@foxmail.com> I read the paper of Nauta with great interest! But I dont have the original pdf for the paper on Stain Technology in 1951 & 1954. Anyone can send me the pdf? I only have the improved method by Fink and Heimer in 1967 published on brain research, giving out two staining procedures. I can send you the pdf if anyone is interested in this. I would like to receive any proved protocol of silver staining for degenerating axons...because we dont have uranyl nitrate or hydroquinone here...I can not repeat the paper by Fink & Heimer. Can anyone provide me a supressive procedure in staining degenerating axons using "common chemicals", just ammonia water, silver nitrate, citric acid, formalin/PFA, sodium hydroxide? I just can not find applicable one with our current facilities! Thanks very much! 2008-12-15 tf ???? John Kiernan ????? 2008-12-15 13:42:09 ???? Susan Bachus ??? Pat Flannery; Weems, Joyce; histonet ??? Silver & ammonia (Was Re: [Histonet] Silly Question? , , ,) Dear Susan, Thank you for bringing Walle Nauta's 1993 review article to Histonetters' attention. I hadn't seen it before, and I agree that it was well worth downloading, reading and filing for future reference. More than 50 years ago Nauta & Gygax published a very easily understood account of ammoniacal silver solutions and the effects of changing their pH. This is fundamental to all techniques involving silver nitrate and ammonia, and the exact information is not in chemistry textbooks. Here's the reference. Nauta WJH, Gygax PA (1951) Silver impregnation of degenerating axon terminals in the central nervous system: (1) Technic. (2) Chemical notes. Stain Technology 26: 5-11. Part 2 was an appendix to their paper on the early (non-suppressive) Nauta-Gygax method, which stains both normal and degenerating axons. Many of the principles explained in Part 2 of this paper apply also to other silver methods. If your library subscribes to journals published by Informa, you can download this Nauta-Gygax paper (and anything else in Stain Technology) as a PDF file from the InformaWorld web site: http://www.informaworld.com/smpp/title~content=t713692932~bb=all The journal's name shows on the present publisher's web site in its present form (Biotechnic & Histochemistry) even though the name was really Stain Technology before about 1990. This is a problem with quite a few journals that have changed their names. The Springer web site, for example, doesn't seem to recognize the name changes of Histochemie --> Histochemistry --> Histochemistry and Cell Biology. Library catalogues usually record name changes faithfully. For journals, changing the name probably isn't always a good move! John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: Susan Bachus Date: Friday, December 12, 2008 20:33 Subject: Re: [Histonet] Silly Question? - Need help quickly! To: rjbuesa@yahoo.com, Pat Flannery , histonet@lists.utsouthwestern.edu, "Weems, Joyce" > I tried earlier to send, for this thread, a wonderful paper by > Nauta, > chronicaling the history his discovery of his tract tracing > method, in which > serendipity and degradation of formalin played critical roles, > not realizing > that the size of the attachment would prevent it from going > through, so I am > trying again with a URL for this paper: > http://www.jneurosci.org/cgi/reprint/13/4/1337 > > Susan > > ----- Original Message ----- > From: "Rene J Buesa" > To: "Pat Flannery" ; > ; > "Weems, Joyce" > Sent: Thursday, December 11, 2008 12:58 PM > Subject: RE: [Histonet] Silly Question? - Need help quickly! > > > Joyce: > Methanal, which is the chemical name of formaldehyde, > polymerizes. If it > forms a polymer of at least 50 molecules or more, it gets solid > = > para-formaldehyde. > Formalin (a trade name as formol is also another trade name)is > the 37-50% > aqueous solution of formaldehyde (with some additiveses to > prevent > polymerization). > You can prepare BNF using the formalin solution or dissolving > the amount of > solid para-formaldehydede to get to the concentrationon you desire. > The chemical in both solutions is the same = methanal or > formaldehyde.Ren? > J. > > --- On Thu, 12/11/08, Weems, Joyce wrote: > > > From: Weems, Joyce > Subject: RE: [Histonet] Silly Question? - Need help quickly! > To: "Pat Flannery" , > histonet@lists.utsouthwestern.eduDate: Thursday, December 11, > 2008, 12:12 PM > > I was just going to post a question regarding paraformaldhyde myself! > Just last week I believe I remember someone saying that > paraformaldehydeand formalin are the same and they had put the > same solution in two > different containers for one of their researchers because they > were so > insistent to have two different solutions. Are they the same? > > Well, today I have a request to put tissue for a researcher in > formalinand paraformaldehyde. So.... Without percentage > required, do I use 10% > NBF? Do I call somewhere and get paraformaldehyde and make 4% > paraformaldehyde? > > I have asked the surgeon twice for the number for the lab so I > can find > out - don't have it yet. I have two fresh adrenals in the > fridge. Help!! > > > Thanks in advance... > Joyce > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 678-843-7376 - Phone > 678-843-7831 - Fax > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Thursday, December 11, 2008 11:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > > Please humor me on this if it's obvious (to everyone but > me): why do we > use paraformaldehyde (which is so inconvenient to make up) > rather than > buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde > (either 2% > or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference > or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've > asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Confidentiality Notice: > This email, including any attachments is the > property of Catholic Health East and is intended > for the sole use of the intended recipient(s). > It may contain information that is privileged and > confidential. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are > not the intended recipient, please reply to the > sender that you have received the message in > error, then delete this 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From ktuttle <@t> umm.edu Mon Dec 15 07:57:10 2008 From: ktuttle <@t> umm.edu (Kimberly Tuttle) Date: Mon Dec 15 07:57:41 2008 Subject: [Histonet] Offended In-Reply-To: <579789926.409031229176017940.JavaMail.root@sz0101a.westchester.pa.mail.comcast.net> References: <5A2BD13465E061429D6455C8D6B40E3907C253440D@IBMB7Exchange.digestivespecialists.com> <579789926.409031229176017940.JavaMail.root@sz0101a.westchester.pa.mail.comcast.net> Message-ID: <49461BE5.90CE.001A.3@umm.edu> I dont think that anyone should assume that a tech is looking for another job because someone called the lab and asked for them. I get telemarketing calls from sales reps and emails and calls from job recruiters because most of them are members of this listserve, and many of our signature blocks contain all of our contact information, including phone number. I would be really offended if my supervisor held me personally responsible for the poor judgement of a recruiter. Maybe thats their angle, because if my boss came at me like Joe, Id start looking for another job. Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax >>> 12/13/2008 8:46 am >>> I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. ----- Original Message ----- From: "Linda Blazek" To: "Charles.Embrey" , "Joe Nocito" , Histonet@lists.utsouthwestern.edu Sent: Friday, December 12, 2008 9:22:11 AM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] Offended Matt may have said no but we still found out! Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lblazek@digestivespecialists.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Friday, December 12, 2008 9:14 AM To: Joe Nocito; Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Offended Joe, I can hardly believe you could be that grouchy :). When I was looking hard for a tech I called my buddy Matt Chase at Children's to try and coax him over to my lab. When he said no, I asked if he would let me chat with his techs and see how they were doing. Alas, he also replied negatively to that. The truth is that techs are becoming harder to find and are going at a premium price. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, December 11, 2008 7:19 PM To: Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Offended here we go and it's almost Friday. I received a call just like that at my last job. Me being me, I gave her a piece of my mind ( not too much, I don't have that much to go around). Told her she had some big testicles (not the word I used, but you get the idea) and hung up. I then called in the tech and had a talk with him. Asked him if he was looking for a job. He told me that he wasn't and didn't know how she got his name. I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door. There are other ways to go about this. Calling a lab is not one of them, but I'm sure some people give their work number for different reasons, trying to use it as leverage for a pay raise, to get the supervisors nervous, because the techs are that brazen, or belong to the id10t club. Let's face it, we are becoming a scarce breed and people have quotas to meet. Some will stop at nothing. That's my story and I'm sticking to it. JTT ----- Original Message ----- From: "Michael LaFriniere" To: Sent: Thursday, December 11, 2008 9:37 AM Subject: [Histonet] Offended I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia _______________________________________________ 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 This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. From marktarango <@t> gmail.com Mon Dec 15 08:01:50 2008 From: marktarango <@t> gmail.com (Mark Tarango) Date: Mon Dec 15 08:02:00 2008 Subject: [Histonet] Mucin blocking reagent for IHC In-Reply-To: References: Message-ID: <5b6eb13e0812150601s14895fdem54eaec0e77fc0e23@mail.gmail.com> Just last week I saw DAB staining mucin for p53 (which should be nuclear staining). We were using Ventana's Iview kit which uses avidin and biotin as links. After we added the a/b block, this unwanted staining completely disappeared. I'd suggest adding a blocking step. Mark On Sun, Dec 14, 2008 at 10:15 PM, John Kiernan wrote: > Please explain "mucin trapping the DAB". Are you sure that this is not > genuine immunohistochemical localization of the antigen? > > At neutral pH DAB cations are attracted to tissue polyanions (in mucus, > cartilage etc). The attracted DAB should not be oxidized to a brown > insoluble polymer in the absence of peroxidase. > > The only working peroxidase molecules in an immunohistochemical preparation > should be those specifically bound at the site recognized by the primary > antibody. Brown staining of mucus suggests the presence of peroxidase in the > DAB-H2O2 final incbation solution. This would catalyze the oxidation of > tissue-bound DAB. More washing after the last HRP-containing reagent might > help. > > John Kiernan > Anatomy, UWO > London, Canada > - - - > ----- Original Message ----- > From: Aprill Watanabe > Date: Friday, December 12, 2008 17:26 > Subject: [Histonet] Mucin blocking reagent for IHC > To: histonet@lists.utsouthwestern.edu > > > I am staining CA19-9 on pancreas and have a good amount of mucin > > trappingthe DAB. Is there a product that either blocks > > mucin or something else that > > will limit the mucin effect? > > > > Aprill Watanabe, B.S. > > Research Associate > > Integrated Cancer Genomics Division > > Tissue Microarray Center (TMA) > > Translational Genomics Research Institute (TGen) > > main: 602-343-8822 > > Fax: 602-343-8840 > > awatanabe@tgen.org > > www.tgen.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 > From Barry.R.Rittman <@t> uth.tmc.edu Mon Dec 15 08:20:04 2008 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Mon Dec 15 08:20:09 2008 Subject: [Histonet] Offended References: <5A2BD13465E061429D6455C8D6B40E3907C253440D@IBMB7Exchange.digestivespecialists.com> <579789926.409031229176017940.JavaMail.root@sz0101a.westchester.pa.mail.comcast.net> <49461BE5.90CE.001A.3@umm.edu> Message-ID: Kimberly I think that it is a sad reflection that both management and outside companies when they hear that an employee is looking at salaries etc., assume that employee is seriously looking for a new job. This is not confined to histotech positions. I know of several instances when individuals were just looking around to compare jobs and salaries. Administration assumed that the individual had already made up their mind to leave and only asked what they could do to retain that employee when it was too late. A simple timely sentence of "what can we do to keep you here" is often all that is needed. In the absence of this, the individuals assumption is that the administration does not care if that individual leaves or not. I do feel that a generic letter, such as we often see on histonet posting available jobs is OK. If you are being bugged by companies then you need to contact them at their highest levels and note that if future correspondence with job offers are directed to you personally then you will ask the administration to look at the business dealings the lab and/or university has with this company. Failing that, I would recommend that you contract this out to Joe - he has a great reputation for getting things done! Barry ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kimberly Tuttle Sent: Mon 12/15/2008 7:57 AM To: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Offended I dont think that anyone should assume that a tech is looking for another job because someone called the lab and asked for them. I get telemarketing calls from sales reps and emails and calls from job recruiters because most of them are members of this listserve, and many of our signature blocks contain all of our contact information, including phone number. I would be really offended if my supervisor held me personally responsible for the poor judgement of a recruiter. Maybe thats their angle, because if my boss came at me like Joe, Id start looking for another job. Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax >>> 12/13/2008 8:46 am >>> I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. ----- Original Message ----- From: "Linda Blazek" To: "Charles.Embrey" , "Joe Nocito" , Histonet@lists.utsouthwestern.edu Sent: Friday, December 12, 2008 9:22:11 AM GMT -05:00 US/Canada Eastern Subject: RE: [Histonet] Offended Matt may have said no but we still found out! Linda Blazek HT (ASCP) Manager/Supervisor GI Pathology of Dayton 7415 Brandt Pike Huber Heights, OH 45424 Phone: (937) 293-4424 ext 7118 Email: lblazek@digestivespecialists.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Friday, December 12, 2008 9:14 AM To: Joe Nocito; Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Offended Joe, I can hardly believe you could be that grouchy :). When I was looking hard for a tech I called my buddy Matt Chase at Children's to try and coax him over to my lab. When he said no, I asked if he would let me chat with his techs and see how they were doing. Alas, he also replied negatively to that. The truth is that techs are becoming harder to find and are going at a premium price. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joe Nocito Sent: Thursday, December 11, 2008 7:19 PM To: Michael LaFriniere; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Offended here we go and it's almost Friday. I received a call just like that at my last job. Me being me, I gave her a piece of my mind ( not too much, I don't have that much to go around). Told her she had some big testicles (not the word I used, but you get the idea) and hung up. I then called in the tech and had a talk with him. Asked him if he was looking for a job. He told me that he wasn't and didn't know how she got his name. I told him that if he was looking for another job to do it on his time and on his dollar. If I ever received another call from a headhunter naming him, he would have to talk to them because he would be shown the door. There are other ways to go about this. Calling a lab is not one of them, but I'm sure some people give their work number for different reasons, trying to use it as leverage for a pay raise, to get the supervisors nervous, because the techs are that brazen, or belong to the id10t club. Let's face it, we are becoming a scarce breed and people have quotas to meet. Some will stop at nothing. That's my story and I'm sticking to it. JTT ----- Original Message ----- From: "Michael LaFriniere" To: Sent: Thursday, December 11, 2008 9:37 AM Subject: [Histonet] Offended I Wanted to obtained opinions of Managers/Supervisors in the histo world and how you handle situations regarding recruiters calling directly into Pathology Labs to specific tech's and stating they are seeking referrals for job placements. I recently had an AP staffing solution company call one of my labs to talk to any "histologist" I felt was to recruit staff. I feel this is an unprofessional practice to call directly into the laboratory and to hide behind the wording "referrals" when actually it appears they are trying to get information from the tech on the phone seeing if they are looking for a "new job". I feel that this is an intrusion as well as inappropriate in our line of business and we are not in the business to give referrals to companies in the staffing solution arena for their monetary gain as well as trying to entice our staff. What the staff does with these types of companies on personal time is their business, however, I think it is offending for companies to demonstrate this behavior. If anyone would like to know which company and person called into my lab I will personally inform you on private email! Michael LaFriniere Executive Director CSM Washington DC/Maryland/Virginia _______________________________________________ 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 This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Mon Dec 15 08:22:34 2008 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Mon Dec 15 08:22:40 2008 Subject: AW: [Histonet] Mucin blocking reagent for IHC In-Reply-To: References: Message-ID: <3F33170DD3F04B879AA638B1EC01EB45@dielangs.at> With the polymer-systems of IHC detection it is often seen, that in some tissueparts an unwanted background occurs. It is said, that the large polymers are trapped in OH-reach tissues like mucin and collagen. And also an additional washing step doesn't erase that. The polymers are labelled with Peroxidase. The trapped peroxidase leads to the colourdevelopement. Gudrun Lang Histolab Linz, Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von John Kiernan Gesendet: Montag, 15. Dezember 2008 07:15 An: Aprill Watanabe Cc: histonet@lists.utsouthwestern.edu Betreff: Re: [Histonet] Mucin blocking reagent for IHC Please explain "mucin trapping the DAB". Are you sure that this is not genuine immunohistochemical localization of the antigen? At neutral pH DAB cations are attracted to tissue polyanions (in mucus, cartilage etc). The attracted DAB should not be oxidized to a brown insoluble polymer in the absence of peroxidase. The only working peroxidase molecules in an immunohistochemical preparation should be those specifically bound at the site recognized by the primary antibody. Brown staining of mucus suggests the presence of peroxidase in the DAB-H2O2 final incbation solution. This would catalyze the oxidation of tissue-bound DAB. More washing after the last HRP-containing reagent might help. John Kiernan Anatomy, UWO London, Canada - - - ----- Original Message ----- From: Aprill Watanabe Date: Friday, December 12, 2008 17:26 Subject: [Histonet] Mucin blocking reagent for IHC To: histonet@lists.utsouthwestern.edu > I am staining CA19-9 on pancreas and have a good amount of mucin > trappingthe DAB. Is there a product that either blocks > mucin or something else that > will limit the mucin effect? > > Aprill Watanabe, B.S. > Research Associate > Integrated Cancer Genomics Division > Tissue Microarray Center (TMA) > Translational Genomics Research Institute (TGen) > main: 602-343-8822 > Fax: 602-343-8840 > awatanabe@tgen.org > www.tgen.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 From micropathlabs <@t> yahoo.com Mon Dec 15 09:42:44 2008 From: micropathlabs <@t> yahoo.com (Sheila Haas) Date: Mon Dec 15 09:42:48 2008 Subject: [Histonet] Data Entry Clerk Needed Message-ID: <935605.16737.qm@web57804.mail.re3.yahoo.com> We?have an immediate opening?for a data entry clerk with extensive medical?terminology (pathology preferred) for?laboratory in Lakeland, FL. Please e-mail me directly if interested. ? Sheila Haas Laboratory Supervisor Micro Path Laboratories From tbraud <@t> holyredeemer.com Mon Dec 15 10:00:37 2008 From: tbraud <@t> holyredeemer.com (Terri Braud) Date: Mon Dec 15 10:00:42 2008 Subject: [Histonet] RE: Recruiting Techs at work In-Reply-To: <778ad86e00002877@HolyRedeemer.com> Message-ID: I agree that if you have a great work environment with competitive pay your staff will stay put, but how many of us at the department supervisory/manager level can actually make the final decision concerning salary and pay scales for a position. I suspect that most of us, including myself, can only make well documented pleas for salary increases. When it comes to spending money, most institutions are reactive rather than proactive. If salaries are low, then you may have to lose a valuable tech before you can make the point that salaries need to be increased. It may be a tough pill to swallow, but I am more than willing to let my folk listen to any pitchman that is trying to get their ear. Any tech worth his/her salt, knows that these calls are out there, so why try to hide it. If a tech can better themselves by a job change, then who am I to stand in their way. We generally discuss these in departmental meetings and I tell techs that I get frequent calls from recruiters and if they would like to correspond with them, may I give the recruiter their personal contact info. I ask in return, that if they do go elsewhere for a better salary, that they please give me the info, so I can use it to help increase the salary of those who remain. This keeps communication up front, honest, and open and I think in the long run, helps to quell murmuring about low salaries which are beyond my control as a supervisor. Just another viewpoint. Terri Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Laboratory Holy Redeemer Hospital and Medical Center 1648 Huntingdon Pike Meadowbrook, PA 19046 (215) 938-3676 phone (215) 938-3689 fax Message: 18 Date: Sat, 13 Dec 2008 13:46:57 +0000 (UTC) From: lee2323@comcast.net Subject: Re: [Histonet] Offended I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. --------------------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. From tbraud <@t> holyredeemer.com Mon Dec 15 10:20:37 2008 From: tbraud <@t> holyredeemer.com (Terri Braud) Date: Mon Dec 15 10:20:41 2008 Subject: [Histonet] RE: Current Regs In-Reply-To: <7861d5c800003728@HolyRedeemer.com> Message-ID: Message: 1 Date: Sat, 13 Dec 2008 09:44:39 -0500 From: thisisann@aol.com Subject: [Histonet] Grosser Requirements To: histonet@lists.utsouthwestern.ed I was recently informed the CAP's grossing requirements changed....I can not find anything about a change on their website. Does anyone have the current regs in reference to educational requirements for Grossing? Ann Hey Ann - There were several changes in the 12/2006 checklist revision, especially in the NOTES sections of the questions, clarifying terms and definitions of the checklist questions. The 9/2007 is the latest revision of the AP questions. The link to the 9/2007 AP checklist questions is http://www.cap.org/apps/docs/laboratory_accreditation/checklists/anatomic_pathology_Sep07.doc What you are looking for is on pages 21-24 of this checklist, and is pasted below: ANP.11600 Phase II N/A YES NO Are all macroscopic tissue examinations performed by a pathologist or pathology resident, or under the supervision of a qualified pathologist? NOTE: Two levels of complexity of macroscopic tissue examination are defined, as follows: 1) Processing is defined as a tissue examination limited to description, inking and cutting of the specimen (if applicable), and submission of the entire specimen to histology. Tissue processing can be performed according to standardized protocols. Processing is generally limited to small specimens (skin ellipses, small biopsies, curettings, etc.) and does not require knowledge of anatomy. 2) Grossing (or gross examination) is defined as a tissue examination requiring a greater exercise of judgment and a knowledge of anatomy. Dissection of the specimen and selection of tissue samples for submission to histology are generally required. The specimen description is not necessarily standardized. Specific requirements for supervision of non-pathologists who process specimens, or assist in grossing specimens, are given below. ANP.11605 Phase II N/A YES NO When individuals other than a pathologist or pathology resident process specimens, or assist in gross examinations, is the extent of their activities (including the types of specimens examined) defined in a documented protocol? NOTE: This protocol must list the specific types of specimens that non-pathologists are permitted to process, and for which non-pathologists are permitted to assist in the gross examination. The laboratory director is responsible for this protocol. And NEW for 2006 altogether: ANP.11665 Phase I N/A YES NO Are there written procedures for processing specimens? NOTE: This question refers to processing as defined in ANP.11600, and applies only to processing of specimens by non-pathologist individuals who are not qualified as high complexity testing personnel under CLIA-88. Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Holy Redeemer Hospital and Medical Center 1648 Huntingdon Pike Meadowbrook, PA 19046 (215) 938-3676 phone (215) 938-3689 fax --------------------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. From slappycraw <@t> yahoo.com Mon Dec 15 10:21:09 2008 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Mon Dec 15 10:21:14 2008 Subject: [Histonet] RE: Recruiting Techs at work References: Message-ID: <979274.59486.qm@web53606.mail.re2.yahoo.com> >>>?If salaries are low, then you may have to lose a valuable tech before you can make the point that salaries need to be increased.<<< 100%?agreed and it's also true for trying to get some other things changed as well.? ? Larry A. Woody Seattle, Wa. ________________________________ From: Terri Braud To: histonet@lists.utsouthwestern.edu Sent: Monday, December 15, 2008 8:00:37 AM Subject: [Histonet] RE: Recruiting Techs at work I agree that if you have a great work environment with competitive pay your staff will stay put, but how many of us at the department supervisory/manager level can actually make the final decision concerning salary and pay scales for a position.? I suspect that most of us, including myself, can only make well documented pleas for salary increases. When it comes to spending money, most institutions are reactive rather than proactive.? If salaries are low, then you may have to lose a valuable tech before you can make the point that salaries need to be increased.? It may be a tough pill to swallow, but I am more than willing to let my folk listen to any pitchman that is trying to get their ear. Any tech worth his/her salt, knows that these calls are out there, so why try to hide it. If a tech can better themselves by a job change, then who am I to stand in their way.? We generally discuss these in departmental meetings and I tell techs that I get frequent calls from recruiters and if they would like to correspond with them, may I give the recruiter their personal contact info.? I ask in return, that if they do go elsewhere for a better salary, that they please give me the info, so I can use it to help increase the salary of those who remain. This keeps communication up front, honest, and open and I think in the long run, helps to quell murmuring about low salaries which are beyond my control as a supervisor. Just another viewpoint.? Terri Terri L. Braud, HT(ASCP) Anatomic Pathology Supervisor Laboratory Holy Redeemer Hospital and Medical Center 1648 Huntingdon Pike Meadowbrook, PA 19046 (215) 938-3676 phone (215) 938-3689 fax Message: 18 Date: Sat, 13 Dec 2008 13:46:57 +0000 (UTC) From: lee2323@comcast.net Subject: Re: [Histonet] Offended I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. --------------------------------------------------------------------------------- CONFIDENTIALITY NOTICE: This E-Mail is intended only for the use of the individual or entity to which it was sent. It may contain information that is privileged and/or confidential, and the use or disclosure of such information may also be restricted under applicable federal and state law. If you received this communication in error, please do not distribute any part of it or retain any copies, and delete the original E-Mail. Please notify the sender of any error by E-Mail. Thank you for your cooperation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From leiker <@t> buffalo.edu Mon Dec 15 11:08:25 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Mon Dec 15 11:08:30 2008 Subject: [Histonet] RE: Recruiting Techs at work In-Reply-To: References: Message-ID: This indeed sounds like the most honest and integrous approach as well as a way to keep positions competitive. I like it. --On Monday, December 15, 2008 11:00 AM -0500 Terri Braud wrote: > I agree that if you have a great work environment with competitive pay > your staff will stay put, but how many of us at the department > supervisory/manager level can actually make the final decision concerning > salary and pay scales for a position. I suspect that most of us, > including myself, can only make well documented pleas for salary > increases. When it comes to spending money, most institutions are > reactive rather than proactive. If salaries are low, then you may have > to lose a valuable tech before you can make the point that salaries need > to be increased. It may be a tough pill to swallow, but I am more than > willing to let my folk listen to any pitchman that is trying to get their > ear. Any tech worth his/her salt, knows that these calls are out there, > so why try to hide it. If a tech can better themselves by a job change, > then who am I to stand in their way. We generally discuss these in > departmental meetings and I tell techs that I get frequent calls from > recruiters and if they would like to correspond with them, may I give the > recruiter their personal contact info. I ask in return, that if they do > go elsewhere for a better salary, that they please give me the info, so I > can use it to help increase the salary of those who remain. This keeps > communication up front, honest, and open and I think in the long run, > helps to quell murmuring about low salaries which are beyond my control > as a supervisor. Just another viewpoint. Terri > > Terri L. Braud, HT(ASCP) > Anatomic Pathology Supervisor > Laboratory > Holy Redeemer Hospital and Medical Center > 1648 Huntingdon Pike > Meadowbrook, PA 19046 > (215) 938-3676 phone > (215) 938-3689 fax > > Message: 18 > Date: Sat, 13 Dec 2008 13:46:57 +0000 (UTC) > From: lee2323@comcast.net > Subject: Re: [Histonet] Offended > > I think if you have a great work environment with competitve pay then you > have nothing to worry about. Your staff will stay put. > > > ------------------------------------------------------------------------- > -------- > > > > CONFIDENTIALITY NOTICE: > > This E-Mail is intended only for the use of the individual or entity to > which it was sent. It may contain information that is privileged and/or > confidential, and the use or disclosure of such information may also be > restricted under applicable federal and state law. If you received this > communication in error, please do not distribute any part of it or retain > any copies, and delete the original E-Mail. Please notify the sender of > any error by E-Mail. > > Thank you for your cooperation. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From joelleweaver <@t> hotmail.com Mon Dec 15 12:29:42 2008 From: joelleweaver <@t> hotmail.com (joelle weaver) Date: Mon Dec 15 12:29:47 2008 Subject: [Histonet] RE: Recruiting Techs at work In-Reply-To: References: <778ad86e00002877@HolyRedeemer.com> Message-ID: Hi Terri I appreciated your post. I think all histotechs know that jobs are plentiful ( at least in the current market), and most of us get lots of postings- But, I really don't even take notice unless there is something that is making me think and wonder if the grass is greener- this only very rarely has to do with $ for me. And, I really couldn't speak for everyone out there, but above a certain dollar amount (needed to buy food and provide shelter) I have personally not been tempted to move to another lab unless other important aspects that I consider important, were missing from my work environment. I do understand that aggressive recruiters could be annoying to management trying to get and retain good techs. I know that managers struggle to get a decent wage for their best employees- I think that is admirable. However, maybe this will make some feel better? The main reasons that I have considered changing employers had to do with work schedule( unreasonable number of hours, all weekends/ no vacation etc), and not feeling valued as an employee-(a big one). Just to give an example of a moral killer- a lab policy states that ALL histotechs, regardless of training, education, or experience, are disqualified for any type of promotion within Histology-or anywhere in the lab. I think that is pretty crappy- and I think that kind of stuff really makes you feel de-valued-as a histotech, even if you are not seeking advancement at the present time. It is just an opinion, but I think that if managers value their techs,and let them see that, they will keep the ones worth keeping! J. Weaver > Date: Mon, 15 Dec 2008 11:00:37 -0500> From: tbraud@holyredeemer.com> To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] RE: Recruiting Techs at work> > I agree that if you have a great work environment with competitive pay your staff will stay put, but how many of us at the department supervisory/manager level can actually make the final decision concerning salary and pay scales for a position. I suspect that most of us, including myself, can only make well documented pleas for salary increases. > When it comes to spending money, most institutions are reactive rather than proactive. If salaries are low, then you may have to lose a valuable tech before you can make the point that salaries need to be increased. > It may be a tough pill to swallow, but I am more than willing to let my folk listen to any pitchman that is trying to get their ear. Any tech worth his/her salt, knows that these calls are out there, so why try to hide it. If a tech can better themselves by a job change, then who am I to stand in their way. We generally discuss these in departmental meetings and I tell techs that I get frequent calls from recruiters and if they would like to correspond with them, may I give the recruiter their personal contact info. I ask in return, that if they do go elsewhere for a better salary, that they please give me the info, so I can use it to help increase the salary of those who remain. This keeps communication up front, honest, and open and I think in the long run, helps to quell murmuring about low salaries which are beyond my control as a supervisor.> Just another viewpoint. Terri> > Terri L. Braud, HT(ASCP)> Anatomic Pathology Supervisor> Laboratory> Holy Redeemer Hospital and Medical Center> 1648 Huntingdon Pike> Meadowbrook, PA 19046> (215) 938-3676 phone> (215) 938-3689 fax> > Message: 18> Date: Sat, 13 Dec 2008 13:46:57 +0000 (UTC)> From: lee2323@comcast.net> Subject: Re: [Histonet] Offended> > I think if you have a great work environment with competitve pay then you have nothing to worry about. Your staff will stay put. > > > ---------------------------------------------------------------------------------> > > > CONFIDENTIALITY NOTICE:> > This E-Mail is intended only for the use of the individual or entity to which> it was sent. It may contain information that is privileged and/or confidential,> and the use or disclosure of such information may also be restricted under applicable> federal and state law. If you received this communication in error, please do not> distribute any part of it or retain any copies, and delete the original E-Mail.> Please notify the sender of any error by E-Mail.> > Thank you for your cooperation.> > _______________________________________________> Histonet mailing list> Histonet@lists.utsouthwestern.edu> http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008 From bakevictoria <@t> gmail.com Mon Dec 15 12:40:47 2008 From: bakevictoria <@t> gmail.com (Victoria Baker) Date: Mon Dec 15 12:41:31 2008 Subject: [Histonet] RE: Recruiting Techs at work In-Reply-To: References: <778ad86e00002877@HolyRedeemer.com> Message-ID: <4f016b690812151040h2d441e26o96898faf5b7a4fcc@mail.gmail.com> Joelle - I am very proud of you and so would Pat. Vikki On Mon, Dec 15, 2008 at 1:29 PM, joelle weaver wrote: > > Hi Terri > I appreciated your post. I think all histotechs know that jobs are > plentiful ( at least in the current market), and most of us get lots of > postings- But, I really don't even take notice unless there is something > that is making me think and wonder if the grass is greener- this only very > rarely has to do with $ for me. And, I really couldn't speak for everyone > out there, but above a certain dollar amount (needed to buy food and provide > shelter) I have personally not been tempted to move to another lab unless > other important aspects that I consider important, were missing from my work > environment. I do understand that aggressive recruiters could be annoying to > management trying to get and retain good techs. I know that managers > struggle to get a decent wage for their best employees- I think that is > admirable. > However, maybe this will make some feel better? The main reasons that I > have considered changing employers had to do with work schedule( > unreasonable number of hours, all weekends/ no vacation etc), and not > feeling valued as an employee-(a big one). Just to give an example of a > moral killer- a lab policy states that ALL histotechs, regardless of > training, education, or experience, are disqualified for any type of > promotion within Histology-or anywhere in the lab. I think that is pretty > crappy- and I think that kind of stuff really makes you feel de-valued-as a > histotech, even if you are not seeking advancement at the present time. It > is just an opinion, but I think that if managers value their techs,and let > them see that, they will keep the ones worth keeping! > J. Weaver > > > > > Date: Mon, 15 Dec 2008 11:00:37 -0500> From: tbraud@holyredeemer.com> > To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] RE: Recruiting > Techs at work> > I agree that if you have a great work environment with > competitive pay your staff will stay put, but how many of us at the > department supervisory/manager level can actually make the final decision > concerning salary and pay scales for a position. I suspect that most of us, > including myself, can only make well documented pleas for salary increases. > > When it comes to spending money, most institutions are reactive rather > than proactive. If salaries are low, then you may have to lose a valuable > tech before you can make the point that salaries need to be increased. > It > may be a tough pill to swallow, but I am more than willing to let my folk > listen to any pitchman that is trying to get their ear. Any tech worth > his/her salt, knows that these calls are out there, so why try to hide it. > If a tech can better themselves by a job change, then who am I to stand in > their way. We generally discuss these in departmental meetings and I tell > techs that I get frequent calls from recruiters and if they would like to > correspond with them, may I give the recruiter their personal contact info. > I ask in return, that if they do go elsewhere for a better salary, that they > please give me the info, so I can use it to help increase the salary of > those who remain. This keeps communication up front, honest, and open and I > think in the long run, helps to quell murmuring about low salaries which are > beyond my control as a supervisor.> Just another viewpoint. Terri> > Terri > L. Braud, HT(ASCP)> Anatomic Pathology Supervisor> Laboratory> Holy Redeemer > Hospital and Medical Center> 1648 Huntingdon Pike> Meadowbrook, PA 19046> > (215) 938-3676 phone> (215) 938-3689 fax> > Message: 18> Date: Sat, 13 Dec > 2008 13:46:57 +0000 (UTC)> From: lee2323@comcast.net> Subject: Re: > [Histonet] Offended> > I think if you have a great work environment with > competitve pay then you have nothing to worry about. Your staff will stay > put. > > > > ---------------------------------------------------------------------------------> > > > > CONFIDENTIALITY NOTICE:> > This E-Mail is intended only for the use of > the individual or entity to which> it was sent. It may contain information > that is privileged and/or confidential,> and the use or disclosure of such > information may also be restricted under applicable> federal and state law. > If you received this communication in error, please do not> distribute any > part of it or retain any copies, and delete the original E-Mail.> Please > notify the sender of any error by E-Mail.> > Thank you for your > cooperation.> > _______________________________________________> Histonet > mailing list> Histonet@lists.utsouthwestern.edu> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________________________________________ > Send e-mail faster without improving your typing skills. > > http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008_______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From tanisha.mcknight <@t> covance.com Mon Dec 15 12:49:40 2008 From: tanisha.mcknight <@t> covance.com (McKnight, Tanisha) Date: Mon Dec 15 12:49:52 2008 Subject: [Histonet] CAP and NYSDOH Regs for Staining Validation Message-ID: <816E3C72F855F14985FC31D7C963AE6F0C4F29EB@indexch03.ent.covance.com> Hello Histonetters: Can anyone share their understanding of CAP and NYSDOH regs for validating staining and fixation procedures (not IHC)? Are the requirements for validation different for Anatomic Pathology than for Clinical Pathology? And if so, how do so of you go about performing and documenting (if you are allowed to share). Thanks, Tanisha Neely, HT(ASCP) AP-Histology/Specimen Management Covance CLS, Indianapolis tanisha.mcknight@covance.com ----------------------------------------------------- Confidentiality Notice: This e-mail transmission may contain confidential or legally privileged information that is intended only for the individual or entity named in the e-mail address. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution, or reliance upon the contents of this e-mail is strictly prohibited. If you have received this e-mail transmission in error, please reply to the sender, so that we can arrange for proper delivery, and then please delete the message from your inbox. Thank you. From CIngles <@t> uwhealth.org Mon Dec 15 13:07:18 2008 From: CIngles <@t> uwhealth.org (Ingles Claire ) Date: Mon Dec 15 13:07:23 2008 Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS References: <746229.92433.qm@web65711.mail.ac4.yahoo.com> Message-ID: <08A0A863637F1349BBFD83A96B27A50A08E609E6@uwhis-xchng3.uwhis.hosp.wisc.edu> We actually do that. We are lazy though, and use the water bath by our microtome(approx 43 F). After cutting is done, of course. That way you don't have anything extra to clean up. (I HOPE everyone cleans their microtome waterbaths daily!) :) We sometimes have to use the waterbath in the fume hood. Keep in mind that the air movement will lower the temp of the bath (especially during Wisconsin winters!). We usually put a small metal tray over the bath leaving an opening just large enough for the coplin jar and monitor the water temp continuously. Claire ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Rene J Buesa Sent: Fri 12/12/2008 3:09 PM To: HISTONET ...; TIM MALLOY; TIMOTHY MALLOY Subject: Re: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS I used the MW oven to heat-boost the slutions but the actual staining was done in a water bath. Why don't you try a water bath at 60?C? ren? J. --- On Fri, 12/12/08, TIMOTHY MALLOY wrote: From: TIMOTHY MALLOY Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS To: "HISTONET ..." , "TIM MALLOY" Date: Friday, December 12, 2008, 4:52 AM From jkiernan <@t> uwo.ca Mon Dec 15 13:30:21 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Dec 15 13:30:24 2008 Subject: AW: [Histonet] Mucin blocking reagent for IHC In-Reply-To: <3F33170DD3F04B879AA638B1EC01EB45@dielangs.at> References: <3F33170DD3F04B879AA638B1EC01EB45@dielangs.at> Message-ID: That's a good point. The original question did not mention whether a polymer-based detection system was being used. John Kiernan Anatomy, UWO London, Canada = = = = ----- Original Message ----- From: Gudrun Lang Date: Monday, December 15, 2008 9:22 Subject: AW: [Histonet] Mucin blocking reagent for IHC To: 'John Kiernan' Cc: histonet@lists.utsouthwestern.edu > With the polymer-systems of IHC detection it is often seen, that > in some > tissueparts an unwanted background occurs. It is said, that the large > polymers are trapped in OH-reach tissues like mucin and > collagen. And also > an additional washing step doesn't erase that. The polymers are > labelledwith Peroxidase. The trapped peroxidase leads to the > colourdevelopement. > Gudrun Lang > Histolab > Linz, Austria > > -----Urspr?ngliche Nachricht----- > Von: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag > von John > Kiernan > Gesendet: Montag, 15. Dezember 2008 07:15 > An: Aprill Watanabe > Cc: histonet@lists.utsouthwestern.edu > Betreff: Re: [Histonet] Mucin blocking reagent for IHC > > Please explain "mucin trapping the DAB". Are you sure that this > is not > genuine immunohistochemical localization of the antigen? > > At neutral pH DAB cations are attracted to tissue polyanions (in > mucus,cartilage etc). The attracted DAB should not be oxidized > to a brown > insoluble polymer in the absence of peroxidase. > > The only working peroxidase molecules in an immunohistochemical > preparationshould be those specifically bound at the site > recognized by the primary > antibody. Brown staining of mucus suggests the presence of > peroxidase in the > DAB-H2O2 final incbation solution. This would catalyze the > oxidation of > tissue-bound DAB. More washing after the last HRP-containing > reagent might > help. > > John Kiernan > Anatomy, UWO > London, Canada > - - - > ----- Original Message ----- > From: Aprill Watanabe > Date: Friday, December 12, 2008 17:26 > Subject: [Histonet] Mucin blocking reagent for IHC > To: histonet@lists.utsouthwestern.edu > > > I am staining CA19-9 on pancreas and have a good amount of > mucin > > trappingthe DAB. Is there a product that either blocks > > mucin or something else that > > will limit the mucin effect? > > > > Aprill Watanabe, B.S. > > Research Associate > > Integrated Cancer Genomics Division > > Tissue Microarray Center (TMA) > > Translational Genomics Research Institute (TGen) > > main: 602-343-8822 > > Fax: 602-343-8840 > > awatanabe@tgen.org > > www.tgen.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 > From carl.hobbs <@t> kcl.ac.uk Mon Dec 15 13:33:26 2008 From: carl.hobbs <@t> kcl.ac.uk (Hobbs, Carl) Date: Mon Dec 15 13:34:31 2008 Subject: [Histonet] pH testing Message-ID: <11D9615B89C10747B1C985966A63D7CA293069EC73@KCL-MAIL04.kclad.ds.kcl.ac.uk> I would be very grateful if John Kiernan had the time to respond to my question. We all talk about pH but, many of us have no idea how to make sure, for eg, that we are actually measuring pH values accurately. How should we do that? I use TRIS buffer for my Immunostaining : I have a TRIS pH electrode. Is this absolutely reqd? I use VWR Std buffers to check my pH meter once a week: I check ph4,7 and 10. Is this neccessary? Respectfully, Carl From sharon.osborn <@t> comcast.net Mon Dec 15 14:44:40 2008 From: sharon.osborn <@t> comcast.net (sharon.osborn@comcast.net) Date: Mon Dec 15 14:44:45 2008 Subject: [Histonet] Embedding center and staining dishes Message-ID: <121520082044.19321.4946C1B8000C466D00004B792215575474029D010D9C01D202019D0E089C@comcast.net> To all histotechs We have a TissueTek II Embedding console in excellent working condition that you can have by paying for the shipping charges. Remember when we were thrilled to have one of these units rather than the coffee pot or ice cream scoop to fill our blocks? We also have a complete set of the glass staining dishes plus covers for the Richard Allan HMS 760 Robotic Stainer. These are also free with you paying the shipping charges. Please let me know your interest in whichever one you wish plus your contact infomtation, shipping information and UPS or Fed Ex shipping number. Sharon Osborn, BS, HT(ASCP) IHC-QC LabVision ThermoFisher Scientific Fremont, CA 510-991-2824 sharon.osborn@thermofisher.com From dusko.trajkovic <@t> pfizer.com Mon Dec 15 16:18:00 2008 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Mon Dec 15 16:18:05 2008 Subject: [Histonet] Need info on Thermo Shandon Pathcenter processors In-Reply-To: Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B2088C1775@lajamrexm01.amer.pfizer.com> Hallo Histonetters, If anyone in the Southern California area has one of these processors, can you please give me a contact person or a company that would be able to service the instrument? Thanks Dusko From tifei <@t> foxmail.com Mon Dec 15 20:00:08 2008 From: tifei <@t> foxmail.com (tf) Date: Mon Dec 15 20:00:29 2008 Subject: [Histonet] Silver staing for degenerating axons Message-ID: <200812161000022633859@foxmail.com> I read the paper of Nauta with great interest! But I dont have the original pdf for the paper on Stain Technology in 1951 & 1954. Anyone can send me the pdf? I only have the improved method by Fink and Heimer in 1967 published on brain research, giving out two staining procedures. I can send you the pdf if anyone is interested in this. I would like to receive any proved protocol of silver staining for degenerating axons...because we dont have uranyl nitrate or hydroquinone here...I can not repeat the paper by Fink & Heimer. Can anyone provide me a supressive procedure in staining degenerating axons using "common chemicals", just ammonia water, silver nitrate, citric acid, formalin/PFA, sodium hydroxide? I just can not find applicable one with our current facilities! Thanks very much! 2008-12-15 tf ???? John Kiernan ????? 2008-12-15 13:42:09 ???? Susan Bachus ??? Pat Flannery; Weems, Joyce; histonet ??? Silver & ammonia (Was Re: [Histonet] Silly Question? , , ,) Dear Susan, Thank you for bringing Walle Nauta's 1993 review article to Histonetters' attention. I hadn't seen it before, and I agree that it was well worth downloading, reading and filing for future reference. More than 50 years ago Nauta & Gygax published a very easily understood account of ammoniacal silver solutions and the effects of changing their pH. This is fundamental to all techniques involving silver nitrate and ammonia, and the exact information is not in chemistry textbooks. Here's the reference. Nauta WJH, Gygax PA (1951) Silver impregnation of degenerating axon terminals in the central nervous system: (1) Technic. (2) Chemical notes. Stain Technology 26: 5-11. Part 2 was an appendix to their paper on the early (non-suppressive) Nauta-Gygax method, which stains both normal and degenerating axons. Many of the principles explained in Part 2 of this paper apply also to other silver methods. If your library subscribes to journals published by Informa, you can download this Nauta-Gygax paper (and anything else in Stain Technology) as a PDF file from the InformaWorld web site: http://www.informaworld.com/smpp/title~content=t713692932~bb=all The journal's name shows on the present publisher's web site in its present form (Biotechnic & Histochemistry) even though the name was really Stain Technology before about 1990. This is a problem with quite a few journals that have changed their names. The Springer web site, for example, doesn't seem to recognize the name changes of Histochemie --> Histochemistry --> Histochemistry and Cell Biology. Library catalogues usually record name changes faithfully. For journals, changing the name probably isn't always a good move! John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: Susan Bachus Date: Friday, December 12, 2008 20:33 Subject: Re: [Histonet] Silly Question? - Need help quickly! To: rjbuesa@yahoo.com, Pat Flannery , histonet@lists.utsouthwestern.edu, "Weems, Joyce" > I tried earlier to send, for this thread, a wonderful paper by > Nauta, > chronicaling the history his discovery of his tract tracing > method, in which > serendipity and degradation of formalin played critical roles, > not realizing > that the size of the attachment would prevent it from going > through, so I am > trying again with a URL for this paper: > http://www.jneurosci.org/cgi/reprint/13/4/1337 > > Susan > > ----- Original Message ----- > From: "Rene J Buesa" > To: "Pat Flannery" ; > ; > "Weems, Joyce" > Sent: Thursday, December 11, 2008 12:58 PM > Subject: RE: [Histonet] Silly Question? - Need help quickly! > > > Joyce: > Methanal, which is the chemical name of formaldehyde, > polymerizes. If it > forms a polymer of at least 50 molecules or more, it gets solid > = > para-formaldehyde. > Formalin (a trade name as formol is also another trade name)is > the 37-50% > aqueous solution of formaldehyde (with some additiveses to > prevent > polymerization). > You can prepare BNF using the formalin solution or dissolving > the amount of > solid para-formaldehydede to get to the concentrationon you desire. > The chemical in both solutions is the same = methanal or > formaldehyde.Ren? > J. > > --- On Thu, 12/11/08, Weems, Joyce wrote: > > > From: Weems, Joyce > Subject: RE: [Histonet] Silly Question? - Need help quickly! > To: "Pat Flannery" , > histonet@lists.utsouthwestern.eduDate: Thursday, December 11, > 2008, 12:12 PM > > I was just going to post a question regarding paraformaldhyde myself! > Just last week I believe I remember someone saying that > paraformaldehydeand formalin are the same and they had put the > same solution in two > different containers for one of their researchers because they > were so > insistent to have two different solutions. Are they the same? > > Well, today I have a request to put tissue for a researcher in > formalinand paraformaldehyde. So.... Without percentage > required, do I use 10% > NBF? Do I call somewhere and get paraformaldehyde and make 4% > paraformaldehyde? > > I have asked the surgeon twice for the number for the lab so I > can find > out - don't have it yet. I have two fresh adrenals in the > fridge. Help!! > > > Thanks in advance... > Joyce > > Joyce Weems > Pathology Manager > Saint Joseph's Hospital > 5665 Peachtree Dunwoody Rd NE > Atlanta, GA 30342 > 678-843-7376 - Phone > 678-843-7831 - Fax > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat > Flannery > Sent: Thursday, December 11, 2008 11:59 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Silly Question? > > Please humor me on this if it's obvious (to everyone but > me): why do we > use paraformaldehyde (which is so inconvenient to make up) > rather than > buffered formalin or just diluted formaldehyde itself? > > It seems that around here, some folks prefer paraformaldehyde > (either 2% > or 4%) and others use formalin, while some others stick to diluted > formaldehyde (I see all 4 on labels for specimens submitted for > histology). Is it mostly a matter of personal preference > or where you > were trained (i.e. force of habit) or is there a valid reason to use > each solution (basically the same chemical once in solution, merely > buffered or not)? The only answer I've gotten when I've > asked is, > "That's what we always use." > > Thanks. > > -Pat Flannery (not a "real" histologist - I just play one in the lab) > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Confidentiality Notice: > This email, including any attachments is the > property of Catholic Health East and is intended > for the sole use of the intended recipient(s). > It may contain information that is privileged and > confidential. Any unauthorized review, use, > disclosure, or distribution is prohibited. If you are > not the intended recipient, please reply to the > sender that you have received the message in > error, then delete this 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From tgenade <@t> gmail.com Tue Dec 16 04:24:38 2008 From: tgenade <@t> gmail.com (Tyrone Genade) Date: Tue Dec 16 04:26:03 2008 Subject: [Histonet] Re: Histonet Digest, Vol 61, Issue 25 In-Reply-To: <49469b39.050bca0a.524c.ffffd630SMTPIN_ADDED@mx.google.com> References: <49469b39.050bca0a.524c.ffffd630SMTPIN_ADDED@mx.google.com> Message-ID: Hi all, > Date: Mon, 15 Dec 2008 18:56:28 +0800 > From: " tf " > Subject: [Histonet] Silver staing for degenerating axons > But I dont have the original pdf for the paper on Stain Technology in 1951 & 1954. Anyone can > send me the pdf? [snip] > Can anyone provide me a supressive procedure in staining degenerating axons using "common > chemicals", just ammonia water, silver nitrate, citric acid, formalin/PFA, sodium hydroxide? I > just can not find applicable one with our current facilities! Have you read Sun et al (2002) Comparative analysis of an improved thioflavin-S stain, Gallas Silver Stain, and immunohistochemistry for neurofibrillay tangle demonstration of the same sections. J. Histochem & Cytochem 50(4):463--472? The reference for Gallays is: Gallays (1971) Silver staining of Alzheimer's neurofibrillary changes by means of physical development. Acta Morphol Acad Sci Hung 19:1--8. Otherwise, you may want to look into FlouroJadeB. If all you are after is visualizing degenerating axons, there are easier ways of going about ot than silver staining. Kind regards -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@freeshell.org tel: +27-84-632-1925 (c) ******************************************************************************** "For there is one God, and there is one mediator between God and men, the man Christ Jesus, who gave himself as a ransom for all." 1 Timothy 2:5-6 From rcharles <@t> state.pa.us Tue Dec 16 07:02:30 2008 From: rcharles <@t> state.pa.us (Charles, Roger) Date: Tue Dec 16 07:02:34 2008 Subject: [Histonet] Paraffin wax, Thank You Message-ID: <1B2F365C5F88A946B7D602E64ACD9CDD044933AB9C@ENHBGMBX04.PA.LCL> Thank you to all who responded to this subject. Your knowledge will help us make our decisions on the type of paraplast to be used in our lab, at least until our contract vendor changes again. Happy Holidays to all Roger Roger Charles Microbiologist Pennsylvania Veterinary Laboratory 2305 N Cameron St Harrisburg, PA 17110 717-787-8808 From cmmathis1 <@t> bellsouth.net Tue Dec 16 09:51:04 2008 From: cmmathis1 <@t> bellsouth.net (cmmathis1@bellsouth.net) Date: Tue Dec 16 09:51:20 2008 Subject: [Histonet] What is the stickest slides Message-ID: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> Hello Histonetters, We are having trouble getting our paraffin processed cow meniscus sections (at 5 microns) to stay on our Superfrost Plus slides. We are drying them overnight in a 58 C oven. The H&Es look great but the immunos wash every time. What is your favorite slide for these hard to stay on samples? Thanks for your time, Cathy From RossS <@t> BaylorHealth.edu Tue Dec 16 09:53:49 2008 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Tue Dec 16 09:54:54 2008 Subject: [Histonet] Silver staing for degenerating axons, thank you for sharing the paper. In-Reply-To: <200812161000022633859@foxmail.com> Message-ID: A great thank you to the person who shared this paper. This was a very interesting read. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu ********************************************************************** This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From slappycraw <@t> yahoo.com Tue Dec 16 10:17:20 2008 From: slappycraw <@t> yahoo.com (Larry Woody) Date: Tue Dec 16 10:17:24 2008 Subject: [Histonet] CD79a Message-ID: <819891.58696.qm@web53608.mail.re2.yahoo.com> Anyone out there in Histonet land using CD79a on Rat tissue with success, I?would appreciate your input. Thanks in advance. ? Larry A. Woody Seattle, Wa. From alexandra.meinl <@t> gmail.com Tue Dec 16 10:23:54 2008 From: alexandra.meinl <@t> gmail.com (Alexandra Meinl) Date: Tue Dec 16 10:23:59 2008 Subject: [Histonet] What is the stickest slides In-Reply-To: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> Message-ID: I like Roti-Bond Adhesion slides for difficult material like bone, you can also use HIER without problems. But these slides are quite expensive and require 2-3 days drying time. Alex 2008/12/16 > Hello Histonetters, > We are having trouble getting our paraffin processed cow meniscus sections > (at 5 microns) to stay on our Superfrost Plus slides. We are drying them > overnight in a 58 C oven. The H&Es look great but the immunos wash every > time. > What is your favorite slide for these hard to stay on samples? > Thanks for your time, > Cathy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From asmith <@t> mail.barry.edu Tue Dec 16 10:47:40 2008 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Tue Dec 16 10:48:06 2008 Subject: [Histonet] What is the stickest slides In-Reply-To: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> Message-ID: I buy Vector Labs' VECTABOND and coat the slides myself. I dry the sections onto the slides for 2 days. I have yet to lose a section. -Allen A. Smith, Ph.D. Professor of Anatomy School of Podiatric Medicine Barry University Miami Shores, Florida -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cmmathis1@bellsouth.net Sent: Tuesday, December 16, 2008 10:51 AM To: histonet Subject: [Histonet] What is the stickest slides Hello Histonetters, We are having trouble getting our paraffin processed cow meniscus sections (at 5 microns) to stay on our Superfrost Plus slides. We are drying them overnight in a 58 C oven. The H&Es look great but the immunos wash every time. What is your favorite slide for these hard to stay on samples? Thanks for your time, Cathy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Tue Dec 16 11:00:05 2008 From: tifei <@t> foxmail.com (tf) Date: Tue Dec 16 11:00:35 2008 Subject: [Histonet] stickest slides for brain sections or how to coat a proper slide for urself References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net>, Message-ID: <200812170100003020790@foxmail.com> I have similar questions... for brain sections. we found that 0.25% gel coated slides are better than 0.5% or 1% gel in sticking 40 um brain sections. just want to share this with you guys. 2008-12-17 tf ???? Alexandra Meinl ????? 2008-12-17 00:27:04 ???? cmmathis1 ??? histonet ??? Re: [Histonet] What is the stickest slides I like Roti-Bond Adhesion slides for difficult material like bone, you can also use HIER without problems. But these slides are quite expensive and require 2-3 days drying time. Alex 2008/12/16 > Hello Histonetters, > We are having trouble getting our paraffin processed cow meniscus sections > (at 5 microns) to stay on our Superfrost Plus slides. We are drying them > overnight in a 58 C oven. The H&Es look great but the immunos wash every > time. > What is your favorite slide for these hard to stay on samples? > Thanks for your time, > Cathy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From Maria.Mejia <@t> ucsf.edu Tue Dec 16 11:24:21 2008 From: Maria.Mejia <@t> ucsf.edu (Mejia, Maria) Date: Tue Dec 16 11:24:32 2008 Subject: [Histonet] stickest slides for brain sections or how to coat aproper slide for urself References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net>, <200812170100003020790@foxmail.com> Message-ID: <6CF686BD6F24A546B85B24FE3B978647049CE8FD@EXVS06.net.ucsf.edu> This is for anyone's interest who work with 40um primate brain section 2x3 inches. We routinely mount our IHC stained sections from working PBS on pre-subbed brain slides from Brain Research Labs. To keep the sections on the slides we use a very fine (#0) brush to brush the section flat on the slide & to remove any PBS that may get trap under the section. We dry overnight. Next day, I put in oven at 37C - 30 minutes to further dry. Regards Maria Bartola Mejia Histology Manager Department of Neurosurgery UCSF San Francisco, CA 94103 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of tf Sent: Tue 12/16/2008 9:00 AM To: Alexandra Meinl; cmmathis1 Cc: histonet Subject: [Histonet] stickest slides for brain sections or how to coat aproper slide for urself I have similar questions... for brain sections. we found that 0.25% gel coated slides are better than 0.5% or 1% gel in sticking 40 um brain sections. just want to share this with you guys. 2008-12-17 tf ???: Alexandra Meinl ????: 2008-12-17 00:27:04 ???: cmmathis1 ??: histonet ??: Re: [Histonet] What is the stickest slides I like Roti-Bond Adhesion slides for difficult material like bone, you can also use HIER without problems. But these slides are quite expensive and require 2-3 days drying time. Alex 2008/12/16 > Hello Histonetters, > We are having trouble getting our paraffin processed cow meniscus sections > (at 5 microns) to stay on our Superfrost Plus slides. We are drying them > overnight in a 58 C oven. The H&Es look great but the immunos wash every > time. > What is your favorite slide for these hard to stay on samples? > Thanks for your time, > Cathy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From carmen_loiselle <@t> hotmail.com Tue Dec 16 11:24:36 2008 From: carmen_loiselle <@t> hotmail.com (carmen loiselle) Date: Tue Dec 16 11:25:46 2008 Subject: [Histonet] Isopentane Message-ID: Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ From mwich <@t> 7thwavelabs.com Tue Dec 16 11:30:56 2008 From: mwich <@t> 7thwavelabs.com (Michele Wich) Date: Tue Dec 16 11:31:01 2008 Subject: [Histonet] at a troubleshooting loss Message-ID: <62A8156F8071C8439080D626DF8C33A602E5B3@wave-mail.7thwave.local> A couple questions for those who are troubleshooting savvy: Other than inadequate fixation, water being introduced during processing, or too much heat, what can cause fuzzy, uneven hematoxylin staining with poor nuclear detail? And secondly, aside from formalin, are there any other common agents of blackish pigment in H & E staining? (The artifact appears on the slides both on and around the tissues.) Any feedback is GREATLY appreciated. This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer From settembr <@t> umdnj.edu Tue Dec 16 12:27:13 2008 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Tue Dec 16 12:28:09 2008 Subject: [Histonet] Isopentane Message-ID: Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SharonC <@t> celligent.net Tue Dec 16 12:38:29 2008 From: SharonC <@t> celligent.net (Sharon Campbell) Date: Tue Dec 16 12:38:35 2008 Subject: [Histonet] Prostate core biopsy submission Message-ID: Hello Histo-netters: My lab is starting to bring in more business with prostate core biopsies. Some of the submitting physicians would like to use a small piece of towel to lay the biopsy on and then place it and the core biopsy into the formalin container. Does anyone have any suggestions for what type of material would be suitable for this? I am afraid that anything cottony will be a problem when removing the biopsy during processing as it may stick. Thank you for your help. Sharon Campbell From jennifer.l.hofecker <@t> Vanderbilt.Edu Tue Dec 16 12:47:41 2008 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Tue Dec 16 12:47:47 2008 Subject: [Histonet] What is the stickest slides In-Reply-To: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> Message-ID: Hi Cathy, I use Starfrost plus slides from Mercedes Medical. We use them for all of our neuropathology specimens, including muscle and nerve at 5 microns, and have no problems. I dry my slides on a hot plate for 10 minutes (after draining thoroughly) and never have a problem with either H&E's or IHC washing off - either by manual or automated staining. Call them for a sample. Mercedes sells different brands of adhesive slides, be sure you ask for the Starfrost Platinum Line, they are wonderful. Have a great day! Jennifer L. Hofecker HT(ASCP) Vanderbilt University Medical Center Division of Neuropathology Nashville, TN ph 615.343.0083 fax 615.343.7089 -----Original Message----- From: cmmathis1@bellsouth.net [mailto:cmmathis1@bellsouth.net] Sent: Tuesday, December 16, 2008 9:51 AM To: histonet Subject: [Histonet] What is the stickest slides Hello Histonetters, We are having trouble getting our paraffin processed cow meniscus sections (at 5 microns) to stay on our Superfrost Plus slides. We are drying them overnight in a 58 C oven. The H&Es look great but the immunos wash every time. What is your favorite slide for these hard to stay on samples? Thanks for your time, Cathy From k.fd <@t> live.com Tue Dec 16 12:49:28 2008 From: k.fd <@t> live.com (Karen Doty) Date: Tue Dec 16 12:49:33 2008 Subject: [Histonet] processing tadpoles Message-ID: Would anyone care to share a paraffin processing protocol for tadpoles? Ideally, we would be processing a whole tadpole, but might be able to remove the head and tip of tail. We have a VIP tissue processor. Thanks Katy _________________________________________________________________ Send e-mail faster without improving your typing skills. http://windowslive.com/Explore/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_speed_122008 From zerfasp <@t> ors.od.nih.gov Tue Dec 16 13:11:35 2008 From: zerfasp <@t> ors.od.nih.gov (Zerfas, Patricia (NIH/OD/ORS) [E]) Date: Tue Dec 16 13:11:51 2008 Subject: [Histonet] autostainer service Message-ID: <424D930E0319E0469DD9C61502A50E3002102052@NIHCESMLBX4.nih.gov> Dear All, I am currently using the DAKOCytomation autostainer that is approximately seven years old. Once the service contract expires in November 2009 DAKOCytomation will not continue the service contract. The equipment is in excellent condition and meets the needs of the laboratory thus I am reluctant to buy a replacement. Would anyone be able to obtain parts as needed and provide us with a service contract once the present contract expires? Thanks, Patricia Zerfas National Institutes of Health Building 28A, Room 112 28 Library Drive Bethesda, MD 20892 ph: (301) 496-4464 fax: (301) 402-1068 From RSRICHMOND <@t> aol.com Tue Dec 16 13:11:15 2008 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Tue Dec 16 13:12:05 2008 Subject: [Histonet] Re: isopentane Message-ID: Carmen Loiselle asks about possible substitutes for Isopentane (2-methylbutane) for freezing muscle. Acetone is sometimes used, but is also a serious explosion hazard. I think that some people are using a fluorocarbon for this purpose, namely 3M? Novec? Engineered Fluid HFE-7100 This product belongs to a class of fluorocarbons called "segregated hydrofluoroethers (HFE's)" According to various MSDS, HFE-7100 is methyl nonafluoroisobutyl ether C4F9-O-CH3 It melts and freezes at -135 C, so that it would remain liquid in the Histobath. (It would freeze solid in liquid nitrogen, however.) It boils at 60 C., and is listed as non-flammable. It cost around $230 a gallon a few years ago - expensive, but it evaporates very slowly. I've posted about the subject on Histonet before. Has anyone on Histonet tried this stuff? Bob Richmond Samurai Pathologist Knoxville TN From JWeems <@t> sjha.org Tue Dec 16 13:16:39 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Tue Dec 16 13:16:44 2008 Subject: [Histonet] Prostate core biopsy submission In-Reply-To: References: Message-ID: <5D64396A0D4A5346BEBC759022AAEAA5208357@ITSSSXM01V6.one.ads.che.org> Telfa pads will work. Be aware that there are new CPT codes coming for Medicare prostates.... j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Campbell Sent: Tuesday, December 16, 2008 1:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Prostate core biopsy submission Hello Histo-netters: My lab is starting to bring in more business with prostate core biopsies. Some of the submitting physicians would like to use a small piece of towel to lay the biopsy on and then place it and the core biopsy into the formalin container. Does anyone have any suggestions for what type of material would be suitable for this? I am afraid that anything cottony will be a problem when removing the biopsy during processing as it may stick. Thank you for your help. Sharon Campbell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From bakevictoria <@t> gmail.com Tue Dec 16 13:27:28 2008 From: bakevictoria <@t> gmail.com (Victoria Baker) Date: Tue Dec 16 13:28:58 2008 Subject: [Histonet] autostainer service In-Reply-To: <424D930E0319E0469DD9C61502A50E3002102052@NIHCESMLBX4.nih.gov> References: <424D930E0319E0469DD9C61502A50E3002102052@NIHCESMLBX4.nih.gov> Message-ID: <4f016b690812161127o2f3797b4kf80e529f7f5d02b0@mail.gmail.com> Patricia, If I'm not mistaken - Michelle Onkowski is your sales representative she may have some options for you to work with. Another source may be Dolbey-Jamesion from Pennsylvania. They do have contracts at NIH and may be able to help you as well. Vikki On Tue, Dec 16, 2008 at 2:11 PM, Zerfas, Patricia (NIH/OD/ORS) [E] < zerfasp@ors.od.nih.gov> wrote: > Dear All, > > I am currently using the DAKOCytomation autostainer that is > approximately seven years old. Once the service contract expires in > November 2009 DAKOCytomation will not continue the service contract. > The equipment is in excellent condition and meets the needs of the > laboratory thus I am reluctant to buy a replacement. Would anyone be > able to obtain parts as needed and provide us with a service contract > once the present contract expires? > > > > Thanks, > > Patricia Zerfas > > National Institutes of Health > > Building 28A, Room 112 > > 28 Library Drive > > Bethesda, MD 20892 > > ph: (301) 496-4464 > > fax: (301) 402-1068 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gu.lang <@t> gmx.at Tue Dec 16 13:44:39 2008 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Dec 16 13:45:51 2008 Subject: AW: [Histonet] Prostate core biopsy submission In-Reply-To: References: Message-ID: There are filterpapers on the market, that have the double size in the length of a cassette and can directly be used for processing. We get corebiopsies of breast on filterpapers in formalin and of prostatae directly on blue sponges in cassettes. The problem with the paper is, that the biopsies stick too much on the dry surface and sometimes they get torn, when moving in the cassettes. We put the biopsies from the paper on sponges before processing. The problem with the sponges as first underground is, that they have to be wetted with formalin previously. And the morbid unfixed tissue can be formed by the hard spongematerial (makes small holes). We also get some corebiopsies "swimming free" in the formalin. I think they are easily fished and between the sponges they get flat enough to be embedded without problems. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Sharon Campbell Gesendet: Dienstag, 16. Dezember 2008 19:38 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Prostate core biopsy submission Hello Histo-netters: My lab is starting to bring in more business with prostate core biopsies. Some of the submitting physicians would like to use a small piece of towel to lay the biopsy on and then place it and the core biopsy into the formalin container. Does anyone have any suggestions for what type of material would be suitable for this? I am afraid that anything cottony will be a problem when removing the biopsy during processing as it may stick. Thank you for your help. Sharon Campbell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Tue Dec 16 16:07:41 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 16 16:07:55 2008 Subject: [Histonet] RE: (reply) silly questions.---PFA In-Reply-To: <200812121635232792765@foxmail.com> Message-ID: tf, =20 Answers as follows: Seems obvious that the 4% formaldehyde (made from PFA) was incorrectly made= . What they did, I have no idea nor did I have the time to show them how to= make it up. My advice: "make it from concentrated 38% formaldehyde (formal= in)". =20 =20 You wrote: "I do think most biomedical labs currently are using PFA to prep= are the fixatives!" - I do not think so. A sweeping statement if ever I hav= e heard one. This s the type of misinformation that we have to deal with. M= y experience (over 30 years) indicates that all pathology labs I have worke= d in as well as those I have visited, as well as several Pathology Colleges= (eg CAP and RCPA) surveys indicate that MOST (if not all) histopathology l= abs prepare their 10% formalin fixative from concentrated formalin not poly= formaldehyde as you have incorrectly stated.=20 =20 Be careful of misinformation. =20 Regards=20 Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)=20 Laboratory Manager & Senior Scientist=20 Tel: 612 9845 3306=20 Fax: 612 9845 3318=20 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: tf [mailto:tifei@foxmail.com]=20 Sent: Friday, 12 December 2008 7:35 PM To: Tony Henwood; Pat Flannery; histonet@lists.utsouthwestern.ed Subject: (reply) silly questions.---PFA =09 =09 "I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!!" =20 Tony: Do you think this is because of inproper preparation of PFA in his l= ab, or the common problem in all researchers using PFA? I do think most biomedical labs currently are using PFA to prepar= e the fixatives! =20 So, anyone has the idea on a correction preparation procedure of 4% PFA? I noticed some of you dissolve PFA powder in NaOH-conditioned alkaline wat= er, then add concentrated PB solution. We here dissolve PFA in concentrated PB solution directly (heat & stir for= 2-3 hours), then adjust pH to 7.4. =20 We dont have big problem in tissue quaility....except when one want to cut= the brain in a cryostat rather sliding microtome. Many times the brain sections from the cryostat have "cheese" like holes/c= avities, which almost never appear on sliding microtome-prepared sections. =20 2008-12-12=20 =09 ________________________________ tf=20 =09 ________________________________ =B7=A2=BC=FE=C8=CB=A3=BA Tony Henwood=20 =B7=A2=CB=CD=CA=B1=BC=E4=A3=BA 2008-12-12 06:18:47=20 =CA=D5=BC=FE=C8=CB=A3=BA Pat Flannery; histonet@lists.utsouthwestern.edu=20 =B3=AD=CB=CD=A3=BA=20 =D6=F7=CC=E2=A3=BA RE: [Histonet] Silly Question?=20 =09 =09 Pat, I agree with you. In a routine diagnostic histopathology laboratory, it makes little difference what you use. Around the world for over 100 years most labs use 10% neutral buffered formalin made from concentrated 38%(or there abouts) formalin (or formaldehyde). Researchers, though, are a different kettle of fish. They will tend to hang on to misinformed, "mystical" methods believing they are being scientific. Funny, you would think that they, as a group, would be the ones pushing the boundaries and critically assessing each step of their research, ensuring that they understand what and why they are doing it. (Disclaimer - not all researchers are like this, thank heavens!!) Using a formaldehyde solution made from polyformaldehyde can cause problems. One researcher used it and wondered why their morphology was sub-optimal and their p53 immunohistochemistry was negative. He assured me that he had fixed small samples of tissue for 6 hours in 4% formaldehyde and then processed them using ethanol, xylene and wax. I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!! This also explains the loss of p53 staining. I gave him some of our routine 10% phosphate buffered fomalin, asked him to fix overnight, and try agin. Low and behold problem solved. How's that for a Friday Flamming!!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead=20 Cnr Hawkesbury Road and Hainsworth Street, Westmead=20 Locked Bag 4001, Westmead NSW 2145, AUSTRALIA=20 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Friday, 12 December 2008 3:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do =20 we use paraformaldehyde (which is so inconvenient to make up) rather =20 than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either =20 2% or 4%) and others use formalin, while some others stick to diluted =20 formaldehyde (I see all 4 on labels for specimens submitted for =20 histology). Is it mostly a matter of personal preference or where you =20 were trained (i.e. force of habit) or is there a valid reason to use =20 each solution (basically the same chemical once in solution, merely =20 buffered or not)? The only answer I've gotten when I've asked is, =20 "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ 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 sen= der. Views expressed in this message and any attachments are those of the indiv= idual sender, and are not necessarily the views of The Children's Hospital = at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, The Children= s Hospital at Westmead accepts no liability for any consequential damage re= sulting from email containing computer viruses. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu ********************************************************************* 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 send= er. Views expressed in this message and any attachments are those of the indivi= dual sender, and are not necessarily the views of The Children's Hospital a= t Westmead This note 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 res= ulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Tue Dec 16 16:11:42 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 16 16:11:47 2008 Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS In-Reply-To: Message-ID: I would suggest the sulphated toluidine blue method as described below. It is quick and demonstrates PC very well (though not quite as good as well-done (or should I say under-done) methenamine silver. Following sulphation, fungi stain metachromatically with toluidine blue. Certain hydroxyl groups are esterified by sulphuric acid to form ester sulphate groups that stain metachromatically with toluidine blue (Smith & Lowrey 1986, J. Histotechn. 9(1):23-24. ). Fixation: 10% buffered formalin Microtomy: 5?m paraffin sections Solutions: 1. Sulphation Solution: Absolute Acetone 1 ml - pre-cooled to 4oC. Slowly add 1 ml concentrated sulphuric acid drop by drop. Allow cooling before use. 2. 3% Acetic Acid. 3. Toluidine Blue O Solution: Toluidine Blue O (CI 52040) 0.01g 3% Acetic Acid 100ml 4. Metanil Yellow Counterstain: Metanil yellow (CI 13065) 0.1g Distilled Water 100ml Glacial Acetic Acid 0.1ml 5. Absolute Acetone. Controls: Fungi containing tissue. Procedure: 1. Deparaffinise and bring to absolute alcohol. 2. Dry briefly, cover with sulphation reagent, 3 minutes. 3. Briefly wash in running water. 4. Place in 3% acetic acid, 1 minute. 5. Stain in Toluidine Blue O solution, 3 minutes. 6. Rinse in 3% acetic acid, 1 minute. 7. Differentiate in absolute acetone, 5 seconds. 8. Rinse in distilled water, 5 seconds. 9. Counterstain in Metanil Yellow solution, 6 seconds. 10. Rinse in distilled water, 5 seconds. 11. Blot, dehydrate rapidly, clear and mount. Results: Fungi stained purple to red against a yellow background. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of TIMOTHY MALLOY Sent: Friday, 12 December 2008 8:52 PM To: HISTONET ...; TIM MALLOY Subject: [Histonet] SPECIAL STAIN PNUEMOCYSTSIS We currently use a stain called grocotts silver methenimine solution for staining pnuemocystis. We would like to use a hot plate for the procedure instead of a unvented microwave. Does anybody have any formula's the would like to share with the hot plate? Timothy G. Malloy, HT ( ASCP ) A.A.S. tmalloy77@hotmail.com This email communication may contain CONFIDENTIAL INFORMATION WHICH ALSO MAY BE LEGALLY PRIVILEGED and is intended only for the use of the intended recipients identified above. If you are not the intended recipient of this communication, you are hereby notified that any unauthorized review, use, dissemination, distribution, downloading, or copying of this communication is strictly prohibited. If you have received this communication in error, please immediately notify us by reply email, delete the communication and destroy all copies._______________________________________________ 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 note 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 Tue Dec 16 16:16:07 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 16 16:17:07 2008 Subject: [Histonet] (reply) silly questions.---PFA In-Reply-To: Message-ID: Kathy, Here is a copy of the email I sent Danielle Danielle, Here is the method we use: Test for validity of formalin concentrations Schiff?s reagent (as used in the PAS stain) is used to detect aldehydes and in this technique it is used to titrate a solution of formalin. The concentration of which is unknown (Jaspers 1987). Solutions: 1. 1 ml of formalin fixative to be tested. 2. 2.4% aqueous sodium bisulphite (sodium metabisulphite) ? prepare fresh. 3. Schiff?s reagent. Procedure: 1. Add 1 ml of formalin to 5 ml of sodium bisulphite. 2. Mix and react for 15 minutes at room temperature stirring from time to time. 3. Add 100?l of Schiff reagent. Results: If solution turns a deep violet than initial concentration of formalin is in excess of 4% (i.e. 1.6% formaldehyde). By only using 0.6ml of 2.4% sodium metabisulphite, you can see whether it is greater than 0.5% formalin Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: Walters, Katherine S [mailto:katherine-walters@uiowa.edu] Sent: Saturday, 13 December 2008 12:48 AM To: tifei@foxmail.com; Tony Henwood; Pat Flannery; histonet@lists.utsouthwestern.ed Subject: RE: [Histonet] (reply) silly questions.---PFA This may be another silly question, but how does one test the concentration of formaldehyde in solution? Thanks, Kathy Notice: This UI Health Care e-mail (including attachments) is covered by the Electronic Communications Privacy Act, 18 U.S.C. 2510-2521, is confidential and may be legally privileged. If you are not the intended recipient, you are hereby notified that any retention, dissemination, distribution, or copying of this communication is strictly prohibited. Please reply to the sender that you have received the message in error, then delete it. Thank you. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of tf Sent: Friday, December 12, 2008 2:35 AM To: Tony Henwood; Pat Flannery; histonet@lists.utsouthwestern.ed Subject: [Histonet] (reply) silly questions.---PFA "I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!!" Tony: Do you think this is because of inproper preparation of PFA in his lab, or the common problem in all researchers using PFA? I do think most biomedical labs currently are using PFA to prepare the fixatives! So, anyone has the idea on a correction preparation procedure of 4% PFA? I noticed some of you dissolve PFA powder in NaOH-conditioned alkaline water, then add concentrated PB solution. We here dissolve PFA in concentrated PB solution directly (heat & stir for 2-3 hours), then adjust pH to 7.4. We dont have big problem in tissue quaility....except when one want to cut the brain in a cryostat rather sliding microtome. Many times the brain sections from the cryostat have "cheese" like holes/cavities, which almost never appear on sliding microtome-prepared sections. 2008-12-12 tf ???? Tony Henwood ????? 2008-12-12 06:18:47 ???? Pat Flannery; histonet@lists.utsouthwestern.edu ??? ??? RE: [Histonet] Silly Question? Pat, I agree with you. In a routine diagnostic histopathology laboratory, it makes little difference what you use. Around the world for over 100 years most labs use 10% neutral buffered formalin made from concentrated 38%(or there abouts) formalin (or formaldehyde). Researchers, though, are a different kettle of fish. They will tend to hang on to misinformed, "mystical" methods believing they are being scientific. Funny, you would think that they, as a group, would be the ones pushing the boundaries and critically assessing each step of their research, ensuring that they understand what and why they are doing it. (Disclaimer - not all researchers are like this, thank heavens!!) Using a formaldehyde solution made from polyformaldehyde can cause problems. One researcher used it and wondered why their morphology was sub-optimal and their p53 immunohistochemistry was negative. He assured me that he had fixed small samples of tissue for 6 hours in 4% formaldehyde and then processed them using ethanol, xylene and wax. I looked at the sections and the cell shrinkage (and prominent spaces between cells and connective tissue) indicated that most of the "fixation" seemed to have occured in the processing ethanols. I asked him for some of the fixative he used, tested the formaldehyde concentration and found it to be less than 0.5%!! This also explains the loss of p53 staining. I gave him some of our routine 10% phosphate buffered fomalin, asked him to fix overnight, and try agin. Low and behold problem solved. How's that for a Friday Flamming!!! Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pat Flannery Sent: Friday, 12 December 2008 3:59 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Silly Question? Please humor me on this if it's obvious (to everyone but me): why do we use paraformaldehyde (which is so inconvenient to make up) rather than buffered formalin or just diluted formaldehyde itself? It seems that around here, some folks prefer paraformaldehyde (either 2% or 4%) and others use formalin, while some others stick to diluted formaldehyde (I see all 4 on labels for specimens submitted for histology). Is it mostly a matter of personal preference or where you were trained (i.e. force of habit) or is there a valid reason to use each solution (basically the same chemical once in solution, merely buffered or not)? The only answer I've gotten when I've asked is, "That's what we always use." Thanks. -Pat Flannery (not a "real" histologist - I just play one in the lab) _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu ********************************************************************* 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 note 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 gp62 <@t> georgetown.edu Tue Dec 16 16:36:46 2008 From: gp62 <@t> georgetown.edu (Guillermo Palchik) Date: Tue Dec 16 16:37:44 2008 Subject: [Histonet] TUNEL on FIXED rat PUPS Message-ID: <2D343A52-018D-4BA8-8ADC-2EDF875EDA9C@georgetown.edu> Dear Histologists... I am currently doing an experiment in which I perform TUNEL on brain slices that were treated with certain drugs, to examine the levels of apoptotic cell death. The rat?s brain is flash frozen (scooped from the skull, directly into cold isopentane) air dried, and then stored into a -80 C freezer. We then cut the brain into 20 um slices using a cryostat and mount them on slides, and store them in a -20 C freezer, where they remain until the TUNEL step. The TUNEL is performed using the Apoptag Plus kit from Chemicon using Peroxidase/DAB (Cat. # S7101) and counterstaining with 0.5% methyl green. This has been done in the lab for quite some time now and we are able to get results? We want to switch over to perfused brains (instead of flash frozen). However, we have had ZERO positive staining once the brains are fixed (in 4% PF). This has been corroborated by other lab members that have tried it for a couple of years already? The protocol that came with the kit has a section for flash frozen and a section for fixed tissue, and since I have done TUNEL using fixed tissue before, I know that it is possible to do TUNEL in fixed tissue, however we cannot get any positive staining whatsoever? Along these lines, since the first step of the Apoptag TUNEL protocol is to fix the tissue with 4% PF (for 10 min), this has led me to believe that the problem is indeed the initial fixation with PF (at the time of perfusion). I should say that we work with rat PUPS for this and that the immature brain is not the same as the mature brain (the immature brain has more fat, for example) and that this might be the cause of the problem?In any case, I was doing some research and I wanted to try using Zinc Formalin as a perfusate and see if this would allow us to do the TUNEL. I would appreciate any comments and suggestions regarding this. I am sorry for the lengthy email, but I wanted to show a more or less complete picture, in case I am overlooking other factors? Guillermo Palchik gp62@georgetown.edu From jcampbell <@t> vdxpathology.com Tue Dec 16 17:06:43 2008 From: jcampbell <@t> vdxpathology.com (Jennifer Campbell) Date: Tue Dec 16 17:07:49 2008 Subject: [Histonet] Any opinions on these microtomes? Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF819E2DE@VDXSERVER01.vdxpathology.local> Hello everyone, Has anyone had any experience with the HM 340 E (Mikrom) or the Rotary Microtome RMTs 40, 50 or 60? We are looking to get a new microtome for paraffin sectioning but, would like one that is suitable for plastics in case we move that way in the future. These were recommended to me by a sales associate but, I would like to hear if anyone has some firsthand experience with any of these and what his or her thoughts are. Thanks a bunch, Jennifer Campbell Vdx Veterinary Diagnostics Davis, CA From amosbrooks <@t> gmail.com Tue Dec 16 17:12:02 2008 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Tue Dec 16 17:13:00 2008 Subject: [Histonet] Speaking of Fluoro Jade Message-ID: <582736990812161512u389aeaf4ke11d0686b3d6584c@mail.gmail.com> Hi, Speaking of Fluoro Jade: I don't do this often and after having done the stain recently I was left with a lot of extra Fluoro Jade stock solution. The data sheet says this is only good for a week or so making it a dang expensive test! For non-diagnostic use of course, could the rest be aliquated and frozen (-80 C) to use later? Thanks, Amos Date: Tue, 16 Dec 2008 12:24:38 +0200 From: "Tyrone Genade" Subject: [Histonet] Re: Histonet Digest, Vol 61, Issue 25 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi all, Otherwise, you may want to look into FlouroJadeB. If all you are after is visualizing degenerating axons, there are easier ways of going about ot than silver staining. Kind regards -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@freeshell.org tel: +27-84-632-1925 (c) From AnthonyH <@t> chw.edu.au Tue Dec 16 17:18:29 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 16 17:19:28 2008 Subject: [Histonet] at a troubleshooting loss In-Reply-To: <62A8156F8071C8439080D626DF8C33A602E5B3@wave-mail.7thwave.local> Message-ID: ?carbon from lead pensils used for labelling slides. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele Wich Sent: Wednesday, 17 December 2008 4:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] at a troubleshooting loss A couple questions for those who are troubleshooting savvy: Other than inadequate fixation, water being introduced during processing, or too much heat, what can cause fuzzy, uneven hematoxylin staining with poor nuclear detail? And secondly, aside from formalin, are there any other common agents of blackish pigment in H & E staining? (The artifact appears on the slides both on and around the tissues.) Any feedback is GREATLY appreciated. This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer _______________________________________________ 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 note 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 Tue Dec 16 17:19:16 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 16 17:20:16 2008 Subject: [Histonet] Isopentane In-Reply-To: Message-ID: Try liquid nitrogen Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Wednesday, 17 December 2008 5:27 AM To: carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ 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 note 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 arose <@t> cellmarque.com Tue Dec 16 17:25:35 2008 From: arose <@t> cellmarque.com (Aaron Rose) Date: Tue Dec 16 17:30:25 2008 Subject: [Histonet] at a troubleshooting loss In-Reply-To: References: <62A8156F8071C8439080D626DF8C33A602E5B3@wave-mail.7thwave.local> Message-ID: <7F2A2AE306CE254DB7279E86A51A74067E12B5@CMROCEX01.cellmarque.local> This may be a silly suggestion but Have you been filtering your hematoxylin? I have seen the sediments that form over time. I'm not sure if this could be the dark regions you're seeing but I figured I'd throw it out there. -Aaron -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: Tuesday, December 16, 2008 3:18 PM To: Michele Wich; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] at a troubleshooting loss ?carbon from lead pensils used for labelling slides. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele Wich Sent: Wednesday, 17 December 2008 4:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] at a troubleshooting loss A couple questions for those who are troubleshooting savvy: Other than inadequate fixation, water being introduced during processing, or too much heat, what can cause fuzzy, uneven hematoxylin staining with poor nuclear detail? And secondly, aside from formalin, are there any other common agents of blackish pigment in H & E staining? (The artifact appears on the slides both on and around the tissues.) Any feedback is GREATLY appreciated. This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Tue Dec 16 17:15:03 2008 From: liz <@t> premierlab.com (Liz Chlipala) Date: Tue Dec 16 17:34:52 2008 Subject: [Histonet] TUNEL on FIXED rat PUPS In-Reply-To: <2D343A52-018D-4BA8-8ADC-2EDF875EDA9C@georgetown.edu> References: <2D343A52-018D-4BA8-8ADC-2EDF875EDA9C@georgetown.edu> Message-ID: Guillermo We have not used that kit specifically we use the one from Roche but we get fine results with the following fixatives, 10% NBF, 4% Paraformaldehyde and Davidisons, its also works fixed and EDTA decalcified samples. You will need to add a proteinase K step to your protocol. You need to order a specific proteinase K the one you may use for IHC may not work. We get ours from Roche here is the catalog number and how we prepare the stock and working solutions, you may need to titer the concentration of proteinase K for your particular project we use it at a concentration of 20ug/ml. I have also added our basic protocol so you can see where the protienase K step figures in. Proteinase K, recombinant, PCR Grade Roche Applied Science 03 115 879 001 Proteinase K Stock Solution - 20mg/ml Proteinase K 1 vial - 100mg Distilled water 5 mls Once the proteinase K lyophilized enzyme has been reconstituted with distilled water then aliquot into RNase free 0.6 ml microtubes a volume of 20 to 40?ls and store at -15 to -25?C. Shelf Life: 12 months Storage: -15 to -25?C Proteinase K Working Solution - 20ug/ml For each slide that needs to be stained you will need to prepare a minimum of 125?ls of proteinase K working solution. 100?ls of working solution will be placed on each slide. The working solution is prepared as follows: Proteinase K Stock Solution 10?ls 10Mm TRIS/HCL pH8.0 990?ls Make fresh prior to use. Remove one aliquot vial of the stock 20mg/ml proteinase K from the freezer and thaw. Unused stock proteinase K may be stored in the refrigerator for 7 days, then discard. Shelf Life: make fresh prior to use, discard unused portion Storage: NA Procedure When performing the TUNEL stain the maximum number of slides that can be run at a time is 12. Within the 12 slides, 10 will be test samples, 1 is a positive control (DNase treated) and 1 is a negative control (Label Solution without Enzyme Solution). All but the rinse steps are performed either in a humidity chamber or in the Dako Hybridizer. 1. Xylene 5 minutes 2. Xylene 5 minutes 3. 100% alcohol 2 minutes 4. 100% alcohol 2 minutes 5. 95% alcohol 1 minute 6. 95% alcohol 1 minute 7. Tap water rinse 1 minute 8. Distilled water rinse 1 minute 9. PBS pH7.4 0.1% tween rinse 1 minute or more 10. Prepare working proteinase K - see solutions 11. Working proteinase K (75 -200?ls/slide) 30 minutes 12. PBS pH7.4 0.1% tween rinse 3 minutes 13. PBS pH7.4 0.1% tween rinse 3 minutes 14. Prepare DNase1 working solution - see solutions 15. DNase1 (75 -150?ls/slide) 10 minutes Note: apply DNase1 to only the positive control sample, the rest of the samples will remain in PBS pH7.4 0.1% tween buffer 16. PBS pH7.4 0.1% tween rinse 3 minutes 17. PBS pH7.4 0.1% tween rinse 3 minutes 18. Prepare TUNEL reaction mixture Note: The TUNEL reaction mixture should be prepared immediately before use. Keep TUNEL reaction mixture on ice until use. Unused reagent may be stored at -70?C for future use. Prepare the TUNEL reaction mixture and negative control solution as follows: a. The TUNEL kit reagents are stored in the -70?C freezer b. Remove one vial 1: enzyme solution and one vial 2: label solution from the freezer c. Remove 100?l of the label solution and place it into a microtube this solution will serve as the negative control solution d. Add the total volume of vial 1: enzyme solution (50?l) to the remaining 450?l in vial 2: label solution and mix well 19. Remove slides from PBS and dry area around sample 20. Add 50 to 100?l of TUNEL reaction mixture 21. Incubate at 37?C in the Dako Hybridizer 60 minutes NOTE: for the negative control add 50 to 75?l of the label solution to each sample. If necessary to ensure a homogenous spread of TUNEL reaction mixture across the sample and to avoid evaporative loss, the samples can be covered with a coverslip during incubation. 22. PBS pH7.4 0.1% tween rinse 3 minutes 23. PBS pH7.4 0.1% tween rinse 3 minutes 24. PBS pH7.4 0.1% tween rinse 1 minute 25. Add 50 to 100?l of Converter-AP (vial 3) to each sample 26. Incubate at 37?C in the Dako Hybridizer 30 minutes NOTE: If necessary to ensure a homogenous spread of TUNEL reaction mixture across the sample and to avoid evaporative loss, the samples can be covered with a coverslip during incubation. 27. PBS pH7.4 0.1% tween rinse 3 minutes or more 28. PBS pH7.4 0.1% tween rinse 3 minutes or more 29. Wash Buffer 1 minute 30. Prepare Vulcan Fast Red Substrate Solution To 2.5 mls of Vulcan Fast Red Buffer add one drop of vial A and one drop of vial b. Use immediately, discard any leftover reagent. 31. Vulcan Fast Red Substrate Solution 10 - 30 minutes 32. Distilled water 2 minutes 33. Distilled water 2 minutes 34. Hematoxylin 3 minutes 35. Distilled water 1 minute 36. Wash Buffer 1 minute 37. Distilled water 1 minute 38. Let slides air dry 39. Xylene 1 minute 40. Mount and coverslip 41. Review positive and negative control slides for appropriate staining Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, CO 80308 phone (303) 682-3949 fax (303) 682-9060 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory, LLC 1567 Skyway Drive Unit E Longmont, CO 80504 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Guillermo Palchik Sent: Tuesday, December 16, 2008 3:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TUNEL on FIXED rat PUPS Dear Histologists... I am currently doing an experiment in which I perform TUNEL on brain slices that were treated with certain drugs, to examine the levels of apoptotic cell death. The rat's brain is flash frozen (scooped from the skull, directly into cold isopentane) air dried, and then stored into a -80 C freezer. We then cut the brain into 20 um slices using a cryostat and mount them on slides, and store them in a -20 C freezer, where they remain until the TUNEL step. The TUNEL is performed using the Apoptag Plus kit from Chemicon using Peroxidase/DAB (Cat. # S7101) and counterstaining with 0.5% methyl green. This has been done in the lab for quite some time now and we are able to get results... We want to switch over to perfused brains (instead of flash frozen). However, we have had ZERO positive staining once the brains are fixed (in 4% PF). This has been corroborated by other lab members that have tried it for a couple of years already... The protocol that came with the kit has a section for flash frozen and a section for fixed tissue, and since I have done TUNEL using fixed tissue before, I know that it is possible to do TUNEL in fixed tissue, however we cannot get any positive staining whatsoever... Along these lines, since the first step of the Apoptag TUNEL protocol is to fix the tissue with 4% PF (for 10 min), this has led me to believe that the problem is indeed the initial fixation with PF (at the time of perfusion). I should say that we work with rat PUPS for this and that the immature brain is not the same as the mature brain (the immature brain has more fat, for example) and that this might be the cause of the problem...In any case, I was doing some research and I wanted to try using Zinc Formalin as a perfusate and see if this would allow us to do the TUNEL. I would appreciate any comments and suggestions regarding this. I am sorry for the lengthy email, but I wanted to show a more or less complete picture, in case I am overlooking other factors... Guillermo Palchik gp62@georgetown.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Tue Dec 16 20:37:50 2008 From: tifei <@t> foxmail.com (tf) Date: Tue Dec 16 20:38:13 2008 Subject: [Histonet] Re: RE: (reply) silly questions.---PFA References: Message-ID: <200812171037451405420@foxmail.com> SGkgVG9ueToNCg0KVGhhbmtzIGEgbG90IGZvciB0aGUgY29ycmVjdGlvbiENCkkgYW0gaW4gcmVz ZWFyY2ggaW5zdGl0dXRlLCBxdWl0ZSBhIGdhcC4uLg0KDQoNCjIwMDgtMTItMTcgDQoNCg0KDQp0 ZiANCg0KDQoNCreivP7Iy6O6IFRvbnkgSGVud29vZCANCreiy83Ksbzko7ogMjAwOC0xMi0xNyAg MDY6MDc6NDggDQrK1bz+yMujuiB0aWZlaUBmb3htYWlsLmNvbTsgUGF0IEZsYW5uZXJ5OyBoaXN0 b25ldEBsaXN0cy51dHNvdXRod2VzdGVybi5lZCANCrOty82juiANCtb3zOKjuiBSRTogKHJlcGx5 KSBzaWxseSBxdWVzdGlvbnMuLS0tUEZBIA0KIA0KdGYsDQoNCkFuc3dlcnMgYXMgZm9sbG93czoN ClNlZW1zIG9idmlvdXMgdGhhdCB0aGUgNCUgZm9ybWFsZGVoeWRlIChtYWRlIGZyb20gUEZBKSB3 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KioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqDQpU aGlzIGVtYWlsIGFuZCBhbnkgZmlsZXMgdHJhbnNtaXR0ZWQgd2l0aCBpdCBhcmUgY29uZmlkZW50 aWFsIGFuZCBpbnRlbmRlZCBzb2xlbHkgZm9yIHRoZSB1c2Ugb2YgdGhlIGluZGl2aWR1YWwgb3Ig ZW50aXR5IHRvIHdob20gdGhleSBhcmUgYWRkcmVzc2VkLiBJZiB5b3UgYXJlIG5vdCB0aGUgaW50 ZW5kZWQgcmVjaXBpZW50LCBwbGVhc2UgZGVsZXRlIGl0IGFuZCBub3RpZnkgdGhlIHNlbmRlci4N Cg0KVmlld3MgZXhwcmVzc2VkIGluIHRoaXMgbWVzc2FnZSBhbmQgYW55IGF0dGFjaG1lbnRzIGFy ZSB0aG9zZSBvZiB0aGUgaW5kaXZpZHVhbCBzZW5kZXIsIGFuZCBhcmUgbm90IG5lY2Vzc2FyaWx5 IHRoZSB2aWV3cyBvZiBUaGUgQ2hpbGRyZW4ncyBIb3NwaXRhbCBhdCBXZXN0bWVhZA0KDQpUaGlz IG5vdGUgYWxzbyBjb25maXJtcyB0aGF0IHRoaXMgZW1haWwgbWVzc2FnZSBoYXMgYmVlbg0Kdmly dXMgc2Nhbm5lZCBhbmQgYWx0aG91Z2ggbm8gY29tcHV0ZXIgdmlydXNlcyB3ZXJlIGRldGVjdGVk LCBUaGUgQ2hpbGRyZW5zIEhvc3BpdGFsIGF0IFdlc3RtZWFkIGFjY2VwdHMgbm8gbGlhYmlsaXR5 IGZvciBhbnkgY29uc2VxdWVudGlhbCBkYW1hZ2UgcmVzdWx0aW5nIGZyb20gZW1haWwgY29udGFp bmluZyBjb21wdXRlciB2aXJ1c2VzLg0KKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioq KioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKioqKg0K From tifei <@t> foxmail.com Tue Dec 16 20:44:35 2008 From: tifei <@t> foxmail.com (=?utf-8?B?dGY=?=) Date: Tue Dec 16 20:44:57 2008 Subject: =?utf-8?B?UmU6IFJFOiBbSGlzdG9uZXRdIHN0aWNrZXN0IHNsaWRlcyBmb3IgYnJhaW4gc2VjdGlvbnMgb3IgaG93IHRvY29hdCBhcHJvcGVyIHNsaWRlIGZvciB1cnNlbGY=?= References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net>, , <200812170100003020790@foxmail.com>, <6CF686BD6F24A546B85B24FE3B978647049CE8FD@EXVS06.net.ucsf.edu> Message-ID: <200812171044296951078@foxmail.com> Thanks for sharing. I do not have big problem in mounted slides from microtome - you pick the sections up from solution. But I did have some sections prepared in cryostat came off during BrdU IHC (citrate buffer 95 degree for 30 min , so harsh~~). I am not sure whether this is because of embedding (O.C.T) ? Or something else. Even you use pre-coated slides. And, As I mentioned in last email, we found the 0.25% gel-slide works as good as pre-coated slides if the sections were mounted from PBS, washed out all embedding materials. 2008-12-17 tf ???? Mejia, Maria ????? 2008-12-17 01:24:31 ???? tifei; Alexandra Meinl; cmmathis1 ??? histonet ??? RE: [Histonet] stickest slides for brain sections or how tocoat aproper slide for urself This is for anyone's interest who work with 40um primate brain section 2x3 inches. We routinely mount our IHC stained sections from working PBS on pre-subbed brain slides from Brain Research Labs. To keep the sections on the slides we use a very fine (#0) brush to brush the section flat on the slide & to remove any PBS that may get trap under the section. We dry overnight. Next day, I put in oven at 37C - 30 minutes to further dry. Regards Maria Bartola Mejia Histology Manager Department of Neurosurgery UCSF San Francisco, CA 94103 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of tf Sent: Tue 12/16/2008 9:00 AM To: Alexandra Meinl; cmmathis1 Cc: histonet Subject: [Histonet] stickest slides for brain sections or how to coat aproper slide for urself I have similar questions... for brain sections. we found that 0.25% gel coated slides are better than 0.5% or 1% gel in sticking 40 um brain sections. just want to share this with you guys. 2008-12-17 tf ???: Alexandra Meinl ????: 2008-12-17 00:27:04 ???: cmmathis1 ??: histonet ??: Re: [Histonet] What is the stickest slides I like Roti-Bond Adhesion slides for difficult material like bone, you can also use HIER without problems. But these slides are quite expensive and require 2-3 days drying time. Alex 2008/12/16 > Hello Histonetters, > We are having trouble getting our paraffin processed cow meniscus sections > (at 5 microns) to stay on our Superfrost Plus slides. We are drying them > overnight in a 58 C oven. The H&Es look great but the immunos wash every > time. > What is your favorite slide for these hard to stay on samples? > Thanks for your time, > Cathy > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From micro <@t> superlink.net Tue Dec 16 20:47:12 2008 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Tue Dec 16 20:47:19 2008 Subject: [Histonet] Isopentane References: Message-ID: <038001c95ff1$c44ce2e0$66893cd1@DJ4VDH31> Isopentane is cooled by liquid N2! To freeze tissue direct in LN2 is not the best way to freeze tissue due to the Leidenfrost effect, creating an insulating gas layer between the tissue and the liquid and delaying the speed of freezing = an increase in ice crystal formation. Regards, Markus F. Meyenhofer ----- Original Message ----- From: "Tony Henwood" To: "Dana Settembre" ; "carmen loiselle" ; "pathology education" Sent: Tuesday, December 16, 2008 6:19 PM Subject: RE: [Histonet] Isopentane Try liquid nitrogen Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Wednesday, 17 December 2008 5:27 AM To: carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Tue Dec 16 21:02:31 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 16 21:03:36 2008 Subject: [Histonet] Isopentane In-Reply-To: <038001c95ff1$c44ce2e0$66893cd1@DJ4VDH31> Message-ID: Depends on how you freeze. When we freeze tissue in liquid nitrogen, the tissue is shaken, ensuring that the gas layer is broken up. And in so doing we have not seen evidence of ice crystal formation. I can send you a reprint of a paper that describes the effect of storage of brain tumours in saline prior to freezing (causing much distortion of the tumour) verses receiving the samples fresh on damp gauze or card. The pics show the evidence. Henwood, A., (2007) "Adverse effect of saline on brain intraoperative (frozen section) Histology" J Histotechnol 30(3):193 Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: Markus F. Meyenhofer [mailto:micro@superlink.net] Sent: Wednesday, 17 December 2008 1:47 PM To: Tony Henwood; Dana Settembre; carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Isopentane is cooled by liquid N2! To freeze tissue direct in LN2 is not the best way to freeze tissue due to the Leidenfrost effect, creating an insulating gas layer between the tissue and the liquid and delaying the speed of freezing = an increase in ice crystal formation. Regards, Markus F. Meyenhofer ----- Original Message ----- From: "Tony Henwood" To: "Dana Settembre" ; "carmen loiselle" ; "pathology education" Sent: Tuesday, December 16, 2008 6:19 PM Subject: RE: [Histonet] Isopentane Try liquid nitrogen Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Wednesday, 17 December 2008 5:27 AM To: carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 note 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 SwainFrancesL <@t> uams.edu Wed Dec 17 07:02:58 2008 From: SwainFrancesL <@t> uams.edu (Swain, Frances L) Date: Wed Dec 17 07:04:15 2008 Subject: [Histonet] What is the stickest slides In-Reply-To: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> References: <121620081551.4776.4947CE6800021B02000012A822243651029B0A02D2089B9A019C04040A0DBFCE9C07089B0E03030C@att.net> Message-ID: I cut MMA sections which are also hard to mount and have them stay on the slides. I have found that Biocare, Surgipath, Newcomer's Supply and Scientific Device Laboratory have what they call HIER slides or superplus slides. All of them are great, the prices are a little steep. Frances L. Swain HT(ASCP) A. A. S. Special Procedures Technician Department of Orthopaedic Surgery Center for Orthopaedic Research Barton Research Building 2R28 4301 West Markham Street Little Rock AR 72205 (501) 686-8739 PHONE (501) 686-8987 FAX swainfrancesl@uams.edu email -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of cmmathis1@bellsouth.net Sent: Tuesday, December 16, 2008 9:51 AM To: histonet Subject: [Histonet] What is the stickest slides Hello Histonetters, We are having trouble getting our paraffin processed cow meniscus sections (at 5 microns) to stay on our Superfrost Plus slides. We are drying them overnight in a 58 C oven. The H&Es look great but the immunos wash every time. What is your favorite slide for these hard to stay on samples? Thanks for your time, Cathy _______________________________________________ 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 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 MAUGER <@t> email.chop.edu Wed Dec 17 07:50:13 2008 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Wed Dec 17 07:51:15 2008 Subject: [Histonet] Isopentane Message-ID: In my experience, liquid nitrogen makes a lot of freezing artifact in muscle tissue. I have found no better substitue for isopentane if you want quality morphology in you specimen. We store it in an explosion proof cabinet. Jo Mauger >>> "Tony Henwood" 12/16/08 6:19 PM >>> Try liquid nitrogen Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Wednesday, 17 December 2008 5:27 AM To: carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWeems <@t> sjha.org Wed Dec 17 08:05:31 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Wed Dec 17 08:08:04 2008 Subject: [Histonet] Isopentane In-Reply-To: References: Message-ID: <5D64396A0D4A5346BEBC759022AAEAA520843B@ITSSSXM01V6.one.ads.che.org> But you can coat the tissue with talc or baby powder and prevent artifact from the nitrogen.. j -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Joanne Mauger Sent: Wednesday, December 17, 2008 8:50 AM To: AnthonyH@chw.edu.au; carmen_loiselle@hotmail.com; histonet@pathology.swmed.edu; settembr@umdnj.edu Subject: RE: [Histonet] Isopentane In my experience, liquid nitrogen makes a lot of freezing artifact in muscle tissue. I have found no better substitue for isopentane if you want quality morphology in you specimen. We store it in an explosion proof cabinet. Jo Mauger >>> "Tony Henwood" 12/16/08 6:19 PM >>> Try liquid nitrogen Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Wednesday, 17 December 2008 5:27 AM To: carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From TMcNemar <@t> lmhealth.org Wed Dec 17 08:44:31 2008 From: TMcNemar <@t> lmhealth.org (Tom McNemar) Date: Wed Dec 17 08:45:13 2008 Subject: [Histonet] autostainer service In-Reply-To: <424D930E0319E0469DD9C61502A50E3002102052@NIHCESMLBX4.nih.gov> Message-ID: <51D5D78FBEDAEA4FBCCD9A9D44211DC505B8E078@lmhsmail.lmhealth.org> My autostainer is about 8 years old. I have heard nothing of the termination of service contracts by Dako. May I ask how you came by this info? I just renewed in August so I have time. Thanks. Tom McNemar, HT(ASCP) Histology Co-ordinator Licking Memorial Health Systems (740) 348-4163 (740) 348-4166 tmcnemar@lmhealth.org www.LMHealth.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Zerfas, Patricia (NIH/OD/ORS) [E] Sent: Tuesday, December 16, 2008 2:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] autostainer service Dear All, I am currently using the DAKOCytomation autostainer that is approximately seven years old. Once the service contract expires in November 2009 DAKOCytomation will not continue the service contract. The equipment is in excellent condition and meets the needs of the laboratory thus I am reluctant to buy a replacement. Would anyone be able to obtain parts as needed and provide us with a service contract once the present contract expires? Thanks, Patricia Zerfas National Institutes of Health Building 28A, Room 112 28 Library Drive Bethesda, MD 20892 ph: (301) 496-4464 fax: (301) 402-1068 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From CHRISH <@t> HEALTHCARESCOUTS.COM Wed Dec 17 09:14:47 2008 From: CHRISH <@t> HEALTHCARESCOUTS.COM (Chris Handrahan) Date: Wed Dec 17 09:24:31 2008 Subject: [Histonet] Holiday Greeting from Healthcare Scouts Message-ID: Wishing you every happiness this holiday season and throughout the Coming Year. With employment opportunities all over the US, we are always looking for qualified and motivated professionals. Recommend a friend or family member, and you could be eligible for up to $1,000 through Healthcare Scouts' referral program. Call us today to learn more! Click below to view our holiday greeting http://www.youtube.com/watch?v=7xV4wWrsuvo Chris Handrahan Managing Director of Allied Health Healthcare Scouts 800-708-0605 office 321-231-5427 cell chrish@healthcarescouts.com www.healthcarescouts.com From carrolpb <@t> umdnj.edu Wed Dec 17 09:32:11 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Wed Dec 17 09:33:47 2008 Subject: [Histonet] Holiday Greeting from Healthcare Scouts In-Reply-To: References: Message-ID: <49491B7B.5070300@umdnj.edu> argh, more spam disguised as a holiday greeting... Chris Handrahan wrote: > Wishing you every happiness this holiday season and throughout the > Coming Year. > > With employment opportunities all over the US, we are always looking for > qualified and motivated professionals. Recommend a friend or family > member, and you could be eligible for up to $1,000 through Healthcare > Scouts' referral program. Call us today to learn more! > > Click below to view our holiday greeting > > http://www.youtube.com/watch?v=7xV4wWrsuvo > > > > > > > Chris Handrahan > > Managing Director of Allied Health > > Healthcare Scouts > > 800-708-0605 office > > 321-231-5427 cell > > chrish@healthcarescouts.com > > > > > > www.healthcarescouts.com > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From greenjumpyone <@t> hotmail.com Wed Dec 17 09:44:04 2008 From: greenjumpyone <@t> hotmail.com (GreenJumpyOne) Date: Wed Dec 17 09:46:59 2008 Subject: [Histonet] Cost Analysis References: Message-ID: I have been asked to do a cost anaylsis on what it would cost to take a single biopsy specimen from the formalin bottle all the way to the H&E slide. I only need a ballpark estimate. Has anyone done this recently? Thanks for any help you can provide. Michelle Carney Bradenton, FL From rjbuesa <@t> yahoo.com Wed Dec 17 10:25:16 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 17 10:25:19 2008 Subject: [Histonet] Cost Analysis In-Reply-To: Message-ID: <331897.20826.qm@web65702.mail.ac4.yahoo.com> Michelle: >From grossing to H&E slide all procedures manual: materials = $2.11 + labor = $3.20 Total = $ 5.31; using automatic stainers and coverslippers: material = $ 2.00 + labor = $ 2.71 TOAL = $ 4.71 Under separate cover I am sending an article I wrote on the subject with costs for many other histology activities. Ren? J. --- On Wed, 12/17/08, GreenJumpyOne wrote: From: GreenJumpyOne Subject: [Histonet] Cost Analysis To: histonet@lists.utsouthwestern.edu Date: Wednesday, December 17, 2008, 10:44 AM I have been asked to do a cost anaylsis on what it would cost to take a single biopsy specimen from the formalin bottle all the way to the H&E slide. I only need a ballpark estimate. Has anyone done this recently? Thanks for any help you can provide. Michelle Carney Bradenton, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rfields <@t> gidocs.net Wed Dec 17 10:32:17 2008 From: rfields <@t> gidocs.net (Rosa Fields) Date: Wed Dec 17 10:34:09 2008 Subject: [Histonet] at a troubleshooting loss Message-ID: <07732CE52EC3174AB891DE1C62DB4D8F43EDA2@GIEXCHANGE.gidocs.net> Black Pigment? Paint "dust" from a slide etcher? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Aaron Rose Sent: Tuesday, December 16, 2008 5:26 PM To: Tony Henwood; Michele Wich; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] at a troubleshooting loss This may be a silly suggestion but Have you been filtering your hematoxylin? I have seen the sediments that form over time. I'm not sure if this could be the dark regions you're seeing but I figured I'd throw it out there. -Aaron -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Tony Henwood Sent: Tuesday, December 16, 2008 3:18 PM To: Michele Wich; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] at a troubleshooting loss ?carbon from lead pensils used for labelling slides. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Michele Wich Sent: Wednesday, 17 December 2008 4:31 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] at a troubleshooting loss A couple questions for those who are troubleshooting savvy: Other than inadequate fixation, water being introduced during processing, or too much heat, what can cause fuzzy, uneven hematoxylin staining with poor nuclear detail? And secondly, aside from formalin, are there any other common agents of blackish pigment in H & E staining? (The artifact appears on the slides both on and around the tissues.) Any feedback is GREATLY appreciated. This communication is intended solely for the use of the addressee and may contain information that is legally privileged, confidential or exempt from disclosure. If you are not the intended recipient, please note that any dissemination, distribution, or copying of this communication is strictly prohibited. Anyone who receives this message in error should notify the sender immediately and delete it from his or her computer _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 cbarone <@t> NEMOURS.ORG Wed Dec 17 10:52:49 2008 From: cbarone <@t> NEMOURS.ORG (Barone, Carol ) Date: Wed Dec 17 10:52:58 2008 Subject: [Histonet] isopentance substitute Message-ID: <37E4BAC017F57141AF64FAA5AEB04CE801471990@wlmmsx01.nemours.org> Histonetters: I have been doing muscle enzyme histochemistry since 1984 and have yet to find anything that works as well, with consistant results, as isopentane. Using it safely under a hood and kept in the explosion proof frig (no exposed electric parts that could arc) can be accomplish. I am still here and have not blown up the building or colleagues, not had liver or kidney damage. That being said...there are still times it cannot be used...( like when you need to freeze in an OR....no iopentance can go there. So we have found, with sugfficient practice, you can get almost as good results by wrapping the sample in foil and placing directly into liquid nitrogen, or dusting well with talc (with aluminum)...like flouring your chicken before frying. It takes some practice to get this to work with little - to no artifact...but, it is possible....and provides an alternative technique for those times when isopentane is just not possible. From hhawkins <@t> utmb.edu Wed Dec 17 11:26:56 2008 From: hhawkins <@t> utmb.edu (Hawkins, Hal K.) Date: Wed Dec 17 11:28:20 2008 Subject: [Histonet] RE: isopentance substitute In-Reply-To: <37E4BAC017F57141AF64FAA5AEB04CE801471990@wlmmsx01.nemours.org> References: <37E4BAC017F57141AF64FAA5AEB04CE801471990@wlmmsx01.nemours.org> Message-ID: <18DADD3ED840AC46AB24616A927CD0DB08CE599096@EXCHANGE2.utmb.edu> We have used the "cryo Jane" freezing device from Instrumedics, which involves dropping a liquid nitrogen-cooled copper block onto the specimen. It takes a little getting used to, but provides very good morphology. We've used it mainly for research tissue. Hal Hawkins, UTMB Galveston ________________________________________ From: histonet-bounces@lists.utsouthwestern.edu [histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Barone, Carol [cbarone@NEMOURS.ORG] Sent: Wednesday, December 17, 2008 10:52 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] isopentance substitute Histonetters: I have been doing muscle enzyme histochemistry since 1984 and have yet to find anything that works as well, with consistant results, as isopentane. Using it safely under a hood and kept in the explosion proof frig (no exposed electric parts that could arc) can be accomplish. I am still here and have not blown up the building or colleagues, not had liver or kidney damage. That being said...there are still times it cannot be used...( like when you need to freeze in an OR....no iopentance can go there. So we have found, with sugfficient practice, you can get almost as good results by wrapping the sample in foil and placing directly into liquid nitrogen, or dusting well with talc (with aluminum)...like flouring your chicken before frying. It takes some practice to get this to work with little - to no artifact...but, it is possible....and provides an alternative technique for those times when isopentane is just not possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jtrynak <@t> hotmail.com Wed Dec 17 12:00:36 2008 From: jtrynak <@t> hotmail.com (jtrynak@hotmail.com) Date: Wed Dec 17 12:00:43 2008 Subject: [Histonet] Vacation reply In-Reply-To: Message-ID:
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From mbisher <@t> Princeton.EDU Wed Dec 17 12:49:50 2008 From: mbisher <@t> Princeton.EDU (Peggy Bisher) Date: Wed Dec 17 12:50:59 2008 Subject: [Histonet] Isopentane In-Reply-To: Message-ID: Liquid nitrogen (-196C) and nitrogen slush (-207) are the coldest cryogens used, but they have the slowest freezing rates. The most effective cryogens are those that are those that stay a liquid when cooled almost to the temperature of liquid nitrogen. Propane (-189.6C mp) or Ethane (183.5C mp) is the cryogen of choice that we use for cryo electron microscopy. Margaret E. Bisher Electron Microscopy & Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbisher@princeton.edu On 12/16/08 10:02 PM, "Tony Henwood" wrote: > Depends on how you freeze. > When we freeze tissue in liquid nitrogen, the tissue is shaken, ensuring > that the gas layer is broken up. > And in so doing we have not seen evidence of ice crystal formation. > > I can send you a reprint of a paper that describes the effect of storage > of brain tumours in saline prior to freezing (causing much distortion of > the tumour) verses receiving the samples fresh on damp gauze or card. > The pics show the evidence. > > Henwood, A., (2007) "Adverse effect of saline on brain intraoperative > (frozen section) Histology" J Histotechnol 30(3):193 > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > > > -----Original Message----- > From: Markus F. Meyenhofer [mailto:micro@superlink.net] > Sent: Wednesday, 17 December 2008 1:47 PM > To: Tony Henwood; Dana Settembre; carmen loiselle; pathology education > Subject: Re: [Histonet] Isopentane > > > Isopentane is cooled by liquid N2! > To freeze tissue direct in LN2 is not the best way to freeze tissue due > to > the Leidenfrost effect, creating an insulating gas layer between the > tissue > and the liquid and delaying the speed of freezing = an increase in ice > crystal formation. > Regards, > Markus F. Meyenhofer > > ----- Original Message ----- > From: "Tony Henwood" > To: "Dana Settembre" ; "carmen loiselle" > ; "pathology education" > > Sent: Tuesday, December 16, 2008 6:19 PM > Subject: RE: [Histonet] Isopentane > > > Try liquid nitrogen > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory > Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana > Settembre > Sent: Wednesday, 17 December 2008 5:27 AM > To: carmen loiselle; pathology education > Subject: Re: [Histonet] Isopentane > > > Hello Carmen, > Sorry I have no alternative. I know most labs use that. > When I worked in a neuropathology lab the isopentane was kept in the > refrigerator - an explosion proof one. I believe that means there's no > lightbulb inside and no inner working of a lightbulb inside either. > > > Dana Settembre, HT ASCP > Immunohistochemistry Lab > UMDNJ - University Hospital > Newark, NJ USA > > >>>> carmen loiselle 12/16/08 12:24 PM >>>> > > Hello , > > I currently work in a neuromuscular lab and all muscle biopsies received > should be cut frozen, no formalin fixed tissues. My concern though, is > to work with isopentane , a well known excellent heat conductor but also > it has the bad reputation of being very explosive. In fact , I've been > told that few years ago, a lab close by of where I work , did explode > because of this solution. Is there , in your knowledge, any other > chemical we can use as replacement and that give as good results. > > Any input will be greatly appreciated > > Merry XMas to all and happy New Year > _________________________________________________________________ > > _______________________________________________ > 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 note 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. > ********************************************************************** > > > _______________________________________________ > 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 note 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. > ********************************************************************** > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Dec 17 13:19:05 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Dec 17 13:19:14 2008 Subject: [Histonet] Speaking of Fluoro Jade Message-ID: Perhaps it need not be too expensive. According to US Patent 6,229,024 (Schmued 2001) Fluoro-Jade is a sodium salt made by suspending 1 g of 5(6)-carboxyfluorescein in 10 ml water, adding NaOH until the pH of the resulting solution is between 7 and 8, and then removing the water by lyophilization or by evaporation in a desiccator to obtain the crystalline salt. You can buy 5(6)-carboxyfluorescein for $37 a gram (e.g. Sigma catalogue no. 21877). Solutions should be stable if protected from light and growth of moulds. Sodium carboxyfluorescein is an anionic xanthene dye, as is eosin Y, which also stains dead cells strongly and is fluorescent. John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: Amos Brooks Date: Tuesday, December 16, 2008 18:13 Subject: [Histonet] Speaking of Fluoro Jade To: tgenade@gmail.com, "histonet@lists.utsouthwestern.edu" > Hi, > Speaking of Fluoro Jade: I don't do this often and > after having done the > stain recently I was left with a lot of extra Fluoro Jade stock > solution.The data sheet says this is only good for a week or so > making it a dang > expensive test! For non-diagnostic use of course, could the rest be > aliquated and frozen (-80 C) to use later? > > Thanks, > Amos > > > Date: Tue, 16 Dec 2008 12:24:38 +0200 > From: "Tyrone Genade" > Subject: [Histonet] Re: Histonet Digest, Vol 61, Issue 25 > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Hi all, > > > > Otherwise, you may want to look into FlouroJadeB. If all you are after > is visualizing degenerating axons, there are easier ways of going > about ot than silver staining. > > Kind regards > -- > Tyrone Genade > http://tgenade.freeshell.org > email: tgenade@freeshell.org > tel: +27-84-632-1925 (c) > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcampbell <@t> vdxpathology.com Wed Dec 17 13:27:29 2008 From: jcampbell <@t> vdxpathology.com (Jennifer Campbell) Date: Wed Dec 17 13:27:33 2008 Subject: [Histonet] In regards to a previous email I sent out.. Message-ID: <5658CBDB9EAE6545ABE50D2563D81BF819E30E@VDXSERVER01.vdxpathology.local> Hi everyone, I sent out an email titled, "Any Opinions on these microtomes?" the other day asking for some opinions and feedback on a few microtomes I was interested in. I wanted to thank those of you who answered my email and gave me some helpful feedback. I have recieved a number of phone calls at work today from sales reps trying to sell me other models that I had not asked about in my email. I have already contacted our own sales rep and are therefore not looking for someone to sell us a new microtome but, rather someone to give us some feedback on the ones we are considering. If phone calls of this matter could be avoided, I would greatly appreciate it. Thank you, Jennifer Campbell From jkiernan <@t> uwo.ca Wed Dec 17 13:35:28 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Dec 17 13:35:32 2008 Subject: [Histonet] at a troubleshooting loss In-Reply-To: <62A8156F8071C8439080D626DF8C33A602E5B3@wave-mail.7thwave.local> References: <62A8156F8071C8439080D626DF8C33A602E5B3@wave-mail.7thwave.local> Message-ID: Others have suggested graphite dust from pencil "lead". I've also seen little bits of ground glass, from writing on slides with a diamond pencil. Another contaminant that we used to get several years ago was dark brown fungal spores, singly or in clusters about the size of a cell's nucleus. Probably the spores came from the building's air ducts (which at the time were lined by mildew several mm thick), and settled on slides that were on the hotplate, waiting to be dewaxed and stained. These spores must have had a strong affinity for tissue because nothing would wash them off. John Kiernan Anatomy, UWO London, Canada = = = ----- From original Message ----- From: Michele Wich mwich@7thwavelabs.com > And secondly, aside from formalin, are there any other common > agents of > blackish pigment in H & E staining? (The artifact appears on the > slidesboth on and around the tissues.) - - - - - From amh-histolab <@t> amh.org Wed Dec 17 13:38:51 2008 From: amh-histolab <@t> amh.org (AMH-HistoLab) Date: Wed Dec 17 13:40:13 2008 Subject: [Histonet] c-erbB-2 clone 5A2 Message-ID: <49490EFE.3C4E.0034.0@amh.org> Our manufacturer (Nova Castra) no longer supplies c-erbB-2. . Does anyone know where I can purchase this antibody. It needs to be clone 5A2. Thank You. Donna AMH Histolab Abington,PA ****************** CONFIDENTIALITY NOTICE ********************** This e-mail contains LEGALLY PRIVILEGED AND CONFIDENTIAL INFORMATION intended only for the use of the recipient named above. If you are not the intended recipient, you are hereby notified that any dissemination or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please notify the transmitting hospital by telephone or e-mail and delete the original e-mail received in error. THIS INFORMATION HAS BEEN DISCLOSED TO YOU FROM RECORDS WHOSE CONFIDENTIALITY IS PROTECTED BY STATE AND FEDERAL LAW. ANY FURTHER DISCLOSURE, COPYING, DISTRIBUTION OR ACTION TAKEN IN RELIANCE ON THE CONTENTS OF THESE DOCUMENTS WITHOUT THE PRIOR WRITTEN CONSENT OF THE PERSON TO WHOM IT PERTAINS IS PROHIBITED. YOU ARE REQUIRED TO DESTROY THE INFORMATION AFTER THE STATED NEED HAS BEEN FULFILLED. From bakevictoria <@t> gmail.com Wed Dec 17 14:23:28 2008 From: bakevictoria <@t> gmail.com (Victoria Baker) Date: Wed Dec 17 14:24:28 2008 Subject: [Histonet] c-erbB-2 clone 5A2 In-Reply-To: <49490EFE.3C4E.0034.0@amh.org> References: <49490EFE.3C4E.0034.0@amh.org> Message-ID: <4f016b690812171223w2635794es67f5298dbbda2ade@mail.gmail.com> have you tried MONOSAN www.monsan.com Catalog number MONX10220 You can try doing Google/Google scholar as well. Hope this is of some assistance. Vikki On Wed, Dec 17, 2008 at 2:38 PM, AMH-HistoLab wrote: > Our manufacturer (Nova Castra) no longer supplies c-erbB-2. . Does anyone > know where I can purchase this antibody. It needs to be clone 5A2. Thank > You. > > Donna > AMH Histolab > Abington,PA > > > > > ****************** CONFIDENTIALITY NOTICE ********************** > > This e-mail contains LEGALLY PRIVILEGED AND CONFIDENTIAL INFORMATION > intended only for the use of the recipient named above. If you are not > the intended recipient, you are hereby notified that any dissemination or > copying of this e-mail is strictly prohibited. If you have received this > e-mail in error, please notify the transmitting hospital by telephone or > e-mail and delete the original e-mail received in error. > > THIS INFORMATION HAS BEEN DISCLOSED TO YOU FROM RECORDS WHOSE > CONFIDENTIALITY IS PROTECTED BY STATE AND FEDERAL LAW. ANY FURTHER > DISCLOSURE, COPYING, DISTRIBUTION OR ACTION TAKEN IN RELIANCE ON THE > CONTENTS OF THESE DOCUMENTS WITHOUT THE PRIOR WRITTEN CONSENT OF THE > PERSON TO WHOM IT PERTAINS IS PROHIBITED. YOU ARE REQUIRED TO DESTROY > THE INFORMATION AFTER THE STATED NEED HAS BEEN FULFILLED. > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From bakevictoria <@t> gmail.com Wed Dec 17 14:31:01 2008 From: bakevictoria <@t> gmail.com (Victoria Baker) Date: Wed Dec 17 14:32:01 2008 Subject: [Histonet] c-erbB-2 clone 5A2 In-Reply-To: <4f016b690812171223w2635794es67f5298dbbda2ade@mail.gmail.com> References: <49490EFE.3C4E.0034.0@amh.org> <4f016b690812171223w2635794es67f5298dbbda2ade@mail.gmail.com> Message-ID: <4f016b690812171231s2f8773cbpa0a60625e0ac200f@mail.gmail.com> Sorry I meant www.monosan.com typo error. Vikki On Wed, Dec 17, 2008 at 3:23 PM, Victoria Baker wrote: > have you tried MONOSAN > www.monsan.com > Catalog number MONX10220 > > You can try doing Google/Google scholar as well. > > Hope this is of some assistance. > > Vikki > > On Wed, Dec 17, 2008 at 2:38 PM, AMH-HistoLab wrote: > >> Our manufacturer (Nova Castra) no longer supplies c-erbB-2. . Does anyone >> know where I can purchase this antibody. It needs to be clone 5A2. Thank >> You. >> >> Donna >> AMH Histolab >> Abington,PA >> >> >> >> >> ****************** CONFIDENTIALITY NOTICE ********************** >> >> This e-mail contains LEGALLY PRIVILEGED AND CONFIDENTIAL INFORMATION >> intended only for the use of the recipient named above. If you are not >> the intended recipient, you are hereby notified that any dissemination or >> copying of this e-mail is strictly prohibited. If you have received this >> e-mail in error, please notify the transmitting hospital by telephone or >> e-mail and delete the original e-mail received in error. >> >> THIS INFORMATION HAS BEEN DISCLOSED TO YOU FROM RECORDS WHOSE >> CONFIDENTIALITY IS PROTECTED BY STATE AND FEDERAL LAW. ANY FURTHER >> DISCLOSURE, COPYING, DISTRIBUTION OR ACTION TAKEN IN RELIANCE ON THE >> CONTENTS OF THESE DOCUMENTS WITHOUT THE PRIOR WRITTEN CONSENT OF THE >> PERSON TO WHOM IT PERTAINS IS PROHIBITED. YOU ARE REQUIRED TO DESTROY >> THE INFORMATION AFTER THE STATED NEED HAS BEEN FULFILLED. >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > From AnthonyH <@t> chw.edu.au Wed Dec 17 15:24:06 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Dec 17 15:25:07 2008 Subject: [Histonet] Isopentane In-Reply-To: Message-ID: Funny, We don't Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: Joanne Mauger [mailto:MAUGER@email.chop.edu] Sent: Thursday, 18 December 2008 12:50 AM To: Tony Henwood; carmen_loiselle@hotmail.com; histonet@pathology.swmed.edu; settembr@umdnj.edu Subject: RE: [Histonet] Isopentane In my experience, liquid nitrogen makes a lot of freezing artifact in muscle tissue. I have found no better substitue for isopentane if you want quality morphology in you specimen. We store it in an explosion proof cabinet. Jo Mauger >>> "Tony Henwood" 12/16/08 6:19 PM >>> Try liquid nitrogen Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Wednesday, 17 December 2008 5:27 AM To: carmen loiselle; pathology education Subject: Re: [Histonet] Isopentane Hello Carmen, Sorry I have no alternative. I know most labs use that. When I worked in a neuropathology lab the isopentane was kept in the refrigerator - an explosion proof one. I believe that means there's no lightbulb inside and no inner working of a lightbulb inside either. Dana Settembre, HT ASCP Immunohistochemistry Lab UMDNJ - University Hospital Newark, NJ USA >>> carmen loiselle 12/16/08 12:24 PM >>> Hello , I currently work in a neuromuscular lab and all muscle biopsies received should be cut frozen, no formalin fixed tissues. My concern though, is to work with isopentane , a well known excellent heat conductor but also it has the bad reputation of being very explosive. In fact , I've been told that few years ago, a lab close by of where I work , did explode because of this solution. Is there , in your knowledge, any other chemical we can use as replacement and that give as good results. Any input will be greatly appreciated Merry XMas to all and happy New Year _________________________________________________________________ _______________________________________________ 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 note 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. ********************************************************************** _______________________________________________ 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 note 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 carrolpb <@t> umdnj.edu Wed Dec 17 15:40:12 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Wed Dec 17 15:40:26 2008 Subject: [Histonet] Isopentane In-Reply-To: References: Message-ID: <494971BC.10506@umdnj.edu> > I have found no better substitue for isopentane ditto with isopentane (methylbutane) here... -160 in liquid form... though it will begin to freeze solid after several minutes... From sharon.osborn <@t> comcast.net Wed Dec 17 17:05:36 2008 From: sharon.osborn <@t> comcast.net (sharon.osborn@comcast.net) Date: Wed Dec 17 17:05:40 2008 Subject: [Histonet] Tissue Tek II embedding center is taken Message-ID: <121720082305.21899.494985C0000242780000558B2216557996029D010D9C01D202019D0E089C@comcast.net> Histonetters... Thanks to all who responded wanting the embedding center. It left for its new home today. It would have been great to have had 12 of these available to fulfill all requests for those who wanted it. Unfortunately, we only had the one. Happy Holidays, Sharon Osborn LabVision, a division of ThermoFisher Scientific Fremont, CA From sharon.osborn <@t> comcast.net Wed Dec 17 17:07:50 2008 From: sharon.osborn <@t> comcast.net (sharon.osborn@comcast.net) Date: Wed Dec 17 17:08:49 2008 Subject: [Histonet] Sorvall JB-4 Microtome Message-ID: <121720082307.26687.49498646000D55F30000683F2216557996029D010D9C01D202019D0E089C@comcast.net> Histonetters, We have available a JB-4 Microtome by Sorvall for whomever is interested. You would need to pay for shipping. It is a good microtome; is minus the chuck. I know there are a few labs that still have use for one or may need spare parts. Please contact me if interested. Sharon Osborn LabVision of ThermoFisher Scientific Fremont, CA 510-991-2824 From jkiernan <@t> uwo.ca Wed Dec 17 22:02:02 2008 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Dec 17 22:02:11 2008 Subject: Histo-Dirt (Was Re: RE: [Histonet] at a troubleshooting loss) Message-ID: Good one Tony. Graphite is quite probable: pencil spray. Fungal spores from the air circulated in the building are another major cause of dirt particles on slides here in SW Ontario. Pencil spray is black. Spores are dark brown, mostly smaller than mammalian cell nuclei, and with internal granulation. Probably we'll get many more descriptions of histo-dirt. Let them come! John Kiernan Anatomy, UWO London, Canada = = = ----- Original Message ----- From: Tony Henwood Date: Tuesday, December 16, 2008 18:20 Subject: RE: [Histonet] at a troubleshooting loss To: Michele Wich , histonet@lists.utsouthwestern.edu > ?carbon from lead pensils used for labelling slides. > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > MicheleWich > Sent: Wednesday, 17 December 2008 4:31 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] at a troubleshooting loss > > > A couple questions for those who are troubleshooting savvy: > Other than > inadequate fixation, water being introduced during processing, > or too > much heat, what can cause fuzzy, uneven hematoxylin staining > with poor > nuclear detail? > > And secondly, aside from formalin, are there any other common > agents of > blackish pigment in H & E staining? (The artifact appears on the > slidesboth on and around the tissues.) > > Any feedback is GREATLY appreciated. > > > > > This communication is intended solely for the use of the > addressee and > may contain information that is legally privileged, confidential or > exempt from disclosure. If you are not the intended > recipient, please > note that any dissemination, distribution, or copying of this > communication is strictly prohibited. Anyone who receives > this message > in error should notify the sender immediately and delete it from > his or > her computer _______________________________________________ > 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 note 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. > ********************************************************************** > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From yourbiomed <@t> cox.net Thu Dec 18 03:11:37 2008 From: yourbiomed <@t> cox.net (Brad YourBiomed.Cox) Date: Thu Dec 18 03:12:52 2008 Subject: [Histonet] autostainer service In-Reply-To: <424D930E0319E0469DD9C61502A50E3002102052@NIHCESMLBX4.nih.gov> References: <424D930E0319E0469DD9C61502A50E3002102052@NIHCESMLBX4.nih.gov> Message-ID: <007e01c960f0$a1ef6c90$e5ce45b0$@net> Dear Patricia and all, My name is Brad and I can service all DAKOCytomation Autostainers. I will contact you directly but would like to inform Histonet that we have all the replacement parts for the stainer and can perform complete PM's (Preventive Maintenances) and repairs. I have almost six years of experience and trained by Dako field service engineers. Our work is guaranteed. We have access to a warehouse full of new and remanufactured parts. I'm confident that you will be pleased with our service and PM's. Please contact: IMEB, Inc Brad Biomedical Engineer Toll free 1-800-543-8496 or email me brad@imebinc.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Zerfas, Patricia (NIH/OD/ORS) [E] Sent: Tuesday, December 16, 2008 11:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] autostainer service Dear All, I am currently using the DAKOCytomation autostainer that is approximately seven years old. Once the service contract expires in November 2009 DAKOCytomation will not continue the service contract. The equipment is in excellent condition and meets the needs of the laboratory thus I am reluctant to buy a replacement. Would anyone be able to obtain parts as needed and provide us with a service contract once the present contract expires? Thanks, Patricia Zerfas National Institutes of Health Building 28A, Room 112 28 Library Drive Bethesda, MD 20892 ph: (301) 496-4464 fax: (301) 402-1068 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dmccaig <@t> ckha.on.ca Thu Dec 18 10:28:47 2008 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Thu Dec 18 10:29:45 2008 Subject: [Histonet] (no subject) Message-ID: What should the pH range be for Scott's Tap Water? From ktuttle <@t> umm.edu Thu Dec 18 12:19:02 2008 From: ktuttle <@t> umm.edu (Kimberly Tuttle) Date: Thu Dec 18 12:19:34 2008 Subject: [Histonet] Which Blocking reagent? Message-ID: <494A4DC5.90CE.001A.3@umm.edu> Im trying to work up a few IHC stains, but Im still learning too. Synaptopodin using a kidney control and Cleaved caspase 3 on tonsil. Im getting a lot of background but there are so many reagents out there I dont know which to try or whats the difference between them. Im using the Dako Envision system. Kimberly C. Tuttle HT (ASCP) Pathology Biorepository and Research Core University of Maryland Room NBW58, UMMC 22 S. Greene St Baltimore, MD 21201 (410) 328-5524 (410) 328-5508 fax This e-mail and any accompanying attachments may be privileged, confidential, contain protected health information about an identified patient or be otherwise protected from disclosure. State and federal law protect the confidentiality of this information. If the reader of this message is not the intended recipient; you are prohibited from using, disclosing, reproducing or distributing this information; you should immediately notify the sender by telephone or e-mail and delete this e-mail. From es144131 <@t> bcm.tmc.edu Thu Dec 18 12:24:39 2008 From: es144131 <@t> bcm.tmc.edu (Stephens, Elizabeth Humes) Date: Thu Dec 18 12:25:42 2008 Subject: [Histonet] Fixing and IHC on PEG hydrgoels Message-ID: Dear Colleagues, I was wondering if anyone has had any experience with fixing and performing immunohistochemistry on cells seeded onto PEG hydrogels. These are very hydrophilic gels and I have heard that, even with a very short fixation time, fixation causes any cells that were growing on the PEG gels to fall off! I am trying to perform IHC on cells seeded onto these PEG hydrogels. Does anyone have any suggestions on how to fix these? Is there some fixative that works very quickly and would maintain the hydrophilicity of the PEG gel? Thank you!! Elizabeth Stephens MD/PhD Student Baylor College of Medicine/Rice University From nyilmaz <@t> mersin.edu.tr Thu Dec 18 13:26:36 2008 From: nyilmaz <@t> mersin.edu.tr (Nejat Yilmaz) Date: Thu Dec 18 13:26:41 2008 Subject: [Histonet] Capillary counting Message-ID: <000d01c96146$8b638550$2101a8c0@nejat1> Hi All, We need a protocol demonstrating capillaries in sceletal muscle tissue other than immunohistochemistry. We're planning to compare number of capillaries between different experiment groups for evaluating angiogenesis. Is there any special stain to suggest for this purpose? Thanks in advance. Necat Yilmaz From NLinke <@t> mednet.ucla.edu Thu Dec 18 13:32:17 2008 From: NLinke <@t> mednet.ucla.edu (Linke, Noelle) Date: Thu Dec 18 13:33:10 2008 Subject: [Histonet] PowerPath Message-ID: <0C96F0BFE078D74C91A1C541D24A6AE435BEDD04@EMGMB1.ad.medctr.ucla.edu> Hi everyone, I am new to Power Path and I have a question I was hoping someone could answer. Is there a way to pull metrics from it (slide totals, stains, etc)?? Any help would be appreciated! Thank you! Noelle No?lle Linke M.S., HTL(ASCP)QIHC Manager, Histology Services Department of Pathology & Laboratory Medicine David Geffen School of Medicine at UCLA Phone: 310-825-7397 nlinke@mednet.ucla.edu ________________________________ IMPORTANT WARNING: This email (and any attachments) is only intended for the use of the person or entity to which it is addressed, and may contain information that is privileged and confidential. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Unauthorized redisclosure or failure to maintain confidentiality may subject you to federal and state penalties. If you are not the intended recipient, please immediately notify us by return email, and delete this message from your computer. From rjbuesa <@t> yahoo.com Thu Dec 18 14:50:26 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 18 14:50:32 2008 Subject: [Histonet] Capillary counting In-Reply-To: <000d01c96146$8b638550$2101a8c0@nejat1> Message-ID: <3742.64276.qm@web65701.mail.ac4.yahoo.com> Nejat: There are several stains for capillaries: 1- Campbell and Alexander (tested by Mallory,1938): black agsinst colorless bbackground. 2- Doherty, Suk and Alexander, 1938: differential staining of blood in capillaries. 3- Pickworth, 1934: blood vessels (capillaries) in thick sections of brain. 4- Slominski and Cunge (tested by Romeis, 1948): differential staining of capillaries, and 5- Ziegler 1945 (tested by Riley, 1945): demonstration of capillaries in wholemounts of cornea. Although developed for other tissues it is very likely that they will work on skeletal muscle. Ren? J. --- On Thu, 12/18/08, Nejat Yilmaz wrote: From: Nejat Yilmaz Subject: [Histonet] Capillary counting To: histonet@lists.utsouthwestern.edu Date: Thursday, December 18, 2008, 2:26 PM Hi All, We need a protocol demonstrating capillaries in sceletal muscle tissue other than immunohistochemistry. We're planning to compare number of capillaries between different experiment groups for evaluating angiogenesis. Is there any special stain to suggest for this purpose? Thanks in advance. Necat Yilmaz _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From KLAPANO1 <@t> hfhs.org Thu Dec 18 14:53:51 2008 From: KLAPANO1 <@t> hfhs.org (Karen Lapanowski) Date: Thu Dec 18 14:54:15 2008 Subject: [Histonet] Mature endothelium marker Message-ID: <494A720F0200002800011CDB@gwia2v.net.hfh.edu> Hello, Does anyone know of a mature endothelium marker other than Factor VIII that can be used in rat brain paraffin sections? Thanks, Karen Lapanowski Henry Ford Health System Radiation Oncology 3065 E & R 313-916-9386 ============================================================================== CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by reply email, delete the email from your computer system and shred any paper copies. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. See our Privacy Policy and Henry Ford My Health at www.henryford.com for more detailed information. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================== From rjbuesa <@t> yahoo.com Thu Dec 18 16:00:29 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 18 16:01:28 2008 Subject: [Histonet] Mature endothelium marker In-Reply-To: <494A720F0200002800011CDB@gwia2v.net.hfh.edu> Message-ID: <715978.97057.qm@web65707.mail.ac4.yahoo.com> Use Ulex europaeus lectins. Ren? J. --- On Thu, 12/18/08, Karen Lapanowski wrote: From: Karen Lapanowski Subject: [Histonet] Mature endothelium marker To: histonet@lists.utsouthwestern.edu Date: Thursday, December 18, 2008, 3:53 PM Hello, Does anyone know of a mature endothelium marker other than Factor VIII that can be used in rat brain paraffin sections? Thanks, Karen Lapanowski Henry Ford Health System Radiation Oncology 3065 E & R 313-916-9386 ============================================================================== CONFIDENTIALITY NOTICE: This email contains information from the sender that may be CONFIDENTIAL, LEGALLY PRIVILEGED, PROPRIETARY or otherwise protected from disclosure. This email is intended for use only by the person or entity to whom it is addressed. If you are not the intended recipient, any use, disclosure, copying, distribution, printing, or any action taken in reliance on the contents of this email, is strictly prohibited. If you received this email in error, please contact the sending party by reply email, delete the email from your computer system and shred any paper copies. Note to Patients: There are a number of risks you should consider before using e-mail to communicate with us. See our Privacy Policy and Henry Ford My Health at www.henryford.com for more detailed information. If you do not believe that our policy gives you the privacy and security protection you need, do not send e-mail or Internet communications to us. ============================================================================== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From disbrc <@t> shands.ufl.edu Thu Dec 18 19:38:05 2008 From: disbrc <@t> shands.ufl.edu (Carrie Disbrow) Date: Thu Dec 18 19:38:37 2008 Subject: [Histonet] dako artisan gram and afb Message-ID: <494AB4AD.72AC.0059.0@shands.ufl.edu> Happy Holidays to all. W ea re using the Dako Artisan stainer and have had problems lately with the AFB and Gram staining inconsistently from day to day. We have had the machine services and the PM done and the problem has returned. Has anyone had the same problem and what did you do? Thanks for all your help. From cristiana.soldani <@t> humanitas.it Fri Dec 19 02:11:12 2008 From: cristiana.soldani <@t> humanitas.it (SOLDANI Cristiana ICH) Date: Fri Dec 19 02:46:37 2008 Subject: [Histonet] Please remove from mailing list In-Reply-To: <4936B0D5.008F.0051.0@jhmi.edu> References: <4936B0D5.008F.0051.0@jhmi.edu> Message-ID: <8D4E4E312BEF164997299EE5B0E12B7C02B8B541@TEHWEX11.techosp.it> Please remove me from the Histonet mailing list. Thanks for the valuable information I've received through this outreach. From heroina <@t> ibiss.bg.ac.yu Fri Dec 19 02:52:32 2008 From: heroina <@t> ibiss.bg.ac.yu (Angelina Subotic) Date: Fri Dec 19 02:52:35 2008 Subject: [Histonet] (no subject) Message-ID: Please remove me from the Histonet mailing list. From borisdobrojevic <@t> gmail.com Fri Dec 19 03:23:09 2008 From: borisdobrojevic <@t> gmail.com (boris dobrojevic) Date: Fri Dec 19 03:24:07 2008 Subject: [Histonet] (no subject) Message-ID: <17e96ca10812190123h794f5e32h357e4310e9aa23e4@mail.gmail.com> Please remove me from the Histonet mailing list. From histonet.nospam <@t> vneubert.com Fri Dec 19 04:17:11 2008 From: histonet.nospam <@t> vneubert.com (V. Neubert) Date: Fri Dec 19 04:17:14 2008 Subject: [Histonet] Removal magic Message-ID: <2a1ed4e803113123f9b63eac6b8e1c89@vneubert.com> Good people, start to read this post and experience the magic which is IN YOUR HANDS! Follow the instructions given on http://lists.utsouthwestern.edu/mailman/listinfo/histonet! YES YOU CAN! Go to the bottom of the page, enter your mail address and follow the wizard :-) From jqb7 <@t> cdc.gov Fri Dec 19 06:33:39 2008 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Fri Dec 19 06:34:28 2008 Subject: [Histonet] dako artisan gram and afb In-Reply-To: <494AB4AD.72AC.0059.0@shands.ufl.edu> References: <494AB4AD.72AC.0059.0@shands.ufl.edu> Message-ID: <1CE1847DFEA0A647B1CCDE4108EA60A70208C6C1@LTA3VS011.ees.hhs.gov> We have not experienced this problem, thank goodness! Have you tried different reagent packs? Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carrie Disbrow Sent: Thursday, December 18, 2008 8:38 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] dako artisan gram and afb Happy Holidays to all. W ea re using the Dako Artisan stainer and have had problems lately with the AFB and Gram staining inconsistently from day to day. We have had the machine services and the PM done and the problem has returned. Has anyone had the same problem and what did you do? Thanks for all your help. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carrolpb <@t> umdnj.edu Fri Dec 19 08:30:28 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Fri Dec 19 08:33:41 2008 Subject: [Histonet] (no subject) In-Reply-To: References: Message-ID: <494BB004.8050006@umdnj.edu> a quick search of the histonet archives brings up many posts about this. most say its pH is 8-9. http://www.histosearch.com/histonet/Jan03/RE.NeedrecipetomakeA.html Diana McCaig wrote: > What should the pH range be for Scott's Tap Water? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From ree3 <@t> leicester.ac.uk Fri Dec 19 08:48:54 2008 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Dec 19 08:49:20 2008 Subject: [Histonet] Speaking of Fluoro Jade In-Reply-To: <582736990812161512u389aeaf4ke11d0686b3d6584c@mail.gmail.com> References: <582736990812161512u389aeaf4ke11d0686b3d6584c@mail.gmail.com> Message-ID: <7722595275A4DD4FA225B92CDBF174A174501D4255@EXC-MBX3.cfs.le.ac.uk> Name sounds familiar, was she not Clint Eastwood's special friend in Wagon Train or was it in Blazing Saddles??. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: 16 December 2008 23:12 To: tgenade@gmail.com; histonet@lists.utsouthwestern.edu Subject: [Histonet] Speaking of Fluoro Jade Hi, Speaking of Fluoro Jade: I don't do this often and after having done the stain recently I was left with a lot of extra Fluoro Jade stock solution. The data sheet says this is only good for a week or so making it a dang expensive test! For non-diagnostic use of course, could the rest be aliquated and frozen (-80 C) to use later? Thanks, Amos Date: Tue, 16 Dec 2008 12:24:38 +0200 From: "Tyrone Genade" Subject: [Histonet] Re: Histonet Digest, Vol 61, Issue 25 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi all, Otherwise, you may want to look into FlouroJadeB. If all you are after is visualizing degenerating axons, there are easier ways of going about ot than silver staining. Kind regards -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@freeshell.org tel: +27-84-632-1925 (c) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From contact <@t> excaliburpathology.com Fri Dec 19 09:08:04 2008 From: contact <@t> excaliburpathology.com (Paula Pierce) Date: Fri Dec 19 09:09:05 2008 Subject: [Histonet] Speaking of Fluoro Jade Message-ID: <305349.72025.qm@web1111.biz.mail.sk1.yahoo.com> Shame on you! Clint Eastwood was in Rawhide. ________________________________ From: "Edwards, R.E." To: Amos Brooks ; "tgenade@gmail.com" ; "histonet@lists.utsouthwestern.edu" Sent: Friday, December 19, 2008 8:48:54 AM Subject: RE: [Histonet] Speaking of Fluoro Jade Name sounds familiar, was she not Clint Eastwood's special friend in Wagon Train or was it in Blazing Saddles??. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amos Brooks Sent: 16 December 2008 23:12 To: tgenade@gmail.com; histonet@lists.utsouthwestern.edu Subject: [Histonet] Speaking of Fluoro Jade Hi, ? Speaking of Fluoro Jade: I don't do this often and after having done the stain recently I was left with a lot of extra Fluoro Jade stock solution. The data sheet says this is only good for a week or so making it a dang expensive test! For non-diagnostic use of course, could the rest be aliquated and frozen (-80 C) to use later? Thanks, Amos Date: Tue, 16 Dec 2008 12:24:38 +0200 From: "Tyrone Genade" Subject: [Histonet] Re: Histonet Digest, Vol 61, Issue 25 To: histonet@lists.utsouthwestern.edu Message-ID: ? ? ? Content-Type: text/plain; charset=ISO-8859-1 Hi all, Otherwise, you may want to look into FlouroJadeB. If all you are after is visualizing degenerating axons, there are easier ways of going about ot than silver staining. Kind regards -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@freeshell.org tel: +27-84-632-1925 (c) _______________________________________________ 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 tifei <@t> foxmail.com Fri Dec 19 09:24:23 2008 From: tifei <@t> foxmail.com (tf) Date: Fri Dec 19 09:24:46 2008 Subject: [Histonet] PSA-NCAM IHC on frozen brain section with BrdU co-staining Message-ID: <200812192324184285251@foxmail.com> Hi everyone, it seems that I lost the PSA-NCAM staining when sections are treated with citrate buffer (breaking the membrane?) For co-staining with BrdU, I treated sections with HCl - however the background is so high for both BrdU staining & PSA-NCAM immunofluroscence~~ Anyone worked on this before? 2008-12-19 tf _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From carrolpb <@t> umdnj.edu Fri Dec 19 09:25:23 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Fri Dec 19 09:25:37 2008 Subject: [Histonet] Speaking of Fluoro Jade In-Reply-To: <305349.72025.qm@web1111.biz.mail.sk1.yahoo.com> References: <305349.72025.qm@web1111.biz.mail.sk1.yahoo.com> Message-ID: <494BBCE3.7080001@umdnj.edu> > Shame on you! Clint Eastwood was in Rawhide. "Every Which Way but Loose" anyone?! > > > > ________________________________ > From: "Edwards, R.E." > To: Amos Brooks ; "tgenade@gmail.com" ; "histonet@lists.utsouthwestern.edu" > Sent: Friday, December 19, 2008 8:48:54 AM > Subject: RE: [Histonet] Speaking of Fluoro Jade > > Name sounds familiar, was she not Clint Eastwood's special friend in Wagon Train or was it in Blazing Saddles??. > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amos Brooks > Sent: 16 December 2008 23:12 > To: tgenade@gmail.com; histonet@lists.utsouthwestern.edu > Subject: [Histonet] Speaking of Fluoro Jade > > Hi, > Speaking of Fluoro Jade: I don't do this often and after having done the > stain recently I was left with a lot of extra Fluoro Jade stock solution. > The data sheet says this is only good for a week or so making it a dang > expensive test! For non-diagnostic use of course, could the rest be > aliquated and frozen (-80 C) to use later? > > Thanks, > Amos > > > Date: Tue, 16 Dec 2008 12:24:38 +0200 > From: "Tyrone Genade" > Subject: [Histonet] Re: Histonet Digest, Vol 61, Issue 25 > To: histonet@lists.utsouthwestern.edu > Message-ID: > > Content-Type: text/plain; charset=ISO-8859-1 > > Hi all, > > > > Otherwise, you may want to look into FlouroJadeB. If all you are after > is visualizing degenerating axons, there are easier ways of going > about ot than silver staining. > > Kind regards > -- > Tyrone Genade > http://tgenade.freeshell.org > email: tgenade@freeshell.org > tel: +27-84-632-1925 (c) > _______________________________________________ > 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 alyssa <@t> alliedsearchpartners.com Fri Dec 19 10:45:46 2008 From: alyssa <@t> alliedsearchpartners.com (Alyssa Peterson) Date: Fri Dec 19 10:46:01 2008 Subject: [Histonet] histology permanent positions in AZ,TN,FL Message-ID: Happy Holidays! As the new year approaches I wanted to pass on some information about the permanent staff histology positions I have available.Please pass the word along to anyone that you may know, and if you are interested, please send questions and resume to: alyssa@alliedsearchpartners.com 1)full time/day shift in Phoenix,AZ 2)part time/day shift in Surprise, AZ 3)full time/day shift in Memphis, TN area 4)full time/5am-1PM in Lakeland/Plant City, FL area.I look forward to hearing from you! -Alyssa alyssa@alliedsearchpartners.com From drvet_anjan <@t> hotmail.com Fri Dec 19 12:19:57 2008 From: drvet_anjan <@t> hotmail.com (anjan kumar) Date: Fri Dec 19 12:20:10 2008 Subject: [Histonet] (no subject) Message-ID: hi, I am Dr. Anjan Kumar working as Junior Research Scientist at Madras veterinary college, chennai, india. currently i am pursuing my PhD in veterinary pathology and i am working on canine mammary tumours for my theses work and i have planned to work on triple immunohistochemistry using CD44, CD24, EPCAM AS ALL these antigens are surface markers of stem like cancer cells and they all are co localized. i am finding it very difficult to select 3 different chromogens. i have planned to procure monoclonals from Thermo - Lab visison i.e the following are mouse monoclonals: CD44: IgG2a CD24: IgM / ? ESA: IgG1 / ? i have planned to use 2 detection system i.e alkaline phosphatase and HRP due to funding constraints i am forced to settle for single goat anti - mouse secondary antibody currently im confused with everything and i am struck here And another doubt ... according to multiple immunostaining methods by Chris Vander loos after double immuno histochemistry we can proceed for one more HIER and perform another single immunohistochemistry so this mounts upto triple immunohistochemistry right! coming to the point, my doubt was after performing HIER WILL the chromogen present earlier will be lost along with the immune complex. i have stopped all my work here and dont know how i can proceed... please help me with your valuable suggestions. can any one plz suggest me established triple immunohistochemistry protocol. regards, Dr anjan. _________________________________________________________________ Find a better job. We have plenty. Visit MSN Jobs http://www.in.msn.com/jobs From drvet_anjan <@t> hotmail.com Fri Dec 19 12:22:08 2008 From: drvet_anjan <@t> hotmail.com (anjan kumar) Date: Fri Dec 19 12:22:22 2008 Subject: [Histonet] RE: triple immunohistochemistry Message-ID: hi, I am Dr. Anjan Kumar working as Junior Research Scientist at Madras veterinary college, chennai, india. currently i am pursuing my PhD in veterinary pathology and i am working on canine mammary tumours for my theses work and i have planned to work on triple immunohistochemistry using CD44, CD24, EPCAM AS ALL these antigens are surface markers of stem like cancer cells and they all are co localized. i am finding it very difficult to select 3 different chromogens. i have planned to procure monoclonals from Thermo - Lab visison i.e the following are mouse monoclonals: CD44: IgG2a CD24: IgM / ? ESA: IgG1 / ? i have planned to use 2 detection system i.e alkaline phosphatase and HRP due to funding constraints i am forced to settle for single goat anti - mouse secondary antibody currently im confused with everything and i am struck here And another doubt ... according to multiple immunostaining methods by Chris Vander loos after double immuno histochemistry we can proceed for one more HIER and perform another single immunohistochemistry so this mounts upto triple immunohistochemistry right! coming to the point, my doubt was after performing HIER WILL the chromogen present earlier will be lost along with the immune complex. i have stopped all my work here and dont know how i can proceed... please help me with your valuable suggestions. can any one plz suggest me established triple immunohistochemistry protocol. regards, Dr anjan. Junior research scientist Department of Veterinary Pathology, Madras veterinary college, chennai, india. Make sure your wardrobe reflects the latest trends and styles in the world of fashion. Try it! _________________________________________________________________ Much more than email - don't miss out on the rest of the Windows LiveT. http://www.microsoft.com/windows/windowslive/events.aspx From michelle.steinkrauss <@t> novartis.com Fri Dec 19 12:39:05 2008 From: michelle.steinkrauss <@t> novartis.com (michelle.steinkrauss@novartis.com) Date: Fri Dec 19 12:39:13 2008 Subject: [Histonet] Michelle Steinkrauss is out of the office. Message-ID: I will be out of the office starting 12/19/2008 and will not return until 12/22/2008. If you require immediate assistance, please contact Michelle Broome at x 47477. From sbreeden <@t> nmda.nmsu.edu Fri Dec 19 13:16:03 2008 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Dec 19 13:17:02 2008 Subject: [Histonet] Felicitations Message-ID: <4D14F0FC9316DD41972D5F03C070908B017E65F7@nmdamailsvr.nmda.ad.nmsu.edu> As I will be in and out ("on call") for most of the next two weeks, I'd like to wish everyone around the world a wonderful and (more than anything) peaceful holiday season. It has been my pleasure to be a part of such a productive and pleasant group of highly-skilled and sometimes humorous (okay, usually humorous) technical-types for the past four years - I am humbled and grateful for all of the answers you've given when I haven't asked the questions; for the assumed acceptance of my attempts to Lighten The Heck Up; and for the wit and wisdom of such a fine group of people. This is a bit more emotion than I would normally indulge in and it's probably all you're going to get, so consider it a gift. Merry and Happy - or whatever you call it - let's hope for good things to come. And now - back to my PAS. Salud. Sally Breeden, HT(ASCP) NM Dept. of Agriculture Veterinary Diagnostic Services PO Box 4700 Albuquerque, NM 87106 505-841-2576 From akemiat3377 <@t> yahoo.com Fri Dec 19 13:18:20 2008 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Fri Dec 19 13:19:18 2008 Subject: [Histonet] Happy Holidays Message-ID: <932272.49637.qm@web31303.mail.mud.yahoo.com> Happy Friday before Christmeas...I couldn't help but share this with you all! Happy Solstice, Happy Hanukkah, Merry Christmas, Happy Kwanza Sorry for the delay?our Legal Department just approved the following Holiday Greeting: Please accept with no obligation, implied or implicit my best wishes for an environmentally conscious, socially responsible, low stress, non-addictive, gender neutral, celebration of the winter solstice holiday (tm), practiced within the most enjoyable traditions of the religious persuasion of your choice, or secular practices of your choice, with respect for the religious/secular persuasions and/or traditions of others, or their choice not to practice religious or secular traditions at all . . . and a fiscally successful, personally fulfilling, and medically uncomplicated recognition of the onset of the generally accepted calendar year 2009, but not without due respect for the calendars of choice of other cultures whose contributions to society have helped make America great, (not to imply that America is necessarily greater than any other country or is the only "AMERICA" in the western hemisphere), and without regard to the race, creed, color, age, physical ability, religious faith, choice of computer platform, or sexual orientation of the wishee. By accepting this greeting, you are accepting these terms: This greeting is subject to clarification or withdrawal. It is freely transferable with no alteration to the original greeting. It implies no promise by the wisher to actually implement any of the wishes for her/himself or others, and is void where prohibited by law, and is revocable at the sole discretion of the wisher. This wish is warranted to perform as expected within the usual application of good tidings for a period of one year, or until the issuance of a subsequent holiday greeting, whichever comes first, and warranty is limited to replacement of this wish or issuance of a new wish at the sole discretion of the wisher. Have a Great 2009: Akemi Allison-Tacha BS, HT (ASCP) HTL President Phoenix Lab Consulting Specializing in Histology, SS, IHC, & TMA Tele: (425) 941-4287 E-Mail: akemiat3377@yahoo.com From pattennj <@t> mail.nih.gov Fri Dec 19 13:38:02 2008 From: pattennj <@t> mail.nih.gov (Patten, Nicole (NIH/NIAAA) [F]) Date: Fri Dec 19 13:39:17 2008 Subject: [Histonet] Myelin Stains for LCM In-Reply-To: References: Message-ID: Hi- I am trying to find a suitable Myelin stain for LCM that maintains RNA integrity. I am currently testing Luxol Fast Blue, Oil Red O and Fluoromyelin but none seem to keep the RNA in tact all that well (although I am still trying to modify these procedures). My ultimate goal is to dissect regions of white matter where demyelination has occurred due to disease. Anyone have any other suggestions? Also, does anyone know of any Oil Red O procedures that do not require formalin fixation of the tissue? I have tried fixing the tissue in isopropanol with poor histological results. Any suggestions? Thanks and happy holidays! -Nicole From leiker <@t> buffalo.edu Fri Dec 19 14:03:49 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Fri Dec 19 14:03:54 2008 Subject: [Histonet] Dark vs. Light Tissue? In-Reply-To: <932272.49637.qm@web31303.mail.mud.yahoo.com> References: <932272.49637.qm@web31303.mail.mud.yahoo.com> Message-ID: Hey all, While embedding rodent tissues I have found that some specimen within the SAME muscle group (such as quadriceps) appear very dark in color while others are very pale. I just recently started noticing this while I've been embedding. (I embed manually; I'm a research tech who does a lot of histological work). Any insights as to why the color difference? Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From hana444 <@t> gmail.com Fri Dec 19 14:03:53 2008 From: hana444 <@t> gmail.com (Hana Peter) Date: Fri Dec 19 14:03:58 2008 Subject: [Histonet] Scott's tap water Message-ID: <68f635b90812191203t2d95dac3r11ce7833a9ffc465@mail.gmail.com> Hello, it seems that Scott's tap water is a very frequent topic here and often is mentioned: Scott SG. On successive double staining for histological purposes (preliminary note). J Path Bact 1911-2;16:390-8. I've been looking for this particular article for some time now. Is there anyone who can send it to me? Thank you very much in advance! Hana Peter From leiker <@t> buffalo.edu Fri Dec 19 16:59:37 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Fri Dec 19 16:59:42 2008 Subject: [Histonet] Dark vs. Light Tissue? In-Reply-To: References: <932272.49637.qm@web31303.mail.mud.yahoo.com> Message-ID: <41EFA375BDE8BFBF2F7FB070@bchwxp2702.ad.med.buffalo.edu> Just to clarify, this is the same rodent muscle. We take out the whole quadriceps from one rodent, the whole quadriceps from another rodent, and both will look different colors while embedding. --On Friday, December 19, 2008 3:03 PM -0500 Merced Leiker wrote: > Hey all, > > While embedding rodent tissues I have found that some specimen within the > SAME muscle group (such as quadriceps) appear very dark in color while > others are very pale. I just recently started noticing this while I've > been embedding. (I embed manually; I'm a research tech who does a lot of > histological work). > > Any insights as to why the color difference? > > Merced M Leiker > Research Technician II > 354 BRB (pkgs) / 140 Farber Hall (letters) > School of Medicine and Biomedical Sciences > State University of New York at Buffalo > 3435 Main St, Buffalo, NY 14214 > Ph: (716) 829-6033 > Fx: (716) 829-2725 > > "Without my flaws I'm really very boring." > - random internet blog commentator > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From sheila_adey <@t> hotmail.com Fri Dec 19 18:39:08 2008 From: sheila_adey <@t> hotmail.com (sheila adey) Date: Fri Dec 19 18:40:06 2008 Subject: [Histonet] dako artisan gram and afb In-Reply-To: <1CE1847DFEA0A647B1CCDE4108EA60A70208C6C1@LTA3VS011.ees.hhs.gov> References: <494AB4AD.72AC.0059.0@shands.ufl.edu> <1CE1847DFEA0A647B1CCDE4108EA60A70208C6C1@LTA3VS011.ees.hhs.gov> Message-ID: Do all the techs prime the packs prior to the first run? Good luckSheila Adey HT MLT Port Huron Hospital Michigan> Date: Fri, 19 Dec 2008 07:33:39 -0500> From: jqb7@cdc.gov> To: disbrc@shands.ufl.edu; histonet@lists.utsouthwestern.edu> Subject: RE: [Histonet] dako artisan gram and afb> CC: > > We have not experienced this problem, thank goodness! Have you tried> different reagent packs? > > > Jeanine Bartlett> Infectious Diseases Pathology Branch> (404) 639-3590 > jeanine.bartlett@cdc.hhs.gov> > > -----Original Message-----> From: histonet-bounces@lists.utsouthwestern.edu> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carrie> Disbrow> Sent: Thursday, December 18, 2008 8:38 PM> To: histonet@lists.utsouthwestern.edu> Subject: [Histonet] dako artisan gram and afb> > Happy Holidays to all.> W ea re using the Dako Artisan stainer and have had problems lately with> the AFB and Gram staining inconsistently from day to day. We have had> the machine services and the PM done and the problem has returned. Has> anyone had the same problem and what did you do?> Thanks for all your 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 _________________________________________________________________ Drag n? drop?Get easy photo sharing with Windows Live? Photos. http://www.microsoft.com/windows/windowslive/photos.aspx From akemiat3377 <@t> yahoo.com Sat Dec 20 15:46:02 2008 From: akemiat3377 <@t> yahoo.com (Akemi Allison-Tacha) Date: Sat Dec 20 15:46:06 2008 Subject: [Histonet] Fw: RE: Holiday Greetings from Chuck Churukian Message-ID: <129706.63931.qm@web31305.mail.mud.yahoo.com> Hi Histonet Subscriber's, The following forwarded e-mail was from Chuck Churukian. I spoke with his wife Irene last evening and she wished to have this Holiday wish forwarded to the histonet world. Sincerely, Akemi Akemi Allison-Tacha BS, HT (ASCP) HTL President Phoenix Lab Consulting Specializing in Histology, SS, IHC, & TMA Tele: (425) 941-4287 E-Mail: akemiat3377@yahoo.com > Subject: RE: Holiday Greetings > To: akemiat3377@yahoo.com > Date: Friday, December 19, 2008, 5:37 PM > Thank you for your Greeting that included every religion, > every circumstance, situation, > sex orientation, etc. etc. It was very humorous. > Here's our greeting.... As simple as > we are....... > > > > > > Names We Hold Dear > > by Vicki J. Kuyper (paraphrased) > > Every name is a touchstone that leads to a place and time, > Where God has used another?s heart to reach out and > touch mine. (ours) > It may have happened years ago or even yesterday, > But every person on our list has changed our lives some > way. > > Through simple conversation, a warm hug or a shared meal, > Every person on our list has helped us grow or in many > ways- heal, > Or laugh or love or learn or smile?..the blessings never > end > As God allows our paths to cross as family and/or friends. > > So please know that this greeting is more than a Christmas > wish. > It?s a ?thank you? to God for putting on our > list------- > Each and everyone whose name we have come to hold dear?. > Those who have shown us Christmas love ? years passed, > even this year. > Have a wonderful holiday..... Get some rest and > relaxation.... > Our wish and prayer for you: > A Happy Healthy Prosperous New Year > Chuck and Irene From tgenade <@t> gmail.com Mon Dec 22 04:29:26 2008 From: tgenade <@t> gmail.com (Tyrone Genade) Date: Mon Dec 22 04:30:36 2008 Subject: [Histonet] Dear Santa, I would like antiibodies for Christmas Message-ID: Hi all, In light of the fact that it is Christmas, the time for giving, I was wondering if any of you histonet subscribers in the EU might be willing to play Santa Clause. I'm looking for test samples (a few microL) of the following antibodies to see if they are active against the protein equivalents in the fish I'm working with: Nothobranchisu guentheri and furzeri. So far we have had good results using antibodies against other evolutionary well conserved epitopes, such as L1, TAG, GFAP, S100, SMI31 etc.. The antibodies I would like to test are those against: BDNF FoxO3A and FoxO1 c-Rel protein posphatase 2A pCREB Phospho-c-Jun (Ser63) phospho-p53 B-catenin and Integrin Beta1 and the TUC-4 antibody for identification of neural stem cells. Any anti-zebrafish leukocyte antibodies would be great too (can't use the biotinylated lectin in the whole mounts because of the back ground). My Prof is in Germany with his family for the next 2 weeks, so if anyone would like to play Santa, please let me know and I will get his address for you. Thanks! -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@freeshell.org tel: +27-84-632-1925 (c) ******************************************************************************** "For there is one God, and there is one mediator between God and men, the man Christ Jesus, who gave himself as a ransom for all." 1 Timothy 2:5-6 From relia1 <@t> earthlink.net Mon Dec 22 08:59:54 2008 From: relia1 <@t> earthlink.net (Pam Barker) Date: Mon Dec 22 08:59:55 2008 Subject: [Histonet] Happy Holidays!! Message-ID: Hi Histonetters, I just wanted to take a moment of your time and say Happy Holidays. I appreciate your acceptance of me as a member of the listserv. I hope everyone has a very Merry Christmas and a Happy New Year. RELIA Solutions offices will be closed from 12/25-12/29. If you need to reach me I will be available by cell phone at 407-353-5070. 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 <> www.myspace.com/pamatrelia From c.m.vanderloos <@t> amc.uva.nl Mon Dec 22 09:31:28 2008 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Mon Dec 22 09:32:37 2008 Subject: [Histonet] RE: triple staining Message-ID: Dear Anjan,Your plans are fine. Don't worry about the following chromogens:DAB / Vector Blue / Liquid Permanent RedThese chromogens easily survive the second HIER procedure without any changes. Take care of the dilutions of the primary antibody after the second HIER: dilutions might be higher as usual (because of two HIER procedures).Lots of success!Chris van der Loos, Amsterdam, Netherlands Date: Fri, 19 Dec 2008 23:49:57 +0530 From: anjan kumar Subject: [Histonet] (no subject) To: histonet@lists.utsouthwestern.edu hi, I am Dr. Anjan Kumar working as Junior Research Scientist at Madras veterinary college, chennai, india. currently i am pursuing my PhD in veterinary pathology and i am working on canine mammary tumours for my theses work and i have planned to work on triple immunohistochemistry using CD44, CD24, EPCAM AS ALL these antigens are surface markers of stem like cancer cells and they all are co localized. i am finding it very difficult to select 3 different chromogens. i have planned to procure monoclonals from Thermo - Lab visison i.e the following are mouse monoclonals: CD44: IgG2a CD24: IgM / ? ESA: IgG1 / ? i have planned to use 2 detection system i.e alkaline phosphatase and HRP due to funding constraints i am forced to settle for single goat anti - mouse secondary antibody currently im confused with everything and i am struck here And another doubt ... according to multiple immunostaining methods by Chris Vander loos after double immuno histochemistry we can proceed for one more HIER and perform another single immunohistochemistry so this mounts upto triple immunohistochemistry right! coming to the point, my doubt was after performing HIER WILL the chromogen present earlier will be lost along with the immune complex. i have stopped all my work here and dont know how i can proceed... please help me with your valuable suggestions. can any one plz suggest me established triple immunohistochemistry protocol. regards, Dr anjan. From melissafphelan <@t> gmail.com Mon Dec 22 09:47:04 2008 From: melissafphelan <@t> gmail.com (Melissa Phelan) Date: Mon Dec 22 09:48:12 2008 Subject: [Histonet] Full Time Permanent Day Shift Histotech Position in Fort Wayne, IN Message-ID: <181bbd170812220747wa8beb42p71f875a643c85854@mail.gmail.com> Hello, My name is Melissa Phelan and I am the* *Director of Lab/Biotech recruitment here at Healthcare Scouts. My company *specializes* in the * permanent/direct* placement of laboratory professionals like you. We are currently partnered with numerous labs, and hospitals to recruit for their *full time/fully befitted* lab and biotech positions nationwide. I have a position in *Fort Wayne, IN*, and it would be a great fit for you and that you uniquely qualify for. I would like to give you the specifics on this position, and I would also like to talk with you in more detail on exactly what you are looking for, so I can help you find the best opportunity for you. I would also like to give you the opportunity to earn our $1000 referral bonus, if we place someone that you know in a position, we would pay you up to $1000 for referring us to that qualified candidate. You can reach me at any time at 800.708.0605, ext 139 and ask for Melissa. -- Melissa Phelan Laboratory and Biotech Phone: 800.708.0605 ext. 139 Cell: 407-697-1175 UCF KNIGHTS!! From micropathlabs <@t> yahoo.com Mon Dec 22 12:00:48 2008 From: micropathlabs <@t> yahoo.com (Sheila Haas) Date: Mon Dec 22 12:01:47 2008 Subject: [Histonet] Basic Flow Cytometry Message-ID: <189485.92560.qm@web57802.mail.re3.yahoo.com> Hi all. I am looking for a basic course in flow cytometry. Does anyone know of one? We're looking to set up testing in our existing lab and I'm looking to get a head start on training.? ? Sheila Haas Laboratory Supervisor Micro Path Laboratories From Erin.Martin <@t> ucsf.edu Mon Dec 22 13:06:22 2008 From: Erin.Martin <@t> ucsf.edu (Martin, Erin) Date: Mon Dec 22 13:08:16 2008 Subject: [Histonet] Slide drying Message-ID: Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology From jqb7 <@t> cdc.gov Mon Dec 22 13:29:11 2008 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/CCID/NCZVED)) Date: Mon Dec 22 13:29:49 2008 Subject: [Histonet] Slide drying In-Reply-To: References: Message-ID: <1CE1847DFEA0A647B1CCDE4108EA60A70402ACD1@LTA3VS011.ees.hhs.gov> I would suggest filing using the spacer coils and then removing them after 4-7 days. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martin, Erin Sent: Monday, December 22, 2008 2:06 PM To: histonet Subject: [Histonet] Slide drying Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Erin.Martin <@t> ucsf.edu Mon Dec 22 13:45:19 2008 From: Erin.Martin <@t> ucsf.edu (Martin, Erin) Date: Mon Dec 22 13:51:20 2008 Subject: [Histonet] Slide drying References: <005301c9646c$9e937800$dbba6800$@com> Message-ID: Hi all, They want them filed that fast because when they order recuts, stains, etc, they want the entire case set up again with the new orders and since we do 400-500 cassettes/day it would take too long for the clerical staff to look through slide flats to fine the original H&Es. Since they are divided between different docs so the flats are would not follow numerical order... I know it's expecting a lot to think that slides would be ready for file so fast. I needed some other opinions to show them. Thanks, Erin ________________________________ From: Michael Mihalik [mailto:mike@pathview.com] Sent: Mon 12/22/2008 11:36 AM To: Martin, Erin Subject: RE: [Histonet] Slide drying Erin, may I ask why your doctors want them filed so quickly? Michael Mihalik PathView Systems | cell: 214.733.7688 | 800.798.3540 | fax: 270.423.0968 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martin, Erin Sent: Monday, December 22, 2008 1:06 PM To: histonet Subject: [Histonet] Slide drying Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Erin.Martin <@t> ucsf.edu Mon Dec 22 13:56:33 2008 From: Erin.Martin <@t> ucsf.edu (Martin, Erin) Date: Mon Dec 22 13:57:32 2008 Subject: [Histonet] Slide drying References: <1CE1847DFEA0A647B1CCDE4108EA60A70402ACD1@LTA3VS011.ees.hhs.gov> Message-ID: Hi Jeanine, What are spacer coils? I don't think I've ever heard of them. Thanks, Erin Martin UCSF Dermatopathology ________________________________ From: Bartlett, Jeanine (CDC/CCID/NCZVED) [mailto:jqb7@cdc.gov] Sent: Mon 12/22/2008 11:29 AM To: Martin, Erin; histonet Subject: RE: [Histonet] Slide drying I would suggest filing using the spacer coils and then removing them after 4-7 days. Jeanine Bartlett Infectious Diseases Pathology Branch (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martin, Erin Sent: Monday, December 22, 2008 2:06 PM To: histonet Subject: [Histonet] Slide drying Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Mon Dec 22 14:03:41 2008 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Mon Dec 22 14:04:49 2008 Subject: [Histonet] Slide drying References: <005301c9646c$9e937800$dbba6800$@com> Message-ID: <5F31F38C96781A4FBE3196EBC22D47807F286C@fhosxchmb006.ADVENTISTCORP.NET> Our Pathologists hold onto the case slides until the cases are complete. If they order stains or recuts, they hold on to the slides until the specials or recuts have been received. We also have a drying system where we have a set of shelves with spaces designated by 100's and sort the slides according to case number by 100s. They dry until the numbers cycle around. When we have to find a case, we only have to look through 20 trays instead of the whole shebang. Toluene based coverslipped slides usually take three days to dry. The xylene based coverslipped slides take about two weeks. Janet Janet L. Bonner, HTL (ASCP) Pathology Laboratory ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Martin, Erin Sent: Mon 12/22/2008 2:45 PM To: Michael Mihalik; histonet Subject: RE: [Histonet] Slide drying Hi all, They want them filed that fast because when they order recuts, stains, etc, they want the entire case set up again with the new orders and since we do 400-500 cassettes/day it would take too long for the clerical staff to look through slide flats to fine the original H&Es. Since they are divided between different docs so the flats are would not follow numerical order... I know it's expecting a lot to think that slides would be ready for file so fast. I needed some other opinions to show them. Thanks, Erin ________________________________ From: Michael Mihalik [mailto:mike@pathview.com] Sent: Mon 12/22/2008 11:36 AM To: Martin, Erin Subject: RE: [Histonet] Slide drying Erin, may I ask why your doctors want them filed so quickly? Michael Mihalik PathView Systems | cell: 214.733.7688 | 800.798.3540 | fax: 270.423.0968 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Martin, Erin Sent: Monday, December 22, 2008 1:06 PM To: histonet Subject: [Histonet] Slide drying Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ 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/or 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 the sender immediately by replying to this message and deleting the material from any computer. ======================================================= From RSRICHMOND <@t> aol.com Mon Dec 22 14:30:01 2008 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Mon Dec 22 14:30:13 2008 Subject: [Histonet] Re: Dear Santa, I would like antiibodies for Christmas Message-ID: Tyrone Genade notes that he's doing research with the killifish Nothobranchius guentheri. I had some of those in an aquarium a long time ago - what a beautiful fish! - never was successful in breeding them - look them up in Wikipedia. What is their particular research interest? Are they now as easily bred and raised as the much-studied zebrafish, Danio (=Brachydanio) rerio? Bob Richmond Samurai Pathologist and onetime aquarist Knoxville TN From JGREWE <@t> OhioHealth.com Mon Dec 22 15:01:51 2008 From: JGREWE <@t> OhioHealth.com (JGREWE@OhioHealth.com) Date: Mon Dec 22 15:01:54 2008 Subject: [Histonet] Jacquelyn Grewe/Staff/OhioHealth is out of the office . Message-ID: I will be out of the office starting 12/22/2008 and will not return until 12/26/2008. From rjbuesa <@t> yahoo.com Mon Dec 22 15:06:32 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 22 15:07:30 2008 Subject: [Histonet] Slide drying In-Reply-To: Message-ID: <162368.18376.qm@web65712.mail.ac4.yahoo.com> One way to allow faster drying is to get a more fluid mounting medium. If you use it as it comes from the manufacturer it will take longer. Even if it is toluene based, if you add some xylene and get a really fluid mounting medium you will be able to file the slides sooner, although perhaps not as soon as your pathologists want. They are always trying for the histotechs to perform miracles and most of their request stem from not knowing how things work in the histology lab. It is a pity. Ren? J. --- On Mon, 12/22/08, Martin, Erin wrote: From: Martin, Erin Subject: [Histonet] Slide drying To: "histonet" Date: Monday, December 22, 2008, 2:06 PM Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWeems <@t> sjha.org Mon Dec 22 15:20:38 2008 From: JWeems <@t> sjha.org (Weems, Joyce) Date: Mon Dec 22 15:21:56 2008 Subject: [Histonet] Slide drying In-Reply-To: <162368.18376.qm@web65712.mail.ac4.yahoo.com> References: <162368.18376.qm@web65712.mail.ac4.yahoo.com> Message-ID: <5D64396A0D4A5346BEBC759022AAEAA5208A04@ITSSSXM01V6.one.ads.che.org> Richard Allen makes some that dries overnight. It is item # 22 050 102 - sold by ThermoScientific. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Monday, December 22, 2008 4:07 PM To: histonet; Martin, Erin Subject: Re: [Histonet] Slide drying One way to allow faster drying is to get a more fluid mounting medium. If you use it as it comes from the manufacturer it will take longer. Even if it is toluene based, if you add some xylene and get a really fluid mounting medium you will be able to file the slides sooner, although perhaps not as soon as your pathologists want. They are always trying for the histotechs to perform miracles and most of their request stem from not knowing how things work in the histology lab. It is a pity. Ren? J. --- On Mon, 12/22/08, Martin, Erin wrote: From: Martin, Erin Subject: [Histonet] Slide drying To: "histonet" Date: Monday, December 22, 2008, 2:06 PM Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ 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: This email, including any attachments is the property of Catholic Health East and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please reply to the sender that you have received the message in error, then delete this message. From mtitford <@t> aol.com Mon Dec 22 16:05:09 2008 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Mon Dec 22 16:06:17 2008 Subject: [Histonet] Christmas/Dr Margraf Message-ID: <8CB32987D7F8C23-898-238@WEBMAIL-DZ07.sysops.aol.com> Christmas greetings from the Heart of Dixie, and thanks to Dr Margraf,?her collegues and?her facility?for hosting the Histonet. I enjoy keeping up with histotechnology through the Histonet. Mike Titford USA Pathology Mobile AL From Tony_Reilly <@t> health.qld.gov.au Mon Dec 22 17:58:00 2008 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Mon Dec 22 17:59:34 2008 Subject: [Histonet] Slide drying In-Reply-To: References: Message-ID: <4950B627.471C.0039.0@health.qld.gov.au> Hi Erin While the request from your pathologists seems unreasonable from my experience the following steps will give the best results. Use a rapid drying mountant many of which are on the market. Consult your local suppliers. Dry in an incubator as you have been doing but when you remove from the incubator allow the slides to cool before filing or even sorting into piles. The mountant may be dry but the heat will make it sticky gluing the slides together. Just think of new bitumen on a very hot day. All the best. Tony Reilly Chief Scientist Anatomical Pathology Pathology Queensland Level 1, Building 15 Princess Alexandra Hospital Ipswich Rd, Woolloongabba Q 4102 Australia Ph: 07 32402412 Fax:07 32402930 tony_reilly@health.qld.gov.au >>> "Martin, Erin" 23/12/2008 5:06 am >>> Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ 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 strictly prohibited. The information contained in this email, including any attachment sent with 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 collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. ********************************************************************************** From alyssa <@t> alliedsearchpartners.com Mon Dec 22 18:16:53 2008 From: alyssa <@t> alliedsearchpartners.com (Alyssa Peterson) Date: Mon Dec 22 18:16:58 2008 Subject: [Histonet] Histology Positions Message-ID: I wanted to send out a few of our new permanent positions: **If you are interested please send updated resume and/or questions to*: Alyssa@alliedsearchpartners.com * Full Time Permanent/Day Shift-7:30am-3:30pm Location: Phoenix, AZ Facility: Anatomical Pathology Laboratory Required Skills/Education: HT/HTL ASCP Part Time Permanent/Day Shift Location: Suprise, AZ Facility: Anatomical Pathology Laboratory Required Skills/Education: HT/HTL ASCP Full Time Permanent/Day Shift Location: Memphis, TN Facility: Anatomical Pathology Laboratory Required Skills/Education: HT/HTL ASCP Full Time Permanent/Day Shift-5am-1pm Location: Lakeland/Plant City, FL Required Skills/Education: HT/HTL ASCP, FL licensed Full Time Permanent/Day Shift-7:30am-3:30pm Location: Conway/Myrtle Beach, SC Facility: Pathology Lab Required Skills/Education: HT/HTL ASCP AA or BS Degree in the Sciences -- Alyssa Allied Search Partners Alyssa@alliedsearchpartners.com From gu.lang <@t> gmx.at Tue Dec 23 05:56:23 2008 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Dec 23 05:57:24 2008 Subject: [Histonet] formaldehyd fixation - ph Message-ID: Hi dear listmembers, Is it generally accepted, that formaldehyd fixation in an acid environment is less effective than in basic? Or in other words: Does formaldehyde bind to proteins in acid solution to a smaller amount than in basic solutions? I have found a publication (Theis, 1944) where the the amount of bound formaldehyde was plotted against pH. And the line increased with the pH. A few years later Gustavson published that Theis' methods of measurement were wrong and also his findings about the reactionpartners of formaldehyde. But I cannot find a newer paper about the effect of pH on formaldehydfixation. Can anyone help me out? Froehliche Weihnachten! Gudrun Lang From rjbuesa <@t> yahoo.com Tue Dec 23 08:19:33 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 23 08:19:36 2008 Subject: [Histonet] formaldehyd fixation - ph In-Reply-To: Message-ID: <579548.33177.qm@web65709.mail.ac4.yahoo.com> Dear Gudrun: In my paper on formaldehyde that I sent you some days ago, I mention the work by Helander (1994) in Table 2 where it is shown that formaldehyde fixation is faster at pH7 than at pH4 using radioactive carbon (Section"4", pp 390 of my article). Hope this will answer your question. Happy holidays. Ren? J. --- On Tue, 12/23/08, Gudrun Lang wrote: From: Gudrun Lang Subject: [Histonet] formaldehyd fixation - ph To: histonet@lists.utsouthwestern.edu Date: Tuesday, December 23, 2008, 6:56 AM Hi dear listmembers, Is it generally accepted, that formaldehyd fixation in an acid environment is less effective than in basic? Or in other words: Does formaldehyde bind to proteins in acid solution to a smaller amount than in basic solutions? I have found a publication (Theis, 1944) where the the amount of bound formaldehyde was plotted against pH. And the line increased with the pH. A few years later Gustavson published that Theis' methods of measurement were wrong and also his findings about the reactionpartners of formaldehyde. But I cannot find a newer paper about the effect of pH on formaldehydfixation. Can anyone help me out? Froehliche Weihnachten! Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From akbitting <@t> geisinger.edu Tue Dec 23 08:26:21 2008 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Tue Dec 23 08:27:00 2008 Subject: [Histonet] Slide drying In-Reply-To: References: Message-ID: <4950AEBD.2B7F.00C9.0@geisinger.edu> We use MM24 from Surgipath. We file our slides within 24 hours. I don't know if it will work in 12 hours, but maybe it's worth trying. 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 No trees were hurt in the sending of this email However many electrons were severly inconvienienced! >>> "Martin, Erin" 12/22/2008 2:06 PM >>> Hello everyone, I was asked to find out how to dry slides quickly. They are glass coverslipped in an automated coverslipper at the reference lab we use and the our docs want them filed in less than 12 hours from the time they are coverslipped. We have been putting them in a 125 degree C convection oven for a few hours but the slides still get all stuck together in the file. They will not consider film coverslipping. Does anyone else file this quickly? I am grateful for any suggestions! Erin Martin UCSF Dermatopathology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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. -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Bitting, Angela TEL;WORK:570-271-6844 ORG:;Histology EMAIL;WORK;PREF;NGW:AKBITTING@geisinger.edu N:Bitting;Angela END:VCARD From Dave.Pizi <@t> carolinashealthcare.org Tue Dec 23 08:55:48 2008 From: Dave.Pizi <@t> carolinashealthcare.org (Pizi, Dave) Date: Tue Dec 23 08:57:21 2008 Subject: [Histonet] Director Position Opportunity Message-ID: <818181F7CDE8B74EA787B8BC628EACE99D392281@CHS-XCHG-VS-04.Carolinas.org> We have an opportunity for an AP Director in Charlotte, NC Director, Carolinas Laboratory Network Located throughout the Charlotte, NC area, Carolinas HealthCare System is a premier, multi-faceted healthcare organization whose facilities make up one of the largest integrated healthcare systems in the United States. A highly visible opportunity exists for a Director in the Anatomic Pathology and Cytology area of Carolinas Laboratory Network to act as a liaison between hospital administration, the department, and the medical staff. Key responsibilities include: administering department policies and procedures; collecting, generating, analyzing, and distributing statistical reports as required; developing a long-range operational plan relative to services, programs, capital, and human resources planning; evaluating and counseling employees relative to performance, conduct, attendance, and adherence to department/hospital rules and regulations; and following up on error correction and establishing corrective action to address identified problems. Qualified candidates must possess a Bachelor's degree, ASCP certification, and a minimum of five years of experience in a medical laboratory with at least two years of supervisory management experience to include personnel, financial, regulatory, and information systems expertise. An advanced degree, preferably in management, or other advanced management training is desirable. For more information about our System and to register online, please visit: www.carolinashealthcare.org/careers or call (704) 444-3041. EOE/AA ----------------------------------------- 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 aajl7979 <@t> hotmail.com Tue Dec 23 11:20:03 2008 From: aajl7979 <@t> hotmail.com (Amanda L) Date: Tue Dec 23 11:20:10 2008 Subject: [Histonet] Shurwave microwave Message-ID: Happy Holiday's Everyone! I was wondering if any of you use a shurwave microprocessor and might happen to have an extra cassett rack i could borrow for a couple of weeks. Our was "accidently" thrown away. also known as tech not paying attention! I am over it! (not really) Any help would be greatly appreciated. Thanks in Advance Amanda LaFlam, HT (ASCP) Pioneer Valley Urology Group _________________________________________________________________ It?s the same Hotmail?. If by ?same? you mean up to 70% faster. http://windowslive.com/online/hotmail?ocid=TXT_TAGLM_WL_hotmail_acq_broad1_122008 From bturdi <@t> gmail.com Tue Dec 23 11:23:40 2008 From: bturdi <@t> gmail.com (subat turdi) Date: Tue Dec 23 11:24:42 2008 Subject: [Histonet] Please unsubscribe Message-ID: <4e61b8250812230923n3fc697derf0f39bf150900724@mail.gmail.com> Please unsubscribe me from the list. From elizabeth.heimrich <@t> bms.com Tue Dec 23 11:31:34 2008 From: elizabeth.heimrich <@t> bms.com (Elizabeth M Heimrich) Date: Tue Dec 23 11:31:49 2008 Subject: [Histonet] Re: Myelin stain Message-ID: <49512076.2050603@bms.com> Nicole, You can try cresyl echt violet. It preserves RNA/ DNA well. Ambion has a kit, AM1935 that we have used before. For LCM we use it on frozen tissue, though. Good luck I hope this helps! ~ Beth From jhaviland <@t> mdanderson.org Tue Dec 23 14:23:19 2008 From: jhaviland <@t> mdanderson.org (Haviland,Joie) Date: Tue Dec 23 14:24:54 2008 Subject: [Histonet] Understained my slides with hematoxylin Message-ID: Hello: I was doing H&E's and it was pointed out that I had understained the hematoxylin. But the eosin was ok. How does one go back and restain the slides and remove the old stain? Or do you use a differentiating solution? Thanks for your help Happy Holidays Joie From HornHV <@t> archildrens.org Tue Dec 23 14:31:45 2008 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Tue Dec 23 14:31:50 2008 Subject: [Histonet] charcot-leyden crystals Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82F66@EMAIL.archildrens.org> Can someone tell me the best stain to use for Charcot Leyden crystals? We are looking for them in feces. Thanks. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall Slot 820 Little Rock, AR 72202 phone 501.364.4240 fax 501.364.3155 visit us on the web at: www.archildrens.org ****************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************** 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 marktarango <@t> gmail.com Tue Dec 23 14:40:23 2008 From: marktarango <@t> gmail.com (Mark Tarango) Date: Tue Dec 23 14:40:27 2008 Subject: [Histonet] Full Time Permanent Day Shift Histotech Position in Fort Wayne, IN In-Reply-To: <181bbd170812220747wa8beb42p71f875a643c85854@mail.gmail.com> References: <181bbd170812220747wa8beb42p71f875a643c85854@mail.gmail.com> Message-ID: <5b6eb13e0812231240n60750e7as5cb3978580d8dc6b@mail.gmail.com> I work with a tech that you, Melissa, said was getting $1,000 after her placement. After five months, multiple phone calls, and letter writing she only got $500. You wouldn't get back to her. Mark On Mon, Dec 22, 2008 at 7:47 AM, Melissa Phelan wrote: > Hello, > > > > My name is Melissa Phelan and I am the* *Director of Lab/Biotech > recruitment > here at Healthcare Scouts. My company *specializes* in the * > permanent/direct* placement of laboratory professionals like you. We are > currently partnered with numerous labs, and hospitals to recruit for > their *full > time/fully befitted* lab and biotech positions nationwide. I have a > position > in *Fort Wayne, IN*, and it would be a great fit for you and that you > uniquely qualify for. I would like to give you the specifics on this > position, and I would also like to talk with you in more detail on exactly > what you are looking for, so I can help you find the best opportunity for > you. I would also like to give you the opportunity to earn our $1000 > referral bonus, if we place someone that you know in a position, we would > pay you up to $1000 for referring us to that qualified candidate. You can > reach me at any time at 800.708.0605, ext 139 and ask for Melissa. > > > -- > Melissa Phelan > > Laboratory and Biotech > > Phone: 800.708.0605 ext. 139 > > Cell: 407-697-1175 > > UCF KNIGHTS!! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From AnthonyH <@t> chw.edu.au Tue Dec 23 16:21:50 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 23 16:22:02 2008 Subject: [Histonet] Understained my slides with hematoxylin In-Reply-To: Message-ID: Joe, I would make sure all the mountant is removed. A good soak in xylene and then a rinse in alcohol, if the mountant has not fully disappeared, you will see a white opacity on the section. Rinse in water and then re-stain as usual. The eosin will tend to leach out in the water rinses and the differentiation solution (after the haematoxylin - if you use it). Check the sections after blueing. If it looks too weak, increase the Hx staining time &/or decrease the differentiation step. Counterstain with eosin as usual and you should be right. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Haviland,Joie Sent: Wednesday, 24 December 2008 7:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Understained my slides with hematoxylin Hello: I was doing H&E's and it was pointed out that I had understained the hematoxylin. But the eosin was ok. How does one go back and restain the slides and remove the old stain? Or do you use a differentiating solution? Thanks for your help Happy Holidays Joie _______________________________________________ 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 note 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 Michelle.Perrins <@t> uct.ac.za Wed Dec 24 00:36:07 2008 From: Michelle.Perrins <@t> uct.ac.za (Michelle Perrins) Date: Wed Dec 24 00:36:22 2008 Subject: [Histonet] merry christmas Message-ID: <4951F476.A704.0070.0@uct.ac.za> To all histonetters around the world, may you have a safe and happy festive holiday. Best wishes for 2009. Regards Michelle from down south in Africa. Michelle Perrins Chief Medical Technologist Forensic Pathology Services Division Forensic Medicine Faculty Health Sciences University of Cape Town tel: +27 21 406 6001 fax: +27 21 448 1249 Email: Michelle.Perrins@uct.ac.za From aj.taylor <@t> blueyonder.co.uk Wed Dec 24 08:01:51 2008 From: aj.taylor <@t> blueyonder.co.uk (alan taylor) Date: Wed Dec 24 08:02:51 2008 Subject: [Histonet] Charcot-Leyden crystals Message-ID: <76E8F29215084951BAA9CDA93FDD2544@merlin> Hazel Charcot-Leyden crystals are frequent findings in faecal samples and sputum submitted for microscopical study, the crystals are easily demonstrated,if present, by making a simple 'wet' prep and adding a drop of Gram's Iodine under the coverslip. Charcot-Leyden crystals in faecal submissions should lead the investigator to seek the presence of parasitic infection from particularly helminthiasis. Note that Charcot-Leyden crystals are virtually absent from samples of bacillary dysentery, but are frequently found in samples of faeces containing the various amoeba spp. Several other intestinal parasitic worm species result in the presence of Charcot-Leyden crystals. Within sputum Charcot-Leyden crystals are found in cases submitted from bronchial asthma sufferers, the crystals are often found within Curschmann's spirals that are a feature of this disease. Lung diseases such as Paragonimus ringeri & P westermani , typically found in the region of the Phillipines and Japan, are fluke diseases that result in the presence of Charcot-Leyden crystals from the haemoptic sputum. The crystals have also been found in pus from amoebic liver abcesses that have found their route of discharge through the lungs. Hope this info is helpful to you. A very happy Christmas & New Year to everybody out there in Histoland. Alan Taylor Microtechnical Services Exeter Devon England From carrolpb <@t> umdnj.edu Wed Dec 24 08:34:40 2008 From: carrolpb <@t> umdnj.edu (Peter Carroll) Date: Wed Dec 24 08:34:51 2008 Subject: [Histonet] Understained my slides with hematoxylin In-Reply-To: References: Message-ID: <49524880.2060806@umdnj.edu> > The eosin will tend to leach out in the water rinses and the differentiation solution a touch of acid alcohol doesnt hurt either :) From gp62 <@t> georgetown.edu Wed Dec 24 09:05:31 2008 From: gp62 <@t> georgetown.edu (Guillermo Palchik) Date: Wed Dec 24 09:06:31 2008 Subject: [Histonet] TUNEL on fixed brain tissue Message-ID: <7814E917-472D-4179-8AF7-C946D4F2F609@georgetown.edu> Dear Histologists, Does anyone have a good protocol for doing TUNEL on fixed brain slides? Thanks Guillermo Palchik gp62@georgetown.edu From tifei <@t> foxmail.com Wed Dec 24 12:48:10 2008 From: tifei <@t> foxmail.com (=?utf-8?B?VEY=?=) Date: Wed Dec 24 12:49:21 2008 Subject: =?utf-8?B?UmU6IFJlOiBbSGlzdG9uZXRdIENhcGlsbGFyeSBjb3VudGluZw==?= References: <3742.64276.qm@web65701.mail.ac4.yahoo.com> Message-ID: <200812250248050584035@foxmail.com> So any one can provide some cell makers for staining of blood vessels? I only know CD31. 2008-12-25 TF ???? Rene J Buesa ????? 2008-12-19 04:53:43 ???? histonet; Nejat Yilmaz ??? ??? Re: [Histonet] Capillary counting Nejat: There are several stains for capillaries: 1- Campbell and Alexander (tested by Mallory,1938): black agsinst colorless bbackground. 2- Doherty, Suk and Alexander, 1938: differential staining of blood in capillaries. 3- Pickworth, 1934: blood vessels (capillaries) in thick sections of brain. 4- Slominski and Cunge (tested by Romeis, 1948): differential staining of capillaries, and 5- Ziegler 1945 (tested by Riley, 1945): demonstration of capillaries in wholemounts of cornea. Although developed for other tissues it is very likely that they will work on skeletal muscle. Ren?J. --- On Thu, 12/18/08, Nejat Yilmaz wrote: From: Nejat Yilmaz Subject: [Histonet] Capillary counting To: histonet@lists.utsouthwestern.edu Date: Thursday, December 18, 2008, 2:26 PM Hi All, We need a protocol demonstrating capillaries in sceletal muscle tissue other than immunohistochemistry. We're planning to compare number of capillaries between different experiment groups for evaluating angiogenesis. Is there any special stain to suggest for this purpose? Thanks in advance. Necat Yilmaz _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From michelle.steinkrauss <@t> novartis.com Wed Dec 24 12:52:07 2008 From: michelle.steinkrauss <@t> novartis.com (michelle.steinkrauss@novartis.com) Date: Wed Dec 24 12:53:11 2008 Subject: [Histonet] Michelle Steinkrauss is out of the office. Message-ID: I will be out of the office starting 12/24/2008 and will not return until 01/05/2009. I will respond to your message when I return. Happy Holidays! From anh2006 <@t> med.cornell.edu Wed Dec 24 13:12:17 2008 From: anh2006 <@t> med.cornell.edu (Andrea Hooper) Date: Wed Dec 24 13:12:30 2008 Subject: [Histonet] Capillary counting In-Reply-To: <200812250248050584035@foxmail.com> References: <3742.64276.qm@web65701.mail.ac4.yahoo.com> <200812250248050584035@foxmail.com> Message-ID: What animal? What organ? In frozen or paraffin? >So any one can provide some cell makers for staining of blood vessels? >I only know CD31. > > >2008-12-25 > > >TF > -- From rjbuesa <@t> yahoo.com Wed Dec 24 13:42:29 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 24 13:43:27 2008 Subject: [Histonet] Capillary counting In-Reply-To: <200812250248050584035@foxmail.com> Message-ID: <68132.27902.qm@web65705.mail.ac4.yahoo.com> Use Ulex europaeus lectins. Ren? J. --- On Wed, 12/24/08, TF wrote: From: TF Subject: Re: Re: [Histonet] Capillary counting To: "rjbuesa" , "histonet" , "Nejat Yilmaz" Date: Wednesday, December 24, 2008, 1:48 PM ? _filtered #yiv1672193670 { font-family:??; } _filtered #yiv1672193670 { font-family:Verdana; } _filtered #yiv1672193670 { } _filtered #yiv1672193670 {margin:72.0pt 90.0pt 72.0pt 90.0pt;} #yiv1672193670 P.MsoNormal { TEXT-JUSTIFY:inter-ideograph;FONT-SIZE:10.5pt;MARGIN:0cm 0cm 0pt;FONT-FAMILY:"Times New Roman";TEXT-ALIGN:justify;} #yiv1672193670 LI.MsoNormal { TEXT-JUSTIFY:inter-ideograph;FONT-SIZE:10.5pt;MARGIN:0cm 0cm 0pt;FONT-FAMILY:"Times New Roman";TEXT-ALIGN:justify;} #yiv1672193670 DIV.MsoNormal { TEXT-JUSTIFY:inter-ideograph;FONT-SIZE:10.5pt;MARGIN:0cm 0cm 0pt;FONT-FAMILY:"Times New Roman";TEXT-ALIGN:justify;} #yiv1672193670 A:link { COLOR:blue;TEXT-DECORATION:underline;} #yiv1672193670 SPAN.MsoHyperlink { COLOR:blue;TEXT-DECORATION:underline;} #yiv1672193670 A:visited { COLOR:purple;TEXT-DECORATION:underline;} #yiv1672193670 SPAN.MsoHyperlinkFollowed { COLOR:purple;TEXT-DECORATION:underline;} #yiv1672193670 SPAN.EmailStyle17 { FONT-WEIGHT:normal;COLOR:windowtext;FONT-STYLE:normal;FONT-FAMILY:Verdana;TEXT-DECORATION:none;} #yiv1672193670 DIV.Section1 { } #yiv1672193670 UNKNOWN { FONT-SIZE:10pt;} #yiv1672193670 BLOCKQUOTE { MARGIN-TOP:0px;MARGIN-BOTTOM:0px;MARGIN-LEFT:2em;} #yiv1672193670 OL { MARGIN-TOP:0px;MARGIN-BOTTOM:0px;} #yiv1672193670 UL { MARGIN-TOP:0px;MARGIN-BOTTOM:0px;} So any one can provide some cell makers for staining of blood vessels? I only know CD31. ? ? 2008-12-25 TF ???? Rene J Buesa ????? 2008-12-19? 04:53:43 ???? histonet; Nejat Yilmaz ??? ??? Re: [Histonet] Capillary counting Nejat: There?are?several?stains?for?capillaries: 1-?Campbell?and?Alexander?(tested?by?Mallory,1938):?black?agsinst?colorless?bbackground. 2-?Doherty,?Suk?and?Alexander,?1938:?differential?staining?of?blood?in?capillaries. 3-?Pickworth,?1934:?blood?vessels?(capillaries)?in?thick?sections?of?brain. 4-?Slominski?and?Cunge?(tested?by?Romeis,?1948):?differential?staining?of?capillaries,?and 5-?Ziegler?1945?(tested?by?Riley,?1945):?demonstration?of?capillaries?in?wholemounts?of?cornea. Although?developed?for?other?tissues?it?is?very?likely?that?they?will?work?on?skeletal?muscle. Ren?J. ---?On?Thu,?12/18/08,?Nejat?Yilmaz??wrote: From:?Nejat?Yilmaz? Subject:?[Histonet]?Capillary?counting To:?histonet@lists.utsouthwestern.edu Date:?Thursday,?December?18,?2008,?2:26?PM Hi?All, We?need?a?protocol?demonstrating?capillaries?in?sceletal?muscle?tissue?other than?immunohistochemistry.?We're?planning?to?compare?number?of?capillaries between?different?experiment?groups?for?evaluating?angiogenesis.?Is?there?any special?stain?to?suggest?for?this?purpose? Thanks?in?advance. Necat?Yilmaz? _______________________________________________ Histonet?mailing?list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ?????? _______________________________________________ Histonet?mailing?list Histonet@lists.utsouthwestern.edu From atunde90 <@t> yahoo.com Wed Dec 24 18:48:32 2008 From: atunde90 <@t> yahoo.com (Ade Tunde) Date: Wed Dec 24 18:48:38 2008 Subject: [Histonet] Re: Understained my slides with hematoxylin Message-ID: <32466.78437.qm@web46110.mail.sp1.yahoo.com> Hi H&E Stain is very important in the Diagnosis. understaining can be caused by many factors, check the age of your hematoxylin or the concentration of your differentiating solution. Then to restain the stained slide .place the stained slide in 1%? acid alcohol for 2-3minutes and rinse well in distilled water. then restain in hematoxylin for the required time.You should be fine . ? ? Tunde? Ajibade? BS, HTL (ASCP) Histology lab Newark beth Israel Medical Center. Newark, NJ ? ________________________________ From: "histonet-request@lists.utsouthwestern.edu" To: histonet@lists.utsouthwestern.edu Sent: Wednesday, December 24, 2008 1:02:18 PM Subject: Histonet Digest, Vol 61, Issue 35 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. Understained my slides with hematoxylin (Haviland,Joie) ? 2. charcot-leyden crystals (Horn, Hazel V) ? 3. Re: Full Time Permanent Day Shift Histotech Position in??? Fort ? ? ? Wayne, IN (Mark Tarango) ? 4. RE: Understained my slides with hematoxylin (Tony Henwood) ? 5. merry christmas (Michelle Perrins) ? 6. Charcot-Leyden crystals (alan taylor) ? 7. Re: Understained my slides with hematoxylin (Peter Carroll) ? 8. TUNEL on fixed brain tissue (Guillermo Palchik) ---------------------------------------------------------------------- Message: 1 Date: Tue, 23 Dec 2008 14:23:19 -0600 From: "Haviland,Joie" Subject: [Histonet] Understained my slides with hematoxylin To: "histonet@lists.utsouthwestern.edu" ??? Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello: I was doing H&E's and it was pointed out that I had understained the hematoxylin. But the eosin was ok. How does one go back and restain the slides and remove the old stain?? Or do you use a differentiating solution? Thanks for your help Happy Holidays Joie ------------------------------ Message: 2 Date: Tue, 23 Dec 2008 14:31:45 -0600 From: "Horn, Hazel V" Subject: [Histonet] charcot-leyden crystals To: Message-ID: ??? <9AE8AA9E1F644B4AA6C155FB6FD51C6317D82F66@EMAIL.archildrens.org> Content-Type: text/plain;??? charset="us-ascii" Can someone tell me the best stain to use for Charcot Leyden crystals? We are looking for them in feces.? Thanks. Hazel Horn Hazel Horn, HT/HTL (ASCP) Supervisor of Histology Arkansas Children's Hospital 800 Marshall? ? Slot 820 Little Rock, AR? 72202 phone? 501.364.4240 fax? ? ? ? 501.364.3155 visit us on the web at:? ? www.archildrens.org ****************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************************** 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.. ------------------------------ Message: 3 Date: Tue, 23 Dec 2008 12:40:23 -0800 From: "Mark Tarango" Subject: Re: [Histonet] Full Time Permanent Day Shift Histotech ??? Position in??? Fort Wayne, IN To: "Melissa Phelan" Cc: histonet@lists.utsouthwestern.edu Message-ID: ??? <5b6eb13e0812231240n60750e7as5cb3978580d8dc6b@mail.gmail.com> Content-Type: text/plain; charset=ISO-8859-1 I work with a tech that you, Melissa, said was getting $1,000 after her placement.? After five months, multiple phone calls, and letter writing she only got $500.? You wouldn't get back to her. Mark On Mon, Dec 22, 2008 at 7:47 AM, Melissa Phelan wrote: > Hello, > > > > My name is Melissa Phelan and I am the* *Director of Lab/Biotech > recruitment > here at Healthcare Scouts.? My company *specializes* in the * > permanent/direct* placement of laboratory professionals like you. We are > currently partnered with numerous labs, and hospitals to recruit for > their *full > time/fully befitted* lab and biotech positions nationwide. I have a > position > in *Fort Wayne, IN*, and it would be a great fit for you and that you > uniquely qualify for. I would like to give you the specifics on this > position, and I would also like to talk with you in more detail on exactly > what you are looking for, so I can help you find the best opportunity for > you. I would also like to give you the opportunity to earn our $1000 > referral bonus, if we place someone that you know in a position, we would > pay you up to $1000 for referring us to that qualified candidate. You can > reach me at any time at 800.708.0605, ext 139 and ask for Melissa. > > > -- > Melissa Phelan > > Laboratory and Biotech > > Phone: 800.708.0605 ext. 139 > > Cell: 407-697-1175 > > UCF KNIGHTS!! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 4 Date: Wed, 24 Dec 2008 09:21:50 +1100 From: "Tony Henwood" Subject: RE: [Histonet] Understained my slides with hematoxylin To: "Haviland,Joie" , ??? Message-ID: Content-Type: text/plain; charset="us-ascii" Joe, I would make sure all the mountant is removed. A good soak in xylene and then a rinse in alcohol, if the mountant has not fully disappeared, you will see a white opacity on the section. Rinse in water and then re-stain as usual. The eosin will tend to leach out in the water rinses and the differentiation solution (after the haematoxylin - if you use it). Check the sections after blueing.. If it looks too weak, increase the Hx staining time &/or decrease the differentiation step. Counterstain with eosin as usual and you should be right. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Haviland,Joie Sent: Wednesday, 24 December 2008 7:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Understained my slides with hematoxylin Hello: I was doing H&E's and it was pointed out that I had understained the hematoxylin. But the eosin was ok. How does one go back and restain the slides and remove the old stain?? Or do you use a differentiating solution? Thanks for your help Happy Holidays Joie _______________________________________________ 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 note 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. ********************************************************************** ------------------------------ Message: 5 Date: Wed, 24 Dec 2008 08:36:07 +0200 From: "Michelle Perrins" Subject: [Histonet] merry christmas To: Message-ID: <4951F476.A704.0070.0@uct.ac.za> Content-Type: text/plain; charset=US-ASCII To all histonetters around the world, may you have a safe and happy festive holiday.. Best wishes for 2009. Regards Michelle from down south in Africa. Michelle Perrins Chief Medical Technologist Forensic Pathology Services Division Forensic Medicine Faculty Health Sciences University of Cape Town tel: +27 21 406 6001 fax: +27 21 448 1249 Email: Michelle.Perrins@uct.ac.za ------------------------------ Message: 6 Date: Wed, 24 Dec 2008 14:01:51 -0000 From: "alan taylor" Subject: [Histonet] Charcot-Leyden crystals To: Message-ID: <76E8F29215084951BAA9CDA93FDD2544@merlin> Content-Type: text/plain;??? charset="iso-8859-1" Hazel Charcot-Leyden crystals are frequent findings in faecal samples and sputum submitted for microscopical study, the crystals are easily demonstrated,if present,? by making a simple 'wet' prep and adding a drop of Gram's Iodine under the coverslip. Charcot-Leyden crystals in faecal submissions should lead the investigator to seek the presence of parasitic infection from particularly helminthiasis. Note that Charcot-Leyden crystals are virtually absent from samples of bacillary dysentery, but are frequently found in samples of faeces containing the various amoeba spp. Several other intestinal parasitic worm species result in the presence of Charcot-Leyden crystals. Within sputum Charcot-Leyden crystals are found in cases submitted from bronchial asthma sufferers, the crystals are often found within Curschmann's spirals that are a feature of this disease. Lung diseases such as Paragonimus ringeri & P westermani , typically found in the region of the Phillipines and Japan, are fluke diseases that result in the presence of Charcot-Leyden crystals from the haemoptic sputum.. The crystals have also been found in pus from amoebic liver abcesses that have found their route of discharge through the lungs. Hope this info is helpful to you. A very happy Christmas & New Year to everybody out there in Histoland. Alan Taylor Microtechnical Services Exeter Devon England ------------------------------ Message: 7 Date: Wed, 24 Dec 2008 09:34:40 -0500 From: Peter Carroll Subject: Re: [Histonet] Understained my slides with hematoxylin Cc: histonet@lists.utsouthwestern.edu Message-ID: <49524880.2060806@umdnj.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed > The eosin will tend to leach out in the water rinses and the differentiation solution a touch of acid alcohol doesnt hurt either :) ------------------------------ Message: 8 Date: Wed, 24 Dec 2008 10:05:31 -0500 From: Guillermo Palchik Subject: [Histonet] TUNEL on fixed brain tissue To: histonet@lists.utsouthwestern.edu Message-ID: <7814E917-472D-4179-8AF7-C946D4F2F609@georgetown.edu> Content-Type: text/plain; charset=US-ASCII; format=flowed Dear Histologists, Does anyone have a good protocol for doing TUNEL on fixed brain slides? Thanks Guillermo Palchik gp62@georgetown.edu ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 61, Issue 35 **************************************** From pruegg <@t> ihctech.net Thu Dec 25 16:12:21 2008 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Dec 25 16:12:32 2008 Subject: [Histonet] Capillary counting In-Reply-To: References: <3742.64276.qm@web65701.mail.ac4.yahoo.com><200812250248050584035@foxmail.com> Message-ID: <83220BA7EB184A1B8CF2D0796E7AD66C@ihctechq9h2qof> Vegf, cd34, sma, factor 8, CD133 can pick up early endothelial cells (stem cells), there are several others I have been working with researchers on, contact me directly after the holidays. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Hooper Sent: Wednesday, December 24, 2008 12:12 PM To: tifei@foxmail.com; Histonet Subject: Re: Re: [Histonet] Capillary counting What animal? What organ? In frozen or paraffin? >So any one can provide some cell makers for staining of blood vessels? >I only know CD31. > > >2008-12-25 > > >TF > -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Dec 25 16:15:05 2008 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Dec 25 16:16:18 2008 Subject: [Histonet] TUNEL on fixed brain tissue In-Reply-To: <7814E917-472D-4179-8AF7-C946D4F2F609@georgetown.edu> References: <7814E917-472D-4179-8AF7-C946D4F2F609@georgetown.edu> Message-ID: <3687D7C608914779B98E863DA122A66A@ihctechq9h2qof> Human brain? I prefer Cleaved Caspase 3 to Tunnel, in my experience Tunnel does not distinguish apoptosis from necrosis but CC3 will only label cells under going apoptosis. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Guillermo Palchik Sent: Wednesday, December 24, 2008 8:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] TUNEL on fixed brain tissue Dear Histologists, Does anyone have a good protocol for doing TUNEL on fixed brain slides? Thanks Guillermo Palchik gp62@georgetown.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nyilmaz <@t> mersin.edu.tr Fri Dec 26 14:39:46 2008 From: nyilmaz <@t> mersin.edu.tr (Nejat Yilmaz) Date: Fri Dec 26 14:40:52 2008 Subject: [Histonet] Thanks for capillary stain Message-ID: <001401c9679a$17624330$2101a8c0@nejat1> Dear All, I'm very grateful for your kindly efforts. We've decided to stain our tissues with PAS. We couldn't perform IHC due to time limitations with the study. Because it usually takes about 2 months obtaining appropriate antibodies here. Fortunately I'm a member of this society. I wish you a merry christmas and happy holidays... Necat Yilmaz From tifei <@t> foxmail.com Fri Dec 26 22:57:02 2008 From: tifei <@t> foxmail.com (=?utf-8?B?VEY=?=) Date: Fri Dec 26 22:58:12 2008 Subject: =?utf-8?B?UmU6IFtIaXN0b25ldF0gVGhhbmtzIGZvciBjYXBpbGxhcnkgc3RhaW4=?= References: <001401c9679a$17624330$2101a8c0@nejat1> Message-ID: <200812271256575182728@foxmail.com> are there any useful histological techniques to stain capillary/blood vessel in brains? and permit co-immunostaining of other cell markers? PAS will stain all connective tissues, especially the myelin, thus you have to interpret your data well....? 2008-12-27 TF ???? Nejat Yilmaz ????? 2008-12-27 04:44:34 ???? histonet@lists.utsouthwestern.edu ??? ??? [Histonet] Thanks for capillary stain Dear All, I'm very grateful for your kindly efforts. We've decided to stain our tissues with PAS. We couldn't perform IHC due to time limitations with the study. Because it usually takes about 2 months obtaining appropriate antibodies here. Fortunately I'm a member of this society. I wish you a merry christmas and happy holidays... Necat Yilmaz _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu From drvet_anjan <@t> hotmail.com Sat Dec 27 02:06:11 2008 From: drvet_anjan <@t> hotmail.com (anjan kumar) Date: Sat Dec 27 02:06:15 2008 Subject: [Histonet] RE:antigen retrieval In-Reply-To: References: Message-ID: hi, can any one tell me which is the best method used for HIER. MICROWAVE OVEN OR PRESSURE COOKER METHOD. What kind of solutions should we use for epitope retrieval? _________________________________________________________________ Find a better job. We have plenty. Visit MSN Jobs http://www.in.msn.com/jobs From tgenade <@t> gmail.com Sat Dec 27 05:29:18 2008 From: tgenade <@t> gmail.com (Tyrone Genade) Date: Sat Dec 27 05:29:31 2008 Subject: [Histonet] Re: Dear Santa, I would like antiibodies for Christmas Message-ID: Hi Bob, > From: "Robert Richmond" > Subject: [Histonet] Re: Dear Santa, I would like antiibodies for > What is their particular research interest? Are they now as easily > bred and raised as the much-studied zebrafish, Danio (=Brachydanio) > rerio? I'm doing aging research. Go to Pubmed and search with "markofsky guentheri" and see what has already been done with these stunning little fish. If you search with "furzeri" you will get some more interesting hits. Two of the hits will be "my" articles. In the Current Biol article we report on the life-prolonging effects of resveratrol on N. furzeri. I've tried to set up the furzeri in my lab here but for some bizaar reason I just can't get them going here. I can hatch and rear every other Nothobranchius species but furzeri---and it isn't like the furzeri are the most difficult to keep and breed that I have. (The biggest joke is that I went to Italy 4 years back specially to run the fishrom there and spawn these guys! Now I can't do it in my own back yard.) In any case, I had hundreds of guentheri and figured I would "play" with them until I can find out what is going wrong with the furzeri. In the mean time the furzeri have been put on the back burner in favour of the guentheri. The guentheri live longer (about 9 months for my strain) and develop other interesting pathologies other than dementia (like furzeri) such as kidney and liver neoplasmas and degeneration. They also show neurodegeneraton, and have got some stunning images with the confocal microscope on whole-mounts. You can watch the nerve tracts go from neatly organized tracts at 12 weeks, to disorganised at 26 weeks, and then a shambles at 34 weeks and the Smi31, instead of showing a solid nerve, shows a string of Smi31 + "beads" where a nerve tract used to be. We are busy getting all our "ducks in a row" in preparation of a publication on this. Next step is to dose with resveratrol and see what changes. This is where those antibodies I mentioned come in. These are against proteins/epitopes which are known to change with age and in response to resveratrol in tissue cutlure. i would like to see if such changes can be visualized in situ. How resveratrol actually works in situ, and just what cellular effects there are, are not really known in a vertebrate lifespan extension context. One important question, coming from some mouse work, is whether resveratrol keeps the organism young or just prevents death. We have some nice markers of a "young" brain which can use to show whether it is keeping the brain young, restoring it to a young state, or just preventing further neurodegeneration. Its effect on muscle loss with age, liver and kidney tumour incidence would also be interesting. Well, that is my story... I would love to get hold of some antibodies to those epitopes to test on guentheri sections. The cost of antibodies here in Sount Africa are astronomical! I can't afford to blow R7000 on an anitbody to see if it works. Kind regards -- Tyrone Genade http://tgenade.freeshell.org email: tgenade@freeshell.org tel: +27-84-632-1925 (c) ******************************************************************************** "For there is one God, and there is one mediator between God and men, the man Christ Jesus, who gave himself as a ransom for all." 1 Timothy 2:5-6 From rjbuesa <@t> yahoo.com Sat Dec 27 09:14:41 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Dec 27 09:14:45 2008 Subject: [Histonet] RE:antigen retrieval In-Reply-To: Message-ID: <61086.23812.qm@web65704.mail.ac4.yahoo.com> There is not such a thing as the "best" method. Microwave, pressure cooker and steamer (the one I prefer) depends on the epitope you are trying to detect and your general protocol. You have to select a method that is good for all your purposes, trying to have different HIER methods for different epitopes is complicated and unpractical for a high volume diagnostic laboratory. As to the solution it should be a buffer and its pH will depend on the epitope you are trying to detect. Go to the DAKO web site and you will find there good methods and protocols. Ren? J. --- On Sat, 12/27/08, anjan kumar wrote: From: anjan kumar Subject: [Histonet] RE:antigen retrieval To: "triple immunohistochem" Date: Saturday, December 27, 2008, 3:06 AM hi, can any one tell me which is the best method used for HIER. MICROWAVE OVEN OR PRESSURE COOKER METHOD. What kind of solutions should we use for epitope retrieval? _________________________________________________________________ Find a better job. We have plenty. Visit MSN Jobs http://www.in.msn.com/jobs _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tifei <@t> foxmail.com Mon Dec 29 02:10:59 2008 From: tifei <@t> foxmail.com (TF) Date: Mon Dec 29 02:11:19 2008 Subject: [Histonet] IgG antibody & IgM antibody Message-ID: <200812291610540665737@foxmail.com> Dear all; Just wonder anyone can tell me the differences between the two kinds of antibodies? 1. Are we mostly using IgG antibodies? 2. Can I use a IgG secondary antibody to visualize the IgM primary antibody? - It works in my case. 2008-12-29 TF From SARAH.REEVES <@t> ekht.nhs.uk Mon Dec 29 05:54:41 2008 From: SARAH.REEVES <@t> ekht.nhs.uk (SARAH REEVES) Date: Mon Dec 29 05:59:21 2008 Subject: [Histonet] Does any body know what causes nuclear bubbling and how it can be avoided? Message-ID: <4958BA81020000B5000035D4@ekhgwia.ekht.nhs.uk> Does any body know what causes nuclear bubbling and how it can be avoided? ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** From Jane.Moose <@t> NewberryHospital.net Mon Dec 29 08:10:33 2008 From: Jane.Moose <@t> NewberryHospital.net (Jane C. Moose) Date: Mon Dec 29 08:15:40 2008 Subject: [Histonet] processor Message-ID: <8BB5FC36DDA373488E60177757BBD698457EC4@ncmhexchbe01.NewberryHospital.net> I know this question repeatedly occurs, but our tissue processor "died" last week. We are a small hospital lab doing about 2500 cases a year (6000 specimens.) Would anyone have a recommendation on a new processor for our size lab? An Excelsior was brought in for demo (not sure what model) but that one was being discontinued and we were waiting to see the new model. We were also waiting on a "new" model VIP to bring in for demo-but that has not happened yet. Any advice would be greatly appreciated. Thanks Jane Jane Moose LIS Coordinator Newberry County Memorial Hospital Newberry, SC 29108 P-803-405-7129 F- 803-405-7474 From godsgalnow <@t> aol.com Mon Dec 29 08:58:19 2008 From: godsgalnow <@t> aol.com (godsgalnow@aol.com) Date: Mon Dec 29 08:58:27 2008 Subject: [Histonet] ethylene glycol Message-ID: <8CB37DD05973C37-122C-1BC9@WEBMAIL-MY22.sysops.aol.com> Does anyone use this in their microwave process and at which point? In my experience, this is used as a fixative.? But does anyone use it after the reagent alcohol and mix it with isopropyl? Roxanne From rjbuesa <@t> yahoo.com Mon Dec 29 09:20:25 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 29 09:21:25 2008 Subject: [Histonet] Does any body know what causes nuclear bubbling and how it can be avoided? In-Reply-To: <4958BA81020000B5000035D4@ekhgwia.ekht.nhs.uk> Message-ID: <618524.82043.qm@web65711.mail.ac4.yahoo.com> Hi Sarah: There is an article published in the JOH some years ago that I am sending to you?under separate cover. They demonstrate that nuclear bubbling?appear when sections that are not completely drained are placed in the over at more than 60?C. The sections have to be completely drained before heating. Regards Ren? J. --- On Mon, 12/29/08, SARAH REEVES wrote: From: SARAH REEVES Subject: [Histonet] Does any body know what causes nuclear bubbling and how it can be avoided? To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 6:54 AM Does any body know what causes nuclear bubbling and how it can be avoided? ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sjromey <@t> bellsouth.net Mon Dec 29 11:59:22 2008 From: sjromey <@t> bellsouth.net (sjromey@bellsouth.net) Date: Mon Dec 29 12:00:21 2008 Subject: [Histonet] H&E slide stainer Message-ID: <122920081759.7243.49590FFA000843D700001C4B22218865869B0A02D2089B9A019C04040A0DBF970A03019D069C@att.net> Looking for a Shandon or Leica slide stainer. ...Used and in good condition. For a new small lab in Gainesville, Florida. From kimtournear <@t> yahoo.com Mon Dec 29 12:15:57 2008 From: kimtournear <@t> yahoo.com (Kim Tournear) Date: Mon Dec 29 12:16:00 2008 Subject: [Histonet] processor In-Reply-To: <8BB5FC36DDA373488E60177757BBD698457EC4@ncmhexchbe01.NewberryHospital.net> Message-ID: <562479.13561.qm@web54202.mail.re2.yahoo.com> Hi Jane, We just bought 2 Peloris processors from Leica. We can process in as little as 1 hour (usually biopsy specimens). Each machine has 2 processing chambers and can process up to 300 blocks per chamber. It was a good choice for us since we do appx 15,000 cases a year...Maintenance is easy....I use the Pathos from Leica (microwave technology) at my part time job processing dermatology specimens (appx 10,000 cases a year) and it works great. Contact Leica for some brochures on their equipment. Hope this helps... ? ~Kim? Tournear ~ Histology Supervisor Tucson Medical Center Tucson, AZ ? ~Don't?let your life end before it begins~ ? OU Rocks!!!! --- On Mon, 12/29/08, Jane C. Moose wrote: From: Jane C. Moose Subject: [Histonet] processor To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 7:10 AM I know this question repeatedly occurs, but our tissue processor "died" last week. We are a small hospital lab doing about 2500 cases a year (6000 specimens.) Would anyone have a recommendation on a new processor for our size lab? An Excelsior was brought in for demo (not sure what model) but that one was being discontinued and we were waiting to see the new model. We were also waiting on a "new" model VIP to bring in for demo-but that has not happened yet. Any advice would be greatly appreciated. Thanks Jane Jane Moose LIS Coordinator Newberry County Memorial Hospital Newberry, SC 29108 P-803-405-7129 F- 803-405-7474 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dwaugh <@t> kent.edu Mon Dec 29 13:19:53 2008 From: dwaugh <@t> kent.edu (David Waugh) Date: Mon Dec 29 13:20:03 2008 Subject: [Histonet] decapod cuticle sections Message-ID: <60220573-CA8F-48CC-93EA-B448FFFD9D56@kent.edu> OK, I'm not a histologist, I'm a paleontologist (&%**'it Jim, I a Doctor not a....). I am trying to make paraffin sections of decalcified decapod cuticle (from modern material, not fossil). The major problem I have been having is that the sections keep falling off the slides during staining.... I was making my own slides with a solution from EMS (3-aminopropyltriethoxysilane in acetone) I lost more than 1/2 of my sections... so I got pre-coated slides (Poly-L- Lysine) from EMS. The sections still fall off! After the section are cut I float them on warm water (48C), they never fully flatten out...? I then slip the slides underneath to pick up the sections. With the slides I subbed, I followed the directions and heated them on a hotplate. The presubbed slides I just air dried. To help the sections stick, I also tried pushing some of them down on the slide with a wet filter paper after removal from the water bath. The slides where then de-waxed in two xylene baths and brought to water in a series of ETOH baths. The sections seem to fall off mostly in the ETOH baths, but sometimes in the xylene. I'm not looking for perfection here, I just need the sections to stick. When a few sections make it through my protocol they look OK. I'm doing this on a low budget, but keep spending more and more on stains, slides etc. I'm getting very frustrated! Any ideas? how long should the slides dry? heat, no heat? Thanks in advance! -David Kent State University Department of Geology Kent, Ohio 44242 dwaugh@kent.edu www.personal.kent.edu/~dwaugh From Charles.Embrey <@t> carle.com Mon Dec 29 14:08:17 2008 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Mon Dec 29 14:09:48 2008 Subject: FW: [Histonet] processor Message-ID: <44780C571F28624DBB446DE55C4D733A1FE61D@EXCHANGEBE1.carle.com> -----Original Message----- From: Charles.Embrey Sent: Monday, December 29, 2008 2:08 PM To: 'kimtournear@yahoo.com' Subject: RE: [Histonet] processor Pathos is made by Milestone not Leica. Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kim Tournear Sent: Monday, December 29, 2008 12:16 PM To: Histonet Subject: Re: [Histonet] processor Hi Jane, We just bought 2 Peloris processors from Leica. We can process in as little as 1 hour (usually biopsy specimens). Each machine has 2 processing chambers and can process up to 300 blocks per chamber. It was a good choice for us since we do appx 15,000 cases a year...Maintenance is easy....I use the Pathos from Leica (microwave technology) at my part time job processing dermatology specimens (appx 10,000 cases a year) and it works great. Contact Leica for some brochures on their equipment. Hope this helps... ? ~Kim? Tournear ~ Histology Supervisor Tucson Medical Center Tucson, AZ ? ~Don't?let your life end before it begins~ ? OU Rocks!!!! --- On Mon, 12/29/08, Jane C. Moose wrote: From: Jane C. Moose Subject: [Histonet] processor To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 7:10 AM I know this question repeatedly occurs, but our tissue processor "died" last week. We are a small hospital lab doing about 2500 cases a year (6000 specimens.) Would anyone have a recommendation on a new processor for our size lab? An Excelsior was brought in for demo (not sure what model) but that one was being discontinued and we were waiting to see the new model. We were also waiting on a "new" model VIP to bring in for demo-but that has not happened yet. Any advice would be greatly appreciated. Thanks Jane Jane Moose LIS Coordinator Newberry County Memorial Hospital Newberry, SC 29108 P-803-405-7129 F- 803-405-7474 _______________________________________________ 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 Carol.Fields <@t> Northside.com Mon Dec 29 14:41:52 2008 From: Carol.Fields <@t> Northside.com (Carol Fields) Date: Mon Dec 29 14:42:22 2008 Subject: [Histonet] Her2 Weekend fixation Message-ID: <8CEB6DA1A3F35743800669D4CFE21F7D072A80C0@NSMXMS04.northside.local> Hi Netters, For those of you doing Her2 and tracking the fixation time of the breast tissue....how do you manage week-ends? Do you have a cut off on Friday afternoons, or what procedure do you follow so techs do not come in on Sunday? Any help with this is appreciated. THX in advance, Carole Fields, HT (ASCP) Histology Supervisor Northside Hospital Atlanta, GA 30342 carol.fields@northside.com CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. From Carol.Fields <@t> Northside.com Mon Dec 29 14:46:33 2008 From: Carol.Fields <@t> Northside.com (Carol Fields) Date: Mon Dec 29 14:47:05 2008 Subject: [Histonet] FW: Her2 Weekend fixation Message-ID: <8CEB6DA1A3F35743800669D4CFE21F7D072A80C7@NSMXMS04.northside.local> From: Carol Fields Sent: Monday, December 29, 2008 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: Her2 Weekend fixation Hi Netters, For those of you doing Her2 and tracking the fixation time of the breast tissue....how do you manage week-ends? Do you have a cut off on Friday afternoons, or what procedure do you follow so techs do not come in on Sunday? Any help with this is appreciated. THX in advance, Carole Fields, HT (ASCP) Histology Supervisor Northside Hospital Atlanta, GA 30342 carol.fields@northside.com CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. From rjbuesa <@t> yahoo.com Mon Dec 29 14:50:17 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 29 14:50:21 2008 Subject: [Histonet] FW: Her2 Weekend fixation In-Reply-To: <8CEB6DA1A3F35743800669D4CFE21F7D072A80C7@NSMXMS04.northside.local> Message-ID: <716643.97147.qm@web65715.mail.ac4.yahoo.com> Carol: It would be better for you to go to HistoNet archives where this topic has been discussed in detail previously with all imaginable solutions. Ren? J. --- On Mon, 12/29/08, Carol Fields wrote: From: Carol Fields Subject: [Histonet] FW: Her2 Weekend fixation To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 3:46 PM From: Carol Fields Sent: Monday, December 29, 2008 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: Her2 Weekend fixation Hi Netters, For those of you doing Her2 and tracking the fixation time of the breast tissue....how do you manage week-ends? Do you have a cut off on Friday afternoons, or what procedure do you follow so techs do not come in on Sunday? Any help with this is appreciated. THX in advance, Carole Fields, HT (ASCP) Histology Supervisor Northside Hospital Atlanta, GA 30342 carol.fields@northside.com CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Dec 29 14:52:28 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 29 14:52:32 2008 Subject: [Histonet] decapod cuticle sections In-Reply-To: <60220573-CA8F-48CC-93EA-B448FFFD9D56@kent.edu> Message-ID: <251046.34004.qm@web65710.mail.ac4.yahoo.com> Under separate cover I am sending some tips on floating paraffin sections that may help you. Ren? J. --- On Mon, 12/29/08, David Waugh wrote: From: David Waugh Subject: [Histonet] decapod cuticle sections To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 2:19 PM OK, I'm not a histologist, I'm a paleontologist (&%**'it Jim, I a Doctor not a....). I am trying to make paraffin sections of decalcified decapod cuticle (from modern material, not fossil). The major problem I have been having is that the sections keep falling off the slides during staining.... I was making my own slides with a solution from EMS (3-aminopropyltriethoxysilane in acetone) I lost more than 1/2 of my sections... so I got pre-coated slides (Poly-L-Lysine) from EMS. The sections still fall off! After the section are cut I float them on warm water (48C), they never fully flatten out...? I then slip the slides underneath to pick up the sections. With the slides I subbed, I followed the directions and heated them on a hotplate. The presubbed slides I just air dried. To help the sections stick, I also tried pushing some of them down on the slide with a wet filter paper after removal from the water bath. The slides where then de-waxed in two xylene baths and brought to water in a series of ETOH baths. The sections seem to fall off mostly in the ETOH baths, but sometimes in the xylene. I'm not looking for perfection here, I just need the sections to stick. When a few sections make it through my protocol they look OK. I'm doing this on a low budget, but keep spending more and more on stains, slides etc. I'm getting very frustrated! Any ideas? how long should the slides dry? heat, no heat? Thanks in advance! -David Kent State University Department of Geology Kent, Ohio 44242 dwaugh@kent.edu www.personal.kent.edu/~dwaugh _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Carol.Fields <@t> Northside.com Mon Dec 29 14:54:08 2008 From: Carol.Fields <@t> Northside.com (Carol Fields) Date: Mon Dec 29 14:55:29 2008 Subject: [Histonet] FW: Her2 Weekend fixation In-Reply-To: <716643.97147.qm@web65715.mail.ac4.yahoo.com> References: <8CEB6DA1A3F35743800669D4CFE21F7D072A80C7@NSMXMS04.northside.local> <716643.97147.qm@web65715.mail.ac4.yahoo.com> Message-ID: <8CEB6DA1A3F35743800669D4CFE21F7D072A80D2@NSMXMS04.northside.local> Great idea! Thank you Rene.....it is appreciated. Carole Carole Fields, HT (ASCP) Histology Supervisor Northside Hospital Atlanta, GA 30342 carol.fields@northside.com ________________________________ From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Monday, December 29, 2008 3:50 PM To: histonet@lists.utsouthwestern.edu; Carol Fields Subject: Re: [Histonet] FW: Her2 Weekend fixation Carol: It would be better for you to go to HistoNet archives where this topic has been discussed in detail previously with all imaginable solutions. Ren? J. --- On Mon, 12/29/08, Carol Fields wrote: From: Carol Fields Subject: [Histonet] FW: Her2 Weekend fixation To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 3:46 PM From: Carol Fields Sent: Monday, December 29, 2008 3:42 PM To: histonet@lists.utsouthwestern.edu Subject: Her2 Weekend fixation Hi Netters, For those of you doing Her2 and tracking the fixation time of the breast tissue....how do you manage week-ends? Do you have a cut off on Friday afternoons, or what procedure do you follow so techs do not come in on Sunday? Any help with this is appreciated. THX in advance, Carole Fields, HT (ASCP) Histology Supervisor Northside Hospital Atlanta, GA 30342 carol.fields@northside.com CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Sheri.Meilus <@t> va.gov Mon Dec 29 15:13:38 2008 From: Sheri.Meilus <@t> va.gov (Meilus, Sheri D.) Date: Mon Dec 29 15:15:30 2008 Subject: [Histonet] Stain for trypanosomes Message-ID: Hi to all you netters! Anybody out there with a suggestion for a good way to demonstrate trypanosomes in tissue? We have an interesting case that we suspect might be these critters but I don't know of a good stain. Thanks! S Sheri Meilus, HT(ASCP)QIHC Anatomic Pathology Supervisor Bay Pines VAMC Bay Pines, FL 33744 727-398-6661 4596 From Terri.Brown <@t> Northside.com Mon Dec 29 15:23:20 2008 From: Terri.Brown <@t> Northside.com (Terri Brown) Date: Mon Dec 29 15:23:48 2008 Subject: [Histonet] HER2 fixation Message-ID: <8CEB6DA1A3F35743800669D4CFE21F7D0689F806@NSMXMS04.northside.local> Can you tell me how you set up the weekend schedule for breast tissue specimens that come in after 3 pm on a Firday? Thanks in advance for your help. Terri H. Brown,, HT (ASCP) Pathology Laboratory Manager Northside Hospital Atlanta terri.brown@northside.com CONFIDENTIALITY NOTICE: This electronic mail transmission has been sent by Northside Hospital. It may contain information that is confidential, privileged, proprietary, or otherwise legally exempt from disclosure. If you are not the intended recipient, you are hereby notified that you are not authorized to read, print, retain, copy or disseminate this message, any part of it, or any attachments. If you have received this message in error, please delete this message and any attachments from your system without reading the content and notify the sender immediately of the inadvertent transmission. There is no intent on the part of the sender to waive any privilege. From rjbuesa <@t> yahoo.com Mon Dec 29 15:28:17 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 29 15:28:22 2008 Subject: [Histonet] Stain for trypanosomes In-Reply-To: Message-ID: <21946.21519.qm@web65714.mail.ac4.yahoo.com> Do Giemsa in tissue. Under separate cover I am sending you an article I wrote about it. Ren? J. --- On Mon, 12/29/08, Meilus, Sheri D. wrote: From: Meilus, Sheri D. Subject: [Histonet] Stain for trypanosomes To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 4:13 PM Hi to all you netters! Anybody out there with a suggestion for a good way to demonstrate trypanosomes in tissue? We have an interesting case that we suspect might be these critters but I don't know of a good stain. Thanks! S Sheri Meilus, HT(ASCP)QIHC Anatomic Pathology Supervisor Bay Pines VAMC Bay Pines, FL 33744 727-398-6661 4596 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pieronelva01 <@t> bigpond.com Tue Dec 30 02:17:06 2008 From: pieronelva01 <@t> bigpond.com (Piero Nelva) Date: Tue Dec 30 02:17:14 2008 Subject: [Histonet] processor References: <562479.13561.qm@web54202.mail.re2.yahoo.com> Message-ID: I agree. THe Peloris is a robust processor that handles all our routine overnight and 2 hour cycles easily. Also love the reagent managment system which assesses reagent purity. It does a good job on our skin specimens. Piero Nelva Anatomical Pathology Monash Medical Centre Australia ----- Original Message ----- From: "Kim Tournear" To: "Histonet" Sent: Tuesday, December 30, 2008 5:15 AM Subject: Re: [Histonet] processor Hi Jane, We just bought 2 Peloris processors from Leica. We can process in as little as 1 hour (usually biopsy specimens). Each machine has 2 processing chambers and can process up to 300 blocks per chamber. It was a good choice for us since we do appx 15,000 cases a year...Maintenance is easy....I use the Pathos from Leica (microwave technology) at my part time job processing dermatology specimens (appx 10,000 cases a year) and it works great. Contact Leica for some brochures on their equipment. Hope this helps... ~Kim Tournear ~ Histology Supervisor Tucson Medical Center Tucson, AZ ~Don't let your life end before it begins~ OU Rocks!!!! --- On Mon, 12/29/08, Jane C. Moose wrote: From: Jane C. Moose Subject: [Histonet] processor To: histonet@lists.utsouthwestern.edu Date: Monday, December 29, 2008, 7:10 AM I know this question repeatedly occurs, but our tissue processor "died" last week. We are a small hospital lab doing about 2500 cases a year (6000 specimens.) Would anyone have a recommendation on a new processor for our size lab? An Excelsior was brought in for demo (not sure what model) but that one was being discontinued and we were waiting to see the new model. We were also waiting on a "new" model VIP to bring in for demo-but that has not happened yet. Any advice would be greatly appreciated. Thanks Jane Jane Moose LIS Coordinator Newberry County Memorial Hospital Newberry, SC 29108 P-803-405-7129 F- 803-405-7474 _______________________________________________ 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 - http://www.avg.com Version: 8.0.176 / Virus Database: 270.10.1/1867 - Release Date: 12/28/2008 2:23 PM From SARAH.REEVES <@t> ekht.nhs.uk Tue Dec 30 09:09:03 2008 From: SARAH.REEVES <@t> ekht.nhs.uk (SARAH REEVES) Date: Tue Dec 30 09:09:44 2008 Subject: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. Message-ID: <495A398F020000B500003627@ekhgwia.ekht.nhs.uk> Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. Thanks in advance Sarah ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** From rjbuesa <@t> yahoo.com Tue Dec 30 09:14:04 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 30 09:14:09 2008 Subject: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. In-Reply-To: <495A398F020000B500003627@ekhgwia.ekht.nhs.uk> Message-ID: <52428.23350.qm@web65704.mail.ac4.yahoo.com> Use acid-fuchsin ponceau instead! Ren? J. --- On Tue, 12/30/08, SARAH REEVES wrote: From: SARAH REEVES Subject: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. To: histonet@lists.utsouthwestern.edu Date: Tuesday, December 30, 2008, 10:09 AM Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. Thanks in advance Sarah ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From SARAH.REEVES <@t> ekht.nhs.uk Tue Dec 30 09:14:14 2008 From: SARAH.REEVES <@t> ekht.nhs.uk (SARAH REEVES) Date: Tue Dec 30 09:17:57 2008 Subject: [Histonet] ELASTIC VAN GIESON Message-ID: <495A3AC6020000B50000362B@ekhgwia.ekht.nhs.uk> We have trouble with the differentiation of our Elastic Van Gieson stain. It seems that after 90 mins in the millers solution BMSs have trouble removing excess millers from the section without pulling colour out of the finer fibres. We would like to remove the 'green' wash from the section. We use alcohol for diffing, what solutions do other labs use? Thanks in advance Sarah ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** From mvlig <@t> burns.nl Tue Dec 30 10:21:34 2008 From: mvlig <@t> burns.nl (Vlig, Marcel) Date: Tue Dec 30 10:16:49 2008 Subject: [Histonet] Buffered formalin solution Message-ID: LS, Recently we discovered that our 37% formalin stock-solution was diluted in ddH2O instead of PBS. Can this affect the fixation properties of formalin, we do not see that much effect on our IHC stainings, but I like to be sure that there are no bad side-effects. Does anyone haven experience with this? Kind regards, Marcel From gu.lang <@t> gmx.at Tue Dec 30 11:19:58 2008 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Dec 30 11:20:07 2008 Subject: AW: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver In-Reply-To: <495A398F020000B500003627@ekhgwia.ekht.nhs.uk> References: <495A398F020000B500003627@ekhgwia.ekht.nhs.uk> Message-ID: The Masson-Trichrome has the PMA/PTA step between acid fuchsin and anilinblue (or light green). You can enhance the brightness of "red", when you prolong the incubation in PMA/PTA. Try some protocolls with 5-20 min. The brightness of "red" can also be a matter due to the washing step after the dyes. Take acidified water and don't wash too long in diluted ethanol. I prefer anilinblue because the contrast is better. If you do a one-step Trichrom stain with the PMA/PTA step before the dye-mixture, the incubation in the acid should be shorter for "more red" and longer for "more blue". Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von SARAH REEVES Gesendet: Dienstag, 30. Dezember 2008 16:09 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver bio Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. Thanks in advance Sarah ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From marjoh3 <@t> telus.net Tue Dec 30 12:19:10 2008 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Tue Dec 30 12:20:07 2008 Subject: [Histonet] Masson Trichrome Counterstain Message-ID: <4714D3D2DF2F493FB18BF62CAA6D9889@VALUED20606295> Aniline Blue for 4 mins. works well as a counterstain for muscle. Good Luck. Marilyn From RSRICHMOND <@t> aol.com Tue Dec 30 13:17:33 2008 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Tue Dec 30 13:17:38 2008 Subject: [Histonet] Re: Masson trichrome stain Message-ID: Sarah Reeves (in the UK) asks: >>Does anyone know how to achieve a perfect Masson trichrome stain? Our counterstain (light green in acetic acid) works well, but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique.<< If you're going to be staining liver biopsy specimens (by far the most common use of the trichrome stain by pathologists today, on paraffin embedded tissue anyhow), your pathologists need a stain with a very strong blue component (aniline blue or similar), since they're going to be semi-quantitating small amounts of fibrosis. I've had pretty good results with commercial trichrome kits, as long as they're not used past their expiration dates. You should use liver tissue - it can be normal, though better with a little fibrosis - as your stain control. Autopsy material is quite suitable. Better talk this over with your pathologists before proceeding. I'll be happy to e-mail with them if it'll help. Bob Richmond Samurai Pathologist Knoxville TN From rjbuesa <@t> yahoo.com Tue Dec 30 13:43:53 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 30 13:44:51 2008 Subject: [Histonet] Buffered formalin solution In-Reply-To: Message-ID: <656320.77113.qm@web65702.mail.ac4.yahoo.com> The stock solution in DW with time tends to acidify and if acid the fixation is less effcetive, that is the use of the buffer, to prevent the acidification. Ren? J. --- On Tue, 12/30/08, Vlig, Marcel wrote: From: Vlig, Marcel Subject: [Histonet] Buffered formalin solution To: histonet@lists.utsouthwestern.edu Date: Tuesday, December 30, 2008, 11:21 AM LS, Recently we discovered that our 37% formalin stock-solution was diluted in ddH2O instead of PBS. Can this affect the fixation properties of formalin, we do not see that much effect on our IHC stainings, but I like to be sure that there are no bad side-effects. Does anyone haven experience with this? Kind regards, Marcel _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Tue Dec 30 14:48:58 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 30 14:49:11 2008 Subject: [Histonet] Re: Masson trichrome stain In-Reply-To: Message-ID: We have had great success with the simple Van Gieson stain. It stains the collagen (& fibrosis) really well. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: Wednesday, 31 December 2008 6:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Masson trichrome stain Sarah Reeves (in the UK) asks: >>Does anyone know how to achieve a perfect Masson trichrome stain? Our >>counterstain (light green in acetic acid) works well, but the >>intensity of the muscle stain is not very bright. Does anyone have any >>suggestions? We will be staining liver biopsies mostly in the near >>future and wish to improve our technique.<< If you're going to be staining liver biopsy specimens (by far the most common use of the trichrome stain by pathologists today, on paraffin embedded tissue anyhow), your pathologists need a stain with a very strong blue component (aniline blue or similar), since they're going to be semi-quantitating small amounts of fibrosis. I've had pretty good results with commercial trichrome kits, as long as they're not used past their expiration dates. You should use liver tissue - it can be normal, though better with a little fibrosis - as your stain control. Autopsy material is quite suitable. Better talk this over with your pathologists before proceeding. I'll be happy to e-mail with them if it'll help. Bob Richmond Samurai Pathologist Knoxville TN _______________________________________________ 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 note 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 RSRICHMOND <@t> aol.com Tue Dec 30 15:12:17 2008 From: RSRICHMOND <@t> aol.com (Robert Richmond) Date: Tue Dec 30 15:12:24 2008 Subject: [Histonet] Re: Masson trichrome stain In-Reply-To: References: Message-ID: Hi Tony Henwood and all! I'm only familiar with the Van Gieson stain as a counterstain for Verhoeff's hematoxylin, the elastic stain. Reproducibility is extremely important in evaluating fibrosis in liver biopsy specimens from hepatitis C patients (most common indication for liver biopsy in many places), since the extent of fibrosis determines whether the patient gets the very arduous year-long treatment or not. And I think the standard, at least in the USA, is the sort of blue trichrome stain standardized a good many years ago by Klatskin and his disciples. Bob Richmond Samurai Pathologist Knoxville TN On Tue, Dec 30, 2008 at 3:48 PM, Tony Henwood wrote: > We have had great success with the simple Van Gieson stain. > > It stains the collagen (& fibrosis) really well. > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) > Laboratory Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert > Richmond > Sent: Wednesday, 31 December 2008 6:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Masson trichrome stain > > > Sarah Reeves (in the UK) asks: > >>>Does anyone know how to achieve a perfect Masson trichrome stain? Our >>>counterstain (light green in acetic acid) works well, but the >>>intensity of the muscle stain is not very bright. Does anyone have any > >>>suggestions? We will be staining liver biopsies mostly in the near >>>future and wish to improve our technique.<< > > If you're going to be staining liver biopsy specimens (by far the most > common use of the trichrome stain by pathologists today, on paraffin > embedded tissue anyhow), your pathologists need a stain with a very > strong blue component (aniline blue or similar), since they're going to > be semi-quantitating small amounts of fibrosis. I've had pretty good > results with commercial trichrome kits, as long as they're not used past > their expiration dates. > > You should use liver tissue - it can be normal, though better with a > little fibrosis - as your stain control. Autopsy material is quite > suitable. > > Better talk this over with your pathologists before proceeding. I'll be > happy to e-mail with them if it'll help. > > Bob Richmond > Samurai Pathologist > Knoxville TN From lpwenk <@t> sbcglobal.net Tue Dec 30 16:20:13 2008 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Dec 30 16:20:19 2008 Subject: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver In-Reply-To: <495A398F020000B500003627@ekhgwia.ekht.nhs.uk> Message-ID: <02599F82B85E4D1EAF3B48F3CB751EFF@HPPav2> Lots of questions, as this is one of those stains that has a lot of places that need to be tweaked: NEW OR OLD? Is this a new problem you are having, or have you always had this problem? MORDANT: Are you still post-mordanting in Bouin, or have you tried to eliminate this or switch over to something else to try to not use picric acid in the lab? What time and temperature? RED DYE: What dye(s) are you using for the red dye for the muscle? Please include name and color index (CI) number, if that's available in the UK. Also, have you switched recently to a new type of red dye, or purchased a replacement when you ran out? What is the dye content percent on the dry powder? PMA/PTA: Are you using Phosphotungstic acid and/or Phospomolybdic acid? How old is it? ONE OR TWO STEP: Just to make certain, is this a true Masson trichrome, where the red dye is applied, and then the PTA/PMA, and then finally the green dye? Or is this the traditional Gomori, where the red and green dyes and the PTA/PMA are all in one solution? TIME IN RED AND GREEN DYES: Are you trying to standardize the time in the red dye and the time in the green dye? This usually doesn't work. Each different type of tissue, and each different different person's tissue, will pick up red dye and green dye at a different rate. TEMPERATURE: Are the dyes at room temp or refrigerator temperature? That can have an influence. MACHINE OR HAND: Are you hand staining, where you have control over the amount of time in each solution, where you can check the progress with a microscope, where you have a choice of dyes? Or are you staining with a kit? Or on a machine? Sorry, need some more information, before we can help you pinpoint the problem. Peggy A. Wenk, HTL(ASCP)SLS Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of SARAH REEVES Sent: Tuesday, December 30, 2008 10:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver bio Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any suggestions? We will be staining liver biopsies mostly in the near future and wish to improve our technique. Thanks in advance Sarah ********************************************************************** 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. Any views or opinions presented are solely those of the author and do not necessarily represent those of East Kent Hospitals University NHS Trust. If you are not the intended recipient, be advised that you have received this email in error and that any use, dissemination, forwarding, printing or copying of this email is strictly prohibited. If you have received this email in error please notify the system manager at the following email address: root.postmaster@ekht.nhs.uk www.kentandmedway.nhs.uk This footnote also confirms that although this email message has been swept by MIMEsweeper for the presence of computer viruses, it is strongly recommended that you carry out your own virus scan of this message and any attachments. www.mimesweeper.com ********************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Tue Dec 30 16:32:15 2008 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 30 16:33:15 2008 Subject: [Histonet] Re: Masson trichrome stain In-Reply-To: Message-ID: A simple stain to do. Stain dewaxed sections with haematoxylin (Iron or celestine blue), stain with Van Gieson solution 5 minutes or more (dependent on the red dye used), blot gently, dehydrate, clear and mount and you are done. Several Pathologists here in Australia are using the Van Gieson stain routinely on liver biopsies and many more are being attracted to it. I recommend you give it a go, you might be surprised Give it a go Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist Tel: 612 9845 3306 Fax: 612 9845 3318 the children's hospital at westmead Cnr Hawkesbury Road and Hainsworth Street, Westmead Locked Bag 4001, Westmead NSW 2145, AUSTRALIA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert Richmond Sent: Wednesday, 31 December 2008 8:12 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: Masson trichrome stain Hi Tony Henwood and all! I'm only familiar with the Van Gieson stain as a counterstain for Verhoeff's hematoxylin, the elastic stain. Reproducibility is extremely important in evaluating fibrosis in liver biopsy specimens from hepatitis C patients (most common indication for liver biopsy in many places), since the extent of fibrosis determines whether the patient gets the very arduous year-long treatment or not. And I think the standard, at least in the USA, is the sort of blue trichrome stain standardized a good many years ago by Klatskin and his disciples. Bob Richmond Samurai Pathologist Knoxville TN On Tue, Dec 30, 2008 at 3:48 PM, Tony Henwood wrote: > We have had great success with the simple Van Gieson stain. > > It stains the collagen (& fibrosis) really well. > > Regards > > Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory > Manager & Senior Scientist > Tel: 612 9845 3306 > Fax: 612 9845 3318 > the children's hospital at westmead > Cnr Hawkesbury Road and Hainsworth Street, Westmead > Locked Bag 4001, Westmead NSW 2145, AUSTRALIA > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robert > Richmond > Sent: Wednesday, 31 December 2008 6:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: Masson trichrome stain > > > Sarah Reeves (in the UK) asks: > >>>Does anyone know how to achieve a perfect Masson trichrome stain? Our >>>counterstain (light green in acetic acid) works well, but the >>>intensity of the muscle stain is not very bright. Does anyone have >>>any > >>>suggestions? We will be staining liver biopsies mostly in the near >>>future and wish to improve our technique.<< > > If you're going to be staining liver biopsy specimens (by far the most > common use of the trichrome stain by pathologists today, on paraffin > embedded tissue anyhow), your pathologists need a stain with a very > strong blue component (aniline blue or similar), since they're going > to be semi-quantitating small amounts of fibrosis. I've had pretty > good results with commercial trichrome kits, as long as they're not > used past their expiration dates. > > You should use liver tissue - it can be normal, though better with a > little fibrosis - as your stain control. Autopsy material is quite > suitable. > > Better talk this over with your pathologists before proceeding. I'll > be happy to e-mail with them if it'll help. > > Bob Richmond > Samurai Pathologist > Knoxville TN _______________________________________________ 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 note 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 nfournier <@t> sasktel.net Tue Dec 30 17:21:59 2008 From: nfournier <@t> sasktel.net (Neil Fournier) Date: Tue Dec 30 17:22:14 2008 Subject: [Histonet] subbing slides Message-ID: <001001c96ad5$6f05d000$e29b2fcf@NEIL> Hi Everyone, Could someone share with me their procedure for cleaning and subbing glass slides with gelantin? I am trying to work out a concentration of gelantin and chromium potassium sulphate that will be optimal for adhering fairly thick rat brain sections (30 to 120 um range depending on applications) Much appreciated Neil E-mail message checked by Spyware Doctor (6.0.0.386) Database version: 5.11440 http://www.pctools.com/en/spyware-doctor-antivirus/ From susanbachus <@t> verizon.net Tue Dec 30 17:31:33 2008 From: susanbachus <@t> verizon.net (Susan Bachus) Date: Tue Dec 30 17:31:49 2008 Subject: [Histonet] subbing slides References: <001001c96ad5$6f05d000$e29b2fcf@NEIL> Message-ID: <9C740F10CC114E63BB6D982B81CB6F87@RESLAPTOP> This always works like a charm for us and costs ~10x less than buying pre-subbed or electrostatic or whatever fancy expensive store-bought slides! Have fun! Susan ----- Original Message ----- From: "Neil Fournier" To: Sent: Tuesday, December 30, 2008 6:21 PM Subject: [Histonet] subbing slides > Hi Everyone, > > Could someone share with me their procedure for cleaning and subbing glass > slides with gelantin? I am trying to work out a concentration of gelantin > and chromium potassium sulphate that will be optimal for adhering fairly > thick rat brain sections (30 to 120 um range depending on applications) > > Much appreciated > > Neil > > > > E-mail message checked by Spyware Doctor (6.0.0.386) > Database version: 5.11440 > http://www.pctools.com/en/spyware-doctor-antivirus/ > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From susanbachus <@t> verizon.net Tue Dec 30 17:34:49 2008 From: susanbachus <@t> verizon.net (Susan Bachus) Date: Tue Dec 30 17:34:57 2008 Subject: [Histonet] subbing slides References: <001001c96ad5$6f05d000$e29b2fcf@NEIL> <9C740F10CC114E63BB6D982B81CB6F87@RESLAPTOP> Message-ID: Well, the attachment seems to have been deleted from what I sent, so here it is pasted below: Gelatin-Subbed slides WEAR GLOVES! (not to protect your hands, but to protect the slides from RNase from your skin) a.. Place slides in racks and soak in hot tap water and soap (e.g. dishwashing soap, or Alconox) for at least 1 hr, but not more than a few hours (as soap could leave residue). b.. Rinse thoroughly in hot tap water until suds are completely gone. c.. Let sit in 80% ethanol for at least one hr. This solution can be re-used until it turns cloudy. Slides can be left overnight in this solution. d.. Rinse in a few changes of deionized water. e.. Subbing solution: Dissolve 3.0 g of gelatin (300 bloom swine) in 150 ml deionized H2O by stirring while heating to 50 degrees C. Add 450 ml deionized water to cool, and then dissolve 0.3 g CrK(SO4)2*12H2O ("chrom alum") in the solution. f.. Filter this solution through Whatman Type I filter paper to remove any air bubbles. This solution is only good for the few hrs it will take for this batch of slides, as it will quickly solidify. g.. Dip the slides into the subbing solution, drain the slides onto a paper towel and allow to air dry for 1 hr. h.. Dip the slides into the subbing solution again. Drain and cover loosely with bench paper and allow to dry for a few hours. i.. When thoroughly dry, heat in oven at ~ 50 degrees C for a few hours, then store the slides in slide boxes in zip-lock bags. As long as they are kept dry, they should be good for years. ----- Original Message ----- From: "Susan Bachus" To: "Neil Fournier" ; Sent: Tuesday, December 30, 2008 6:31 PM Subject: Re: [Histonet] subbing slides > This always works like a charm for us and costs ~10x less than buying > pre-subbed or electrostatic or whatever fancy expensive store-bought > slides! > Have fun! Susan > ----- Original Message ----- > From: "Neil Fournier" > To: > Sent: Tuesday, December 30, 2008 6:21 PM > Subject: [Histonet] subbing slides > > >> Hi Everyone, >> >> Could someone share with me their procedure for cleaning and subbing >> glass >> slides with gelantin? I am trying to work out a concentration of gelantin >> and chromium potassium sulphate that will be optimal for adhering fairly >> thick rat brain sections (30 to 120 um range depending on applications) >> >> Much appreciated >> >> Neil >> >> >> >> E-mail message checked by Spyware Doctor (6.0.0.386) >> Database version: 5.11440 >> http://www.pctools.com/en/spyware-doctor-antivirus/ >> _______________________________________________ >> 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 tifei <@t> foxmail.com Tue Dec 30 20:17:18 2008 From: tifei <@t> foxmail.com (=?utf-8?B?VEY=?=) Date: Tue Dec 30 20:17:32 2008 Subject: =?utf-8?B?UmU6IFJlOiBbSGlzdG9uZXRdIHN1YmJpbmcgc2xpZGVz?= References: <001001c96ad5$6f05d000$e29b2fcf@NEIL>, <9C740F10CC114E63BB6D982B81CB6F87@RESLAPTOP>, Message-ID: <200812311017130627983@foxmail.com> If you are talking about very thick brain sections, I have used 40-50 um sections only. However my collegues noticed that 0.25% gel coated slides are much better than 0.5% gel coated slides. We are performing BrdU staining-including the step of citrate buffer heat at 95 degree for 30 min. The 0.25% gel stands out from this test. 2008-12-31 TF ???? Susan Bachus ????? 2008-12-31 07:42:23 ???? Neil Fournier; histonet@lists.utsouthwestern.edu ??? ??? Re: [Histonet] subbing slides Well, the attachment seems to have been deleted from what I sent, so here it is pasted below: Gelatin-Subbed slides WEAR GLOVES! (not to protect your hands, but to protect the slides from RNase from your skin) a.. Place slides in racks and soak in hot tap water and soap (e.g. dishwashing soap, or Alconox) for at least 1 hr, but not more than a few hours (as soap could leave residue). b.. Rinse thoroughly in hot tap water until suds are completely gone. c.. Let sit in 80% ethanol for at least one hr. This solution can be re-used until it turns cloudy. Slides can be left overnight in this solution. d.. Rinse in a few changes of deionized water. e.. Subbing solution: Dissolve 3.0 g of gelatin (300 bloom swine) in 150 ml deionized H2O by stirring while heating to 50 degrees C. Add 450 ml deionized water to cool, and then dissolve 0.3 g CrK(SO4)2*12H2O ("chrom alum") in the solution. f.. Filter this solution through Whatman Type I filter paper to remove any air bubbles. This solution is only good for the few hrs it will take for this batch of slides, as it will quickly solidify. g.. Dip the slides into the subbing solution, drain the slides onto a paper towel and allow to air dry for 1 hr. h.. Dip the slides into the subbing solution again. Drain and cover loosely with bench paper and allow to dry for a few hours. i.. When thoroughly dry, heat in oven at ~ 50 degrees C for a few hours, then store the slides in slide boxes in zip-lock bags. As long as they are kept dry, they should be good for years. ----- Original Message ----- From: "Susan Bachus" To: "Neil Fournier" ; Sent: Tuesday, December 30, 2008 6:31 PM Subject: Re: [Histonet] subbing slides > This always works like a charm for us and costs ~10x less than buying > pre-subbed or electrostatic or whatever fancy expensive store-bought > slides! > Have fun! Susan > ----- Original Message ----- > From: "Neil Fournier" > To: > Sent: Tuesday, December 30, 2008 6:21 PM > Subject: [Histonet] subbing slides > > >> Hi Everyone, >> >> Could someone share with me their procedure for cleaning and subbing >> glass >> slides with gelantin? I am trying to work out a concentration of gelantin >> and chromium potassium sulphate that will be optimal for adhering fairly >> thick rat brain sections (30 to 120 um range depending on applications) >> >> Much appreciated >> >> Neil >> >> >> >> E-mail message checked by Spyware Doctor (6.0.0.386) >> Database version: 5.11440 >> http://www.pctools.com/en/spyware-doctor-antivirus/ >> _______________________________________________ >> 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 From pruegg <@t> ihctech.net Wed Dec 31 10:48:28 2008 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Wed Dec 31 10:48:37 2008 Subject: [Histonet] Does anyone know how to achieve a perfect Masson Trichrome stain. Our counterstain (light green in acetic acid) works well but the intensity of the muscle stain is not very bright. Does anyone have any =?UTF-8?Q?suggestions=3F=20We=20will=20be=20staining=20liver?= Message-ID: <20081231094828.f86bd30e73b823f57b516b5451216a98.8feb0248b5.wbe@email.secureserver.net> i have a protocol from Bryan Hewlett that is called "the fail pro Masons Trichrome" Patsy -------- Original Message -------- Subject: RE: [Histonet] Does Masson Trichrome stain. Our counter acid) works well but the intensity of the m Does anyone have any suggestions? We wil From: "Lee & Peggy Wenk" Lots of questions, as this is one of those stains that has a lot of p laces that need to be tweaked: NEW OR OLD? Is this a new problem had this problem? MORDANT: Are eliminate this picric acid in the l RED DYE: What dye(s) are you using fo Please include name and color index (CI) n UK. Also, have you switched recently t purchased a replacement when you ran out? Wh dry powder? PMA/PTA: Are you acid? How old is it? ONE OR TWO STEP: Just to make certain, is this a true Masson trichrom e, where the red dye is applied, and then the PTA/PMA, and then finally the green dye? Or is this the traditional Gomori, where the red and gree n dyes and the PTA/PMA are all in one solution? TIME IN RED AND G in the red dye and the Each different type o tissue, will pick up red TEMPERATURE: Are the dyes a That can have an influence. MACHINE OR HAND: Are you hand staining, where you have control over the amount of time in each solution, where you can check the progress with microscope, where you have a choice of dyes? Or are you staining with Or on a machine? Sorry, need some more information, before the problem. Peggy A. Wenk, HTL(ASCP)SL Beaumont Hospital Royal Oak, MI 48073 -----Original Message- From: histonet-bounces@lists.utsouthwestern.edu [[1]mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf O SARAH REEVES Sent: Tuesday, December 30, 2008 10:09 AM To: histonet Subject: [Histonet] Does anyone know how to ac Trichrome stain. Our counterstain (light green in well but the intensity of the muscle stain is not ve anyone have any suggestions? We will be staining liver b Does anyone know how to achieve a perfect Masson Trichrome stain. counterstain (light green in acetic acid) works well but the intens ity of the muscle stain is not very bright. Does anyone have any suggest ions? We will be staining liver biopsies mostly in the near future and w to improve our technique. Thanks in advance Sarah < ******************************************************************** ** This email and any files transmitted with it are confidential and int ended solely for the use of the individual or entity to whom they are ad dressed. Any views or opinions presented are solely those of the author do not necessarily represent those of East Kent Hospitals University Trust. If you are not the intended recipient, be advised that you h received this email in error and that any use, dissemination, forwar or copying of this email is strictly prohibited. I manager at the following email address: root.postmaster@ekht.nhs.uk [2]www.kentandmedway.nhs.uk This footnote al been swept by MIMEsweep recommended that and any attachments.< [3]www.mimesweeper.com ********************* ************************************************* ______________ Histonet mailing list Histonet@list [4]http://lists.utsouthwestern.edu/mailman/list ______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu [5] References 1. 3D"http://email.secureserver.net/pcompose 2. ="http://www.kentandmedway.nhs.uk"/ 3. 3D"http://www.mimesweeper.com"/ 4. 3D"http://lists.utsouthwestern.edu/mailman/ 5. ="http://lists.utsouthwestern.edu/mailman/listinfo/histonet" From amber.mckenzie <@t> gastrodocs.net Wed Dec 31 12:43:51 2008 From: amber.mckenzie <@t> gastrodocs.net (Amber McKenzie) Date: Wed Dec 31 12:44:53 2008 Subject: [Histonet] pH strips In-Reply-To: <20081231094828.f86bd30e73b823f57b516b5451216a98.8feb0248b5.wbe@email.secureserver.net> References: <20081231094828.f86bd30e73b823f57b516b5451216a98.8feb0248b5.wbe@email.secureserver.net> Message-ID: <03C921A1EAF7F541B16543F6EC6A4B37021832EF@giamail2.Gia.com> Do pH strips go bad after a certain time period? From Jackie.O'Connor <@t> abbott.com Wed Dec 31 12:57:57 2008 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Dec 31 12:59:25 2008 Subject: [Histonet] pH strips In-Reply-To: <03C921A1EAF7F541B16543F6EC6A4B37021832EF@giamail2.Gia.com> Message-ID: I heard of some that went on a graffiti rampage in downtown Chicago last year . . . . . Happy New Year. "Amber McKenzie" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/31/2008 12:43 PM To cc Subject [Histonet] pH strips Do pH strips go bad after a certain time period? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From leiker <@t> buffalo.edu Wed Dec 31 13:49:17 2008 From: leiker <@t> buffalo.edu (Merced Leiker) Date: Wed Dec 31 13:50:25 2008 Subject: [Histonet] pH strips In-Reply-To: References: Message-ID: <05F18E040D8D4D45FC8CF6F9@bchwxp2702.ad.med.buffalo.edu> LOL, Jackie!!! I've used 10-year old strips that seemed to have worked fine. --On Wednesday, December 31, 2008 12:57 PM -0600 Jackie M O'Connor wrote: > I heard of some that went on a graffiti rampage in downtown Chicago last > year . . . . . > > > Happy New Year. > > > > "Amber McKenzie" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 12/31/2008 12:43 PM > > To > > cc > > Subject > [Histonet] pH strips > > > > > > > Do pH strips go bad after a certain time period? > > > _______________________________________________ > 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 > Merced M Leiker Research Technician II 354 BRB (pkgs) / 140 Farber Hall (letters) School of Medicine and Biomedical Sciences State University of New York at Buffalo 3435 Main St, Buffalo, NY 14214 Ph: (716) 829-6033 Fx: (716) 829-2725 "Without my flaws I'm really very boring." - random internet blog commentator From ploykasek <@t> phenopath.com Wed Dec 31 13:59:22 2008 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Dec 31 14:00:30 2008 Subject: [Histonet] pH strips In-Reply-To: <03C921A1EAF7F541B16543F6EC6A4B37021832EF@giamail2.Gia.com> Message-ID: I got a great laugh from Jackie's comment. However, I don't feel so clever today. I would like to add that our pH strips have an expiration date on the package. I find it hard to believe the strips will quit working on a certain date, but we don't use expired reagents in the lab! Patti > Do pH strips go bad after a certain time period? > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From rjbuesa <@t> yahoo.com Wed Dec 31 14:11:03 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 31 14:12:01 2008 Subject: [Histonet] pH strips In-Reply-To: <03C921A1EAF7F541B16543F6EC6A4B37021832EF@giamail2.Gia.com> Message-ID: <898015.6031.qm@web65716.mail.ac4.yahoo.com> pH strips are made of a "filter paper like" base paper?with pH reacting reagents for certain pH ranges and, yes, they react improperly after?the date set by the manufacturer. Ren? J.? --- On Wed, 12/31/08, Amber McKenzie wrote: From: Amber McKenzie Subject: [Histonet] pH strips To: histonet@lists.utsouthwestern.edu Date: Wednesday, December 31, 2008, 1:43 PM Do pH strips go bad after a certain time period? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From katelin <@t> cuttingedgehistology.com Wed Dec 31 14:13:14 2008 From: katelin <@t> cuttingedgehistology.com (Katelin Lester) Date: Wed Dec 31 14:13:21 2008 Subject: [Histonet] SAFE-CLEAR Xylene Substitute Message-ID: Our pathologist has agreed to let us try out SAFE-CLEAR xylene substitute in our VIP processor. We currently have 2 changes of xylene at 30 minutes each and process mostly small surgical specimens. Do I need to increase the time the tissue spends in the SAFE-CLEAR? I have searched the Histonet Archives for answers about xylene substitutes, and have mostly found what I am looking for, but am unclear on whether or not I need to increase the time during processing. Thank you all, Katelin Lester Cutting Edge Histology Services, LLC (503) 443-2157 From rjbuesa <@t> yahoo.com Wed Dec 31 15:15:02 2008 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 31 15:16:01 2008 Subject: [Histonet] SAFE-CLEAR Xylene Substitute In-Reply-To: Message-ID: <558860.8118.qm@web65715.mail.ac4.yahoo.com> Katelin: Safe-Clear, as you probably have found in HistoNet archives, comes in 2 "versions": Safe-Clear that is nothing but Stoddard Solvent (or White Spirits also known as Mineral Spirits) and Safe Clear II that has naphthenic hydrocarbons added. These reagents are the ones usually?employed in "dry cleaning" and are less effective than xylene and cannot be used to dewax sections or clean the tissue processor. I would ask the representative of Thermo Fisher Scientific about the time but I think that the clearing time should be increased to at least 45 minutes per station (instead of the 30 minutes you have now with xylene). They cost between 1.72 (Safe-Clear) and 1.91 (Safe-Clear II) the price of wylene but being what they really are, that is overpricing. You could get the same product cheaper in any HomeDepot store. Happy New Year Ren? J. --- On Wed, 12/31/08, Katelin Lester wrote: From: Katelin Lester Subject: [Histonet] SAFE-CLEAR Xylene Substitute To: histonet@lists.utsouthwestern.edu Date: Wednesday, December 31, 2008, 3:13 PM Our pathologist has agreed to let us try out SAFE-CLEAR xylene substitute in our VIP processor. We currently have 2 changes of xylene at 30 minutes each and process mostly small surgical specimens. Do I need to increase the time the tissue spends in the SAFE-CLEAR? I have searched the Histonet Archives for answers about xylene substitutes, and have mostly found what I am looking for, but am unclear on whether or not I need to increase the time during processing. Thank you all, Katelin Lester Cutting Edge Histology Services, LLC (503) 443-2157 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet