From Diane.Gladney <@t> se.amedd.army.mil Thu Dec 1 06:05:28 2005 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Thu Dec 1 06:02:09 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: <4D55B2E997EFAE4DA6081DDE100B83029AA7BC@amedmlsermc133.amed.ds.army.mil> I, too, was a fan of Quincy and now am a fan of CSI. But the episode of CSI where I saw the pathologist cutting a block on the microtome and turning the fly wheel backwards sent me screaming at the TV...."Are you trying to strip every gear in that microtome!!!" I absolutely cringed when I saw that. But the biggest laugh that I got out of the episode was seeing a pathologist cutting blocks. Not to put down pathologists but it is very rare to see one cutting blocks in a clinical environment. The CSI consultants also need to get with the latest technology in histology and not have someone drying slides over an open flame. If anyone saw this episode, you know what I mean. Hollywood, you gotta love them; they hire consultants then refuse to listen to them. I still am a huge fan of CSI and the program has sparked an interest in the medical laboratory field including histology. Hey, we need as much exposure as possible. Histology is the greatest of laboratory professions. I have no regrets of working for more than 30 years in this profession and look forward to working many more. Diane Diane C. Gladney, HT (ASCP) Supervisor, Histology/Cytology Moncrief Army Community Hospital Dept. of Pathology 4500 Stuart Street P.O. Box 484 Ft. Jackson, SC 29207 Email: diane.gladney@se.amedd.army.mil Phone: 803-751-2530 FAX: 803-751-7829 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Monfils, Paul Sent: Wednesday, November 30, 2005 3:17 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Histotechs: born or made? Yes we have come a long way since one of the pathologists on one of my first jobs referred to histotechs as "glorified salami slicers". Our supervisor however was not one to take any baloney, and promptly told the good doctor that without our salami his diagnoses would be baloney. The only line I remember from Quincy, after watching it for several years, is "Sam, get me a PAS on this - STAT!" But CSI, which I also watch frequently, really is a mix of science and science fiction. I wish we did have machines that could do everything their machines do. Some of them are really way out there. Like the episode where two people had a conversation near a pottery wheel, and the clay picked up the vibrations of their conversation, which were subsequently scanned by a laser and turned back into audible voices. :-) I even noticed a technical error related to my avocation - malacology. In one episode a dealer in illegally harvested abalone (a type of shellfish) is shot. The medical examiner says "serves him right for selling an endangered bivalve". As you may remember from your invertebrate zoology class, abalone are gastropods, not bivalves. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Thu Dec 1 06:02:32 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Thu Dec 1 06:06:10 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: Isn't it odd that there usually is no one else in the labs except the one or two investigators working on the one or two cases going on that week? Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. 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 Shirley Powell Sent: Wednesday, November 30, 2005 1:45 PM To: king.laurie@marshfieldclinic.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? Yep, that is when I stopped watching Quincy. Made me want to fight too. I do watch CSI but usually not for accuracy, mostly for the scenery. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of king.laurie@marshfieldclinic.org Sent: Wednesday, November 30, 2005 2:30 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? I used to love that show, until the episode where Quincy complained that Sam was demoted to work in histology, who were, after all, only a bunch of "bottle labelers". Them's fightin' words! Laurie King ------Original Message------ From: "Bartlett, Jeanine" Date: Wed Nov 30, 2005 -- 01:15:00 PM To: "Pam Marcum" , "Ford Royer" , "Histonet" Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet >From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 _______________________________________________ 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 mauger <@t> email.chop.edu Thu Dec 1 07:30:34 2005 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Thu Dec 1 07:31:13 2005 Subject: [Histonet] Aspergillus antibody Message-ID: Hi Bill, We use a monoclonal anti- aspergillus from Dako #M3564. For us, it works well at 1:200 after retrieval in high pH buffer also from Dako #S3307. Jo Mauger >>> "Bill Sinai" 11/30/05 8:55 PM >>> Dear All, Is there anyone out there using an antibody to aspergillus species? If so how do you do it (dilutions, manually or by machine)? Do any antibody work better than others? Is it available for FFPE specimens. Any info would be greatly appreciated. Thanks Bill Sinai Chief Hospital Scientist Tissue Pathology ICPMR Westmead Hospital Westmead NSW 2145 Phone 02 9845 7774 Fax 02 9687 2330 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BMolinari <@t> heart.thi.tmc.edu Thu Dec 1 07:46:32 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Thu Dec 1 07:50:10 2005 Subject: [Histonet] Paraffin sections Message-ID: Hi, I have heard that paraffin sections that have been deparaffinized and run down to DH20 but for some reason cannot be stained that day, can be left to air dry and then be placed back into H2O when they are ready for staining. Does anyone have any experience with this technique? I usually just leave them in DH2O till I can get to them, usually the next day. Thanks. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) From Catherine.Dreanno <@t> newcastle.ac.uk Thu Dec 1 08:17:21 2005 From: Catherine.Dreanno <@t> newcastle.ac.uk (NCD20) Date: Thu Dec 1 08:17:35 2005 Subject: [Histonet] Immunogold Message-ID: <4381454D@webmail.ncl.ac.uk> Dear Histonetters, I am??trying to developp a protocol for??immunogold on??crustacea larvae, but I am??new??in this??technic....??I am looking at the cuticular??tissues.?? I will be gratefull if someone??can??share with me any protocols, advices set??up for arthropods , plankton, ... . Thank you in advance for any help ! Cheers, Catherine ????????????????????????????????????????????????????????????? MBA - Marine Biological Association of the UK???????><> The Laboratory????????????<><>??????><> Citadel Hill???????????????????????????????????????????????<>< Plymouth, PL1 2PB??????UK??????<><> ???????><> ><>????????????????????????????????????????<>< <>< _____\|/_____\|/_______\|/______\|/_________\|/_____ From Sionagh.Smith <@t> ed.ac.uk Thu Dec 1 08:30:40 2005 From: Sionagh.Smith <@t> ed.ac.uk (Sionagh Smith) Date: Thu Dec 1 08:30:46 2005 Subject: [Histonet] (no subject) Message-ID: <00b201c5f683$cde56a00$cc7ed781@AHW126204> Can anyone help me with a recipe for Jore's or Klotz fixative solutions? I have used both in the past for retaining colour and consistency of post mortem specimens for teaching veterinary students. I have been unable to find an actual recipe since someone else always prepared them! I wish I'd paid more attention now.... Any pointers would be much appreciated. Sionagh Smith Dept. of Veterinary Pathology Royal (Dick) School of Veterinary Studies Easter Bush Veterinary Centre Easter Bush Midlothian EH25 9RG Tel 0131 650 8787 From ryaskovich <@t> dir.nidcr.nih.gov Thu Dec 1 08:48:58 2005 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Thu Dec 1 08:49:06 2005 Subject: [Histonet] deOlmos Amino Cupric Silver Message-ID: Does anyone have a procedure for this Disintegrative Degeneration Stain (deOlmos Amino Cupric Silver)? Thanks so much in advance Ruth Yaskovich National Institutes of Health National Institute of Dental and Crainiofacial Research Histochemistry Drug Discovery From rjbuesa <@t> yahoo.com Thu Dec 1 08:49:45 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 1 08:49:54 2005 Subject: [Histonet] (no subject) In-Reply-To: <00b201c5f683$cde56a00$cc7ed781@AHW126204> Message-ID: <20051201144945.67214.qmail@web61213.mail.yahoo.com> Hi Sionagh: Jore's fixative (1896): Water dist. ---100 mL + 40% formaldehyde--- 10 mL + sodium sulfate --- 2g + magnesium sulfate ---2 g + sodium chloride --- 1 g Klotz's: there are 3 formulas: 2 by Klotz & Coburn (1916) and 1 by Klotz & MacLachan (1915) The 1915 formula (original) is as follows: Water dist.---100 mL + 40% formaldehyde --- 0.5 mL + chloral hydrate --- 3 g + sodium sulfate --- 0.7 g + sodium bicarbonate --- 0.6 g + sodium chloride --- 0.55 g + potassium sulfate --- 0.06 g + potassium nitrate --- 1.2 g Rene J. Sionagh Smith wrote: Can anyone help me with a recipe for Jore's or Klotz fixative solutions? I have used both in the past for retaining colour and consistency of post mortem specimens for teaching veterinary students. I have been unable to find an actual recipe since someone else always prepared them! I wish I'd paid more attention now.... Any pointers would be much appreciated. Sionagh Smith Dept. of Veterinary Pathology Royal (Dick) School of Veterinary Studies Easter Bush Veterinary Centre Easter Bush Midlothian EH25 9RG Tel 0131 650 8787 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From b003046 <@t> nf.au.dk Thu Dec 1 08:51:56 2005 From: b003046 <@t> nf.au.dk (b003046@nf.au.dk) Date: Thu Dec 1 08:52:07 2005 Subject: [Histonet] c-kit immunohistochemical staining Message-ID: <1133448716.438f0e0c80ce2@webmail.nf.au.dk> Dear all, Have any experience with c-kit (CD117/C-19) immunohistochemical staining in rat paraffin-embeded tissue? I have tryed different antigen retrieval methods, and still cant get it to work! Any info would be greatly appreciated. Thank you Mette K. Hagensen Department of Cardiology B, Research Unit Aarhus University Hospital, Skejby Hospital Brendstrupgaardsvej 100 8200 Aarhus N Denmark Mail: b003046@nf.au.dk From TJJ <@t> Stowers-Institute.org Thu Dec 1 08:55:46 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Dec 1 08:56:19 2005 Subject: [Histonet] Re: Aspirgillus antibody Message-ID: Can someone enlighten me why it would be advantageous to use an antibody against aspirgillus when the silver and PAS techniques work so well? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From histosci <@t> shentel.net Thu Dec 1 09:02:38 2005 From: histosci <@t> shentel.net (HSRL) Date: Thu Dec 1 09:03:01 2005 Subject: [Histonet] RE: Data, block/slide wet tissue storage Message-ID: <007b01c5f688$455b6590$0200a8c0@HSRLMAIN> Dear Laura, We have a histopath lab, but also have a short/long term archiving facility for data, wet tissue, slides/blocks and refrigerated and frozen specimens. We have an FM-200 fire suppression system which basically has taken the place of Halon because of its ozone depleting properties. We also have humidity and temperature recorders in all vaults and refrigerators/freezers that monitor the temps and humidity 24/7. We have alarm contacts on them as well. This is in place in case anything goes below or above the set temperatures and humidity, our alarm system will sound immediately and we will be called automatically. In short, we have to keep precise records as to how and when everything is maintained. If anyone would like to talk further about such matters, please feel free to call Sam Jeffrey or Ed Galati at 540-477-4441. Either of them would be happy to discuss this with you. Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jones, Laura Sent: Wednesday, November 30, 2005 2:03 PM To: Histonet (E-mail) Subject: [Histonet] Paraffin Block Storage Greetings to everyone in Histoland! We are hoping you all can help us with a question. We store our paraffin blocks in a small closet/room (5' x 10', we're guessing) and have had a Halon fire extinguishing system in there for years. We have apparently been cited by the PA Dept. of Health (?) for this system not being adequate. So, the question is, what type of system does everyone else have? We realize that regulations may differ from state to state, but would appreciate any input, as our safety officer seems to be puzzled as well. Thanks in advance to all the experts. Laura Jones _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From andrae <@t> u.washington.edu Thu Dec 1 09:59:58 2005 From: andrae <@t> u.washington.edu (A. Erickson) Date: Thu Dec 1 10:00:09 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: References: Message-ID: And they only run one sample at a time, and only work on one case at a time!!!! andra On Thu, 1 Dec 2005, Molinari, Betsy wrote: > Isn't it odd that there usually is no one else in the labs except the > one or two investigators working on the one or two cases going on that > week? > > Betsy Molinari HT (ASCP) > Texas Heart Institute > Cardiovascular Pathology > 6770 Bertner Ave. > 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 Shirley > Powell > Sent: Wednesday, November 30, 2005 1:45 PM > To: king.laurie@marshfieldclinic.org; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > Yep, that is when I stopped watching Quincy. Made me want to fight too. > I > do watch CSI but usually not for accuracy, mostly for the scenery. > > Shirley > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > king.laurie@marshfieldclinic.org > Sent: Wednesday, November 30, 2005 2:30 PM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > > I used to love that show, until the episode where Quincy complained that > Sam > was demoted to work in histology, who were, after all, only a bunch of > "bottle labelers". Them's fightin' words! > > Laurie King > > > ------Original Message------ > From: "Bartlett, Jeanine" > Date: Wed Nov 30, 2005 -- 01:15:00 PM > To: "Pam Marcum" , "Ford Royer" > , "Histonet" > Subject: RE: [Histonet] Histotechs: born or made? > > And wasn't it amazing how much Sam and Quincy got done in one hour!? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam > Marcum > Sent: Wednesday, November 30, 2005 1:22 PM > To: Ford Royer; 'Histonet' > Subject: RE: [Histonet] Histotechs: born or made? > > Ford, > > I don't think we are allowed to tell the new people in the field how > much > fun we had in the old days. I have always loved my job however, > sometimes I > have to watch my sense of humor in non-histologist/medical company as we > don't ususally see things the same way they do. > > Heck, I thought Qunicy and now CSI were sit coms at first as it was so > far > from what we were doing and really funny for mistakes. All my first > boss in > histology (and also a city/county coroner) wanted for several years was > a > Sam like Qunicy had with all the equipment of course. He figured he > could > rid of at least 5 people in the lab with one Sam. > > Pam Marcum > > >> When I was a practicing laboratory scientist (27 years ago), we would >> have some of the wildest lab parties and everyone seemed to be on the >> same page as far as having a weird sense of humor. A work day didn't >> go by without some form of laughter in our lab. Non-laboratory people > >> often asked me why this was. The only thing that I could come up with > >> is it was how we dealt with the profession that we chose. I won't go >> into details or give examples. We all know what I am talking about. >> It does take a special kind of person to this sometimes morbid (some >> would say hideous) work and I for one am glad that there are these >> types of persons to take it on. Thank you all for your dedication to >> your profession and the people that you serve - mankind. >> >> ~ Ford >> >> Ford M. Royer, MT(ASCP) >> Minneapolis, MN >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> Morken, Tim >> - Labvision >> Sent: Wednesday, November 30, 2005 11:11 AM >> To: 'Ingles Claire'; Histonet >> Subject: [Histonet] Histotechs: born or made? >> >> The first time I walked into a histology lab it was the day after the >> 4th of July and there were 4 blackened fingers sitting on the grossing > >> bench (one guess how they got there - and it's nothing to do with >> anything Cajun!). My first thought was : 'this is going to be a >> strange job.' I've seen much stranger things since, so I think >> histotechs become strange due to exposure to unnatural sights (among >> other things!). And, of course, the pathology staff of any hospital is > infamous for their "gross" humor. >> >> >> Tim Morken >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > >> Claire >> Sent: Wednesday, November 30, 2005 8:55 AM >> To: Histonet >> Subject: RE: [Histonet] Re: 70% from NBF >> >> >> I have wondered the same thing many times myself. Whether it was >> naturally me or the addition of the chemicals that made me a bit >> strange. I think it may be partly both. I usually blame it on the >> chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison >> WI >> >> ________________________________ >> >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan >> Llewellyn >> Sent: Tue 11/29/2005 11:15 AM >> To: Histonet >> Subject: Re: [Histonet] Re: 70% from NBF >> >> >> >> I have often wondered whether I became a histotech because I was born >> strange, or whether I became strange because of the time I spent >> training in a place like that! >> >> Bryan Llewellyn >> >> >> ----- Original Message ----- >> From: "Gayle Callis" >> To: >> Sent: Tuesday, November 29, 2005 8:23 AM >> Subject: [Histonet] Re: 70% from NBF >> >> >>> Joseph made some excellent points here >>> >>> Chloroform is an excellent clearing agent (used it back in the 60's >>> in open dip and dunk processors - O.K. so I'm old!) but no one >>> warned us about its carcinogenic nature and there were no safety >>> issue regulations then. Take his advice! >> >> >> _______________________________________________ >> 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >> From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From froyer <@t> bitstream.net Thu Dec 1 10:12:33 2005 From: froyer <@t> bitstream.net (Ford Royer) Date: Thu Dec 1 10:12:58 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: Message-ID: <002001c5f692$0a2e6d00$0e00a8c0@fords> I always got a kick out of Quincy when he would get personally involved with a victim's family... even to the extent of going to their homes and sitting down with them in their family room. Great stuff, and very touching, but I just could not see any of our pathologists doing this. Not that they might not have wanted to at times, they were a great bunch of guys & gals... but a "house call" by the chief pathologist? I don't THINK so... ~ Ford Ford M. Royer, MT(ASCP) Minnesota Medical Specialists Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of A. Erickson Sent: Thursday, December 01, 2005 10:00 AM To: Molinari, Betsy Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? And they only run one sample at a time, and only work on one case at a time!!!! andra On Thu, 1 Dec 2005, Molinari, Betsy wrote: > Isn't it odd that there usually is no one else in the labs except the > one or two investigators working on the one or two cases going on that > week? > > Betsy Molinari HT (ASCP) > Texas Heart Institute > Cardiovascular Pathology > 6770 Bertner Ave. > 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 Shirley > Powell > Sent: Wednesday, November 30, 2005 1:45 PM > To: king.laurie@marshfieldclinic.org; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > Yep, that is when I stopped watching Quincy. Made me want to fight too. > I > do watch CSI but usually not for accuracy, mostly for the scenery. > > Shirley > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > king.laurie@marshfieldclinic.org > Sent: Wednesday, November 30, 2005 2:30 PM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > > I used to love that show, until the episode where Quincy complained that > Sam > was demoted to work in histology, who were, after all, only a bunch of > "bottle labelers". Them's fightin' words! > > Laurie King > > > ------Original Message------ > From: "Bartlett, Jeanine" > Date: Wed Nov 30, 2005 -- 01:15:00 PM > To: "Pam Marcum" , "Ford Royer" > , "Histonet" > Subject: RE: [Histonet] Histotechs: born or made? > > And wasn't it amazing how much Sam and Quincy got done in one hour!? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam > Marcum > Sent: Wednesday, November 30, 2005 1:22 PM > To: Ford Royer; 'Histonet' > Subject: RE: [Histonet] Histotechs: born or made? > > Ford, > > I don't think we are allowed to tell the new people in the field how > much > fun we had in the old days. I have always loved my job however, > sometimes I > have to watch my sense of humor in non-histologist/medical company as we > don't ususally see things the same way they do. > > Heck, I thought Qunicy and now CSI were sit coms at first as it was so > far > from what we were doing and really funny for mistakes. All my first > boss in > histology (and also a city/county coroner) wanted for several years was > a > Sam like Qunicy had with all the equipment of course. He figured he > could > rid of at least 5 people in the lab with one Sam. > > Pam Marcum > > >> When I was a practicing laboratory scientist (27 years ago), we would >> have some of the wildest lab parties and everyone seemed to be on the >> same page as far as having a weird sense of humor. A work day didn't >> go by without some form of laughter in our lab. Non-laboratory people > >> often asked me why this was. The only thing that I could come up with > >> is it was how we dealt with the profession that we chose. I won't go >> into details or give examples. We all know what I am talking about. >> It does take a special kind of person to this sometimes morbid (some >> would say hideous) work and I for one am glad that there are these >> types of persons to take it on. Thank you all for your dedication to >> your profession and the people that you serve - mankind. >> >> ~ Ford >> >> Ford M. Royer, MT(ASCP) >> Minneapolis, MN >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> Morken, Tim >> - Labvision >> Sent: Wednesday, November 30, 2005 11:11 AM >> To: 'Ingles Claire'; Histonet >> Subject: [Histonet] Histotechs: born or made? >> >> The first time I walked into a histology lab it was the day after the >> 4th of July and there were 4 blackened fingers sitting on the grossing > >> bench (one guess how they got there - and it's nothing to do with >> anything Cajun!). My first thought was : 'this is going to be a >> strange job.' I've seen much stranger things since, so I think >> histotechs become strange due to exposure to unnatural sights (among >> other things!). And, of course, the pathology staff of any hospital is > infamous for their "gross" humor. >> >> >> Tim Morken >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > >> Claire >> Sent: Wednesday, November 30, 2005 8:55 AM >> To: Histonet >> Subject: RE: [Histonet] Re: 70% from NBF >> >> >> I have wondered the same thing many times myself. Whether it was >> naturally me or the addition of the chemicals that made me a bit >> strange. I think it may be partly both. I usually blame it on the >> chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison >> WI >> >> ________________________________ >> >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan >> Llewellyn >> Sent: Tue 11/29/2005 11:15 AM >> To: Histonet >> Subject: Re: [Histonet] Re: 70% from NBF >> >> >> >> I have often wondered whether I became a histotech because I was born >> strange, or whether I became strange because of the time I spent >> training in a place like that! >> >> Bryan Llewellyn >> >> >> ----- Original Message ----- >> From: "Gayle Callis" >> To: >> Sent: Tuesday, November 29, 2005 8:23 AM >> Subject: [Histonet] Re: 70% from NBF >> >> >>> Joseph made some excellent points here >>> >>> Chloroform is an excellent clearing agent (used it back in the 60's >>> in open dip and dunk processors - O.K. so I'm old!) but no one >>> warned us about its carcinogenic nature and there were no safety >>> issue regulations then. Take his advice! >> >> >> _______________________________________________ >> 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >> From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 > > > > > > _______________________________________________ > 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 gu.lang <@t> gmx.at Thu Dec 1 10:16:51 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Dec 1 10:16:55 2005 Subject: AW: [Histonet] Paraffin sections In-Reply-To: Message-ID: We did the dry method a couple of times and never heard any complaints. We even put the dry slides directly into the staining solutions. Gudrun -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Molinari, Betsy Gesendet: Donnerstag, 01. Dezember 2005 14:47 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Paraffin sections Hi, I have heard that paraffin sections that have been deparaffinized and run down to DH20 but for some reason cannot be stained that day, can be left to air dry and then be placed back into H2O when they are ready for staining. Does anyone have any experience with this technique? I usually just leave them in DH2O till I can get to them, usually the next day. Thanks. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Dec 1 10:23:42 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Dec 1 10:24:23 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: I am a big fan of "ER" - but it cracks me up when the ER residents go to the lab to diagnose a biopsy or FNA they took in the ER. "Ford Royer" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/01/2005 10:12 AM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] Histotechs: born or made? I always got a kick out of Quincy when he would get personally involved with a victim's family... even to the extent of going to their homes and sitting down with them in their family room. Great stuff, and very touching, but I just could not see any of our pathologists doing this. Not that they might not have wanted to at times, they were a great bunch of guys & gals... but a "house call" by the chief pathologist? I don't THINK so... ~ Ford Ford M. Royer, MT(ASCP) Minnesota Medical Specialists Minneapolis, MN -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of A. Erickson Sent: Thursday, December 01, 2005 10:00 AM To: Molinari, Betsy Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? And they only run one sample at a time, and only work on one case at a time!!!! andra On Thu, 1 Dec 2005, Molinari, Betsy wrote: > Isn't it odd that there usually is no one else in the labs except the > one or two investigators working on the one or two cases going on that > week? > > Betsy Molinari HT (ASCP) > Texas Heart Institute > Cardiovascular Pathology > 6770 Bertner Ave. > 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 Shirley > Powell > Sent: Wednesday, November 30, 2005 1:45 PM > To: king.laurie@marshfieldclinic.org; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > Yep, that is when I stopped watching Quincy. Made me want to fight too. > I > do watch CSI but usually not for accuracy, mostly for the scenery. > > Shirley > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > king.laurie@marshfieldclinic.org > Sent: Wednesday, November 30, 2005 2:30 PM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > > I used to love that show, until the episode where Quincy complained that > Sam > was demoted to work in histology, who were, after all, only a bunch of > "bottle labelers". Them's fightin' words! > > Laurie King > > > ------Original Message------ > From: "Bartlett, Jeanine" > Date: Wed Nov 30, 2005 -- 01:15:00 PM > To: "Pam Marcum" , "Ford Royer" > , "Histonet" > Subject: RE: [Histonet] Histotechs: born or made? > > And wasn't it amazing how much Sam and Quincy got done in one hour!? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam > Marcum > Sent: Wednesday, November 30, 2005 1:22 PM > To: Ford Royer; 'Histonet' > Subject: RE: [Histonet] Histotechs: born or made? > > Ford, > > I don't think we are allowed to tell the new people in the field how > much > fun we had in the old days. I have always loved my job however, > sometimes I > have to watch my sense of humor in non-histologist/medical company as we > don't ususally see things the same way they do. > > Heck, I thought Qunicy and now CSI were sit coms at first as it was so > far > from what we were doing and really funny for mistakes. All my first > boss in > histology (and also a city/county coroner) wanted for several years was > a > Sam like Qunicy had with all the equipment of course. He figured he > could > rid of at least 5 people in the lab with one Sam. > > Pam Marcum > > >> When I was a practicing laboratory scientist (27 years ago), we would >> have some of the wildest lab parties and everyone seemed to be on the >> same page as far as having a weird sense of humor. A work day didn't >> go by without some form of laughter in our lab. Non-laboratory people > >> often asked me why this was. The only thing that I could come up with > >> is it was how we dealt with the profession that we chose. I won't go >> into details or give examples. We all know what I am talking about. >> It does take a special kind of person to this sometimes morbid (some >> would say hideous) work and I for one am glad that there are these >> types of persons to take it on. Thank you all for your dedication to >> your profession and the people that you serve - mankind. >> >> ~ Ford >> >> Ford M. Royer, MT(ASCP) >> Minneapolis, MN >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> Morken, Tim >> - Labvision >> Sent: Wednesday, November 30, 2005 11:11 AM >> To: 'Ingles Claire'; Histonet >> Subject: [Histonet] Histotechs: born or made? >> >> The first time I walked into a histology lab it was the day after the >> 4th of July and there were 4 blackened fingers sitting on the grossing > >> bench (one guess how they got there - and it's nothing to do with >> anything Cajun!). My first thought was : 'this is going to be a >> strange job.' I've seen much stranger things since, so I think >> histotechs become strange due to exposure to unnatural sights (among >> other things!). And, of course, the pathology staff of any hospital is > infamous for their "gross" humor. >> >> >> Tim Morken >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > >> Claire >> Sent: Wednesday, November 30, 2005 8:55 AM >> To: Histonet >> Subject: RE: [Histonet] Re: 70% from NBF >> >> >> I have wondered the same thing many times myself. Whether it was >> naturally me or the addition of the chemicals that made me a bit >> strange. I think it may be partly both. I usually blame it on the >> chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison >> WI >> >> ________________________________ >> >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan >> Llewellyn >> Sent: Tue 11/29/2005 11:15 AM >> To: Histonet >> Subject: Re: [Histonet] Re: 70% from NBF >> >> >> >> I have often wondered whether I became a histotech because I was born >> strange, or whether I became strange because of the time I spent >> training in a place like that! >> >> Bryan Llewellyn >> >> >> ----- Original Message ----- >> From: "Gayle Callis" >> To: >> Sent: Tuesday, November 29, 2005 8:23 AM >> Subject: [Histonet] Re: 70% from NBF >> >> >>> Joseph made some excellent points here >>> >>> Chloroform is an excellent clearing agent (used it back in the 60's >>> in open dip and dunk processors - O.K. so I'm old!) but no one >>> warned us about its carcinogenic nature and there were no safety >>> issue regulations then. Take his advice! >> >> >> _______________________________________________ >> 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >> From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sluhisto <@t> yahoo.com Thu Dec 1 10:30:43 2005 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Thu Dec 1 10:30:53 2005 Subject: [Histonet] Detecting contrast media-Iodine in Human Kidney Tissue Message-ID: <20051201163043.34361.qmail@web51001.mail.yahoo.com> Hello All: I am in a meeting with two renal pathologists and need some help quickly, if possible. Does anyone know of a way to detect contrast media specifically iodine in human kidney biopsy tissue? Thanks in advance. Susan SLU --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From jfish <@t> gladstone.ucsf.edu Thu Dec 1 10:34:56 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Thu Dec 1 10:35:11 2005 Subject: [Histonet] Histotechs: born or made? In-Reply-To: Message-ID: <001101c5f695$2a111610$8903010a@JFISH> I haven't seen an episode of CSI in a while, but when I did watch I noticed that they always had everything backlit with NO direct lighting. You could see the light shining up into their faces, but never onto their work surface. I'd go blind working in those conditions! Jo Dee -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Molinari, Betsy Sent: Thursday, December 01, 2005 4:03 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? Isn't it odd that there usually is no one else in the labs except the one or two investigators working on the one or two cases going on that week? Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. 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 Shirley Powell Sent: Wednesday, November 30, 2005 1:45 PM To: king.laurie@marshfieldclinic.org; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? Yep, that is when I stopped watching Quincy. Made me want to fight too. I do watch CSI but usually not for accuracy, mostly for the scenery. Shirley -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of king.laurie@marshfieldclinic.org Sent: Wednesday, November 30, 2005 2:30 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? I used to love that show, until the episode where Quincy complained that Sam was demoted to work in histology, who were, after all, only a bunch of "bottle labelers". Them's fightin' words! Laurie King ------Original Message------ From: "Bartlett, Jeanine" Date: Wed Nov 30, 2005 -- 01:15:00 PM To: "Pam Marcum" , "Ford Royer" , "Histonet" Subject: RE: [Histonet] Histotechs: born or made? And wasn't it amazing how much Sam and Quincy got done in one hour!? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam Marcum Sent: Wednesday, November 30, 2005 1:22 PM To: Ford Royer; 'Histonet' Subject: RE: [Histonet] Histotechs: born or made? Ford, I don't think we are allowed to tell the new people in the field how much fun we had in the old days. I have always loved my job however, sometimes I have to watch my sense of humor in non-histologist/medical company as we don't ususally see things the same way they do. Heck, I thought Qunicy and now CSI were sit coms at first as it was so far from what we were doing and really funny for mistakes. All my first boss in histology (and also a city/county coroner) wanted for several years was a Sam like Qunicy had with all the equipment of course. He figured he could rid of at least 5 people in the lab with one Sam. Pam Marcum > When I was a practicing laboratory scientist (27 years ago), we would > have some of the wildest lab parties and everyone seemed to be on the > same page as far as having a weird sense of humor. A work day didn't > go by without some form of laughter in our lab. Non-laboratory people > often asked me why this was. The only thing that I could come up with > is it was how we dealt with the profession that we chose. I won't go > into details or give examples. We all know what I am talking about. > It does take a special kind of person to this sometimes morbid (some > would say hideous) work and I for one am glad that there are these > types of persons to take it on. Thank you all for your dedication to > your profession and the people that you serve - mankind. > > ~ Ford > > Ford M. Royer, MT(ASCP) > Minneapolis, MN > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Morken, Tim > - Labvision > Sent: Wednesday, November 30, 2005 11:11 AM > To: 'Ingles Claire'; Histonet > Subject: [Histonet] Histotechs: born or made? > > The first time I walked into a histology lab it was the day after the > 4th of July and there were 4 blackened fingers sitting on the grossing > bench (one guess how they got there - and it's nothing to do with > anything Cajun!). My first thought was : 'this is going to be a > strange job.' I've seen much stranger things since, so I think > histotechs become strange due to exposure to unnatural sights (among > other things!). And, of course, the pathology staff of any hospital is infamous for their "gross" humor. > > > Tim Morken > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > Claire > Sent: Wednesday, November 30, 2005 8:55 AM > To: Histonet > Subject: RE: [Histonet] Re: 70% from NBF > > > I have wondered the same thing many times myself. Whether it was > naturally me or the addition of the chemicals that made me a bit > strange. I think it may be partly both. I usually blame it on the > chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison > WI > > ________________________________ > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan > Llewellyn > Sent: Tue 11/29/2005 11:15 AM > To: Histonet > Subject: Re: [Histonet] Re: 70% from NBF > > > > I have often wondered whether I became a histotech because I was born > strange, or whether I became strange because of the time I spent > training in a place like that! > > Bryan Llewellyn > > > ----- Original Message ----- > From: "Gayle Callis" > To: > Sent: Tuesday, November 29, 2005 8:23 AM > Subject: [Histonet] Re: 70% from NBF > > > > Joseph made some excellent points here > > > > Chloroform is an excellent clearing agent (used it back in the 60's > > in open dip and dunk processors - O.K. so I'm old!) but no one > > warned us about its carcinogenic nature and there were no safety > > issue regulations then. Take his advice! > > > _______________________________________________ > 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet >From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 _______________________________________________ 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 mauger <@t> email.chop.edu Thu Dec 1 10:38:16 2005 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Thu Dec 1 10:39:20 2005 Subject: [Histonet] aspergillus ihc Message-ID: All, We use the aspergillus antibody stain to distinguish it from other fungi or yeast like candida, which also stain positively with the silver stain. There is a specific treatment available for aspergillus infections, so we want to be sure thats what it is! Jo Mauger From tahseen <@t> brain.net.pk Thu Dec 1 02:38:25 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Thu Dec 1 10:40:32 2005 Subject: [Histonet] c-kit immunohistochemical staining References: <1133448716.438f0e0c80ce2@webmail.nf.au.dk> Message-ID: <001f01c5f652$99897200$972bfea9@m7c0y4> Dear Mette K. Hagensen, We are using citrate buffer pH 6 for 15 min and get good result. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: To: Sent: Friday, December 02, 2005 3:51 AM Subject: [Histonet] c-kit immunohistochemical staining > Dear all, > Have any experience with c-kit (CD117/C-19) immunohistochemical staining in rat > paraffin-embeded tissue? I have tryed different antigen retrieval methods, and > still cant get it to work! Any info would be greatly appreciated. > Thank you > > Mette K. Hagensen > Department of Cardiology B, Research Unit > Aarhus University Hospital, Skejby Hospital > Brendstrupgaardsvej 100 > 8200 Aarhus N > Denmark > > Mail: b003046@nf.au.dk > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Dec 1 10:56:43 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 1 10:56:57 2005 Subject: [Histonet] Paraffin sections In-Reply-To: References: Message-ID: <6.0.0.22.1.20051201094601.01b5d880@gemini.msu.montana.edu> Betsy, I have never left a deparaffinized section in distilled water for staining next day - personally, doing the deparaffinization doesn't really take that long in the first place. I'm not very trusting of leaving sections in distilled water, even though Distilled H2) is supposed to be "pure", I worry about some funky contaminant growing overnight, maybe this is a silly worry. However, a little trick I learned from Nate Brinn (when he gave workshops on staining years ago) was deparaffinize through xylenes - just to remove paraffin and air dry at that point. We rinsed these sections with 95% alcohol, quick 70% then distilled water before staining protocols. I have shipped slides that were deparaffinized a week in advance of an H&E wet workshop and also a decalcified bone staining workshop, did this little technic and we had excellent staining results. I don't think I would want to store these deparaffinzed sections for a long period of time particularly if IHC is needed nor some other stain that may be losing protection of paraffin or exposure to air that changes how the stain works. At 06:46 AM 12/1/2005, you wrote: >Hi, > > I have heard that paraffin sections that have been deparaffinized and >run down to DH20 but for some reason cannot be stained that day, can be >left to air dry and then be placed back into H2O when they are ready for >staining. Does anyone have any experience with this technique? I usually >just leave them in DH2O till I can get to them, usually the next day. > >Thanks. > > > >Betsy Molinari HT (ASCP) > >Texas Heart Institute > >Cardiovascular Pathology > >6770 Bertner Ave. > >Houston,TX 77030 > >832-355-6524 > >832-355-6812 (fax) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cforster <@t> umn.edu Thu Dec 1 10:56:05 2005 From: cforster <@t> umn.edu (Colleen Forster) Date: Thu Dec 1 10:56:58 2005 Subject: [Histonet] Paraffin sections In-Reply-To: References: Message-ID: <438F2B25.4090701@umn.edu> I personally don't like to do that at all. I think you loose staining after they have sat, doesn't matter if it is in dh2o,buffer or sir dried(ouch!). Maybe others have had success. You would be better off leaving them until the next day. Colleen Forster U of MN Molinari, Betsy wrote: >Hi, > > I have heard that paraffin sections that have been deparaffinized and >run down to DH20 but for some reason cannot be stained that day, can be >left to air dry and then be placed back into H2O when they are ready for >staining. Does anyone have any experience with this technique? I usually >just leave them in DH2O till I can get to them, usually the next day. > >Thanks. > > > >Betsy Molinari HT (ASCP) > >Texas Heart Institute > >Cardiovascular Pathology > >6770 Bertner Ave. > >Houston,TX 77030 > >832-355-6524 > >832-355-6812 (fax) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >. > > > From gcallis <@t> montana.edu Thu Dec 1 11:08:10 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 1 11:08:26 2005 Subject: [Histonet] RE: Histotechs: born or made? In-Reply-To: <001101c5f695$2a111610$8903010a@JFISH> References: <001101c5f695$2a111610$8903010a@JFISH> Message-ID: <6.0.0.22.1.20051201100041.01b0b3d0@gemini.msu.montana.edu> Jo, Never, you need to buy one of the trusty, tidy little flashlights they use for everything!!! Even in broad daylight!! Gayle Callis At 09:34 AM 12/1/2005, you wrote: >I haven't seen an episode of CSI in a while, but when I did watch I noticed >that they always had everything backlit with NO direct lighting. You could >see the light shining up into their faces, but never onto their work >surface. I'd go blind working in those conditions! >Jo Dee From dpahisto <@t> yahoo.com Thu Dec 1 11:51:54 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Thu Dec 1 11:52:03 2005 Subject: [Histonet] Microtomy Message-ID: <20051201175154.83738.qmail@web33408.mail.mud.yahoo.com> This is a question for those techs who cut on a manual Reichert-Jung 2030. I have one tech who insists on completely taking apart her knife holder everyday to clean it. She removes the springs and all. This person is also the only tech who has trouble cutting and is constantly having to recut a specimen or nearly cutting through needle biopsies. I maintain that handling the knife holder spring so much causes the problems (knife won't clamp tight evenly, chatter and skipping). The other techs, including myself only dismantle and clean the knife holder about once a month. We have had to purchase new spring and bolts for this microtome about every 6 months, whereas the other microtomes have never had to be replaced. How do I go about telling this person to quit cleaning so much. I have had the knife holder and microtome serviced to make sure there is nothing else wrong. I have cut on that machine and find it cuts beautifully as long as you don't take it apart after every 10 blocks to clean behind the blade. Any suggestions would be greatly appreciated, Cindy DuBois Integrated Pathology Assoc. Stockton CA --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Try Yahoo! Personals From Rcartun <@t> harthosp.org Thu Dec 1 11:56:07 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Dec 1 11:56:52 2005 Subject: [Histonet] Aspergillus antibody Message-ID: Hi Bill: Yes, I do IHC for aspergillus on human tissue and have obtained excellent results. Unfortunately, Dako no longer sells the monoclonal antibody that I was using. The clone (Mab-WF-AF-1), as well as other anti-aspergillus antibodies, may be available elsewhere. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Bill Sinai" 11/30/05 08:55PM >>> Dear All, Is there anyone out there using an antibody to aspergillus species? If so how do you do it (dilutions, manually or by machine)? Do any antibody work better than others? Is it available for FFPE specimens. Any info would be greatly appreciated. Thanks Bill Sinai Chief Hospital Scientist Tissue Pathology ICPMR Westmead Hospital Westmead NSW 2145 Phone 02 9845 7774 Fax 02 9687 2330 __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From asmith <@t> mail.barry.edu Thu Dec 1 11:59:17 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Thu Dec 1 12:01:56 2005 Subject: [Histonet] Re: 70% from NBF Message-ID: <5D2189E74151CC42BEC02906BA8996322B9132@exchsrv01.barrynet.barry.edu> Phosgene dates from World War I, in which it was the principal Allied war gas. It is not a neurotoxin, but a pulmonary toxin. It has a deceptively pleasant smell, like new-mown hay. About an hour after exposure, it causes pulmonary edema, which is often fatal. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Tuesday, November 29, 2005 12:15 PM To: Histonet Subject: Re: [Histonet] Re: 70% from NBF Talking about chloroform use in the past. I trained as a lab tech in the early 60s, and the hospital where I did histology specialist training in London used chloroform exclusively. The pathologist apparently thought that xylene made tissues too brittle. I have many times dunked my hand in a litre container of it to get a block out. You wouldn't believe how much it stings, apparently by slightly defatting the skin! I was told this after I had done it a few times, of course. We not only used chloroform without any safety concerns at all, but we also redistilled it to save money. We had an upright, water cooled still with a Liebig type condensor. It was heated with an electrical coil in a sand bath, which surrounded a 2 litre round bottom flask. There was no automatic cut off for the electricity, and we had to keep an eye on it and switch it off when the flask was almost empty. Of course, being a very attentive teenager at the time I missed it more often than not. This was usually announced by a disgusting smell and billows of white smoke. I was never sure whether the smoke was burning fat and stuff, or phosgene, which is produced from chloroform. For those not aware, phosgene is one of the gases used during World War II as a nerve gas. I have often wondered whether I became a histotech because I was born strange, or whether I became strange because of the time I spent training in a place like that! Bryan Llewellyn ----- Original Message ----- From: "Gayle Callis" To: Sent: Tuesday, November 29, 2005 8:23 AM Subject: [Histonet] Re: 70% from NBF > Joseph made some excellent points here > > Chloroform is an excellent clearing agent (used it back in the 60's in > open dip and dunk processors - O.K. so I'm old!) but no one warned us > about its carcinogenic nature and there were no safety issue regulations > then. Take his advice! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From Rcartun <@t> harthosp.org Thu Dec 1 12:04:05 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Dec 1 12:04:40 2005 Subject: [Histonet] Re: Aspirgillus antibody Message-ID: Immunohistochemistry is more sensitive (and easier to interpret) than histochemical stains for many infectious disease agents. I have seen a few cases where the GMS stain was non-diagnostic, but the immunoperoxidase stain for aspergillus revealed unequivocal immunoreactivity. I have also seen cases where the fungus was diagnosed as aspergillus, but the immunoperoxidase stain was negative, and was subsequently proven to be mucor, once again by immunoperoxidase staining. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Johnson, Teri" 12/01/05 09:55AM >>> Can someone enlighten me why it would be advantageous to use an antibody against aspirgillus when the silver and PAS techniques work so well? Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From Luis.Chiriboga <@t> med.nyu.edu Thu Dec 1 12:13:58 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Thu Dec 1 12:11:52 2005 Subject: [Histonet] Question: Inventory Message-ID: Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis From gagnone <@t> KGH.KARI.NET Thu Dec 1 12:11:46 2005 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Thu Dec 1 12:11:54 2005 Subject: [Histonet] Microtomy Message-ID: Having used 2030's and 2035's, I find that not taking the knife holder apart very often works for me. If a minimal amount of wax gets into the knife holder, it seems to me that it won't have to be cleaned/removed later. I find that a "dust gun" (canned air) works well to remove bits of wax around the hard-to-reach areas where the blade is placed in or removed from the knife holder. Maybe receiving responses like this will add weight to your argument, and show that the microtome can indeed work well with minimal knife holder cleaning. Hope this helps, Eric Gagnon MLT Kingston General Hospital Kingston, Ontario From rjbuesa <@t> yahoo.com Thu Dec 1 12:24:48 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 1 12:24:57 2005 Subject: [Histonet] Microtomy In-Reply-To: <20051201175154.83738.qmail@web33408.mail.mud.yahoo.com> Message-ID: <20051201182448.51452.qmail@web61215.mail.yahoo.com> Hi Cindy: First it is not necessary to take appart a knife holder daily. For what you describe this techs acts as some do, thinking that they are like "independent contractors" within the lab setting able to do anything the want regardless of general rules. Do you have an SOP that addresses this cleaning procedure? If you don't you can write this aspect and have everybody to adhere to it. With some statistics about recuts required for bad technique you can also try to convince her. You could also get advise from the sales representative regarding this practice. If everything fails you will have to impose authority on the issue. Hope you will be able to reign in this stubborn tech. Rene J. Cindy DuBois wrote: This is a question for those techs who cut on a manual Reichert-Jung 2030. I have one tech who insists on completely taking apart her knife holder everyday to clean it. She removes the springs and all. This person is also the only tech who has trouble cutting and is constantly having to recut a specimen or nearly cutting through needle biopsies. I maintain that handling the knife holder spring so much causes the problems (knife won't clamp tight evenly, chatter and skipping). The other techs, including myself only dismantle and clean the knife holder about once a month. We have had to purchase new spring and bolts for this microtome about every 6 months, whereas the other microtomes have never had to be replaced. How do I go about telling this person to quit cleaning so much. I have had the knife holder and microtome serviced to make sure there is nothing else wrong. I have cut on that machine and find it cuts beautifully as long as you don't take it apart after every 10 blocks to clean behind the blade. Any suggestions would be greatly appreciated, Cindy DuBois Integrated Pathology Assoc. Stockton CA --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Try Yahoo! Personals _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From rjbuesa <@t> yahoo.com Thu Dec 1 12:30:15 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 1 12:30:26 2005 Subject: [Histonet] Question: Inventory In-Reply-To: Message-ID: <20051201183015.57199.qmail@web61221.mail.yahoo.com> Hi Luis: A good managerial practice is to register all the reagents as they arrive. That is the best way to check delivery versus order. Once they are going to be used you should write date opened on each reagenet and this will allow you to calculate usage rate. Hope this will help! Rene J. Luis Chiriboga wrote: Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Music Unlimited - Access over 1 million songs. Try it free. From JOliver <@t> asterand.com Thu Dec 1 12:31:26 2005 From: JOliver <@t> asterand.com (Jeff Oliver) Date: Thu Dec 1 12:31:35 2005 Subject: [Histonet] Modified Schmidt's Hematoxylin Message-ID: <1B0B60D56CA6A440A240E866BA3458D8101025@ATL1VEXC017.usdom004.tco.tc> Hello, I'm looking for a recipe for a modified Schmidt's Hematoxylin. The only references I have found to it are from a couple papers that did not include a recipe. Until a couple days ago I had no idea this version existed. Does anyone have a recipe or know a vendor for this? Any help would be greatly appreciated. Thanks! -Jeff From vazquezr <@t> ohsu.edu Thu Dec 1 12:32:42 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Dec 1 12:33:11 2005 Subject: [Histonet] moh's surgery Message-ID: I am a histo tech working as a Mohs tech. The qualifications is a BS or equivalent of. Robyn OHSU From vazquezr <@t> ohsu.edu Thu Dec 1 12:35:44 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Dec 1 12:36:27 2005 Subject: [Histonet] moh's surgery Message-ID: That is correct, you don't have to be a histo tech, I have trained 3 non-histo tech to do Mohs surgery. And our surgeons read their slides. Robyn OHSU From lrichey <@t> u.washington.edu Thu Dec 1 13:17:33 2005 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Thu Dec 1 13:17:42 2005 Subject: [Histonet] c-kit immunohistochemical staining In-Reply-To: <1133448716.438f0e0c80ce2@webmail.nf.au.dk> References: <1133448716.438f0e0c80ce2@webmail.nf.au.dk> Message-ID: <438F4C4D.9070101@u.washington.edu> We use C-kit on human tissue. Antigen Retrieval for our rabbit Ckit is 15 microwave in EDTA pH8 b003046@nf.au.dk wrote: >Dear all, >Have any experience with c-kit (CD117/C-19) immunohistochemical staining in rat >paraffin-embeded tissue? I have tryed different antigen retrieval methods, and >still cant get it to work! Any info would be greatly appreciated. >Thank you > >Mette K. Hagensen >Department of Cardiology B, Research Unit >Aarhus University Hospital, Skejby Hospital >Brendstrupgaardsvej 100 >8200 Aarhus N >Denmark > >Mail: b003046@nf.au.dk > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From BMolinari <@t> heart.thi.tmc.edu Thu Dec 1 13:14:39 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Thu Dec 1 13:18:17 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: And..they must get paid A LOT more than me to wear those clothes! Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: A. Erickson [mailto:andrae@u.washington.edu] Sent: Thursday, December 01, 2005 10:00 AM To: Molinari, Betsy Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Histotechs: born or made? And they only run one sample at a time, and only work on one case at a time!!!! andra On Thu, 1 Dec 2005, Molinari, Betsy wrote: > Isn't it odd that there usually is no one else in the labs except the > one or two investigators working on the one or two cases going on that > week? > > Betsy Molinari HT (ASCP) > Texas Heart Institute > Cardiovascular Pathology > 6770 Bertner Ave. > 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 Shirley > Powell > Sent: Wednesday, November 30, 2005 1:45 PM > To: king.laurie@marshfieldclinic.org; histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > Yep, that is when I stopped watching Quincy. Made me want to fight too. > I > do watch CSI but usually not for accuracy, mostly for the scenery. > > Shirley > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > king.laurie@marshfieldclinic.org > Sent: Wednesday, November 30, 2005 2:30 PM > To: histonet@lists.utsouthwestern.edu > Subject: RE: [Histonet] Histotechs: born or made? > > > I used to love that show, until the episode where Quincy complained that > Sam > was demoted to work in histology, who were, after all, only a bunch of > "bottle labelers". Them's fightin' words! > > Laurie King > > > ------Original Message------ > From: "Bartlett, Jeanine" > Date: Wed Nov 30, 2005 -- 01:15:00 PM > To: "Pam Marcum" , "Ford Royer" > , "Histonet" > Subject: RE: [Histonet] Histotechs: born or made? > > And wasn't it amazing how much Sam and Quincy got done in one hour!? > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pam > Marcum > Sent: Wednesday, November 30, 2005 1:22 PM > To: Ford Royer; 'Histonet' > Subject: RE: [Histonet] Histotechs: born or made? > > Ford, > > I don't think we are allowed to tell the new people in the field how > much > fun we had in the old days. I have always loved my job however, > sometimes I > have to watch my sense of humor in non-histologist/medical company as we > don't ususally see things the same way they do. > > Heck, I thought Qunicy and now CSI were sit coms at first as it was so > far > from what we were doing and really funny for mistakes. All my first > boss in > histology (and also a city/county coroner) wanted for several years was > a > Sam like Qunicy had with all the equipment of course. He figured he > could > rid of at least 5 people in the lab with one Sam. > > Pam Marcum > > >> When I was a practicing laboratory scientist (27 years ago), we would >> have some of the wildest lab parties and everyone seemed to be on the >> same page as far as having a weird sense of humor. A work day didn't >> go by without some form of laughter in our lab. Non-laboratory people > >> often asked me why this was. The only thing that I could come up with > >> is it was how we dealt with the profession that we chose. I won't go >> into details or give examples. We all know what I am talking about. >> It does take a special kind of person to this sometimes morbid (some >> would say hideous) work and I for one am glad that there are these >> types of persons to take it on. Thank you all for your dedication to >> your profession and the people that you serve - mankind. >> >> ~ Ford >> >> Ford M. Royer, MT(ASCP) >> Minneapolis, MN >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> Morken, Tim >> - Labvision >> Sent: Wednesday, November 30, 2005 11:11 AM >> To: 'Ingles Claire'; Histonet >> Subject: [Histonet] Histotechs: born or made? >> >> The first time I walked into a histology lab it was the day after the >> 4th of July and there were 4 blackened fingers sitting on the grossing > >> bench (one guess how they got there - and it's nothing to do with >> anything Cajun!). My first thought was : 'this is going to be a >> strange job.' I've seen much stranger things since, so I think >> histotechs become strange due to exposure to unnatural sights (among >> other things!). And, of course, the pathology staff of any hospital is > infamous for their "gross" humor. >> >> >> Tim Morken >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles > >> Claire >> Sent: Wednesday, November 30, 2005 8:55 AM >> To: Histonet >> Subject: RE: [Histonet] Re: 70% from NBF >> >> >> I have wondered the same thing many times myself. Whether it was >> naturally me or the addition of the chemicals that made me a bit >> strange. I think it may be partly both. I usually blame it on the >> chemical fumes though. :) Claire Ingles Mohs Clinic, UW Hosp. Madison >> WI >> >> ________________________________ >> >> From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bryan >> Llewellyn >> Sent: Tue 11/29/2005 11:15 AM >> To: Histonet >> Subject: Re: [Histonet] Re: 70% from NBF >> >> >> >> I have often wondered whether I became a histotech because I was born >> strange, or whether I became strange because of the time I spent >> training in a place like that! >> >> Bryan Llewellyn >> >> >> ----- Original Message ----- >> From: "Gayle Callis" >> To: >> Sent: Tuesday, November 29, 2005 8:23 AM >> Subject: [Histonet] Re: 70% from NBF >> >> >>> Joseph made some excellent points here >>> >>> Chloroform is an excellent clearing agent (used it back in the 60's >>> in open dip and dunk processors - O.K. so I'm old!) but no one >>> warned us about its carcinogenic nature and there were no safety >>> issue regulations then. Take his advice! >> >> >> _______________________________________________ >> 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 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >> From histonet-bounces@lists.utsouthwestern.edu Wed Nov 30 13:03:55 2005 > > > > > > _______________________________________________ > 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 anh2006 <@t> med.cornell.edu Thu Dec 1 13:31:09 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Dec 1 13:25:55 2005 Subject: [Histonet] Microtomy In-Reply-To: <20051201182448.51452.qmail@web61215.mail.yahoo.com> References: <20051201182448.51452.qmail@web61215.mail.yahoo.com> Message-ID: Although I do not use this kind of microtome in particular, I do dismantle and clean my blade holder after every use. I find it needs it and long term maintenance as well as section quality is better for my machine. If someone who knows what they are doing does the cleaning then there should be no problem over time -- to ensure this I completely orient new users to every aspect of the machine. That being said, this tech should follow whatever the working procedure is in your lab. I agree with Rene J in that a written SOP should solve your problems. Write one for the lab as a whole or for each brand of microtome and insist that the techs follow it. At 10:24 AM -0800 12/1/05, Rene J Buesa wrote: >Hi Cindy: > First it is not necessary to take appart a knife holder daily. For >what you describe > this techs acts as some do, thinking that they are like >"independent contractors" > within the lab setting able to do anything the want regardless of >general rules. > Do you have an SOP that addresses this cleaning procedure? If you don't you > can write this aspect and have everybody to adhere to it. > With some statistics about recuts required for bad technique you >can also try > to convince her. > You could also get advise from the sales representative regarding >this practice. > If everything fails you will have to impose authority on the issue. > Hope you will be able to reign in this stubborn tech. > Rene J. > >Cindy DuBois wrote: > This is a question for those techs who cut on a manual Reichert-Jung 2030. >I have one tech who insists on completely taking apart her knife >holder everyday to clean it. She removes the springs and all. This >person is also the only tech who has trouble cutting and is >constantly having to recut a specimen or nearly cutting through >needle biopsies. >I maintain that handling the knife holder spring so much causes the >problems (knife won't clamp tight evenly, chatter and skipping). The >other techs, including myself only dismantle and clean the knife >holder about once a month. We have had to purchase new spring and >bolts for this microtome about every 6 months, whereas the other >microtomes have never had to be replaced. How do I go about telling >this person to quit cleaning so much. >I have had the knife holder and microtome serviced to make sure >there is nothing else wrong. I have cut on that machine and find it >cuts beautifully as long as you don't take it apart after every 10 >blocks to clean behind the blade. > >Any suggestions would be greatly appreciated, > >Cindy DuBois >Integrated Pathology Assoc. >Stockton CA > -- From jkiernan <@t> uwo.ca Thu Dec 1 13:33:02 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Thu Dec 1 13:33:02 2005 Subject: [Histonet] Modified Schmidt's Hematoxylin References: <1B0B60D56CA6A440A240E866BA3458D8101025@ATL1VEXC017.usdom004.tco.tc> Message-ID: <438F4FEE.D95C0426@uwo.ca> At http://stainsfile.info/stain/hemindex.htm Bryan Llewellyn gives 84 haematoxylin recipes, but Schnidt's (even unmodified) isn't among them. Perhaps one of the others would do instead. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Jeff Oliver wrote: > > Hello, > > I'm looking for a recipe for a modified Schmidt's Hematoxylin. The only > references I have found to it are from a couple papers that did not > include a recipe. Until a couple days ago I had no idea this version > existed. Does anyone have a recipe or know a vendor for this? Any help > would be greatly appreciated. > > Thanks! > > -Jeff > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ploykasek <@t> phenopath.com Thu Dec 1 13:41:34 2005 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Thu Dec 1 13:42:04 2005 Subject: [Histonet] RE: Histotechs: born or made? In-Reply-To: <6.0.0.22.1.20051201100041.01b0b3d0@gemini.msu.montana.edu> Message-ID: >From all my quips about CSI at work I received a great 'gag' gift from a co-worker last year - an itty-bitty mag lite. It's even in my favorite color - purple. I haven't yet convinced anyone to turn down the lights & let me use it at work. Actually it did come in handy when I was trying to match up some paraffin blocks & H&Es to make TMAs. I held the light between my knees, put the paraffin block on top & had the H&E nearby to place on the block for comparison (don't try this in a skirt!) Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > Jo, > > Never, you need to buy one of the trusty, tidy little flashlights they use > for everything!!! Even in broad daylight!! > > Gayle Callis At 09:34 AM 12/1/2005, you wrote: >> I haven't seen an episode of CSI in a while, but when I did watch I noticed >> that they always had everything backlit with NO direct lighting. You could >> see the light shining up into their faces, but never onto their work >> surface. I'd go blind working in those conditions! >> Jo Dee > > > _______________________________________________ > 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 RRA <@t> Stowers-Institute.org Thu Dec 1 13:55:11 2005 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Thu Dec 1 13:55:38 2005 Subject: [Histonet] RE: Histotechs: born or made? Message-ID: What about the show House? I love it when all the residents draw blood, take it and culture's to the lab and personally perform all the tests! They do all the x-rays and any other test you can think of. Yet non-medical people think this is complete reality. Then they go to the patient's house to find anything they might have poisoned themselves with from the bathroom to inside the walls. Rhonda Allen Stowers Institute 1000 E. 50th street Kansas City, Missouri 64110 rra@stowers-institute.org From mucram11 <@t> comcast.net Thu Dec 1 14:01:51 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Thu Dec 1 14:02:15 2005 Subject: [Histonet] RE: Histotechs: born or made? Message-ID: <120120052001.3519.438F56AF0003F47200000DBF2207020953CECE030E9D0C9A03@comcast.net> Thanks Rhonda. I actually like House mainly because he is so far out and says some things I think we would all love to say at times to some of the Docs we know. It is by far the most outrageous of the ones people believe are real. We had CSI day at the Region 2 NSH meeting and it was packed. Two friends from the New Jersey State Police Lab came and talked about how different and boring their work is comapred to to TV which as they said very clearly "THIS IS NOT WHAT WE DO -- wE RARELY EVEN GO TO CRIME SCENES" Pam Marcum > What about the show House? I love it when all the residents draw blood, > take it and culture's to the lab and personally perform all the tests! > They do all the x-rays and any other test you can think of. Yet > non-medical people think this is complete reality. Then they go to the > patient's house to find anything they might have poisoned themselves > with from the bathroom to inside the walls. > > Rhonda Allen > Stowers Institute > 1000 E. 50th street > Kansas City, Missouri 64110 > rra@stowers-institute.org > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Thu Dec 1 14:02:06 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Thu Dec 1 14:02:18 2005 Subject: [Histonet] RE: Histotechs: born or made? Message-ID: <120120052002.3873.438F56BE0001FC1F00000F212207020953CECE030E9D0C9A03@comcast.net> Thanks Rhonda. I actually like House mainly because he is so far out and says some things I think we would all love to say at times to some of the Docs we know. It is by far the most outrageous of the ones people believe are real. We had CSI day at the Region 2 NSH meeting and it was packed. Two friends from the New Jersey State Police Lab came and talked about how different and boring their work is comapred to to TV which as they said very clearly "THIS IS NOT WHAT WE DO -- wE RARELY EVEN GO TO CRIME SCENES" Pam Marcum > What about the show House? I love it when all the residents draw blood, > take it and culture's to the lab and personally perform all the tests! > They do all the x-rays and any other test you can think of. Yet > non-medical people think this is complete reality. Then they go to the > patient's house to find anything they might have poisoned themselves > with from the bathroom to inside the walls. > > Rhonda Allen > Stowers Institute > 1000 E. 50th street > Kansas City, Missouri 64110 > rra@stowers-institute.org > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mucram11 <@t> comcast.net Thu Dec 1 14:01:57 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Thu Dec 1 14:02:20 2005 Subject: [Histonet] RE: Histotechs: born or made? Message-ID: <120120052001.3679.438F56B50002418400000E5F2207020953CECE030E9D0C9A03@comcast.net> Thanks Rhonda. I actually like House mainly because he is so far out and says some things I think we would all love to say at times to some of the Docs we know. It is by far the most outrageous of the ones people believe are real. We had CSI day at the Region 2 NSH meeting and it was packed. Two friends from the New Jersey State Police Lab came and talked about how different and boring their work is comapred to to TV which as they said very clearly "THIS IS NOT WHAT WE DO -- wE RARELY EVEN GO TO CRIME SCENES" Pam Marcum > What about the show House? I love it when all the residents draw blood, > take it and culture's to the lab and personally perform all the tests! > They do all the x-rays and any other test you can think of. Yet > non-medical people think this is complete reality. Then they go to the > patient's house to find anything they might have poisoned themselves > with from the bathroom to inside the walls. > > Rhonda Allen > Stowers Institute > 1000 E. 50th street > Kansas City, Missouri 64110 > rra@stowers-institute.org > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Dec 1 14:02:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 1 14:03:00 2005 Subject: [Histonet] Microtomy In-Reply-To: References: <20051201182448.51452.qmail@web61215.mail.yahoo.com> Message-ID: <6.0.0.22.1.20051201125940.00bb5df8@gemini.msu.montana.edu> If one writes an SOP for microtome cleaning, I strongly suggest you find out the manufacturer recommendations on what they use the job. Removal of lubricants or not replacing them can create problems - something I learned from Leica. At 12:31 PM 12/1/2005, you wrote: >Although I do not use this kind of microtome in particular, I do dismantle >and clean my blade holder after every use. I find it needs it and long >term maintenance as well as section quality is better for my machine. If >someone who knows what they are doing does the cleaning then there should >be no problem over time -- to ensure this I completely orient new users to >every aspect of the machine. > >That being said, this tech should follow whatever the working procedure is >in your lab. I agree with Rene J in that a written SOP should solve your >problems. Write one for the lab as a whole or for each brand of microtome >and insist that the techs follow it. > > > > >At 10:24 AM -0800 12/1/05, Rene J Buesa wrote: >>Hi Cindy: >> First it is not necessary to take appart a knife holder daily. For >> what you describe >> this techs acts as some do, thinking that they are like "independent >> contractors" >> within the lab setting able to do anything the want regardless of >> general rules. >> Do you have an SOP that addresses this cleaning procedure? If you >> don't you >> can write this aspect and have everybody to adhere to it. >> With some statistics about recuts required for bad technique you can >> also try >> to convince her. >> You could also get advise from the sales representative regarding this >> practice. >> If everything fails you will have to impose authority on the issue. >> Hope you will be able to reign in this stubborn tech. >> Rene J. >> >>Cindy DuBois wrote: >> This is a question for those techs who cut on a manual Reichert-Jung 2030. >>I have one tech who insists on completely taking apart her knife holder >>everyday to clean it. She removes the springs and all. This person is >>also the only tech who has trouble cutting and is constantly having to >>recut a specimen or nearly cutting through needle biopsies. >>I maintain that handling the knife holder spring so much causes the >>problems (knife won't clamp tight evenly, chatter and skipping). The >>other techs, including myself only dismantle and clean the knife holder >>about once a month. We have had to purchase new spring and bolts for this >>microtome about every 6 months, whereas the other microtomes have never >>had to be replaced. How do I go about telling this person to quit >>cleaning so much. >>I have had the knife holder and microtome serviced to make sure there is >>nothing else wrong. I have cut on that machine and find it cuts >>beautifully as long as you don't take it apart after every 10 blocks to >>clean behind the blade. >> >>Any suggestions would be greatly appreciated, >> >>Cindy DuBois >>Integrated Pathology Assoc. >>Stockton CA > >-- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From mucram11 <@t> comcast.net Thu Dec 1 14:07:04 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Thu Dec 1 14:07:24 2005 Subject: [Histonet] RE: Histotechs: born or made? Message-ID: <120120052007.10426.438F57E80006704B000028BA2207020953CECE030E9D0C9A03@comcast.net> Sorry the enter button stuck and sent it three times. Pam > Thanks Rhonda. I actually like House mainly because he is so far out and says > some things I think we would all love to say at times to some of the Docs we > know. It is by far the most outrageous of the ones people believe are real. > > We had CSI day at the Region 2 NSH meeting and it was packed. Two friends from > the New Jersey State Police Lab came and talked about how different and boring > their work is comapred to to TV which as they said very clearly "THIS IS NOT > WHAT WE DO -- wE RARELY EVEN GO TO CRIME SCENES" > > Pam Marcum > > > > What about the show House? I love it when all the residents draw blood, > > take it and culture's to the lab and personally perform all the tests! > > They do all the x-rays and any other test you can think of. Yet > > non-medical people think this is complete reality. Then they go to the > > patient's house to find anything they might have poisoned themselves > > with from the bathroom to inside the walls. > > > > Rhonda Allen > > Stowers Institute > > 1000 E. 50th street > > Kansas City, Missouri 64110 > > rra@stowers-institute.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 Jackie.O'Connor <@t> abbott.com Thu Dec 1 14:09:40 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Dec 1 14:10:17 2005 Subject: [Histonet] RE: Histotechs: born or made? Message-ID: That's show biz. I love House. I wish I could find a doctor like him. I was shocked to hear that actor's real accent - he's a Brit!! "Allen, Rhonda" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/01/2005 01:55 PM To: "Patti Loykasek" , "histonet" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] RE: Histotechs: born or made? What about the show House? I love it when all the residents draw blood, take it and culture's to the lab and personally perform all the tests! They do all the x-rays and any other test you can think of. Yet non-medical people think this is complete reality. Then they go to the patient's house to find anything they might have poisoned themselves with from the bathroom to inside the walls. Rhonda Allen Stowers Institute 1000 E. 50th street Kansas City, Missouri 64110 rra@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lr_hsvlle2 <@t> yahoo.com.au Thu Dec 1 14:18:39 2005 From: lr_hsvlle2 <@t> yahoo.com.au (Lorraine Rolston) Date: Thu Dec 1 14:18:49 2005 Subject: [Histonet] Re:70% to NBF Message-ID: <20051201201839.24002.qmail@web30615.mail.mud.yahoo.com> A great big thank you to all of you for all of those very helpful answers!!! I too have been around in histology for some time,however chiefly in the diagnostic/clinical arena and I am consistently challenged to expand my histological knowledge, both about that which I think I already know and that which I know I do not know. Here in this other world at the university I am surrounded by teaching and research, a great place to be but I stumble lots................or maybe that's the chloroform!! Cheers, Lorraine. --------------------------------- Do you Yahoo!? Messenger 7.0: Free worldwide PC to PC calls From anh2006 <@t> med.cornell.edu Thu Dec 1 14:30:30 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Dec 1 14:25:15 2005 Subject: [Histonet] Microtomy In-Reply-To: <6.0.0.22.1.20051201125940.00bb5df8@gemini.msu.montana.edu> References: <20051201182448.51452.qmail@web61215.mail.yahoo.com> <6.0.0.22.1.20051201125940.00bb5df8@gemini.msu.montana.edu> Message-ID: I couldn't agree more. All this info and more should be included. Often times cleaning with nothing more than paper towels does the trick and won't remove all the essential lubrication. Anything else removed can be replaced with oil in the weekly maintenance. At 1:02 PM -0700 12/1/05, Gayle Callis wrote: >If one writes an SOP for microtome cleaning, I strongly suggest you >find out the manufacturer recommendations on what they use the job. >Removal of lubricants or not replacing them can create problems - >something I learned from Leica. -- From Charles.Embrey <@t> carle.com Thu Dec 1 14:27:33 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Thu Dec 1 14:27:45 2005 Subject: FW: [Histonet] Microtomy Message-ID: -----Original Message----- From: Charles.Embrey Sent: Thursday, December 01, 2005 12:09 PM To: 'Cindy DuBois' Subject: RE: [Histonet] Microtomy This is really such a sad question and highlights a growing problem in many labs. People are so afraid to "step on toes" that they have forgotten how to be in charge. The answer to this question is so simple, TELL HER TO STOP DOING IT. You don't have to get into a huge debate just tell her to stop. If she refuses then she can look into dismantling another lab's microtome. If it were merely an increased maintenance cost problem that would be one thing but from what you say she is putting patient's tissue and diagnosis at risk and that cannot be tolerated. Charles Embrey, PA(ASCP) Histology Manager Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cindy DuBois Sent: Thursday, December 01, 2005 11:52 AM To: Histonet Subject: [Histonet] Microtomy This is a question for those techs who cut on a manual Reichert-Jung 2030. I have one tech who insists on completely taking apart her knife holder everyday to clean it. She removes the springs and all. This person is also the only tech who has trouble cutting and is constantly having to recut a specimen or nearly cutting through needle biopsies. I maintain that handling the knife holder spring so much causes the problems (knife won't clamp tight evenly, chatter and skipping). The other techs, including myself only dismantle and clean the knife holder about once a month. We have had to purchase new spring and bolts for this microtome about every 6 months, whereas the other microtomes have never had to be replaced. How do I go about telling this person to quit cleaning so much. I have had the knife holder and microtome serviced to make sure there is nothing else wrong. I have cut on that machine and find it cuts beautifully as long as you don't take it apart after every 10 blocks to clean behind the blade. Any suggestions would be greatly appreciated, Cindy DuBois Integrated Pathology Assoc. Stockton CA --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Try Yahoo! Personals _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From llewllew <@t> shaw.ca Thu Dec 1 14:27:09 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Thu Dec 1 14:27:52 2005 Subject: [Histonet] Modified Schmidt's Hematoxylin References: <1B0B60D56CA6A440A240E866BA3458D8101025@ATL1VEXC017.usdom004.tco.tc> <438F4FEE.D95C0426@uwo.ca> Message-ID: <001a01c5f6b5$9abe8da0$690a4246@yourlk4rlmsu> Thanks for the plug, but the url is actually http://stainsfile.info/StainsFile/stain/hemindex.htm I used several search engines for this formula, and came up with three references but no details. One of the references included a comment that it was useful when an aqueous mounting medium was used. If anyone does locate it, I would appreciate details so I can include it on StainsFile. Bryan ----- Original Message ----- From: "John A. Kiernan" To: "Jeff Oliver" Cc: Sent: Thursday, December 01, 2005 11:33 AM Subject: Re: [Histonet] Modified Schmidt's Hematoxylin > At http://stainsfile.info/stain/hemindex.htm > Bryan Llewellyn gives 84 haematoxylin recipes, > but Schnidt's (even unmodified) isn't among > them. Perhaps one of the others would do > instead. > -- > ------------------------------- > John A. Kiernan > Department of Anatomy and Cell Biology > The University of Western Ontario > London, Canada N6A 5C1 > kiernan[AT]uwo.ca > http://publish.uwo.ca/~jkiernan/ > http://instruct.uwo.ca/anatomy/530/index.htm > _______________________________ > Jeff Oliver wrote: >> >> Hello, >> >> I'm looking for a recipe for a modified Schmidt's Hematoxylin. The only >> references I have found to it are from a couple papers that did not >> include a recipe. Until a couple days ago I had no idea this version >> existed. Does anyone have a recipe or know a vendor for this? Any help >> would be greatly appreciated. >> >> Thanks! >> >> -Jeff >> >> _______________________________________________ >> 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 llewllew <@t> shaw.ca Thu Dec 1 14:42:14 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Thu Dec 1 14:42:29 2005 Subject: [Histonet] Microtomy References: Message-ID: <003801c5f6b7$b68c6730$690a4246@yourlk4rlmsu> I would imagine this takes her some time to do, perhaps an hour or so. What would she be doing if she were not cleaning her knife holder. Possibly she does not feel comfortable doing that other work and this may be a way to avoid it. If so, an answer could be to increase her confidence level for the other work. It could also be she is not comfortable sectioning and the cleaning is a distractor to avoid facing the issue of poor quality, in which case further training would be needed. Most employers find that assisting an employee to overcome problems is more effective than replacing one. However, the bottom line is that the work must be done properly. Bryan Llewellyn ----- Original Message ----- From: "Charles.Embrey" To: Sent: Thursday, December 01, 2005 12:27 PM Subject: FW: [Histonet] Microtomy -----Original Message----- From: Charles.Embrey Sent: Thursday, December 01, 2005 12:09 PM To: 'Cindy DuBois' Subject: RE: [Histonet] Microtomy This is really such a sad question and highlights a growing problem in many labs. People are so afraid to "step on toes" that they have forgotten how to be in charge. The answer to this question is so simple, TELL HER TO STOP DOING IT. You don't have to get into a huge debate just tell her to stop. If she refuses then she can look into dismantling another lab's microtome. If it were merely an increased maintenance cost problem that would be one thing but from what you say she is putting patient's tissue and diagnosis at risk and that cannot be tolerated. Charles Embrey, PA(ASCP) Histology Manager Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cindy DuBois Sent: Thursday, December 01, 2005 11:52 AM To: Histonet Subject: [Histonet] Microtomy This is a question for those techs who cut on a manual Reichert-Jung 2030. I have one tech who insists on completely taking apart her knife holder everyday to clean it. She removes the springs and all. This person is also the only tech who has trouble cutting and is constantly having to recut a specimen or nearly cutting through needle biopsies. I maintain that handling the knife holder spring so much causes the problems (knife won't clamp tight evenly, chatter and skipping). The other techs, including myself only dismantle and clean the knife holder about once a month. We have had to purchase new spring and bolts for this microtome about every 6 months, whereas the other microtomes have never had to be replaced. How do I go about telling this person to quit cleaning so much. I have had the knife holder and microtome serviced to make sure there is nothing else wrong. I have cut on that machine and find it cuts beautifully as long as you don't take it apart after every 10 blocks to clean behind the blade. Any suggestions would be greatly appreciated, Cindy DuBois Integrated Pathology Assoc. Stockton CA --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Try Yahoo! Personals _______________________________________________ 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 BRIAN.CHELACK <@t> usask.ca Thu Dec 1 15:04:22 2005 From: BRIAN.CHELACK <@t> usask.ca (Brian Chelack) Date: Thu Dec 1 15:28:13 2005 Subject: [Histonet] Anyone running IHC on Cyclin A1 Cyclin E or CREB-binding protein?? Message-ID: <438F6556.409@sask.usask.ca> Hello All; Someone has asked me if it is possible to stain for Cyclin A1, Cyclin E or CREB-Binding proteins in formalin fixed paraffin embedded tissues. I've run a couple of quick searches with no sign it is possible. Can anyone prove me wrong? Thanks, Brian Chelack Prairie Diagnostic Services ps. - just to add another level of complexity, they would like to do this on bovine tissues, but I would be interested to know if it is possible on human tissues first, then I would put efforts into trying to demonstrate it in bovine. bjc From Charles.Embrey <@t> carle.com Thu Dec 1 15:36:20 2005 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Thu Dec 1 15:36:30 2005 Subject: [Histonet] Microtomy Message-ID: You are 100% right that helping a technician is much more effective than replacing one. The big problem here is that while you are busy psychoanalyzing her problems she is cutting through needle core biopsies that may contain needed tissue for correct diagnosis. I trained a PA student last year from a nearby University and she had a real problem with obsessive compulsive behavior when it came to cleaning between cases on the grossing board. She actually got the shakes when I took her cleaning solution away but she was able to work through it and her grossing speed doubled. I fully believe in working through problems as long as the patient is not in jeopardy but that is where I have to draw the line. Charles Embrey PA(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Thursday, December 01, 2005 2:42 PM To: Histonet Subject: Re: [Histonet] Microtomy I would imagine this takes her some time to do, perhaps an hour or so. What would she be doing if she were not cleaning her knife holder. Possibly she does not feel comfortable doing that other work and this may be a way to avoid it. If so, an answer could be to increase her confidence level for the other work. It could also be she is not comfortable sectioning and the cleaning is a distractor to avoid facing the issue of poor quality, in which case further training would be needed. Most employers find that assisting an employee to overcome problems is more effective than replacing one. However, the bottom line is that the work must be done properly. Bryan Llewellyn ----- Original Message ----- From: "Charles.Embrey" To: Sent: Thursday, December 01, 2005 12:27 PM Subject: FW: [Histonet] Microtomy -----Original Message----- From: Charles.Embrey Sent: Thursday, December 01, 2005 12:09 PM To: 'Cindy DuBois' Subject: RE: [Histonet] Microtomy This is really such a sad question and highlights a growing problem in many labs. People are so afraid to "step on toes" that they have forgotten how to be in charge. The answer to this question is so simple, TELL HER TO STOP DOING IT. You don't have to get into a huge debate just tell her to stop. If she refuses then she can look into dismantling another lab's microtome. If it were merely an increased maintenance cost problem that would be one thing but from what you say she is putting patient's tissue and diagnosis at risk and that cannot be tolerated. Charles Embrey, PA(ASCP) Histology Manager Carle Clinic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cindy DuBois Sent: Thursday, December 01, 2005 11:52 AM To: Histonet Subject: [Histonet] Microtomy This is a question for those techs who cut on a manual Reichert-Jung 2030. I have one tech who insists on completely taking apart her knife holder everyday to clean it. She removes the springs and all. This person is also the only tech who has trouble cutting and is constantly having to recut a specimen or nearly cutting through needle biopsies. I maintain that handling the knife holder spring so much causes the problems (knife won't clamp tight evenly, chatter and skipping). The other techs, including myself only dismantle and clean the knife holder about once a month. We have had to purchase new spring and bolts for this microtome about every 6 months, whereas the other microtomes have never had to be replaced. How do I go about telling this person to quit cleaning so much. I have had the knife holder and microtome serviced to make sure there is nothing else wrong. I have cut on that machine and find it cuts beautifully as long as you don't take it apart after every 10 blocks to clean behind the blade. Any suggestions would be greatly appreciated, Cindy DuBois Integrated Pathology Assoc. Stockton CA --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Try Yahoo! Personals _______________________________________________ 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 sharon.osborn <@t> dnax.org Thu Dec 1 16:11:54 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Thu Dec 1 16:12:15 2005 Subject: [Histonet] deparaffinized sections to air dry or not to air dry? Message-ID: <29B25753F6B1D51196110002A589D444032C4900@PALMSG30.us.schp.com> Betsy, A possible problem you could have with allowing the slides to air dry after deparafinization is what we old techs know as "cornflake" effect. I spent years doing both cytology and histology and saw quite a bit of cornflaking on the cytology slides when the fixative was inadequately sprayed. (This was back when the scrape and smear technic was method of choice--before the mono-layer technology. Cornflake appearance looks just like a piece of cornflake cereal was dropped on the slide. Loose cells exhibit it more than tissue sections; however, inadequate hydration will produce similar brown artifact effect in tissue sections. If you do airdry, you need to be certain the sections are fully hydrated before continuing the staining process. Then, you may still have some sections that exhibit the dark artifacts of not being fully hydrated or stained. sharon osborn DNAX S-P BioPharma Palo Alto, CA An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Paraffin sections Hi, I have heard that paraffin sections that have been deparaffinized and run down to DH20 but for some reason cannot be stained that day, can be left to air dry and then be placed back into H2O when they are ready for staining. Does anyone have any experience with this technique? I usually just leave them in DH2O till I can get to them, usually the next day. Thanks. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From vazquezr <@t> ohsu.edu Thu Dec 1 16:22:04 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Dec 1 16:22:32 2005 Subject: [Histonet] Histotechs: born or made? Message-ID: I love the CSI's, but what got me was one of the NY episodes was that someone was in the lab and none of the bottles were labeled...that sent me into a frenzi and I almost emailed the producers and ask them what are you guys thinking. I have noticed some bottles labeled now. And I am better now... Robyn From scoop <@t> mail.nih.gov Thu Dec 1 16:33:09 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Thu Dec 1 16:33:19 2005 Subject: [Histonet] looking for electron microscopy services Message-ID: Dear Histonet, I am looking for a company or person who would do electron microscopy on our samples. We are interested in examining mitochondrial morphology in brain and possibly other tissues of mice. If there's anyone who does that kind of work and would be interested, please contact me. I would also appreciate referrals if anyone knows of someone appropriate. Thanks! Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From sharon.osborn <@t> dnax.org Thu Dec 1 16:53:35 2005 From: sharon.osborn <@t> dnax.org (Osborn, Sharon) Date: Thu Dec 1 16:54:14 2005 Subject: [Histonet] RE: microtome cleaning Message-ID: <29B25753F6B1D51196110002A589D444032C4901@PALMSG30.us.schp.com> Years ago, when I worked in a lab that used the Leica 1512, we were required to clean the knife holder every day. The microtomes were shared, but they decided to purchase a separate knife holder for each tech. We did as your tech is doing, take the knife holder apart at the end or our cutting shift and cleaned it. We also had to lubricate it. There is a technic to doing it properly and handling the springs properly for repositioning to ensure it goes back together. We also had ball bearings we had to clean and replace properly. It seemed that pieces of paraffin or tissue would get inside and could create a problem. Because of the cleaning requirement (don't remember if required by the lab or suggested by Leica) and techs who wanted their own knifeholder because they did not trust others to clean theirs the correct way, etc., the supr decided to purchase a knife holder for each tech and have them responsible for their own. Oh, you did not dare use anothers knife holder, even to cut one or two blocks! So, did the tech come from a lab that had requirement to clean every day? If so, these technics sometimes die hard for the tech may have been given reasons it was required daily; therefore the reframing as to why it is not now necessary needs to replace the old habit. Or, is this something the tech feels important? Have you talked with the tech about their reasoning for doing the process? Gayle Callis has the best suggestion; ask the manufacturer what they recommend;--ask Jan Minshew at Leica in Chicago since she is their expert histotech. Then do an inservice (with a rep or other qualified person) with the SOP you write about the proper maintenance for the instrument. And,then you can require everyone to comply with the SOP. Sharon Osborn DNAX Palo Alto, CA Sent: Thursday, December 01, 2005 11:52 AM To: Histonet Subject: [Histonet] Microtomy This is a question for those techs who cut on a manual Reichert-Jung 2030. I have one tech who insists on completely taking apart her knife holder everyday to clean it. She removes the springs and all. This person is also the only tech who has trouble cutting and is constantly having to recut a specimen or nearly cutting through needle biopsies. I maintain that handling the knife holder spring so much causes the problems (knife won't clamp tight evenly, chatter and skipping). The other techs, including myself only dismantle and clean the knife holder about once a month. We have had to purchase new spring and bolts for this microtome about every 6 months, whereas the other microtomes have never had to be replaced. How do I go about telling this person to quit cleaning so much. I have had the knife holder and microtome serviced to make sure there is nothing else wrong. I have cut on that machine and find it cuts beautifully as long as you don't take it apart after every 10 blocks to clean behind the blade. Any suggestions would be greatly appreciated, Cindy DuBois Integrated Pathology Assoc. Stockton CA ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From scoop <@t> mail.nih.gov Thu Dec 1 18:17:37 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Thu Dec 1 18:17:51 2005 Subject: [Histonet] looking for electron microscopy services Message-ID: Dear Histonet, I am looking for a company or person who would do electron microscopy on our samples. We are interested in examining mitochondrial morphology in brain and possibly other tissues of mice. If there's anyone who does that kind of work and would be interested, please contact me. I would also appreciate referrals if anyone knows of someone appropriate. Thanks! Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From c.m.vanderloos <@t> amc.uva.nl Fri Dec 2 02:52:13 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Dec 2 02:52:30 2005 Subject: [Histonet] RE: Paraffin sections Message-ID: <18660851861bf9.1861bf91866085@amc.uva.nl> Betsy, For my NSH wet-workshops I tested the option of going up to endogenous PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. Next day we started at the protein blocking step and then the rest. Staining was as good as running the whole procedure in one day. Hope this info helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Thu, 1 Dec 2005 07:46:32 -0600 From: "Molinari, Betsy" Subject: [Histonet] Paraffin sections To: Hi, I have heard that paraffin sections that have been deparaffinized and run down to DH20 but for some reason cannot be stained that day, can be left to air dry and then be placed back into H2O when they are ready for staining. Does anyone have any experience with this technique? I usually just leave them in DH2O till I can get to them, usually the next day. Thanks. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 From ROrr <@t> enh.org Fri Dec 2 07:35:00 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Dec 2 07:35:17 2005 Subject: [Histonet] Goat polymer Message-ID: Hello everyone, Just recently there was a posting on how to run a rabbit polymer with goat antibodies...or something to that nature, using an anti rabbit secondary and then the Dako polymer...could that person send me an email please? I had a quick question about blocking... Also, does anyone have the name of the company that makes a goat polymer? I know there's a company out there...I've seen the ads but can't locate it... Many thanks and have a great weekend! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From brett_connolly <@t> merck.com Fri Dec 2 08:27:28 2005 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Dec 2 08:28:05 2005 Subject: [Histonet] looking for electron microscopy services Message-ID: <355C35514FEAC9458F75947F5270974D079731@usctmx1103.merck.com> Sharon, In your area, check out Paragon Bioservices in Baltimore http://www.paragonbioservices.com/ Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sharon Cooperman Sent: Thursday, December 01, 2005 7:18 PM To: Histonet@pathology.swmed.edu Subject: [Histonet] looking for electron microscopy services Dear Histonet, I am looking for a company or person who would do electron microscopy on our samples. We are interested in examining mitochondrial morphology in brain and possibly other tissues of mice. If there's anyone who does that kind of work and would be interested, please contact me. I would also appreciate referrals if anyone knows of someone appropriate. Thanks! Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From GoodwinD <@t> pahosp.com Fri Dec 2 08:49:11 2005 From: GoodwinD <@t> pahosp.com (Goodwin, Diana) Date: Fri Dec 2 08:49:23 2005 Subject: [Histonet] MSH-2 and MLH-1 Message-ID: <80CDD9C3FEEAFD4982B114C4A6DFD00E256F77@uphsmbx2.UPHS.PENNHEALTH.PRV> Greetings, Histonet. I anyone running paraffin IHC with these Abs for human colo-rectal ca successfully on the Ventana Benchmark, and if so, which clone, vendor, dilution, etc. Thanks in advance, Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com From anh2006 <@t> med.cornell.edu Fri Dec 2 08:56:52 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Fri Dec 2 08:51:37 2005 Subject: [Histonet] RE: Paraffin sections In-Reply-To: <18660851861bf9.1861bf91866085@amc.uva.nl> References: <18660851861bf9.1861bf91866085@amc.uva.nl> Message-ID: Very interesting Chris! I have found sometimes this works and sometimes it doesn't and you get complete loss of staining - frustrating when one was just trying to save time to begin with!! I imagine it's antigen dependent but I have not had time to check. In my opinion Betsy, if you can avoid it (because you are just doing it to save time instead of setting up a workshop) then avoid it ... it simply isn't worth the risk IMHO. At 9:52 AM +0100 12/2/05, C.M. van der Loos wrote: > Betsy, > > For my NSH wet-workshops I tested the option of going up to endogenous > PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. > Next day we started at the protein blocking step and then the rest. > Staining was as good as running the whole procedure in one day. > > Hope this info helps. > > Chris van der Loos, PhD > Dept. of Pathology > Academic Medical Center M2-230 > Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands -- From Diego.RodriguezGil <@t> yale.edu Fri Dec 2 09:41:35 2005 From: Diego.RodriguezGil <@t> yale.edu (Diego J. =?iso-8859-1?b?Um9kcu1ndWV6?= Gil) Date: Fri Dec 2 09:42:02 2005 Subject: [Histonet] Extracellular matrix proteins fixation. Message-ID: <1133538095.43906b2f20c86@webmail.med.yale.edu> Hi everybody: I am brand new on the list, so I do not know if this question was posted before. I apologize for this. I am trying to characterize a family of secreted proteins in the Nervous System by immunohistochemistry, and with the regular fixation protocol that we used (intracardiac perfusion with 4% PFA), I am not getting any good results. Not even after some antigen-retrieval procedures. Before trying random things, does anybody have a good protocol to get extracellular proteins well fixed? It doesn't matter if it is set up for some other tissue, I would be glad to try other protocol(s) if there are any available. Thank you very much for your help, Best regards, Diego. -- Diego J. Rodr?guez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. E-mail: diego.rodriguezgil@yale.edu From gcallis <@t> montana.edu Fri Dec 2 10:03:11 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Dec 2 10:03:26 2005 Subject: [Histonet] May I ask what kind of knife holder? RE: microtome cleaning In-Reply-To: <29B25753F6B1D51196110002A589D444032C4901@PALMSG30.us.schp. com> References: <29B25753F6B1D51196110002A589D444032C4901@PALMSG30.us.schp.com> Message-ID: <6.0.0.22.1.20051202085352.01b369e8@gemini.msu.montana.edu> Dear Sharon et al, What knife holder were you using? The only knife holder I ever had to take apart and tediously clean after every use was the Miles AccuEdge Disposable Blade Insert for low profile disposable blades.using manufacturers cleaning kit/instructions, and kept a repair kit on hand at all times. Fortunately, we only used this holder for thin (1 and 2 um) paraffin sections i.e. kidney biopsies on an old AO 820 microtome. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From mcauliff <@t> umdnj.edu Fri Dec 2 10:32:43 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Dec 2 10:40:13 2005 Subject: [Histonet] Extracellular matrix proteins fixation. In-Reply-To: <1133538095.43906b2f20c86@webmail.med.yale.edu> References: <1133538095.43906b2f20c86@webmail.med.yale.edu> Message-ID: <4390772B.2030404@umdnj.edu> Hi Diego: First off, "well fixed" and a strong immuno reaction are often incompatible. The first thing to do is a PubMed search for the exact ECM molecules you are interested in. That way you will have peer-reviewed material from which to begin work. Or you could try a lot of suggestions from the list which may or may not be applicable to your animal, ECM molecule, antibody, detection system, etc. Perhaps frozen sections are required, or an alcohol-based fixative, etc. Good luck! Geoff Diego J. Rodr?guez Gil wrote: >Hi everybody: > > I am brand new on the list, so I do not know if this question was posted >before. I apologize for this. > I am trying to characterize a family of secreted proteins in the Nervous >System by immunohistochemistry, and with the regular fixation protocol that we >used (intracardiac perfusion with 4% PFA), I am not getting any good results. >Not even after some antigen-retrieval procedures. Before trying random things, >does anybody have a good protocol to get extracellular proteins well fixed? It >doesn't matter if it is set up for some other tissue, I would be glad to try >other protocol(s) if there are any available. > Thank you very much for your help, > Best regards, > Diego. > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From pruegg <@t> ihctech.net Fri Dec 2 11:12:46 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Dec 2 11:13:28 2005 Subject: [Histonet] deparaffinized sections to air dry or not to air dry? In-Reply-To: <29B25753F6B1D51196110002A589D444032C4900@PALMSG30.us.schp.com> Message-ID: <200512021713.jB2HCeN6016176@chip.viawest.net> I use the dry after deparaffinization routinely for IHC so that I can pap pen the slides while they are dry and also I have found that hard to adhere sections like brain stay on better if you airdry after 95% alcohol. True you must carefully rehydrate before staining and never let the sections dry during IHC except after chromogen/substrate it doesn't seem to hurt anything. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Osborn, Sharon Sent: Thursday, December 01, 2005 3:12 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] deparaffinized sections to air dry or not to air dry? Betsy, A possible problem you could have with allowing the slides to air dry after deparafinization is what we old techs know as "cornflake" effect. I spent years doing both cytology and histology and saw quite a bit of cornflaking on the cytology slides when the fixative was inadequately sprayed. (This was back when the scrape and smear technic was method of choice--before the mono-layer technology. Cornflake appearance looks just like a piece of cornflake cereal was dropped on the slide. Loose cells exhibit it more than tissue sections; however, inadequate hydration will produce similar brown artifact effect in tissue sections. If you do airdry, you need to be certain the sections are fully hydrated before continuing the staining process. Then, you may still have some sections that exhibit the dark artifacts of not being fully hydrated or stained. sharon osborn DNAX S-P BioPharma Palo Alto, CA An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] Paraffin sections Hi, I have heard that paraffin sections that have been deparaffinized and run down to DH20 but for some reason cannot be stained that day, can be left to air dry and then be placed back into H2O when they are ready for staining. Does anyone have any experience with this technique? I usually just leave them in DH2O till I can get to them, usually the next day. Thanks. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From drgrant <@t> cmh.edu Fri Dec 2 11:31:12 2005 From: drgrant <@t> cmh.edu (Grant, Debra, R) Date: Fri Dec 2 11:38:12 2005 Subject: [Histonet] Specail stains Negative controls Message-ID: <6F434E27D5DE944B9E898984CADDD149043982@exchmail.CMH.Internal> Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 From maria <@t> ski.org Fri Dec 2 12:16:05 2005 From: maria <@t> ski.org (Maria Mejia) Date: Fri Dec 2 12:16:22 2005 Subject: [Histonet] RE: Paraffin sections References: <18660851861bf9.1861bf91866085@amc.uva.nl> Message-ID: <43908F65.6010808@ski.org> This type of experimenting would be interesting to check out especially with lower percentages of BSA. Andrea, I would agree that this overnight storage step probably is antigen dependent (Chris, that's what I meant by too much) and it would require a whole gamut testing of antibodies to see which respond positively and which would not...if one is inclined to do so. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Andrea T. Hooper wrote: > Very interesting Chris! I have found sometimes this works and > sometimes it doesn't and you get complete loss of staining - > frustrating when one was just trying to save time to begin with!! I > imagine it's antigen dependent but I have not had time to check. > > In my opinion Betsy, if you can avoid it (because you are just doing > it to save time instead of setting up a workshop) then avoid it ... it > simply isn't worth the risk IMHO. > > > > At 9:52 AM +0100 12/2/05, C.M. van der Loos wrote: > >> Betsy, >> >> For my NSH wet-workshops I tested the option of going up to endogenous >> PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. >> Next day we started at the protein blocking step and then the rest. >> Staining was as good as running the whole procedure in one day. >> >> Hope this info helps. >> >> Chris van der Loos, PhD >> Dept. of Pathology >> Academic Medical Center M2-230 >> Meibergdreef 9 >> NL-1105 AZ Amsterdam >> The Netherlands > > From Diego.RodriguezGil <@t> yale.edu Fri Dec 2 12:55:04 2005 From: Diego.RodriguezGil <@t> yale.edu (Diego J. =?iso-8859-1?b?Um9kcu1ndWV6?= Gil) Date: Fri Dec 2 13:01:41 2005 Subject: [Histonet] Extracellular matrix proteins fixation. Details Message-ID: <1133549704.43909888f401e@webmail.med.yale.edu> Hi: Thanks a lot to all of you who answer!. Since some of you asked, I am sending some more details. I am working on mice (CD-1), and the molecules I am trying to stain are named Wnt (Wingless-Int). Antibodies are from R&D systems (most of them are goats), and there are no previous works reporting the use of these antibodies. I am only making the assumption that they work, based on the datasheet the company provides. I could see some very faint positive stain for some of them, but I am sure I losing some because of fixation. Besides that, I know they are expressed based on RT-PCR and in situ hybridization experiments. Brief protocol is: Perfusion with 4% PFA, and then overnight deep in 4% PFA at 4 C. Transfer to PBS, then 30% sucrose til they sink, freeze on OCT and then section on cryostat at -18C. The antigen retrieval protocols that I tried were steaming (2, 5 and 10 min with sodium citrate) or pepsin in HCl. None of them gave me any better stainings. Staining protocol: Wash in TBS and then add 2% BSA in TBS-T, for 30 min. Primary antibody (on 2% BSA) Over night . Wash 3 x TBS-T. Add secondary antibody on 2% BSA for an hour. Wash 2 x TBS-T. Wash TBS and mount with Crystal Mount. Thanks again! Best regards, Diego. -- Diego J. Rodr?guez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. From Jeffery-Smith <@t> ouhsc.edu Fri Dec 2 13:23:19 2005 From: Jeffery-Smith <@t> ouhsc.edu (Smith, Jeffery D. (HSC)) Date: Fri Dec 2 13:28:17 2005 Subject: [Histonet] Specail stains Negative controls Message-ID: Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Fri Dec 2 14:23:28 2005 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Dec 2 14:25:06 2005 Subject: [Histonet] Specail stains Negative controls References: Message-ID: <002001c5f77e$786f0990$6400a8c0@mainbox> Debby and Jeff, Negative control slides are useful for GMS, Gram and AFB in order to eliminate false positives due to contamination from the reagents, washing, carry-over etc. Regards, Bryan ----- Original Message ----- From: "Smith, Jeffery D. (HSC)" To: "Grant, Debra, R" ; Sent: Friday, December 02, 2005 2:23 PM Subject: RE: [Histonet] Specail stains Negative controls Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ 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 Fri Dec 2 14:39:43 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Fri Dec 2 14:40:09 2005 Subject: [Histonet] Goat polymer In-Reply-To: Message-ID: <200512022039.jB2KdkXX005415@pro12.abac.com> Becky, Invitrogen is the company who makes the goat labeled polymer. As for blocking, I use serum free protein block before the primary and then before the rab. Anti-goat secondary and then again before the rabbit labeled polymer. If I still encounter non-specific staining I put some serum (from the host of the secondary, about 10%) in with the protein block. This method is very sensitive and you can bring up lots of non-specific BG so be careful to optimize your dilutions and times for the antibody and for the detection reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, December 02, 2005 6:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Goat polymer Hello everyone, Just recently there was a posting on how to run a rabbit polymer with goat antibodies...or something to that nature, using an anti rabbit secondary and then the Dako polymer...could that person send me an email please? I had a quick question about blocking... Also, does anyone have the name of the company that makes a goat polymer? I know there's a company out there...I've seen the ads but can't locate it... Many thanks and have a great weekend! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From arunams <@t> interchange.ubc.ca Fri Dec 2 16:04:38 2005 From: arunams <@t> interchange.ubc.ca (arunams) Date: Fri Dec 2 16:04:47 2005 Subject: [Histonet] Knife holder Message-ID: <5641228.1133561078869.JavaMail.myubc2@portal9.itservices.ubc.ca> Hi All Does eny one know where to find a Tungston Carbide Knive holder for a Sorvall JB-4 Microtome. I really need one urgently. Thanks for your help Aruna Somasiri From cfavara <@t> niaid.nih.gov Fri Dec 2 16:35:26 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Fri Dec 2 16:35:35 2005 Subject: [Histonet] Question: Inventory Message-ID: I do not do high volume consistently but have some weeks where I am doing 200-300 stains per week. I do not register my reagents as the come into the lab. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Thursday, December 01, 2005 11:14 AM To: Histonet Subject: [Histonet] Question: Inventory Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Sat Dec 3 01:52:40 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Dec 3 01:52:42 2005 Subject: AW: [Histonet] Question: Inventory In-Reply-To: Message-ID: We register the bulk-reagents and prep-kits in the Ventana System just before use. The antibodies are registered, when they arrive, to have them on the list. Gudrun Lang Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ 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 nina.owen <@t> tiscali.co.uk Sat Dec 3 14:18:21 2005 From: nina.owen <@t> tiscali.co.uk (nina.owen@tiscali.co.uk) Date: Sat Dec 3 14:18:37 2005 Subject: [Histonet] Problems with cutting frozen sections of skin punch biopsies Message-ID: <43858E3C00036BBA@mk-cpfrontend-1.mail.uk.tiscali.com> Hi all! Just wondered if anyone could help us with the following problem... Our histology lab occasionally receives fresh skin punch biopsies for direct immunofluorescence from the hospital's dermatology department. When obtaining frozen sections from these specimens we don't experience any problems during cutting them. However, we are increasingly receiving punch biospies from other hospitals for DIF staining; these are sent in Michael's medium for preservation on transit. We seem to have a lot of difficulty cutting these! The problem is that the sections seem to crease up immediately, or if we get a decent section the tissue ends up sticking to the anti-roll plate. Thus, when the anti-roll plate is lifted to allow the section to be picked up onto a slide, the tissue is lifted out of the OCT mountant (Shandon) leaving a hole! We have tried a couple of things to try and stop this from happening. If we manage to get the specimen into the deep freeze asap and leave it for a few days it is slightly better. Also, leaving the tissue mounted on a chuck in the cryostat to cool down for about an hour prior to attempting to cut it stops the tissue from sticking to the anti roll plate, but only long enough for about two sections to be obtained. Has anyoneelse had this problem with fresh specimens that have been in Michael's medium? If so, how did you sort it? Thanks Histonetters! Nina Owen Dewsbury and District Hospital, Mid Yorkshire NHS Trust ___________________________________________________________ Tiscali Broadband from 14.99 with free setup! http://www.tiscali.co.uk/products/broadband/ Christmas gift ideas and festive features - visit the Tiscali Christmas microsite. http://www.tiscali.co.uk/christmas From katri <@t> cogeco.ca Sat Dec 3 19:55:07 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Sat Dec 3 19:55:08 2005 Subject: [Histonet] Problems with cutting frozen sections of skin punchbiopsies References: <43858E3C00036BBA@mk-cpfrontend-1.mail.uk.tiscali.com> Message-ID: <001201c5f875$c07851e0$6a9a9618@Katri> Hi Nina, My understanding is, that Michel's is a transport medium and the specimen needs to be washed through certain buffers to remove the salts before you are able to get good sections. I don't have the buffer recipe on hand, but maybe other histonetters can help you there. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: To: Sent: Saturday, December 03, 2005 3:18 PM Subject: [Histonet] Problems with cutting frozen sections of skin punchbiopsies Hi all! Just wondered if anyone could help us with the following problem... Our histology lab occasionally receives fresh skin punch biopsies for direct immunofluorescence from the hospital's dermatology department. When obtaining frozen sections from these specimens we don't experience any problems during cutting them. However, we are increasingly receiving punch biospies from other hospitals for DIF staining; these are sent in Michael's medium for preservation on transit. We seem to have a lot of difficulty cutting these! The problem is that the sections seem to crease up immediately, or if we get a decent section the tissue ends up sticking to the anti-roll plate. Thus, when the anti-roll plate is lifted to allow the section to be picked up onto a slide, the tissue is lifted out of the OCT mountant (Shandon) leaving a hole! We have tried a couple of things to try and stop this from happening. If we manage to get the specimen into the deep freeze asap and leave it for a few days it is slightly better. Also, leaving the tissue mounted on a chuck in the cryostat to cool down for about an hour prior to attempting to cut it stops the tissue from sticking to the anti roll plate, but only long enough for about two sections to be obtained. Has anyoneelse had this problem with fresh specimens that have been in Michael's medium? If so, how did you sort it? Thanks Histonetters! Nina Owen Dewsbury and District Hospital, Mid Yorkshire NHS Trust ___________________________________________________________ Tiscali Broadband from 14.99 with free setup! http://www.tiscali.co.uk/products/broadband/ Christmas gift ideas and festive features - visit the Tiscali Christmas microsite. http://www.tiscali.co.uk/christmas _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Steven2146 <@t> aol.com Sun Dec 4 12:47:17 2005 From: Steven2146 <@t> aol.com (Steven2146@aol.com) Date: Sun Dec 4 12:47:35 2005 Subject: [Histonet] Re: Histonet Digest, Vol 25, Issue 6 Message-ID: <126.6adb39ca.30c493b5@aol.com> Hi Fellow Histonetters... Anyone that works with Mohs surgeons needs to look at this new web site....it's the first one I've found....www.mohstechstaffing.com. From conniegrubaugh <@t> hotmail.com Sun Dec 4 13:35:27 2005 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Sun Dec 4 13:35:38 2005 Subject: [Histonet] Bone marrow smears Message-ID: I would appreciate it if anyone has any methods that they use when staining bone marrow smears to have less precipitate. Right now I'm trying a meth alcoh and dist. water 50/50 for 30 sec. after the buffer. Thanks Connie Grubaugh From GDawson <@t> dynacaremilwaukee.com Mon Dec 5 07:48:15 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Mon Dec 5 07:50:05 2005 Subject: [Histonet] Problems with cutting frozen sections of skin punc h biopsies Message-ID: Nina, Place your outside skin punch into a beaker (I use a 100 ml. beaker) filled 3/4 full with something like a pH 7.6 Tris buffer (if you have no buffer, DI water will work better than nothing). Put your beaker on a stirring plate & add a stir bar to the buffer that now contains the bx. Stir (I stir at about 50% speed) for ten minutes. If the bx. is especially large repeat the previous steps again with fresh Tris. Get your cryostat chuck ready, place your bx on a paper towel to soak off the excess buffer, mount & cut. Unless you rinse out the Michel's transport medium, you will mainly get an empty hole where the tissue was. You'll be amazed at how much better it will cut after a good rinse. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: nina.owen@tiscali.co.uk [mailto:nina.owen@tiscali.co.uk] Sent: Saturday, December 03, 2005 2:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Problems with cutting frozen sections of skin punch biopsies Hi all! Just wondered if anyone could help us with the following problem... Our histology lab occasionally receives fresh skin punch biopsies for direct immunofluorescence from the hospital's dermatology department. When obtaining frozen sections from these specimens we don't experience any problems during cutting them. However, we are increasingly receiving punch biospies from other hospitals for DIF staining; these are sent in Michael's medium for preservation on transit. We seem to have a lot of difficulty cutting these! The problem is that the sections seem to crease up immediately, or if we get a decent section the tissue ends up sticking to the anti-roll plate. Thus, when the anti-roll plate is lifted to allow the section to be picked up onto a slide, the tissue is lifted out of the OCT mountant (Shandon) leaving a hole! We have tried a couple of things to try and stop this from happening. If we manage to get the specimen into the deep freeze asap and leave it for a few days it is slightly better. Also, leaving the tissue mounted on a chuck in the cryostat to cool down for about an hour prior to attempting to cut it stops the tissue from sticking to the anti roll plate, but only long enough for about two sections to be obtained. Has anyoneelse had this problem with fresh specimens that have been in Michael's medium? If so, how did you sort it? Thanks Histonetters! Nina Owen Dewsbury and District Hospital, Mid Yorkshire NHS Trust ___________________________________________________________ Tiscali Broadband from 14.99 with free setup! http://www.tiscali.co.uk/products/broadband/ Christmas gift ideas and festive features - visit the Tiscali Christmas microsite. http://www.tiscali.co.uk/christmas _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From billingconsultants <@t> yahoo.com Mon Dec 5 10:06:44 2005 From: billingconsultants <@t> yahoo.com (Billing Consultants, LLC) Date: Mon Dec 5 10:06:54 2005 Subject: [Histonet] Histology Position Athens Georgia Message-ID: <20051205160644.94435.qmail@web54205.mail.yahoo.com> Hi Everyone, There is an opening for a histology technician in a private lab in Athens, Georgia. Great pay and working conditions If interested, please call 706-546-4884 or fax your resume to 706-546-4084. Thanks, Louri --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Yahoo! Personals From tennyjin <@t> gmail.com Mon Dec 5 10:14:26 2005 From: tennyjin <@t> gmail.com (tenny jin) Date: Mon Dec 5 10:14:34 2005 Subject: [Histonet] active Caspase-3 antibody and TUNEL kit Message-ID: Dear Histonetter, I would like hear some advice on good antibody against active Caspase-3. Prefer it is working in different species. Also, could anyone here recommend an apoptosis dectection kit (TUNEL assay)? We used to try the one from PerkinElmer. I am working with guinea pig tissue. Any input is greatly appreciated! /Tenny KI,stockholm From gcallis <@t> montana.edu Mon Dec 5 10:23:34 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Dec 5 10:23:43 2005 Subject: [Histonet] Rinsing Michels transport media from skin biopsies Message-ID: <6.0.0.22.1.20051205090750.01b5b8c8@gemini.msu.montana.edu> We always used the Citrate Stock/Rinse buffer made specifically for Michels transport media Stock Citrate Buffer 1M potassium citrate buffer pH 7 2.5 ml 0.1M Magnesium sulfate 5 ml 0.1M N-ethyl malemide 5 ml distilled water 87.5 ml Working solution i.e. Michels transport medium is made up in the same buffer with a salt added ammonium sulfate 55 g Stock buffer 100 ml The ammonium sulfate precipitates the immunoglobulins you want to stain, and this precipitation is reversed by rinsing the tissue with the Stock buffer at least 3 changes and with larger pieces of tissue, rinse longer - we did 3 changes/10 minutes each change - the suggested agitation is a good idea, on a rocker or with stirring, then blot and preferrably snap freeze to prevent excessive freezing artifact. Go to this website: www.zeusscientific.com to read up on this fixative and how it is used. We much preferred to buy the buffer ready made as it was a pain to make up all the time. Barry Rittman made an excellent suggestion is to embed in the OCT (Sakura Finetek) and let sit for a short time before snap freezing. This sometimes gives a better interface of cryoembedding media and the tissue. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cfavara <@t> niaid.nih.gov Mon Dec 5 10:58:40 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID)) Date: Mon Dec 5 10:58:50 2005 Subject: [Histonet] Question: Inventory Message-ID: I am replying to this just to let people know how easily things can be misinterpreted. I take responsibility for my statements and apologize for any misinformation I have given. The original question was "For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use?" My reply was "not do high volume consistently but have some weeks where I am doing 200-300 stains per week. I do not register my reagents as the come into the lab." What I should have said is that I do not register my Ventana Reagents on my Ventana Nexus Stainer as they arrive in the lab. I do register my Ventana Reagents on my Ventana Nexus as I am going to use them. The response to me was "If you are GLP or ISO complaint (even not registered) you must keep inventory records and record which reagents are used for making solutions. It is a pain and they think this will insure we do not use any expired or improper materials." I feel that someone thinks I do not follow "Good Laboratory Practice" This is a trivial point but I think it is interesting to note how different people respond to statements made not only here but everywhere. I feel it a wonder that I am understood at all. Perhaps I need to slow down and respond more carefully. Some people make very good precise statements and I will try and follow suit. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] Sent: Monday, December 05, 2005 7:34 AM To: Favara, Cynthia (NIH/NIAID) Subject: RE: [Histonet] Question: Inventory Hi Cynthia, If you are GLP or ISO complaint (even not registered) you must keep inventory records and record which reagents are used for making solutions. It is a pain and they think this will insure we do not use any expired or improper materials. Pam Marcum At 05:35 PM 12/2/2005, you wrote: >I do not do high volume consistently but have some weeks where I am >doing 200-300 stains per week. I do not register my reagents as the come >into the lab. >c > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is >confidential and may contain sensitive information. It should not be >used by anyone who is not the original intended recipient. If you have >received this e-mail in error please inform the sender and delete it >from your mailbox or any other storage devices. National Institute of >Allergy and Infectious Diseases shall not accept liability for any >statements made that are sender's own and not expressly made on behalf >of the NIAID by one of its representatives > >-----Original Message----- >From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] >Sent: Thursday, December 01, 2005 11:14 AM >To: Histonet >Subject: [Histonet] Question: Inventory > >Hi All >For those of you who are in high volume labs using ventana stainers, do >you >register all your reagents when they arrive? or do you register as you >use? >Thanks in advance >Luis >_______________________________________________ >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 Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From pmarcum <@t> vet.upenn.edu Mon Dec 5 11:25:18 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Mon Dec 5 11:25:38 2005 Subject: [Histonet] Question: Inventory In-Reply-To: References: Message-ID: <6.1.1.1.2.20051205121418.019afe28@mail.vet.upenn.edu> Cynthia, I apologize if that sounded like I thought you did not use GLP rules. Many people do not list or use the full practice of recording all SOPs and reagents with someone reviewing and signing off on them quarterly. That was what I meant. If you are fully GLP and are registered as such, which is not the same as following the rules with out full compliance it is still GLP just not full reporting reviews. I always followed GLP rules without a full reporting structure and only recently had to begin all of the record keeping and filing. It is a lot of work and I should have explained it better as we now have all of the SOPs and records files and it was very time consuming and different from what I had done before. We did not list all inventory as it came in on a viewable record. We have someone come in every three months and check all of those records and files now and sign off on them. Sorry for the misunderstanding and I did not mean you did not do good laboratory practices. I was questioning whether you had the reporting structure or not and if not you might not need to list all inventory the same way. Pam Marcum At 11:58 AM 12/5/2005, Favara, Cynthia \(NIH/NIAID\) wrote: >I am replying to this just to let people know how easily things can be >misinterpreted. I take responsibility for my statements and apologize >for any misinformation I have given. > >The original question was "For those of you who are in high volume labs >using ventana stainers, do you register all your reagents when they >arrive? or do you register as you use?" > >My reply was "not do high volume consistently but have some weeks where >I am doing 200-300 stains per week. I do not register my reagents as the >come into the lab." > >What I should have said is that I do not register my Ventana Reagents on >my Ventana Nexus Stainer as they arrive in the lab. I do register my >Ventana Reagents on my Ventana Nexus as I am going to use them. > >The response to me was "If you are GLP or ISO complaint (even not >registered) you must keep inventory records and record which reagents >are used for making solutions. It is a pain and they think this will >insure we do not use any expired or improper materials." > >I feel that someone thinks I do not follow "Good Laboratory Practice" > >This is a trivial point but I think it is interesting to note how >different people respond to statements made not only here but >everywhere. I feel it a wonder that I am understood at all. > >Perhaps I need to slow down and respond more carefully. Some people make >very good precise statements and I will try and follow suit. > >c > > >Cynthia Favara >NIAID/NIH/RML/LPVD >903 South 4th Street >Hamilton, MT 59840 >406-363-9317 > >Disclaimer: >The information in this e-mail and any of its attachments is >confidential and may contain sensitive information. It should not be >used by anyone who is not the original intended recipient. If you have >received this e-mail in error please inform the sender and delete it >from your mailbox or any other storage devices. National Institute of >Allergy and Infectious Diseases shall not accept liability for any >statements made that are sender's own and not expressly made on behalf >of the NIAID by one of its representatives > > >-----Original Message----- >From: Pamela Marcum [mailto:pmarcum@vet.upenn.edu] >Sent: Monday, December 05, 2005 7:34 AM >To: Favara, Cynthia (NIH/NIAID) >Subject: RE: [Histonet] Question: Inventory > >Hi Cynthia, > >If you are GLP or ISO complaint (even not registered) you must keep >inventory records and record which reagents are used for making >solutions. It is a pain and they think this will insure we do not use >any >expired or improper materials. > >Pam Marcum > >At 05:35 PM 12/2/2005, you wrote: > >I do not do high volume consistently but have some weeks where I am > >doing 200-300 stains per week. I do not register my reagents as the >come > >into the lab. > >c > > > >Cynthia Favara > >NIAID/NIH/RML/LPVD > >903 South 4th Street > >Hamilton, MT 59840 > >406-363-9317 > > > >Disclaimer: > >The information in this e-mail and any of its attachments is > >confidential and may contain sensitive information. It should not be > >used by anyone who is not the original intended recipient. If you have > >received this e-mail in error please inform the sender and delete it > >from your mailbox or any other storage devices. National Institute of > >Allergy and Infectious Diseases shall not accept liability for any > >statements made that are sender's own and not expressly made on behalf > >of the NIAID by one of its representatives > > > >-----Original Message----- > >From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] > >Sent: Thursday, December 01, 2005 11:14 AM > >To: Histonet > >Subject: [Histonet] Question: Inventory > > > >Hi All > >For those of you who are in high volume labs using ventana stainers, do > >you > >register all your reagents when they arrive? or do you register as you > >use? > >Thanks in advance > >Luis > >_______________________________________________ > >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 > >Best Regards, > >Pamela A Marcum >Manager, Histology Special Procedures >University of Pennsylvania >School of Veterinary Medicine >R.S. Reynolds Jr. CORL >New Bolton Center >382 West Street Road >Kennett Square, PA 19348 > >Phone - 610-925-6278 >Fax - 610-925-8120 >E-mail - pmarcum@vet.upenn.edu Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From pathrm35 <@t> adelphia.net Mon Dec 5 12:32:14 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Mon Dec 5 12:32:24 2005 Subject: [Histonet] Looking for Dr. Valerie Ray Message-ID: <5120670.1133807534399.JavaMail.root@web6.mail.adelphia.net> I am trying to contact Dr. Valerie Anne Yantsos Ray from the Tampa Bay, Florida area. If anyone knows her, please forward this email to her as I would like to speak with her. Thanks in advance. Ron Martin 561-721-2400 From Melissa.Gonzalez <@t> cellgenesys.com Mon Dec 5 12:46:33 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Mon Dec 5 12:47:00 2005 Subject: [Histonet] RE: active Caspase-3 antibody and TUNEL kit Message-ID: Tenny, Try the TUNEL kit from Chemicon, and active Caspase3 from R&D Systems (not sure if it will x-react with g.p). I used to use the Roche TUNEL kit, but have not been happy with the consistency of the kit in the past few years. Good luck, Melissa -----Original Message----- Dear Histonetter, I would like hear some advice on good antibody against active Caspase-3. Prefer it is working in different species. Also, could anyone here recommend an apoptosis dectection kit (TUNEL assay)? We used to try the one from PerkinElmer. I am working with guinea pig tissue. Any input is greatly appreciated! /Tenny KI,stockholm From EJones <@t> Ventanamed.com Mon Dec 5 13:38:11 2005 From: EJones <@t> Ventanamed.com (Emma JONES) Date: Mon Dec 5 13:40:51 2005 Subject: [Histonet] re:MSH-2 and MLH-1 Message-ID: Emma Jones =20 =20 Hi Diana,=20 Have you asked your Ventana rep, for info on these antibodies. They now sell both these primaries, pre-diluted and with recommended = protocol. MSH 2 Cat # 760-4265 MLH 1 Cat # 760-4264=20 Regards Emma Message: 12 Date: Fri, 2 Dec 2005 09:49:11 -0500 From: "Goodwin, Diana" Subject: [Histonet] MSH-2 and MLH-1 To: "Histonet \(E-mail\)" Message-ID: <80CDD9C3FEEAFD4982B114C4A6DFD00E256F77@uphsmbx2.UPHS.PENNHEALTH.PRV> Content-Type: text/plain; charset=3D"iso-8859-1" Greetings, Histonet. =20 I anyone running paraffin IHC with these Abs for human colo-rectal ca = successfully on the Ventana Benchmark, and if so, which clone, vendor, = dilution, etc. =20 Thanks in advance, =20 Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C =20 ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com =20 --=20 No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.362 / Virus Database: 267.13.11/191 - Release Date: = 02/12/2005 =20 From jzeliadt <@t> msn.com Mon Dec 5 15:27:23 2005 From: jzeliadt <@t> msn.com (JEFF TATUM ZELIADT) Date: Mon Dec 5 15:27:35 2005 Subject: [Histonet] specific Ig detection Message-ID: I am a student taking an immunobiology course. I was wondering if someone could help point me in the right direction. I have an unknown infections agent, I know that it produces anaphlylaxis and will produce IgE and IgA. If I isolate the IgE's from the serum I will get all IgE's even those not specific to the bug. How can I seperate the two and get the specific antigen? Can I use a protein which would be specific to the bug but how do I know what protein? From gcallis <@t> montana.edu Mon Dec 5 16:02:18 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Dec 5 16:02:28 2005 Subject: [Histonet] specific Ig detection In-Reply-To: References: Message-ID: <6.0.0.22.1.20051205144633.01b2e820@gemini.msu.montana.edu> You say "I know that it produces anaphlylaxis and will produce IgE and IgA so isn't this called the Host response. In other words the host infected by the "bug" responds to this by producing IgE (an allergic response as is anaphylaxis along with IgA) I don't think the bug is producing these Ig's. I suggest you try to get your "bug" identified via some microbiological means, such as a diagnostic culturing protocol. The serum of an animal infected with this infectious agent should contain antiserum to the agent, hence antiserum i.e. antibodies to the bug should be present in the serum of the infected host. So if you infected mouse cells with the bug, then the serum of the mouse will contain mouse antiBug antibodies - not necessarily IgA. I guess I am confused on what you are trying to do here. At 02:27 PM 12/5/2005, you wrote: >I am a student taking an immunobiology course. I was wondering if someone >could help point me in the right direction. I have an unknown infections >agent, >If I isolate the IgE's from the serum I will get all IgE's even those not >specific to the bug. How can I seperate the two and get the specific >antigen? Can I use a protein which would be specific to the bug but how >do I know what protein? > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From mikael.niku <@t> helsinki.fi Tue Dec 6 01:08:59 2005 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Tue Dec 6 01:09:18 2005 Subject: [Histonet] Anti-DIG peroxidase problem Message-ID: <4395390B.8040003@helsinki.fi> Dear Histonetters, I'm trying to set up tyramide amplification for my in situ hybridizations with digoxigenin probes. Unfortunately it seems as if the anti-DIG-POD by Roche doesn't work as well as the anti-DIG-AP I'm normally using. Basically, I get a weaker signal with anti-DIG-POD + tyramide amplification than with unamplified anti-DIG-AP. I have also tried biotinylated probes to avoid using anti-DIG-POD altogether, but so far I haven't got any signal at all with them. Any ideas, anyone? With best regards, Mikael Niku -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// From gmondragon <@t> gsopath.com Tue Dec 6 02:29:46 2005 From: gmondragon <@t> gsopath.com (Gus Mondragon) Date: Tue Dec 6 02:30:01 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 Message-ID: <03B63DE44B5C014D84F9A9D8A968B5174C4D3E@lithium.corp.gsopath.com> A positive and a negative control is required by CLIA in all AFB stains (Leprosy and TB). A known positive control validates your staining procedure; a negative control eliminates the presence of artifact in your sections Gus Mondragon gmondragon@gsopath.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, December 03, 2005 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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. Re: RE: Paraffin sections (Maria Mejia) 2. Extracellular matrix proteins fixation. Details (Diego J. Rodr?guez Gil) 3. RE: Specail stains Negative controls (Smith, Jeffery D. (HSC)) 4. Re: Specail stains Negative controls (Bryan Hewlett) 5. RE: Goat polymer (pruegg@ihctech.net) 6. Knife holder (arunams) 7. RE: Question: Inventory (Favara, Cynthia (NIH/NIAID)) 8. AW: [Histonet] Question: Inventory (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Fri, 02 Dec 2005 10:16:05 -0800 From: Maria Mejia Subject: Re: [Histonet] RE: Paraffin sections To: "Andrea T. Hooper" Cc: histonet@lists.utsouthwestern.edu, "C.M. van der Loos" , BMolinari@heart.thi.tmc.edu Message-ID: <43908F65.6010808@ski.org> Content-Type: text/plain; charset=us-ascii; format=flowed This type of experimenting would be interesting to check out especially with lower percentages of BSA. Andrea, I would agree that this overnight storage step probably is antigen dependent (Chris, that's what I meant by too much) and it would require a whole gamut testing of antibodies to see which respond positively and which would not...if one is inclined to do so. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Andrea T. Hooper wrote: > Very interesting Chris! I have found sometimes this works and > sometimes it doesn't and you get complete loss of staining - > frustrating when one was just trying to save time to begin with!! I > imagine it's antigen dependent but I have not had time to check. > > In my opinion Betsy, if you can avoid it (because you are just doing > it to save time instead of setting up a workshop) then avoid it ... it > simply isn't worth the risk IMHO. > > > > At 9:52 AM +0100 12/2/05, C.M. van der Loos wrote: > >> Betsy, >> >> For my NSH wet-workshops I tested the option of going up to endogenous >> PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. >> Next day we started at the protein blocking step and then the rest. >> Staining was as good as running the whole procedure in one day. >> >> Hope this info helps. >> >> Chris van der Loos, PhD >> Dept. of Pathology >> Academic Medical Center M2-230 >> Meibergdreef 9 >> NL-1105 AZ Amsterdam >> The Netherlands > > ------------------------------ Message: 2 Date: Fri, 02 Dec 2005 13:55:04 -0500 From: "Diego J. Rodr?guez Gil" Subject: [Histonet] Extracellular matrix proteins fixation. Details To: histonet@lists.utsouthwestern.edu Message-ID: <1133549704.43909888f401e@webmail.med.yale.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi: Thanks a lot to all of you who answer!. Since some of you asked, I am sending some more details. I am working on mice (CD-1), and the molecules I am trying to stain are named Wnt (Wingless-Int). Antibodies are from R&D systems (most of them are goats), and there are no previous works reporting the use of these antibodies. I am only making the assumption that they work, based on the datasheet the company provides. I could see some very faint positive stain for some of them, but I am sure I losing some because of fixation. Besides that, I know they are expressed based on RT-PCR and in situ hybridization experiments. Brief protocol is: Perfusion with 4% PFA, and then overnight deep in 4% PFA at 4 C. Transfer to PBS, then 30% sucrose til they sink, freeze on OCT and then section on cryostat at -18C. The antigen retrieval protocols that I tried were steaming (2, 5 and 10 min with sodium citrate) or pepsin in HCl. None of them gave me any better stainings. Staining protocol: Wash in TBS and then add 2% BSA in TBS-T, for 30 min. Primary antibody (on 2% BSA) Over night . Wash 3 x TBS-T. Add secondary antibody on 2% BSA for an hour. Wash 2 x TBS-T. Wash TBS and mount with Crystal Mount. Thanks again! Best regards, Diego. -- Diego J. Rodr?guez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. ------------------------------ Message: 3 Date: Fri, 2 Dec 2005 13:23:19 -0600 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] Specail stains Negative controls To: "Grant, Debra, R" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 2 Dec 2005 15:23:28 -0500 From: "Bryan Hewlett" Subject: Re: [Histonet] Specail stains Negative controls To: "Smith, Jeffery D. (HSC)" , "Grant, Debra, R" , Message-ID: <002001c5f77e$786f0990$6400a8c0@mainbox> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Debby and Jeff, Negative control slides are useful for GMS, Gram and AFB in order to eliminate false positives due to contamination from the reagents, washing, carry-over etc. Regards, Bryan ----- Original Message ----- From: "Smith, Jeffery D. (HSC)" To: "Grant, Debra, R" ; Sent: Friday, December 02, 2005 2:23 PM Subject: RE: [Histonet] Specail stains Negative controls Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ 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: 5 Date: Fri, 2 Dec 2005 13:39:43 -0700 From: pruegg@ihctech.net Subject: RE: [Histonet] Goat polymer To: "'Orr, Rebecca'" , Message-ID: <200512022039.jB2KdkXX005415@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" Becky, Invitrogen is the company who makes the goat labeled polymer. As for blocking, I use serum free protein block before the primary and then before the rab. Anti-goat secondary and then again before the rabbit labeled polymer. If I still encounter non-specific staining I put some serum (from the host of the secondary, about 10%) in with the protein block. This method is very sensitive and you can bring up lots of non-specific BG so be careful to optimize your dilutions and times for the antibody and for the detection reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, December 02, 2005 6:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Goat polymer Hello everyone, Just recently there was a posting on how to run a rabbit polymer with goat antibodies...or something to that nature, using an anti rabbit secondary and then the Dako polymer...could that person send me an email please? I had a quick question about blocking... Also, does anyone have the name of the company that makes a goat polymer? I know there's a company out there...I've seen the ads but can't locate it... Many thanks and have a great weekend! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Fri, 02 Dec 2005 14:04:38 -0800 (PST) From: arunams Subject: [Histonet] Knife holder To: histonet@lists.utsouthwestern.edu Message-ID: <5641228.1133561078869.JavaMail.myubc2@portal9.itservices.ubc.ca> Content-Type: text/plain; charset=us-ascii Hi All Does eny one know where to find a Tungston Carbide Knive holder for a Sorvall JB-4 Microtome. I really need one urgently. Thanks for your help Aruna Somasiri ------------------------------ Message: 7 Date: Fri, 2 Dec 2005 17:35:26 -0500 From: "Favara, Cynthia \(NIH/NIAID\)" Subject: RE: [Histonet] Question: Inventory To: "Luis Chiriboga" , "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" I do not do high volume consistently but have some weeks where I am doing 200-300 stains per week. I do not register my reagents as the come into the lab. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Thursday, December 01, 2005 11:14 AM To: Histonet Subject: [Histonet] Question: Inventory Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Sat, 3 Dec 2005 08:52:40 +0100 From: "Gudrun Lang" Subject: AW: [Histonet] Question: Inventory To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We register the bulk-reagents and prep-kits in the Ventana System just before use. The antibodies are registered, when they arrive, to have them on the list. Gudrun Lang Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ 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 End of Histonet Digest, Vol 25, Issue 5 *************************************** From Tiffany.L.Sheffield <@t> uth.tmc.edu Tue Dec 6 08:11:03 2005 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Sheffield, Tiffany L) Date: Tue Dec 6 08:11:10 2005 Subject: [Histonet] Bone EM Question Message-ID: Hi Fellow Histonetters! All you EM pro's, I have a question for you...What has been your experience with bone and EM? What is the largest sample size you have worked with (of course decaled)? I have been under the impression that only very small pieces(or very young bone that is not very calcified yet) that are very de-caled yield acceptable results. I am hearing conflicting accounts as to sample size and whether it has to be decaled or not. It is my understanding that it does have to be decaled and very small if it has a shot of working. I have only worked with soft tissue samples for EM. Any help would be greatly appreciated. Thanks! Tiffany Sheffield-Lopez, HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery UTHSC-Houston Medical School 6431 Fannin, Suite 6.144 MSB Houston , TX 77030 713-500-6803 WK 713-500-0729 Fax From GDawson <@t> dynacaremilwaukee.com Tue Dec 6 11:03:34 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Tue Dec 6 11:05:23 2005 Subject: [Histonet] FISH for HER2 testing Message-ID: All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI From osteffe <@t> online.no Tue Dec 6 11:06:29 2005 From: osteffe <@t> online.no (ole) Date: Tue Dec 6 11:06:38 2005 Subject: [Histonet] Tropheryma whippleii Message-ID: <000c01c5fa87$66d214a0$0100000a@ole> Anyone know of a commercially available Anti-Tropheryma Wippleii antibody that works on FFPET? Regards Ole J Steffensen Lab scientist Dep of Pathology Aalesund Norway From rjbuesa <@t> yahoo.com Tue Dec 6 11:48:57 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 6 11:49:05 2005 Subject: [Histonet] FISH for HER2 testing In-Reply-To: Message-ID: <20051206174857.93672.qmail@web61216.mail.yahoo.com> Hi Glen: I cannot recommend you a reference lab because we used to do our own tests for the DAKO and the FISH (from VYSIS). The results agreed and we used to run the DAKO tests on Thursdays and the FISH on Fridays on the cases requested from Thursday of one week to Wednesdays of the following. Rene J. "Dawson, Glen" wrote: All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Yahoo! Personals From tahseen <@t> brain.net.pk Tue Dec 6 04:24:19 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Tue Dec 6 12:24:39 2005 Subject: [Histonet] software for SOP Message-ID: <009101c5fa4f$3924ca80$972bfea9@m7c0y4> Hi All, I am looking software for my SOP, which cover the CAP requirement as per ANP.02888 NOTE 4. Thanks, Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. From HParker <@t> Skaggs.Net Tue Dec 6 12:27:23 2005 From: HParker <@t> Skaggs.Net (Parker, Helayne) Date: Tue Dec 6 12:27:46 2005 Subject: [Histonet] Service Contracts on Finesse ME Microtomes Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9D3@mail1-schc.skaggs.net> Hello, I was wondering if anyone could tell me if they have or have not purchased service contracts (or extended warranties) on the Finesse ME Microtomes ? I have been asked to weigh out whether it would be more cost effective off just paying per visit and repair or having the "plans". Our units have been here just under a year's time. I am new here and do not know much about the history of them- except that one was a demo and another brand new and one has had a defective knife holder replaced. Any input would be greatly appreciated. Helayne Parker Skaggs Community Health Center Branson, Missouri From HParker <@t> Skaggs.Net Tue Dec 6 12:34:08 2005 From: HParker <@t> Skaggs.Net (Parker, Helayne) Date: Tue Dec 6 12:34:31 2005 Subject: [Histonet] Mar-Med Bone Saws (Extenders) Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9D4@mail1-schc.skaggs.net> Hello again, I saw in the achieves that there is an "extender" that can be purchased for the Mar-Med saw. The drawback to the saw is that is only had a 3 inch clearance. With the extender how much more clearance is offered ? Again thank you for your input in advance. Sincerely, Helayne Parker, HT (A.S.C.P.) Histology Section Head Skaggs Community Health Center Branson, Missouri From pex0220 <@t> yahoo.com.cn Tue Dec 6 12:44:00 2005 From: pex0220 <@t> yahoo.com.cn (pex) Date: Tue Dec 6 12:44:12 2005 Subject: [Histonet] antibodies of bone marker Message-ID: <20051206184400.34928.qmail@web15509.mail.cnb.yahoo.com> Dear all, I would like to do immunostaing in mouse bone paraffin-embedded sections or mouse primary osteoblasts, so firstly I need some good antibodies of bone marker( such as alkaline phosphatase, osteocalcin, osteopontin, bone sialoprotein). But I have no ideas about them . If you have much more experience about them in immunostaining, can you do me a favor? Tell me some information about antibodies( company, catalogue number, or lab, etc) Thanks a lot Guofeng --------------------------------- ÎÞÏÞÈÝÁ¿ÑÅ»¢Ïà²á£¬Ô­Í¼µÈ´óÏÂÔØ£¬³¬¿ìËٶȣ¬¸Ï¿ìÇÀ×¢£¡ From la.sebree <@t> hosp.wisc.edu Tue Dec 6 12:58:06 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Tue Dec 6 12:58:56 2005 Subject: [Histonet] FISH for HER2 testing Message-ID: Hey Glen, that's way too long. We send our slides to the State Lab of Hygiene in Madison and they do FISH Her/2-neu at least twice per week as far as I know. Phone number: (608)265-4604. A little farther afield, you might also check into US Labs, (800)710-1800. We haven't used them for FISH but we outsource IHC to them and get excellent TAT and quality. Good luck, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Tuesday, December 06, 2005 11:04 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] FISH for HER2 testing All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.m.vanderloos <@t> amc.uva.nl Tue Dec 6 13:15:01 2005 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Tue Dec 6 13:15:19 2005 Subject: [Histonet] RE: Anti-DIG peroxidase problem Message-ID: <19df38c19ddd3a.19ddd3a19df38c@amc.uva.nl> Dear Mikael, Reading your message I have two remarks that might explain your strange findings: 1. The used chromogens are of stong influence to the final sensitivity/efficiency of a detection system. In your case I guess that with anti-DIG/AP, NBT/BCIP was used which is one of the most sensitive/efficient chromogens available. Far better than most peroxidase chromogens. Your remark you wasn't successful detecting bio-probes with peroxidase points a bit into this direction. Have you tried for example TrueBlue or Vector NovaRed as peroxidase chromogens. Both are very sensitive/efficient! 2. Obviously you are not an enthusiastic kit-user (just like me...). What did you do in the tyramide amplification step? Especially the peroxide concentration should be very very low here, otherwise you will get and adverse effect and certainly no amplification! Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 06 Dec 2005 09:08:59 +0200 From: Mikael Niku Subject: [Histonet] Anti-DIG peroxidase problem To: histonet@lists.utsouthwestern.edu Dear Histonetters, I'm trying to set up tyramide amplification for my in situ hybridizations with digoxigenin probes. Unfortunately it seems as if the anti-DIG-POD by Roche doesn't work as well as the anti-DIG-AP I'm normally using. Basically, I get a weaker signal with anti-DIG-POD + tyramide amplification than with unamplified anti-DIG-AP. I have also tried biotinylated probes to avoid using anti-DIG-POD altogether, but so far I haven't got any signal at all with them. Any ideas, anyone? From Nancy.Temple <@t> ssfhs.org Tue Dec 6 13:22:07 2005 From: Nancy.Temple <@t> ssfhs.org (Temple Nancy) Date: Tue Dec 6 13:22:16 2005 Subject: [Histonet] FISH for HER2 testing Message-ID: We send our FISH HER2 to CAD (Center for Advanced Diagnostics, an AmeriPath Lab)in Orlando, Fla. Our turnaround time is about 1 week. Nancy Temple Histology St. Francis Hospital Indianapolis, Indiana -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dawson, Glen Sent: Tuesday, December 06, 2005 12:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] FISH for HER2 testing All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. From O.M.Sandifort <@t> lumc.nl Fri Dec 2 07:23:29 2005 From: O.M.Sandifort <@t> lumc.nl (Sandifort, O.M. (PATH)) Date: Tue Dec 6 13:49:32 2005 Subject: [Histonet] defatting Message-ID: <40964D91251A18469CB08B6E96FB30BA226257@mailc.lumcnet.prod.intern> Hello, I'm looking for a method to defat breast tissue and while searching the web I found you question on the internet and now I'm wondering if somebody has answered you of if you found the answer to your question. Regards Orchid Sandifort, pathology, labworker Hospital Leiden, the Netherlands (LUMC) Defatting << Previous Message | Next Message >> From: "Badley, Carmen M SrA LRMC" To: "'histonet@pathology.swmed.edu'" Reply-To: Date: Thu, 16 Sep 1999 15:31:29 +0200 Content-Type: text/plain; charset="iso-8859-1" I was wondering if anyone has a good procedure on defatting breast tissue? Thank you Carmen Badley-Histology Landstuhl Regional Medical Center, GE From Michael.Rice <@t> holy-cross.com Tue Dec 6 13:49:34 2005 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Tue Dec 6 13:50:19 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 8 Message-ID: <3BC92F29BE821745AB15E04C98EE028D6D38DB@HCH2KMAIL.holy-cross.com> Hi Glen, We are using USLABS and getting results back in 4-5 days. We also use them for flow and have results back in 24-48 hours. They are great to work with and no, I do not having any financial interest in them Mike rice Holy cross hospital Ft lauderdale -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, December 06, 2005 1:12 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, Issue 8 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. Looking for Dr. Valerie Ray (pathrm35@adelphia.net) 2. RE: active Caspase-3 antibody and TUNEL kit (Melissa Gonzalez) 3. re:MSH-2 and MLH-1 (Emma JONES) 4. specific Ig detection (JEFF TATUM ZELIADT) 5. Re: specific Ig detection (Gayle Callis) 6. Anti-DIG peroxidase problem (Mikael Niku) 7. RE: Histonet Digest, Vol 25, Issue 5 (Gus Mondragon) 8. Bone EM Question (Sheffield, Tiffany L) 9. FISH for HER2 testing (Dawson, Glen) 10. Tropheryma whippleii (ole) 11. Re: FISH for HER2 testing (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Mon, 5 Dec 2005 13:32:14 -0500 From: Subject: [Histonet] Looking for Dr. Valerie Ray To: histonet@lists.utsouthwestern.edu Message-ID: <5120670.1133807534399.JavaMail.root@web6.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 I am trying to contact Dr. Valerie Anne Yantsos Ray from the Tampa Bay, Florida area. If anyone knows her, please forward this email to her as I would like to speak with her. Thanks in advance. Ron Martin 561-721-2400 ------------------------------ Message: 2 Date: Mon, 5 Dec 2005 10:46:33 -0800 From: "Melissa Gonzalez" Subject: [Histonet] RE: active Caspase-3 antibody and TUNEL kit To: Cc: tenny jin Message-ID: Content-Type: text/plain; charset="us-ascii" Tenny, Try the TUNEL kit from Chemicon, and active Caspase3 from R&D Systems (not sure if it will x-react with g.p). I used to use the Roche TUNEL kit, but have not been happy with the consistency of the kit in the past few years. Good luck, Melissa -----Original Message----- Dear Histonetter, I would like hear some advice on good antibody against active Caspase-3. Prefer it is working in different species. Also, could anyone here recommend an apoptosis dectection kit (TUNEL assay)? We used to try the one from PerkinElmer. I am working with guinea pig tissue. Any input is greatly appreciated! /Tenny KI,stockholm ------------------------------ Message: 3 Date: Mon, 5 Dec 2005 20:38:11 +0100 From: "Emma JONES" Subject: [Histonet] re:MSH-2 and MLH-1 To: Message-ID: Content-Type: text/plain; charset="windows-1250" Emma Jones Hi Diana, Have you asked your Ventana rep, for info on these antibodies. They now sell both these primaries, pre-diluted and with recommended protocol. MSH 2 Cat # 760-4265 MLH 1 Cat # 760-4264 Regards Emma Message: 12 Date: Fri, 2 Dec 2005 09:49:11 -0500 From: "Goodwin, Diana" Subject: [Histonet] MSH-2 and MLH-1 To: "Histonet \(E-mail\)" Message-ID: <80CDD9C3FEEAFD4982B114C4A6DFD00E256F77@uphsmbx2.UPHS.PENNHEALTH.PRV> Content-Type: text/plain; charset="iso-8859-1" Greetings, Histonet. I anyone running paraffin IHC with these Abs for human colo-rectal ca successfully on the Ventana Benchmark, and if so, which clone, vendor, dilution, etc. Thanks in advance, Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com -- No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.362 / Virus Database: 267.13.11/191 - Release Date: 02/12/2005 ------------------------------ Message: 4 Date: Mon, 05 Dec 2005 21:27:23 +0000 From: "JEFF TATUM ZELIADT" Subject: [Histonet] specific Ig detection To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed I am a student taking an immunobiology course. I was wondering if someone could help point me in the right direction. I have an unknown infections agent, I know that it produces anaphlylaxis and will produce IgE and IgA. If I isolate the IgE's from the serum I will get all IgE's even those not specific to the bug. How can I seperate the two and get the specific antigen? Can I use a protein which would be specific to the bug but how do I know what protein? ------------------------------ Message: 5 Date: Mon, 05 Dec 2005 15:02:18 -0700 From: Gayle Callis Subject: Re: [Histonet] specific Ig detection To: "JEFF TATUM ZELIADT" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051205144633.01b2e820@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed You say "I know that it produces anaphlylaxis and will produce IgE and IgA so isn't this called the Host response. In other words the host infected by the "bug" responds to this by producing IgE (an allergic response as is anaphylaxis along with IgA) I don't think the bug is producing these Ig's. I suggest you try to get your "bug" identified via some microbiological means, such as a diagnostic culturing protocol. The serum of an animal infected with this infectious agent should contain antiserum to the agent, hence antiserum i.e. antibodies to the bug should be present in the serum of the infected host. So if you infected mouse cells with the bug, then the serum of the mouse will contain mouse antiBug antibodies - not necessarily IgA. I guess I am confused on what you are trying to do here. At 02:27 PM 12/5/2005, you wrote: >I am a student taking an immunobiology course. I was wondering if someone >could help point me in the right direction. I have an unknown infections >agent, >If I isolate the IgE's from the serum I will get all IgE's even those not >specific to the bug. How can I seperate the two and get the specific >antigen? Can I use a protein which would be specific to the bug but how >do I know what protein? > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 6 Date: Tue, 06 Dec 2005 09:08:59 +0200 From: Mikael Niku Subject: [Histonet] Anti-DIG peroxidase problem To: histonet@lists.utsouthwestern.edu Message-ID: <4395390B.8040003@helsinki.fi> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Dear Histonetters, I'm trying to set up tyramide amplification for my in situ hybridizations with digoxigenin probes. Unfortunately it seems as if the anti-DIG-POD by Roche doesn't work as well as the anti-DIG-AP I'm normally using. Basically, I get a weaker signal with anti-DIG-POD + tyramide amplification than with unamplified anti-DIG-AP. I have also tried biotinylated probes to avoid using anti-DIG-POD altogether, but so far I haven't got any signal at all with them. Any ideas, anyone? With best regards, Mikael Niku -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// ------------------------------ Message: 7 Date: Tue, 6 Dec 2005 03:29:46 -0500 From: "Gus Mondragon" Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 To: Message-ID: <03B63DE44B5C014D84F9A9D8A968B5174C4D3E@lithium.corp.gsopath.com> Content-Type: text/plain; charset="iso-8859-1" A positive and a negative control is required by CLIA in all AFB stains (Leprosy and TB). A known positive control validates your staining procedure; a negative control eliminates the presence of artifact in your sections Gus Mondragon gmondragon@gsopath.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, December 03, 2005 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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. Re: RE: Paraffin sections (Maria Mejia) 2. Extracellular matrix proteins fixation. Details (Diego J. Rodr?guez Gil) 3. RE: Specail stains Negative controls (Smith, Jeffery D. (HSC)) 4. Re: Specail stains Negative controls (Bryan Hewlett) 5. RE: Goat polymer (pruegg@ihctech.net) 6. Knife holder (arunams) 7. RE: Question: Inventory (Favara, Cynthia (NIH/NIAID)) 8. AW: [Histonet] Question: Inventory (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Fri, 02 Dec 2005 10:16:05 -0800 From: Maria Mejia Subject: Re: [Histonet] RE: Paraffin sections To: "Andrea T. Hooper" Cc: histonet@lists.utsouthwestern.edu, "C.M. van der Loos" , BMolinari@heart.thi.tmc.edu Message-ID: <43908F65.6010808@ski.org> Content-Type: text/plain; charset=us-ascii; format=flowed This type of experimenting would be interesting to check out especially with lower percentages of BSA. Andrea, I would agree that this overnight storage step probably is antigen dependent (Chris, that's what I meant by too much) and it would require a whole gamut testing of antibodies to see which respond positively and which would not...if one is inclined to do so. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Andrea T. Hooper wrote: > Very interesting Chris! I have found sometimes this works and > sometimes it doesn't and you get complete loss of staining - > frustrating when one was just trying to save time to begin with!! I > imagine it's antigen dependent but I have not had time to check. > > In my opinion Betsy, if you can avoid it (because you are just doing > it to save time instead of setting up a workshop) then avoid it ... it > simply isn't worth the risk IMHO. > > > > At 9:52 AM +0100 12/2/05, C.M. van der Loos wrote: > >> Betsy, >> >> For my NSH wet-workshops I tested the option of going up to endogenous >> PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. >> Next day we started at the protein blocking step and then the rest. >> Staining was as good as running the whole procedure in one day. >> >> Hope this info helps. >> >> Chris van der Loos, PhD >> Dept. of Pathology >> Academic Medical Center M2-230 >> Meibergdreef 9 >> NL-1105 AZ Amsterdam >> The Netherlands > > ------------------------------ Message: 2 Date: Fri, 02 Dec 2005 13:55:04 -0500 From: "Diego J. Rodr?guez Gil" Subject: [Histonet] Extracellular matrix proteins fixation. Details To: histonet@lists.utsouthwestern.edu Message-ID: <1133549704.43909888f401e@webmail.med.yale.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi: Thanks a lot to all of you who answer!. Since some of you asked, I am sending some more details. I am working on mice (CD-1), and the molecules I am trying to stain are named Wnt (Wingless-Int). Antibodies are from R&D systems (most of them are goats), and there are no previous works reporting the use of these antibodies. I am only making the assumption that they work, based on the datasheet the company provides. I could see some very faint positive stain for some of them, but I am sure I losing some because of fixation. Besides that, I know they are expressed based on RT-PCR and in situ hybridization experiments. Brief protocol is: Perfusion with 4% PFA, and then overnight deep in 4% PFA at 4 C. Transfer to PBS, then 30% sucrose til they sink, freeze on OCT and then section on cryostat at -18C. The antigen retrieval protocols that I tried were steaming (2, 5 and 10 min with sodium citrate) or pepsin in HCl. None of them gave me any better stainings. Staining protocol: Wash in TBS and then add 2% BSA in TBS-T, for 30 min. Primary antibody (on 2% BSA) Over night . Wash 3 x TBS-T. Add secondary antibody on 2% BSA for an hour. Wash 2 x TBS-T. Wash TBS and mount with Crystal Mount. Thanks again! Best regards, Diego. -- Diego J. Rodr?guez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. ------------------------------ Message: 3 Date: Fri, 2 Dec 2005 13:23:19 -0600 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] Specail stains Negative controls To: "Grant, Debra, R" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 2 Dec 2005 15:23:28 -0500 From: "Bryan Hewlett" Subject: Re: [Histonet] Specail stains Negative controls To: "Smith, Jeffery D. (HSC)" , "Grant, Debra, R" , Message-ID: <002001c5f77e$786f0990$6400a8c0@mainbox> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Debby and Jeff, Negative control slides are useful for GMS, Gram and AFB in order to eliminate false positives due to contamination from the reagents, washing, carry-over etc. Regards, Bryan ----- Original Message ----- From: "Smith, Jeffery D. (HSC)" To: "Grant, Debra, R" ; Sent: Friday, December 02, 2005 2:23 PM Subject: RE: [Histonet] Specail stains Negative controls Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ 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: 5 Date: Fri, 2 Dec 2005 13:39:43 -0700 From: pruegg@ihctech.net Subject: RE: [Histonet] Goat polymer To: "'Orr, Rebecca'" , Message-ID: <200512022039.jB2KdkXX005415@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" Becky, Invitrogen is the company who makes the goat labeled polymer. As for blocking, I use serum free protein block before the primary and then before the rab. Anti-goat secondary and then again before the rabbit labeled polymer. If I still encounter non-specific staining I put some serum (from the host of the secondary, about 10%) in with the protein block. This method is very sensitive and you can bring up lots of non-specific BG so be careful to optimize your dilutions and times for the antibody and for the detection reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, December 02, 2005 6:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Goat polymer Hello everyone, Just recently there was a posting on how to run a rabbit polymer with goat antibodies...or something to that nature, using an anti rabbit secondary and then the Dako polymer...could that person send me an email please? I had a quick question about blocking... Also, does anyone have the name of the company that makes a goat polymer? I know there's a company out there...I've seen the ads but can't locate it... Many thanks and have a great weekend! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Fri, 02 Dec 2005 14:04:38 -0800 (PST) From: arunams Subject: [Histonet] Knife holder To: histonet@lists.utsouthwestern.edu Message-ID: <5641228.1133561078869.JavaMail.myubc2@portal9.itservices.ubc.ca> Content-Type: text/plain; charset=us-ascii Hi All Does eny one know where to find a Tungston Carbide Knive holder for a Sorvall JB-4 Microtome. I really need one urgently. Thanks for your help Aruna Somasiri ------------------------------ Message: 7 Date: Fri, 2 Dec 2005 17:35:26 -0500 From: "Favara, Cynthia \(NIH/NIAID\)" Subject: RE: [Histonet] Question: Inventory To: "Luis Chiriboga" , "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" I do not do high volume consistently but have some weeks where I am doing 200-300 stains per week. I do not register my reagents as the come into the lab. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Thursday, December 01, 2005 11:14 AM To: Histonet Subject: [Histonet] Question: Inventory Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Sat, 3 Dec 2005 08:52:40 +0100 From: "Gudrun Lang" Subject: AW: [Histonet] Question: Inventory To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We register the bulk-reagents and prep-kits in the Ventana System just before use. The antibodies are registered, when they arrive, to have them on the list. Gudrun Lang Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ 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 End of Histonet Digest, Vol 25, Issue 5 *************************************** ------------------------------ Message: 8 Date: Tue, 6 Dec 2005 08:11:03 -0600 From: "Sheffield, Tiffany L" Subject: [Histonet] Bone EM Question To: Message-ID: Content-Type: text/plain; charset="us-ascii" Hi Fellow Histonetters! All you EM pro's, I have a question for you...What has been your experience with bone and EM? What is the largest sample size you have worked with (of course decaled)? I have been under the impression that only very small pieces(or very young bone that is not very calcified yet) that are very de-caled yield acceptable results. I am hearing conflicting accounts as to sample size and whether it has to be decaled or not. It is my understanding that it does have to be decaled and very small if it has a shot of working. I have only worked with soft tissue samples for EM. Any help would be greatly appreciated. Thanks! Tiffany Sheffield-Lopez, HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery UTHSC-Houston Medical School 6431 Fannin, Suite 6.144 MSB Houston , TX 77030 713-500-6803 WK 713-500-0729 Fax ------------------------------ Message: 9 Date: Tue, 6 Dec 2005 11:03:34 -0600 From: "Dawson, Glen" Subject: [Histonet] FISH for HER2 testing To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI ------------------------------ Message: 10 Date: Tue, 6 Dec 2005 18:06:29 +0100 From: "ole" Subject: [Histonet] Tropheryma whippleii To: Message-ID: <000c01c5fa87$66d214a0$0100000a@ole> Content-Type: text/plain; charset="iso-8859-1" Anyone know of a commercially available Anti-Tropheryma Wippleii antibody that works on FFPET? Regards Ole J Steffensen Lab scientist Dep of Pathology Aalesund Norway ------------------------------ Message: 11 Date: Tue, 6 Dec 2005 09:48:57 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] FISH for HER2 testing To: "Dawson, Glen" , Histonet@lists.utsouthwestern.edu Message-ID: <20051206174857.93672.qmail@web61216.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi Glen: I cannot recommend you a reference lab because we used to do our own tests for the DAKO and the FISH (from VYSIS). The results agreed and we used to run the DAKO tests on Thursdays and the FISH on Fridays on the cases requested from Thursday of one week to Wednesdays of the following. Rene J. "Dawson, Glen" wrote: All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Personals Single? There's someone we'd like you to meet. Lots of someones, actually. Yahoo! Personals ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 25, Issue 8 *************************************** ---------------------- Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. 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. ---------------------- From tmmrosla <@t> healtheast.org Tue Dec 6 14:10:06 2005 From: tmmrosla <@t> healtheast.org (Mrosla, Tina M) Date: Tue Dec 6 14:23:40 2005 Subject: [Histonet] RE: FISH for Her-2 testing Message-ID: <22E6C4C32AFB3E44B7056C7EC3E1F2040531268E@HECLUSTER.HEALTHEAST.LOC> Have you tried Mayo Clinic in Rochester, MN? Their lab phone is 800-533-1710. We get our results in about a week. Tina Mrosla St. Joseph's Hospital St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. From AnthonyH <@t> chw.edu.au Tue Dec 6 14:51:55 2005 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Dec 6 14:52:13 2005 Subject: [Histonet] Anti-DIG peroxidase problem Message-ID: Try using a FITC labelled probe, followed by a mouse or rabbit anti-FITC. Then use your usual immunoperoxidase demonstration technique (ABC or polymer - I use a peroxidase-labelled polymer) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mikael Niku Sent: Tuesday, 6 December 2005 6:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Anti-DIG peroxidase problem Dear Histonetters, I'm trying to set up tyramide amplification for my in situ hybridizations with digoxigenin probes. Unfortunately it seems as if the anti-DIG-POD by Roche doesn't work as well as the anti-DIG-AP I'm normally using. Basically, I get a weaker signal with anti-DIG-POD + tyramide amplification than with unamplified anti-DIG-AP. I have also tried biotinylated probes to avoid using anti-DIG-POD altogether, but so far I haven't got any signal at all with them. Any ideas, anyone? With best regards, Mikael Niku -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// _______________________________________________ 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 jennifer.l.hofecker <@t> Vanderbilt.Edu Tue Dec 6 15:01:12 2005 From: jennifer.l.hofecker <@t> Vanderbilt.Edu (Hofecker, Jennifer L) Date: Tue Dec 6 15:01:23 2005 Subject: [Histonet] CD133 on Paraffin Message-ID: <898D946569A27444B65667A49C074052852B01@mailbe06.mc.vanderbilt.edu> Hi to all, Has anybody successfully used any antibodies to CD133 on FFPE tissues (formalin fixed, paraffin embedded)? I have searched the archives as well as several other sites, and I haven't found anything that looks promising. Hopefully somebody can point me in the right direction. A researcher asked me to find one. I am assuming it will be on human tissues (maybe mouse/rat) and most likely brain tissue. Will get those answeres once I know there is an antibody out there that may work in paraffin. Thanks in advance, Jennifer Jennifer Hofecker, HT (ASCP) Vanderbilt University Medical Center Division of Neuropathology (615) 343-0083 (615) 343-7089 fax From FAILM <@t> musc.edu Tue Dec 6 17:00:26 2005 From: FAILM <@t> musc.edu (Mildred Fail) Date: Tue Dec 6 17:00:56 2005 Subject: [Histonet] control issue Message-ID: I have been asked to put the patient tissue on the control slides. I readily see the benefits in doing so, but I need some advice on how to make the transition. We have seventy controls for IHC and 30 for SS 131 Antibodies I would like input from anyone who has had to make this transition. How did you get enough controls cut to make the transition? Who cuts your controls? How do you manage to keep up with the volume necessary to manage this task? Any advice would be greatly appreciated? Rena Fail Rena Fail From vanann702 <@t> skmc.gov.ae Tue Dec 6 22:04:56 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Tue Dec 6 22:02:10 2005 Subject: [Histonet] FISH for HER2 testing Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E0445CD5B@SKMCEMAIL.skmc.gov.ae> We use the Mayo Clinic - get our results within a week Annieinarabia -----Original Message----- From: Dawson, Glen [mailto:GDawson@dynacaremilwaukee.com] Sent: Tuesday, December 06, 2005 9:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] FISH for HER2 testing All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> sbcglobal.net Wed Dec 7 04:08:32 2005 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Wed Dec 7 04:10:16 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 In-Reply-To: <03B63DE44B5C014D84F9A9D8A968B5174C4D3E@lithium.corp.gsopath.com> Message-ID: I would like to pursue the comment that CLIA requires a positive and negative control for AFB. I went looking under CLIA (see below). I'll put my comments/questions in CAPS after each section I could find that somewhat pertained. (So, no, I'm not shouting. Just trying to differentiate between CLIA regs and my comments/questions.) - - - - Sec. 493.1256 Standard: Control procedures (a) For each test system, the laboratory is responsible for having control procedures that monitor the accuracy and precision of the complete analytical process. (b) The laboratory must establish the number, type, and frequency of testing control materials using, if applicable, the performance specifications verified d) Unless CMS approves a procedure, specified in Appendix C of the State Operations Manual (CMS Pub. 7), that provides equivalent quality testing, the laboratory must- (1) Perform control procedures as defined in this section unless otherwise specified in the additional specialty and subspecialty requirements at Sec. Sec. 493.1261 through 493.1278. (2) For each test system, perform control procedures using the number and frequency specified by the manufacturer or established by the laboratory when they meet or exceed the requirements in paragraph (d)(3) of this section. (3) At least once each day patient specimens are assayed or examined perform the following for-- (i) Each quantitative procedure, include two control materials of different concentrations; (ii) Each qualitative procedure, include a negative and positive control material; - - - - SO FOR QUALITATIVE PROCEDURES IN ALL LABS, THEY MUST HAVE NEGATIVE AND POSITIVE CONTROL MATERIAL - DOES THAT MEAN ALL OUR HISTOCHEMICAL SPECIAL STAINS NEED A POSITIVE AND NEGATIVE CONTROL , NOT JUST AFB?, CONTINUING WITH CLIA, UNDER MYCOBACTERIOLOGY: - - - - Sec. 493.1262 Standard: Mycobacteriology (a) Each day of use, the laboratory must check all reagents or test procedures used for mycobacteria identification with at least one acid-fast organism that produces a positive reaction and an acid-fast organism that produces a negative reaction. - - - - - SINCE THIS SECTION IS SPECIFIC FOR MYCOBACTERIOLOGY, DOES IT APPLY TO HISTOLOGY'S AFB HISTOCHEMICAL STAINS? THE STANDARD FOR THE HISTOLOGY LAB STATES: - - - - - Sec. 493.1273 Standard: Histopathology (a) Fluorescent and immunohistochemical stains must be checked for positive and negative reactivity each time of use. For all other differential or special stains, a control slide of known reactivity must be stained with each patient slide or group of patient slides. Reaction(s) of the control slide with each special stain must be documented. (b) The laboratory must retain stained slides, specimen blocks, and tissue remnants as specified in Sec. 493.1105. The remnants of tissue specimens must be maintained in a manner that ensures proper preservation of the tissue specimens until the portions submitted for microscopic examination have been examined and a diagnosis made by anindividual qualified under Sec. Sec. 493.1449(b), (l), or (m). (c) An individual who has successfully completed a training program in neuromuscular pathology approved by HHS may examine and provide reports for neuromuscular pathology. (d) Tissue pathology reports must be signed by an individual qualified as specified in paragraph (b) or, as appropriate, paragraph (c) of this section. If a computer report is generated with an electronic signature, it must be authorized by the individual who performed the examination and made the diagnosis. (e) The laboratory must use acceptable terminology of a recognized system of disease nomenclature in reporting results. (f) The laboratory must document all control procedures performed, as specified in this section. - - - - POSITIVE AND NEGATIVE CONTROLS ARE MENTIONED FOR IHC AND FLUORESCENT STAINS, BUT FOR SPECIAL STAINS, ONLY A KNOWN POSITIVE CONTROL IS REQUIRED. DOES THIS SENTENCE NEGATE THE TWO STANDARDS ABOVE (ALL LABS AND MYCOBACTERIOLOGY), WHEN IT PERTAINS TO HISTOLOGY SPECIAL STAINS? I WOULD SAY YES. BUT I WOULD APPRECIATE VERY MUCH, IF SOMEONE WITH MORE EXPERTISE IN CLIA REGS WOULD COMMENT ON POSITIVE/NEGATIVE CONTROLS IN HISTOCHEMICAL SPECIAL STAINS. THANK YOU. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gus Mondragon Sent: Tuesday, December 06, 2005 3:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 A positive and a negative control is required by CLIA in all AFB stains (Leprosy and TB). A known positive control validates your staining procedure; a negative control eliminates the presence of artifact in your sections Gus Mondragon gmondragon@gsopath.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, December 03, 2005 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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. Re: RE: Paraffin sections (Maria Mejia) 2. Extracellular matrix proteins fixation. Details (Diego J. Rodr?guez Gil) 3. RE: Specail stains Negative controls (Smith, Jeffery D. (HSC)) 4. Re: Specail stains Negative controls (Bryan Hewlett) 5. RE: Goat polymer (pruegg@ihctech.net) 6. Knife holder (arunams) 7. RE: Question: Inventory (Favara, Cynthia (NIH/NIAID)) 8. AW: [Histonet] Question: Inventory (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Fri, 02 Dec 2005 10:16:05 -0800 From: Maria Mejia Subject: Re: [Histonet] RE: Paraffin sections To: "Andrea T. Hooper" Cc: histonet@lists.utsouthwestern.edu, "C.M. van der Loos" , BMolinari@heart.thi.tmc.edu Message-ID: <43908F65.6010808@ski.org> Content-Type: text/plain; charset=us-ascii; format=flowed This type of experimenting would be interesting to check out especially with lower percentages of BSA. Andrea, I would agree that this overnight storage step probably is antigen dependent (Chris, that's what I meant by too much) and it would require a whole gamut testing of antibodies to see which respond positively and which would not...if one is inclined to do so. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Andrea T. Hooper wrote: > Very interesting Chris! I have found sometimes this works and > sometimes it doesn't and you get complete loss of staining - > frustrating when one was just trying to save time to begin with!! I > imagine it's antigen dependent but I have not had time to check. > > In my opinion Betsy, if you can avoid it (because you are just doing > it to save time instead of setting up a workshop) then avoid it ... it > simply isn't worth the risk IMHO. > > > > At 9:52 AM +0100 12/2/05, C.M. van der Loos wrote: > >> Betsy, >> >> For my NSH wet-workshops I tested the option of going up to endogenous >> PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. >> Next day we started at the protein blocking step and then the rest. >> Staining was as good as running the whole procedure in one day. >> >> Hope this info helps. >> >> Chris van der Loos, PhD >> Dept. of Pathology >> Academic Medical Center M2-230 >> Meibergdreef 9 >> NL-1105 AZ Amsterdam >> The Netherlands > > ------------------------------ Message: 2 Date: Fri, 02 Dec 2005 13:55:04 -0500 From: "Diego J. Rodr?guez Gil" Subject: [Histonet] Extracellular matrix proteins fixation. Details To: histonet@lists.utsouthwestern.edu Message-ID: <1133549704.43909888f401e@webmail.med.yale.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi: Thanks a lot to all of you who answer!. Since some of you asked, I am sending some more details. I am working on mice (CD-1), and the molecules I am trying to stain are named Wnt (Wingless-Int). Antibodies are from R&D systems (most of them are goats), and there are no previous works reporting the use of these antibodies. I am only making the assumption that they work, based on the datasheet the company provides. I could see some very faint positive stain for some of them, but I am sure I losing some because of fixation. Besides that, I know they are expressed based on RT-PCR and in situ hybridization experiments. Brief protocol is: Perfusion with 4% PFA, and then overnight deep in 4% PFA at 4 C. Transfer to PBS, then 30% sucrose til they sink, freeze on OCT and then section on cryostat at -18C. The antigen retrieval protocols that I tried were steaming (2, 5 and 10 min with sodium citrate) or pepsin in HCl. None of them gave me any better stainings. Staining protocol: Wash in TBS and then add 2% BSA in TBS-T, for 30 min. Primary antibody (on 2% BSA) Over night . Wash 3 x TBS-T. Add secondary antibody on 2% BSA for an hour. Wash 2 x TBS-T. Wash TBS and mount with Crystal Mount. Thanks again! Best regards, Diego. -- Diego J. Rodr?guez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. ------------------------------ Message: 3 Date: Fri, 2 Dec 2005 13:23:19 -0600 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] Specail stains Negative controls To: "Grant, Debra, R" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 2 Dec 2005 15:23:28 -0500 From: "Bryan Hewlett" Subject: Re: [Histonet] Specail stains Negative controls To: "Smith, Jeffery D. (HSC)" , "Grant, Debra, R" , Message-ID: <002001c5f77e$786f0990$6400a8c0@mainbox> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Debby and Jeff, Negative control slides are useful for GMS, Gram and AFB in order to eliminate false positives due to contamination from the reagents, washing, carry-over etc. Regards, Bryan ----- Original Message ----- From: "Smith, Jeffery D. (HSC)" To: "Grant, Debra, R" ; Sent: Friday, December 02, 2005 2:23 PM Subject: RE: [Histonet] Specail stains Negative controls Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ 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: 5 Date: Fri, 2 Dec 2005 13:39:43 -0700 From: pruegg@ihctech.net Subject: RE: [Histonet] Goat polymer To: "'Orr, Rebecca'" , Message-ID: <200512022039.jB2KdkXX005415@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" Becky, Invitrogen is the company who makes the goat labeled polymer. As for blocking, I use serum free protein block before the primary and then before the rab. Anti-goat secondary and then again before the rabbit labeled polymer. If I still encounter non-specific staining I put some serum (from the host of the secondary, about 10%) in with the protein block. This method is very sensitive and you can bring up lots of non-specific BG so be careful to optimize your dilutions and times for the antibody and for the detection reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, December 02, 2005 6:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Goat polymer Hello everyone, Just recently there was a posting on how to run a rabbit polymer with goat antibodies...or something to that nature, using an anti rabbit secondary and then the Dako polymer...could that person send me an email please? I had a quick question about blocking... Also, does anyone have the name of the company that makes a goat polymer? I know there's a company out there...I've seen the ads but can't locate it... Many thanks and have a great weekend! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Fri, 02 Dec 2005 14:04:38 -0800 (PST) From: arunams Subject: [Histonet] Knife holder To: histonet@lists.utsouthwestern.edu Message-ID: <5641228.1133561078869.JavaMail.myubc2@portal9.itservices.ubc.ca> Content-Type: text/plain; charset=us-ascii Hi All Does eny one know where to find a Tungston Carbide Knive holder for a Sorvall JB-4 Microtome. I really need one urgently. Thanks for your help Aruna Somasiri ------------------------------ Message: 7 Date: Fri, 2 Dec 2005 17:35:26 -0500 From: "Favara, Cynthia \(NIH/NIAID\)" Subject: RE: [Histonet] Question: Inventory To: "Luis Chiriboga" , "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" I do not do high volume consistently but have some weeks where I am doing 200-300 stains per week. I do not register my reagents as the come into the lab. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Thursday, December 01, 2005 11:14 AM To: Histonet Subject: [Histonet] Question: Inventory Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Sat, 3 Dec 2005 08:52:40 +0100 From: "Gudrun Lang" Subject: AW: [Histonet] Question: Inventory To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We register the bulk-reagents and prep-kits in the Ventana System just before use. The antibodies are registered, when they arrive, to have them on the list. Gudrun Lang Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ 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 End of Histonet Digest, Vol 25, Issue 5 *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Wed Dec 7 06:21:18 2005 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Wed Dec 7 06:22:10 2005 Subject: [Histonet] re CD133 Message-ID: <002e01c5fb28$ba414330$112b5c9f@Carlos> Hi, I use Abcam's ab16518 rabbit poly on rat FFPW sections after HIER. I just put a a pic up on here http://www.immunoportal.com/index.php From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Dec 7 06:52:16 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Wed Dec 7 06:56:01 2005 Subject: [Histonet] Re: control tissues Message-ID: We have a range of blocks both for immunos and special stains. We cut test and positive controls at the same time; case sections are mounted at the top of the slide; section from the control block goes at the bottom. We never pre-cut control sections. Also, we make a multiblock from 2 or 3 types of tissue (ie appendix+tonsil+colon all in one block; appendix+pancreas+carcinoid are another, as is testis+appendix) as this covers quite a wide range of immunos and, if one of the tissue samples isn't as good as the last for, example, CEA in colons (they do vary from case to case), then the other backs it up. It also means that you can cut a long ribbon of just the one block so it's a lot quicker than using many blocks, and it means that as little as 3 mulitblocks can be used for many of the immunos ranging from cytokeratin markers and lymphomas to some neuroendocrine markers. As for case ID, each sample of appendix or colon or tonsil etc taken is logged onto a specific sheet with the case number and number of blocks - example - first case sampled with 3 blocks taken would be 1-(1,2,3); the second case with 5 blocks 2-(1,2,3,4,5) etc. When the multiblock is made, these "1-(1,2,3)" etc details for all of the tissues are logged on its own sheet - example: "tonsil-colon-appendix multiblock 1" could consist of "colon block 1-1 + appendix block 3-4 + tonsil block 2-7"; multiblock 2 could be the same, depending on how many blocks there were for each tissue type. I test each positive control block for suitability (where appropriate) and keep the slides as a record plus log it down on its sheet with date and staff initials. I am now trying to create a digital image gallery file so if CPA ever ask us what is the source of Mr Blogg's positive control tissue, we can not only identify the actual case used but supply photo evidence that it has passes its suitability test. Does that sound complicated? Isn't really when you get used to it. Jacqui Malam Lancaster DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From julien_lambreydesouza <@t> uqar.qc.ca Wed Dec 7 07:17:32 2005 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Wed Dec 7 07:15:46 2005 Subject: [Histonet] RE: Service Contracts on Finesse ME Microtomes Message-ID: <1dc7f2878e697cb011e21616820369dd@uqar.qc.ca> Hello, We have had the Finesse ME for three years now. We took the warranty extension but never had to use it. However, it was an ease of mind to have it because you never know what might happen. Even if these microtomes are good quality machines, there is always an exception to the rule. If your budget is not a problem, I would consider taking to extension, but if money is limited, you might want to seriously think about it. It also depends on who is using the microtome (whether they are experienced people or not) and the usage rate (does it work 24H/day, 7 days/week?). I wish I could be of more help, Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 From rjbuesa <@t> yahoo.com Wed Dec 7 07:58:14 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 7 07:58:22 2005 Subject: [Histonet] control issue In-Reply-To: Message-ID: <20051207135814.74313.qmail@web61222.mail.yahoo.com> Hi Mildred: Case and control together in the same slide is now almost the norm and for laboratories with high volume of HC and IHC is cumbersome to cut controls and cases simultaneously. Also there are specimens (like liver and bone marrow biopsies) that the pathologists want the HC or IHC on the unstained slides in a series; in these cases the control cannot be placed in the same slide. Our transition went slowly because we first used up our controls before beginning to cut a new series of controls. We use the Fisher's + charged slides that have an area dedicated to the control, at the top of the slide (the case is put at the bottom). We cut our IHC controls beforehand BUT just a few because when the controls have some time in storage (about 1-2 weeks) the IHC reaction weakens. This weakening is not an "idea" or just a subjective percention. I published a paper in the J. of Histotechnology [28(2):89-97, 2005] where it is demonstrated that the weakening is statistically highly significant, i.e. not due to chance. Due to that we tried to use tissues that were good controls for several antibodies and cut only those that were going to be used during 1 week. Among the tissues that show weakening more easily are: muscle, colon, tonsil and prostate. Hope this will help you! Rene J. Mildred Fail wrote: I have been asked to put the patient tissue on the control slides. I readily see the benefits in doing so, but I need some advice on how to make the transition. We have seventy controls for IHC and 30 for SS 131 Antibodies I would like input from anyone who has had to make this transition. How did you get enough controls cut to make the transition? Who cuts your controls? How do you manage to keep up with the volume necessary to manage this task? Any advice would be greatly appreciated? Rena Fail Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Personals Skip the bars and set-ups and start using Yahoo! Personals for free From la.sebree <@t> hosp.wisc.edu Wed Dec 7 08:10:36 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Wed Dec 7 08:10:47 2005 Subject: [Histonet] control slides Message-ID: We routinely stockpile our control slides (Fisher "red box" control slides) and store those that are for antibodies that decrease in antigenicity over time in a -20 degree C freezer. Others that have no such problem we store at RT. We have used control slides that have been frozen for 2-3 years with no problem whatsoever; the antigenicity is the same as the stained slide we keep on file. As for who cuts them, we cut them if we have time, otherwise we ask one of the histotechs in Surgical Pathology to cut a batch for us in their slow time. Hope this helps, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 From sa.drew <@t> hosp.wisc.edu Wed Dec 7 08:52:38 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Wed Dec 7 08:52:50 2005 Subject: [Histonet] control issue Message-ID: One thing I would heartily recommend if storing control slides in the freezer is to make sure the sections are thoroughly dry prior to freezing-you don't want the tissues to lift off. You will also want to make sure the freezer is not the self-defrosting sort. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mildred Fail Sent: Tuesday, December 06, 2005 5:00 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] control issue I have been asked to put the patient tissue on the control slides. I readily see the benefits in doing so, but I need some advice on how to make the transition. We have seventy controls for IHC and 30 for SS 131 Antibodies I would like input from anyone who has had to make this transition. How did you get enough controls cut to make the transition? Who cuts your controls? How do you manage to keep up with the volume necessary to manage this task? Any advice would be greatly appreciated? Rena Fail Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From HParker <@t> Skaggs.Net Wed Dec 7 08:59:00 2005 From: HParker <@t> Skaggs.Net (Parker, Helayne) Date: Wed Dec 7 08:59:24 2005 Subject: [Histonet] Tap Water in Staining Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9D6@mail1-schc.skaggs.net> In using tap water for an H & E stain is there any documentation or testing that needs to be done to confirm that the tap water is ok for use. They have been using DI water here and there were complaints of slides being too pink from the Pathologist. I put the tap water into the regimen instead as I have always used tap for H & E staining -color is now fine - no problems. But I was asked by the other tech is that was acceptable. Having never been a supervisor I am not aware of any documentation needed. Can anyone advise. Helayne Parker, HT (A.S.C.P.) Histology Section Head Skaggs Community Health Center Branson, MO From bwhitaker <@t> brownpathology.com Wed Dec 7 09:47:18 2005 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Wed Dec 7 09:42:42 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 In-Reply-To: <200512070413.aa20305@MessagEX.iapc.net> Message-ID: <000601c5fb45$817634d0$3601a8c0@brownpathology.net> My thoughts on this are that the general lab standard and the mycobacteriology standards do not apply because the staining is not "testing" under CLIA rules. Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lee & Peggy Wenk Sent: Wednesday, December 07, 2005 4:09 AM To: 'Gus Mondragon'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 I would like to pursue the comment that CLIA requires a positive and negative control for AFB. I went looking under CLIA (see below). I'll put my comments/questions in CAPS after each section I could find that somewhat pertained. (So, no, I'm not shouting. Just trying to differentiate between CLIA regs and my comments/questions.) - - - - Sec. 493.1256 Standard: Control procedures (a) For each test system, the laboratory is responsible for having control procedures that monitor the accuracy and precision of the complete analytical process. (b) The laboratory must establish the number, type, and frequency of testing control materials using, if applicable, the performance specifications verified d) Unless CMS approves a procedure, specified in Appendix C of the State Operations Manual (CMS Pub. 7), that provides equivalent quality testing, the laboratory must- (1) Perform control procedures as defined in this section unless otherwise specified in the additional specialty and subspecialty requirements at Sec. Sec. 493.1261 through 493.1278. (2) For each test system, perform control procedures using the number and frequency specified by the manufacturer or established by the laboratory when they meet or exceed the requirements in paragraph (d)(3) of this section. (3) At least once each day patient specimens are assayed or examined perform the following for-- (i) Each quantitative procedure, include two control materials of different concentrations; (ii) Each qualitative procedure, include a negative and positive control material; - - - - SO FOR QUALITATIVE PROCEDURES IN ALL LABS, THEY MUST HAVE NEGATIVE AND POSITIVE CONTROL MATERIAL - DOES THAT MEAN ALL OUR HISTOCHEMICAL SPECIAL STAINS NEED A POSITIVE AND NEGATIVE CONTROL , NOT JUST AFB?, CONTINUING WITH CLIA, UNDER MYCOBACTERIOLOGY: - - - - Sec. 493.1262 Standard: Mycobacteriology (a) Each day of use, the laboratory must check all reagents or test procedures used for mycobacteria identification with at least one acid-fast organism that produces a positive reaction and an acid-fast organism that produces a negative reaction. - - - - - SINCE THIS SECTION IS SPECIFIC FOR MYCOBACTERIOLOGY, DOES IT APPLY TO HISTOLOGY'S AFB HISTOCHEMICAL STAINS? THE STANDARD FOR THE HISTOLOGY LAB STATES: - - - - - Sec. 493.1273 Standard: Histopathology (a) Fluorescent and immunohistochemical stains must be checked for positive and negative reactivity each time of use. For all other differential or special stains, a control slide of known reactivity must be stained with each patient slide or group of patient slides. Reaction(s) of the control slide with each special stain must be documented. (b) The laboratory must retain stained slides, specimen blocks, and tissue remnants as specified in Sec. 493.1105. The remnants of tissue specimens must be maintained in a manner that ensures proper preservation of the tissue specimens until the portions submitted for microscopic examination have been examined and a diagnosis made by anindividual qualified under Sec. Sec. 493.1449(b), (l), or (m). (c) An individual who has successfully completed a training program in neuromuscular pathology approved by HHS may examine and provide reports for neuromuscular pathology. (d) Tissue pathology reports must be signed by an individual qualified as specified in paragraph (b) or, as appropriate, paragraph (c) of this section. If a computer report is generated with an electronic signature, it must be authorized by the individual who performed the examination and made the diagnosis. (e) The laboratory must use acceptable terminology of a recognized system of disease nomenclature in reporting results. (f) The laboratory must document all control procedures performed, as specified in this section. - - - - POSITIVE AND NEGATIVE CONTROLS ARE MENTIONED FOR IHC AND FLUORESCENT STAINS, BUT FOR SPECIAL STAINS, ONLY A KNOWN POSITIVE CONTROL IS REQUIRED. DOES THIS SENTENCE NEGATE THE TWO STANDARDS ABOVE (ALL LABS AND MYCOBACTERIOLOGY), WHEN IT PERTAINS TO HISTOLOGY SPECIAL STAINS? I WOULD SAY YES. BUT I WOULD APPRECIATE VERY MUCH, IF SOMEONE WITH MORE EXPERTISE IN CLIA REGS WOULD COMMENT ON POSITIVE/NEGATIVE CONTROLS IN HISTOCHEMICAL SPECIAL STAINS. THANK YOU. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gus Mondragon Sent: Tuesday, December 06, 2005 3:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 5 A positive and a negative control is required by CLIA in all AFB stains (Leprosy and TB). A known positive control validates your staining procedure; a negative control eliminates the presence of artifact in your sections Gus Mondragon gmondragon@gsopath.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Saturday, December 03, 2005 1:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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. Re: RE: Paraffin sections (Maria Mejia) 2. Extracellular matrix proteins fixation. Details (Diego J. Rodr?guez Gil) 3. RE: Specail stains Negative controls (Smith, Jeffery D. (HSC)) 4. Re: Specail stains Negative controls (Bryan Hewlett) 5. RE: Goat polymer (pruegg@ihctech.net) 6. Knife holder (arunams) 7. RE: Question: Inventory (Favara, Cynthia (NIH/NIAID)) 8. AW: [Histonet] Question: Inventory (Gudrun Lang) ---------------------------------------------------------------------- Message: 1 Date: Fri, 02 Dec 2005 10:16:05 -0800 From: Maria Mejia Subject: Re: [Histonet] RE: Paraffin sections To: "Andrea T. Hooper" Cc: histonet@lists.utsouthwestern.edu, "C.M. van der Loos" , BMolinari@heart.thi.tmc.edu Message-ID: <43908F65.6010808@ski.org> Content-Type: text/plain; charset=us-ascii; format=flowed This type of experimenting would be interesting to check out especially with lower percentages of BSA. Andrea, I would agree that this overnight storage step probably is antigen dependent (Chris, that's what I meant by too much) and it would require a whole gamut testing of antibodies to see which respond positively and which would not...if one is inclined to do so. Yours Maria Bartola Mejia Smith-Kettlewell Eye Research Institute San Francisco, CA 94115 Email: maria@ski.org Phone: (415)-345-2185 Andrea T. Hooper wrote: > Very interesting Chris! I have found sometimes this works and > sometimes it doesn't and you get complete loss of staining - > frustrating when one was just trying to save time to begin with!! I > imagine it's antigen dependent but I have not had time to check. > > In my opinion Betsy, if you can avoid it (because you are just doing > it to save time instead of setting up a workshop) then avoid it ... it > simply isn't worth the risk IMHO. > > > > At 9:52 AM +0100 12/2/05, C.M. van der Loos wrote: > >> Betsy, >> >> For my NSH wet-workshops I tested the option of going up to endogenous >> PO blocking, HIER and then overnight storage at 4C in PBS with 1% BSA. >> Next day we started at the protein blocking step and then the rest. >> Staining was as good as running the whole procedure in one day. >> >> Hope this info helps. >> >> Chris van der Loos, PhD >> Dept. of Pathology >> Academic Medical Center M2-230 >> Meibergdreef 9 >> NL-1105 AZ Amsterdam >> The Netherlands > > ------------------------------ Message: 2 Date: Fri, 02 Dec 2005 13:55:04 -0500 From: "Diego J. Rodr?guez Gil" Subject: [Histonet] Extracellular matrix proteins fixation. Details To: histonet@lists.utsouthwestern.edu Message-ID: <1133549704.43909888f401e@webmail.med.yale.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi: Thanks a lot to all of you who answer!. Since some of you asked, I am sending some more details. I am working on mice (CD-1), and the molecules I am trying to stain are named Wnt (Wingless-Int). Antibodies are from R&D systems (most of them are goats), and there are no previous works reporting the use of these antibodies. I am only making the assumption that they work, based on the datasheet the company provides. I could see some very faint positive stain for some of them, but I am sure I losing some because of fixation. Besides that, I know they are expressed based on RT-PCR and in situ hybridization experiments. Brief protocol is: Perfusion with 4% PFA, and then overnight deep in 4% PFA at 4 C. Transfer to PBS, then 30% sucrose til they sink, freeze on OCT and then section on cryostat at -18C. The antigen retrieval protocols that I tried were steaming (2, 5 and 10 min with sodium citrate) or pepsin in HCl. None of them gave me any better stainings. Staining protocol: Wash in TBS and then add 2% BSA in TBS-T, for 30 min. Primary antibody (on 2% BSA) Over night . Wash 3 x TBS-T. Add secondary antibody on 2% BSA for an hour. Wash 2 x TBS-T. Wash TBS and mount with Crystal Mount. Thanks again! Best regards, Diego. -- Diego J. Rodr?guez Gil Postdoctoral Associate Department of Neurosurgery Yale University School of Medicine PO Box 208082 333 Cedar St, FMB-430 New Haven CT, 06520-8082 U.S.A. ------------------------------ Message: 3 Date: Fri, 2 Dec 2005 13:23:19 -0600 From: "Smith, Jeffery D. \(HSC\)" Subject: RE: [Histonet] Specail stains Negative controls To: "Grant, Debra, R" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Fri, 2 Dec 2005 15:23:28 -0500 From: "Bryan Hewlett" Subject: Re: [Histonet] Specail stains Negative controls To: "Smith, Jeffery D. (HSC)" , "Grant, Debra, R" , Message-ID: <002001c5f77e$786f0990$6400a8c0@mainbox> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Debby and Jeff, Negative control slides are useful for GMS, Gram and AFB in order to eliminate false positives due to contamination from the reagents, washing, carry-over etc. Regards, Bryan ----- Original Message ----- From: "Smith, Jeffery D. (HSC)" To: "Grant, Debra, R" ; Sent: Friday, December 02, 2005 2:23 PM Subject: RE: [Histonet] Specail stains Negative controls Debby, Routine special stains only require a positive control as you are looking to see if the stain worked or not. Specimen is either positive or negative. No need for negative control. IHC uses both pos. and neg. controls because you are looking for varying intensities of staining as well as pos. or neg. Best of Luck, Jeff ________________________________ Hi Histonetters, Does anyone run negative control slides with their GMS, AFB, and Gram special stains, and if so why? Kind Regards, Debby R. Grant HT(ASCP) Histology Coordinator The Children's Mercy Hospital & Clinics 2401 Gillham Rd. Kansas City, Missouri 64108 lab (816)234-3827 fax (816) 802-1492 _______________________________________________ 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: 5 Date: Fri, 2 Dec 2005 13:39:43 -0700 From: pruegg@ihctech.net Subject: RE: [Histonet] Goat polymer To: "'Orr, Rebecca'" , Message-ID: <200512022039.jB2KdkXX005415@pro12.abac.com> Content-Type: text/plain; charset="us-ascii" Becky, Invitrogen is the company who makes the goat labeled polymer. As for blocking, I use serum free protein block before the primary and then before the rab. Anti-goat secondary and then again before the rabbit labeled polymer. If I still encounter non-specific staining I put some serum (from the host of the secondary, about 10%) in with the protein block. This method is very sensitive and you can bring up lots of non-specific BG so be careful to optimize your dilutions and times for the antibody and for the detection reagents. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, December 02, 2005 6:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Goat polymer Hello everyone, Just recently there was a posting on how to run a rabbit polymer with goat antibodies...or something to that nature, using an anti rabbit secondary and then the Dako polymer...could that person send me an email please? I had a quick question about blocking... Also, does anyone have the name of the company that makes a goat polymer? I know there's a company out there...I've seen the ads but can't locate it... Many thanks and have a great weekend! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Fri, 02 Dec 2005 14:04:38 -0800 (PST) From: arunams Subject: [Histonet] Knife holder To: histonet@lists.utsouthwestern.edu Message-ID: <5641228.1133561078869.JavaMail.myubc2@portal9.itservices.ubc.ca> Content-Type: text/plain; charset=us-ascii Hi All Does eny one know where to find a Tungston Carbide Knive holder for a Sorvall JB-4 Microtome. I really need one urgently. Thanks for your help Aruna Somasiri ------------------------------ Message: 7 Date: Fri, 2 Dec 2005 17:35:26 -0500 From: "Favara, Cynthia \(NIH/NIAID\)" Subject: RE: [Histonet] Question: Inventory To: "Luis Chiriboga" , "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" I do not do high volume consistently but have some weeks where I am doing 200-300 stains per week. I do not register my reagents as the come into the lab. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Luis Chiriboga [mailto:Luis.Chiriboga@med.nyu.edu] Sent: Thursday, December 01, 2005 11:14 AM To: Histonet Subject: [Histonet] Question: Inventory Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Sat, 3 Dec 2005 08:52:40 +0100 From: "Gudrun Lang" Subject: AW: [Histonet] Question: Inventory To: "Histonetliste \(Histonetliste\)" Message-ID: Content-Type: text/plain; charset="us-ascii" We register the bulk-reagents and prep-kits in the Ventana System just before use. The antibodies are registered, when they arrive, to have them on the list. Gudrun Lang Hi All For those of you who are in high volume labs using ventana stainers, do you register all your reagents when they arrive? or do you register as you use? Thanks in advance Luis _______________________________________________ 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 End of Histonet Digest, Vol 25, Issue 5 *************************************** _______________________________________________ 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 Luis.Chiriboga <@t> med.nyu.edu Wed Dec 7 09:58:18 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Wed Dec 7 09:55:40 2005 Subject: [Histonet] Thanks Message-ID: Thanks to all who replied to my inventory question, Happy Holidays!! LC From mucram11 <@t> comcast.net Wed Dec 7 09:57:11 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Wed Dec 7 09:57:22 2005 Subject: [Histonet] Tap Water in Staining Message-ID: <120720051557.5366.4397065700075F1A000014F62202888744CECE030E9D0C9A03@comcast.net> Helayne, Tap water is fine in most cases. It is wise to know a baseline pH so you have something to look at if it changes over time, and it can. Tap water can change with the amount of chlorine added at different times of the year. When we have heavy rains here and when I lived in St Louis years ago it could mean the slides would literally bleach out due to higher chlorine levels in running water. Then we had to change again when they reduced it. One area of California has tap water at 8.0 to 8.5 pH which can over blued or change the colour with long rinses. Hope this helps. Pam Marcum -------------- Original message ---------------------- From: "Parker, Helayne" > In using tap water for an H & E stain is there any documentation or > testing that needs to be done to confirm that the tap water is ok for > use. They have been using DI water here and there were complaints of > slides being too pink from the Pathologist. I put the tap water into > the regimen instead as I have always used tap for H & E staining -color > is now fine - no problems. But I was asked by the other tech is that > was acceptable. Having never been a supervisor I am not aware of any > documentation needed. Can anyone advise. > > Helayne Parker, HT (A.S.C.P.) > Histology Section Head > Skaggs Community Health Center > Branson, MO > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caron_fournier <@t> yahoo.ca Wed Dec 7 10:22:18 2005 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Wed Dec 7 10:22:59 2005 Subject: [Histonet] Re: help needed with sheep spine Message-ID: <20051207162218.94147.qmail@web36315.mail.mud.yahoo.com> Hi All: being new to hard tissue processing I would like to ask for some help with a problem....I have some sheep spines that need to be embedded for sectioning and grinding but they have a pmma implant in them and I'm not sure how to process them without causing some problem with the existing PMMA and still be able to get good results on the cancellous bone in the vertebrae. Any suggestions are welcome. Caron histonet-request@lists.utsouthwestern.edu wrote: 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. software for SOP (Muhammad Tahseen) 2. Service Contracts on Finesse ME Microtomes (Parker, Helayne) 3. Mar-Med Bone Saws (Extenders) (Parker, Helayne) 4. antibodies of bone marker (pex) 5. RE: FISH for HER2 testing (Sebree Linda A.) 6. RE: Anti-DIG peroxidase problem (C.M. van der Loos) 7. RE: FISH for HER2 testing (Temple Nancy) 8. NBF fixed tissue and subsequent first step for dehydration. (Bruce Abaloz) 9. Mart 1 (karen johnson) 10. defatting (Sandifort, O.M. (PATH)) 11. RE: Histonet Digest, Vol 25, Issue 8 (Rice, Michael) 12. RE: FISH for Her-2 testing (Mrosla, Tina M) 13. RE: Anti-DIG peroxidase problem (Tony Henwood) 14. CD133 on Paraffin (Hofecker, Jennifer L) 15. control issue (Mildred Fail) 16. RE: FISH for HER2 testing (Anne Van Binsbergen) 17. RE: RE: Histonet Digest, Vol 25, Issue 5 (Lee & Peggy Wenk) ---------------------------------------------------------------------- Message: 1 Date: Tue, 6 Dec 2005 23:24:19 +1300 From: "Muhammad Tahseen" Subject: [Histonet] software for SOP To: Message-ID: <009101c5fa4f$3924ca80$972bfea9@m7c0y4> Content-Type: text/plain; charset="iso-8859-1" Hi All, I am looking software for my SOP, which cover the CAP requirement as per ANP.02888 NOTE 4. Thanks, Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ------------------------------ Message: 2 Date: Tue, 6 Dec 2005 12:27:23 -0600 From: "Parker, Helayne" Subject: [Histonet] Service Contracts on Finesse ME Microtomes To: Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9D3@mail1-schc.skaggs.net> Content-Type: text/plain; charset="us-ascii" Hello, I was wondering if anyone could tell me if they have or have not purchased service contracts (or extended warranties) on the Finesse ME Microtomes ? I have been asked to weigh out whether it would be more cost effective off just paying per visit and repair or having the "plans". Our units have been here just under a year's time. I am new here and do not know much about the history of them- except that one was a demo and another brand new and one has had a defective knife holder replaced. Any input would be greatly appreciated. Helayne Parker Skaggs Community Health Center Branson, Missouri ------------------------------ Message: 3 Date: Tue, 6 Dec 2005 12:34:08 -0600 From: "Parker, Helayne" Subject: [Histonet] Mar-Med Bone Saws (Extenders) To: Message-ID: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9D4@mail1-schc.skaggs.net> Content-Type: text/plain; charset="us-ascii" Hello again, I saw in the achieves that there is an "extender" that can be purchased for the Mar-Med saw. The drawback to the saw is that is only had a 3 inch clearance. With the extender how much more clearance is offered ? Again thank you for your input in advance. Sincerely, Helayne Parker, HT (A.S.C.P.) Histology Section Head Skaggs Community Health Center Branson, Missouri ------------------------------ Message: 4 Date: Wed, 7 Dec 2005 02:44:00 +0800 (CST) From: pex Subject: [Histonet] antibodies of bone marker To: Histonet@lists.utsouthwestern.edu Message-ID: <20051206184400.34928.qmail@web15509.mail.cnb.yahoo.com> Content-Type: text/plain; charset=gb2312 Dear all, I would like to do immunostaing in mouse bone paraffin-embedded sections or mouse primary osteoblasts, so firstly I need some good antibodies of bone marker( such as alkaline phosphatase, osteocalcin, osteopontin, bone sialoprotein). But I have no ideas about them . If you have much more experience about them in immunostaining, can you do me a favor? Tell me some information about antibodies( company, catalogue number, or lab, etc) Thanks a lot Guofeng --------------------------------- ???????????????????­???????????????????????????????? ------------------------------ Message: 5 Date: Tue, 6 Dec 2005 12:58:06 -0600 From: "Sebree Linda A." Subject: RE: [Histonet] FISH for HER2 testing To: "Dawson, Glen" Cc: Histonet Message-ID: Content-Type: text/plain; charset="us-ascii" Hey Glen, that's way too long. We send our slides to the State Lab of Hygiene in Madison and they do FISH Her/2-neu at least twice per week as far as I know. Phone number: (608)265-4604. A little farther afield, you might also check into US Labs, (800)710-1800. We haven't used them for FISH but we outsource IHC to them and get excellent TAT and quality. Good luck, Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Tuesday, December 06, 2005 11:04 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] FISH for HER2 testing All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Tue, 06 Dec 2005 20:15:01 +0100 From: "C.M. van der Loos" Subject: [Histonet] RE: Anti-DIG peroxidase problem To: histonet@lists.utsouthwestern.edu Cc: mikael.niku@helsinki.fi Message-ID: <19df38c19ddd3a.19ddd3a19df38c@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Dear Mikael, Reading your message I have two remarks that might explain your strange findings: 1. The used chromogens are of stong influence to the final sensitivity/efficiency of a detection system. In your case I guess that with anti-DIG/AP, NBT/BCIP was used which is one of the most sensitive/efficient chromogens available. Far better than most peroxidase chromogens. Your remark you wasn't successful detecting bio-probes with peroxidase points a bit into this direction. Have you tried for example TrueBlue or Vector NovaRed as peroxidase chromogens. Both are very sensitive/efficient! 2. Obviously you are not an enthusiastic kit-user (just like me...). What did you do in the tyramide amplification step? Especially the peroxide concentration should be very very low here, otherwise you will get and adverse effect and certainly no amplification! Hope this helps. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 06 Dec 2005 09:08:59 +0200 From: Mikael Niku Subject: [Histonet] Anti-DIG peroxidase problem To: histonet@lists.utsouthwestern.edu Dear Histonetters, I'm trying to set up tyramide amplification for my in situ hybridizations with digoxigenin probes. Unfortunately it seems as if the anti-DIG-POD by Roche doesn't work as well as the anti-DIG-AP I'm normally using. Basically, I get a weaker signal with anti-DIG-POD + tyramide amplification than with unamplified anti-DIG-AP. I have also tried biotinylated probes to avoid using anti-DIG-POD altogether, but so far I haven't got any signal at all with them. Any ideas, anyone? ------------------------------ Message: 7 Date: Tue, 6 Dec 2005 14:22:07 -0500 From: "Temple Nancy" Subject: RE: [Histonet] FISH for HER2 testing To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" We send our FISH HER2 to CAD (Center for Advanced Diagnostics, an AmeriPath Lab)in Orlando, Fla. Our turnaround time is about 1 week. Nancy Temple Histology St. Francis Hospital Indianapolis, Indiana -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dawson, Glen Sent: Tuesday, December 06, 2005 12:04 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] FISH for HER2 testing All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this e-mail and any accompanying documents is intended for the sole use of the recipient to whom it is addressed, and may contain information that is privileged, confidential, and prohibited from disclosure under applicable law. If you are not the intended recipient, or authorized to receive this on behalf of the recipient, you are hereby notified that any review, use, disclosure, copying, or distribution is prohibited. If you are not the intended recipients(s), please contact the sender by e-mail and destroy all copies of the original message. Thank you. ------------------------------ Message: 8 Date: Wed, 30 Nov 2005 12:15:35 +1100 From: Bruce Abaloz Subject: [Histonet] NBF fixed tissue and subsequent first step for dehydration. To: Lorraine Rolston Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Lorraine, In our Lab. we fix in 70% ETOH without any detrimental effects to morphology & then start processing in this also.....am happy to help in any way. Cheers, Bruce in OZ -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA. AUSTRALIA 3010 Nobody Can Make You Feel Inferior Without YOUR Permission - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING I am happy and content because I think I am. Q: What's a specimen? A: An Italian astronaut. ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. ------------------------------ Message: 9 Date: Wed, 30 Nov 2005 17:16:50 -0800 (PST) From: karen johnson Subject: [Histonet] Mart 1 To: histonet@lists.utsouthwestern.edu Message-ID: <20051201011650.86833.qmail@web52305.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I work in a small dermatology lab and want to start to do MART-1 on frozen sections especially for lentigo maligna. I need a procedure that is easy, and pretty quick. Any help would be appreciated. THANKS __________________________________ Start your day with Yahoo! - Make it your home page! http://www.yahoo.com/r/hs ------------------------------ Message: 10 Date: Fri, 2 Dec 2005 14:23:29 +0100 From: "Sandifort, O.M. \(PATH\)" Subject: [Histonet] defatting To: Message-ID: <40964D91251A18469CB08B6E96FB30BA226257@mailc.lumcnet.prod.intern> Content-Type: text/plain; charset="iso-8859-1" Hello, I'm looking for a method to defat breast tissue and while searching the web I found you question on the internet and now I'm wondering if somebody has answered you of if you found the answer to your question. Regards Orchid Sandifort, pathology, labworker Hospital Leiden, the Netherlands (LUMC) Defatting << Previous Message | Next Message >> From: "Badley, Carmen M SrA LRMC" To: "'histonet@pathology.swmed.edu'" Reply-To: Date: Thu, 16 Sep 1999 15:31:29 +0200 Content-Type: text/plain; charset="iso-8859-1" I was wondering if anyone has a good procedure on defatting breast tissue? Thank you Carmen Badley-Histology Landstuhl Regional Medical Center, GE ------------------------------ Message: 11 Date: Tue, 6 Dec 2005 14:49:34 -0500 From: "Rice, Michael" Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 8 To: Message-ID: <3BC92F29BE821745AB15E04C98EE028D6D38DB@HCH2KMAIL.holy-cross.com> Content-Type: text/plain; charset="utf-8" Hi Glen, We are using USLABS and getting results back in 4-5 days. We also use them for flow and have results back in 24-48 hours. They are great to work with and no, I do not having any financial interest in them Mike rice Holy cross hospital Ft lauderdale -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, December 06, 2005 1:12 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, Issue 8 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. Looking for Dr. Valerie Ray (pathrm35@adelphia.net) 2. RE: active Caspase-3 antibody and TUNEL kit (Melissa Gonzalez) 3. re:MSH-2 and MLH-1 (Emma JONES) 4. specific Ig detection (JEFF TATUM ZELIADT) 5. Re: specific Ig detection (Gayle Callis) 6. Anti-DIG peroxidase problem (Mikael Niku) 7. RE: Histonet Digest, Vol 25, Issue 5 (Gus Mondragon) 8. Bone EM Question (Sheffield, Tiffany L) 9. FISH for HER2 testing (Dawson, Glen) 10. Tropheryma whippleii (ole) 11. Re: FISH for HER2 testing (Rene J Buesa) ---------------------------------------------------------------------- Message: 1 Date: Mon, 5 Dec 2005 13:32:14 -0500 From: Subject: [Histonet] Looking for Dr. Valerie Ray To: histonet@lists.utsouthwestern.edu Message-ID: <5120670.1133807534399.JavaMail.root@web6.mail.adelphia.net> Content-Type: text/plain; charset=utf-8 I am trying to contact Dr. Valerie Anne Yantsos Ray from the Tampa Bay, Florida area. If anyone knows her, please forward this email to her as I would like to speak with her. Thanks in advance. Ron Martin 561-721-2400 ------------------------------ Message: 2 Date: Mon, 5 Dec 2005 10:46:33 -0800 From: "Melissa Gonzalez" Subject: [Histonet] RE: active Caspase-3 antibody and TUNEL kit To: Cc: tenny jin Message-ID: Content-Type: text/plain; charset="us-ascii" Tenny, Try the TUNEL kit from Chemicon, and active Caspase3 from R&D Systems (not sure if it will x-react with g.p). I used to use the Roche TUNEL kit, but have not been happy with the consistency of the kit in the past few years. Good luck, Melissa -----Original Message----- Dear Histonetter, I would like hear some advice on good antibody against active Caspase-3. Prefer it is working in different species. Also, could anyone here recommend an apoptosis dectection kit (TUNEL assay)? We used to try the one from PerkinElmer. I am working with guinea pig tissue. Any input is greatly appreciated! /Tenny KI,stockholm ------------------------------ Message: 3 Date: Mon, 5 Dec 2005 20:38:11 +0100 From: "Emma JONES" Subject: [Histonet] re:MSH-2 and MLH-1 To: Message-ID: Content-Type: text/plain; charset="windows-1250" Emma Jones Hi Diana, Have you asked your Ventana rep, for info on these antibodies. They now sell both these primaries, pre-diluted and with recommended protocol. MSH 2 Cat # 760-4265 MLH 1 Cat # 760-4264 Regards Emma Message: 12 Date: Fri, 2 Dec 2005 09:49:11 -0500 From: "Goodwin, Diana" Subject: [Histonet] MSH-2 and MLH-1 To: "Histonet \(E-mail\)" Message-ID: <80CDD9C3FEEAFD4982B114C4A6DFD00E256F77@uphsmbx2.UPHS.PENNHEALTH.PRV> Content-Type: text/plain; charset="iso-8859-1" Greetings, Histonet. I anyone running paraffin IHC with these Abs for human colo-rectal ca successfully on the Ventana Benchmark, and if so, which clone, vendor, dilution, etc. Thanks in advance, Diana Goodwin Supervisor, Anatomic Pathology Pennsylvania Hospital, Preston 655-C ph: 215-829-6532 fax: 215-829-7564 e-mail: goodwind@pahosp.com -- No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.362 / Virus Database: 267.13.11/191 - Release Date: 02/12/2005 ------------------------------ Message: 4 Date: Mon, 05 Dec 2005 21:27:23 +0000 From: "JEFF TATUM ZELIADT" Subject: [Histonet] specific Ig detection To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format=flowed I am a student taking an immunobiology course. I was wondering if someone could help point me in the right direction. I have an unknown infections agent, I know that it produces anaphlylaxis and will produce IgE and IgA. If I isolate the IgE's from the serum I will get all IgE's even those not specific to the bug. How can I seperate the two and get the specific === message truncated === Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos From Rcartun <@t> harthosp.org Wed Dec 7 11:42:53 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Dec 7 11:43:26 2005 Subject: [Histonet] FISH for HER2 testing Message-ID: I used PhenoPath (Seattle, WA) before we set-up the test in our Department. They were excellent. We received a faxed copy of the report in one week or less. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Dawson, Glen" 12/06/05 12:03PM >>> All, I have a pathologist who is looking for good turn around time on FISH HER2 testing. Their current turn around time is 3 weeks and the clinicians are clamoring for something better. If anyone knows of a reference lab that can do good FISH HER2 testing in a timely fashion, please let me know. On a side note, has anyone experienced any problems with the Dako FDA approved HercepTest kit when using it on breast core bx's? My Docs are not confident using this kit (especially on our breast cores). Has anyone run side by side comparisons between this kit and FISH HER2 testing? If someone can suggest a good FISH HER2 reference lab, the second set of questions are moot. Thank-you In Advance, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From cforster <@t> umn.edu Wed Dec 7 11:54:23 2005 From: cforster <@t> umn.edu (Colleen Forster) Date: Wed Dec 7 11:54:35 2005 Subject: [Histonet] looking for Wanda Jones Message-ID: <439721CF.1030602@umn.edu> HEllo histonetters, Could someone please give me contact information for Wanda Jones??? Thanks. Colleen Forster U of MN From Don.Birgerson <@t> leica-microsystems.com Wed Dec 7 14:10:10 2005 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Wed Dec 7 14:10:24 2005 Subject: [Histonet] Histology equipment sales position available Phoenix Message-ID: Leica Microsystems has an immediate opening for a Sales Representative of it's Histology Specimen Preparation Division. The representative will ideally be based in Phoenix area as the territory includes the entire states of Arizona and New Mexico, as well as Las Vegas, Nevada and West Texas. About Leica Leica Microsystems Inc. is an internationally renowned manufacturer and distributor of scientific instrumentation. We offer a competitive benefits package (including medical, prescription, dental, vision, life insurance, STD, LTD, 401K, long term care). Key Duties We are looking for a dynamic individual to sell and technically support our histology product line in the assigned territory. Responsibilities include achieving monthly, quarterly, annual and strategic product mix sales goals for the territory. Plan and schedule face-to-face account calls to current and potential end-users of Leica histology products. Perform instrument demonstrations. Install/set-up instrumentation in customer laboratories. Technically support and train customers on the use of histology instruments. Prepare monthly territory reports to Area Sales Director as well as other routine administrative duties. The ability to travel is required and is targeted at 2-3 nights per week. Knowledge and Background Requirements Qualified candidates must possess a BA/BS in Life Sciences or Associates Degree with 1-3 years Capital Equipment sales experience and demonstrated success in achieving quota are preferred. Understanding of the histology marketplace or a related discipline is preferred. Must be computer literate and have a basic understanding of Microsoft Office applications. Please forward your resume and salary requirements to: Beth Farrow Leica Microsystems Inc. HR Department 2345 Waukegan Road Bannockburn, IL 60015 Resumes submitted without salary requirements will not be considered. EOE M/F/D/V ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From sjchtascp <@t> yahoo.com Wed Dec 7 14:31:10 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Dec 7 14:31:18 2005 Subject: [Histonet] Brdu staining fixed cultured cells. Message-ID: <20051207203110.26572.qmail@web90205.mail.scd.yahoo.com> Good afternoon everyone, I'm working witha scientist who is attempting to stain previously fixed cultured cells in "wells" and cover slips with anti-Brdu. He has fixed the cells 1st in methanol, air dryed, then in 10%formalin/PBS and again air dryed. Our anti-BrdU/DNAase works great on tissue but not on the fixed/air dryed cultured cells. We tried a 30 minute 0.1% Saponin step to help permeablize the cells but w/o success. Any ideas. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Gifts at Yahoo! Shopping From Kristopher.Kalleberg <@t> unilever.com Wed Dec 7 15:41:52 2005 From: Kristopher.Kalleberg <@t> unilever.com (Kristopher Kalleberg) Date: Wed Dec 7 15:41:45 2005 Subject: [Histonet] Surgipath Ready-Vial Message-ID: Can anyone tell where I can buy Surgipath's Ready-Vial 10% Neutral Buffered Formalin. These are pre-filled vials with about 6mL of 10% NBF. I haven't bought a bag of them in a while and cannot seem to find them anywhere now. Thanks. From ander093 <@t> tc.umn.edu Wed Dec 7 15:50:09 2005 From: ander093 <@t> tc.umn.edu (LuAnn Anderson) Date: Wed Dec 7 15:50:22 2005 Subject: [Histonet] Surgipath Ready-Vial In-Reply-To: References: Message-ID: <6.2.3.4.0.20051207154840.0388f130@ander093.email.umn.edu> From Surgipath ;o) LuAnn oh yeah........their number is 1-800-225-3035 At 03:41 PM 12/7/2005, Kristopher Kalleberg wrote: >Can anyone tell where I can buy Surgipath's Ready-Vial 10% Neutral Buffered >Formalin. These are pre-filled vials with about 6mL of 10% NBF. I haven't >bought a bag of them in a while and cannot seem to find them anywhere now. >Thanks. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcolclefa <@t> aol.com Wed Dec 7 21:10:54 2005 From: jcolclefa <@t> aol.com (John PJ Coleman) Date: Wed Dec 7 21:11:11 2005 Subject: [Histonet] Re: Histonet Digest, Vol 25, Issue 9 In-Reply-To: <200512070511.1df4396b54ce9@rly-yc05.mail.aol.com> References: <200512070511.1df4396b54ce9@rly-yc05.mail.aol.com> Message-ID: Her2 by fish and her 2 by IHC like in Dako Herceptest are not testing for the same item. Fish lights up the genes, IHC labels the cell surface proteins. Herceptin is a drug based on surface expression of protein, not presence of multiple gene copies. The assumption by Vysis ( the FISH pushers) is that multiple gene copies = protein presence at cell surface and that IHC labs are not competent. What if the fish is called negative because of a 1:1 ratio but that multiple copies of gene and centromere are present within a set of cells that are 3+ according to IHC because of accelerated protein production? Deny therapy in a potentially responsive patient? But I digress. Herceptest is not recommended for core biopsies- i run i high volume IHC lab (518 slides yesterday) and we found that core bxs score higher by IHC than average. We also do FISH weekly, and we have found that 1+ ihc= always neg fish, 2+ ihc = 96% or so Neg fish, and 3+ = 85 or so pos fish. What specifically are your issues with the cores? like I said - we get potentially higher IHC +, fish neg on cores. Also- in IHC negative controls are used to tell if the secondary reagents are causing a chromogenic reaction on the slides independent of the primary antibody. Really has nothing to do with the varying intensity of the reaction in the test slides. It's just to avoid reporting out a false positive result. JPJC-757 335-2159 http://journals.aol.com/jcolclefa/Waytoomuchboringcraptosayusually/ From CrochiereSteve <@t> aol.com Wed Dec 7 21:21:23 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Wed Dec 7 21:21:38 2005 Subject: [Histonet] ht position Message-ID: Hello, I will soon have an opening for a histo tech in my laboratory located in Springfield, MA. 20,000 surgical cases/year 800 IHC/month (automated) range from 400 to 900 slides/day great work environment, friendly, helpful coworkers HT(ASCP) with 2 years experience preferred Send resume to: Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew Street Springfield, MA 01104 From Rcartun <@t> harthosp.org Wed Dec 7 15:46:27 2005 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Dec 8 04:54:36 2005 Subject: [Histonet] CD123 and TCL1 Message-ID: Is anyone doing paraffin section immunohistochemistry for CD123 and TCL1? If so, would you mind identifying your antibody source? Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ----------------------------------------------- Confidentiality Notice This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential or proprietary information which is legally privileged. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please promptly contact the sender by reply e-mail and destroy all copies of the original message. From jkiernan <@t> uwo.ca Thu Dec 8 08:38:40 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Thu Dec 8 08:38:50 2005 Subject: [Histonet] Tap Water in Staining References: <2FABC6145388F24EBA8CBD6EC165BCCB02AFB9D6@mail1-schc.skaggs.net> Message-ID: <43984570.DBF3D8EF@uwo.ca> Distilled or deionized ("pure") water nearly always has a pH between 5 and 6 because of dissolved carbon dioxide (carbonic acid) from the air. To change the colour of haemalum-stained nuclei from red to blue the pH should be 7 or higher. Hard tap water is neutral or slightly alkaline, and the dissolved calcium bicarbonate can serve as a buffer. A drop of anything alkaline will bring the pH of pure water to a little over 7. I use limewater (saturated, that's about 0.3%, aqueous calcium hydroxide) because Ca(OH)2 is harmless and cheap. John Kiernan Anatomy, UWO London, Canada ------------------------------------ "Parker, Helayne" wrote: > > In using tap water for an H & E stain is there any documentation or > testing that needs to be done to confirm that the tap water is ok for > use. They have been using DI water here and there were complaints of > slides being too pink from the Pathologist. I put the tap water into > the regimen instead as I have always used tap for H & E staining -color > is now fine - no problems. But I was asked by the other tech is that > was acceptable. Having never been a supervisor I am not aware of any > documentation needed. Can anyone advise. > > Helayne Parker, HT (A.S.C.P.) > Histology Section Head > Skaggs Community Health Center > Branson, MO > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fawn <@t> cs.cmu.edu Thu Dec 8 09:03:55 2005 From: fawn <@t> cs.cmu.edu (Fawn Jones) Date: Thu Dec 8 09:02:35 2005 Subject: [Histonet] fixation in 70% alcohol Message-ID: <43984B5B.9080903@cs.cmu.edu> I just started working in a new lab and they use 70% Ethanol for fixation, however I do not know how long to keep the tissues in 70% to ensure adequate fixation. I am working with animal bones (mice, rats, sheep). Could anyone give me suggestions on fixation times? Thank you Fawn Jones From cfavara <@t> niaid.nih.gov Thu Dec 8 10:03:35 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Thu Dec 8 10:03:45 2005 Subject: [Histonet] coverslipping bubbles Message-ID: I was wondering if anyone has tried a wetting agent in the distilled water prior to coverslipping with aqueous mounting media? If so what did you use and at what concentration. I just thought it might help with my constant bubble problem on immunos. Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From llewllew <@t> shaw.ca Thu Dec 8 10:18:43 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Thu Dec 8 10:19:34 2005 Subject: [Histonet] fixation in 70% alcohol References: <43984B5B.9080903@cs.cmu.edu> Message-ID: <001901c5fc13$0f787d60$690a4246@yourlk4rlmsu> 70% ethanol is not considered a fixative. Ethanol fixes by dehydration, so at a minimum you should be using 95% ethanol, and preferably absolute. 70% can be used as a preservative after fixation. You have unfixed tissue if they are put into it immediately after harvest. Since it is not a fixative, the length of time is not really a factor. What is most likely happening is that you then dehydrate with higher concentrations of ethanol and the tissue is fixed at that time. Ethanol alone is considered to be a very poor fixative, although it may be necessary depending on your application. I suggest you either use a proper fixative such as a formalin variant, preferably overnight, or put your tissue directly into absolute ethanol for a few hours. Bryan Llewellyn ----- Original Message ----- From: "Fawn Jones" To: Sent: Thursday, December 08, 2005 7:03 AM Subject: [Histonet] fixation in 70% alcohol >I just started working in a new lab and they use 70% Ethanol for fixation, >however I do not know how long to keep the tissues in 70% to ensure >adequate fixation. I am working with animal bones (mice, rats, sheep). >Could anyone give me suggestions on fixation times? > Thank you > Fawn Jones > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From pruegg <@t> ihctech.net Thu Dec 8 10:28:26 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Dec 8 10:28:25 2005 Subject: [Histonet] coverslipping bubbles In-Reply-To: Message-ID: <200512081628.jB8GSDN1000304@chip.viawest.net> Good idea, I will have to try that with something like tween. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Favara, Cynthia (NIH/NIAID) [E] Sent: Thursday, December 08, 2005 9:04 AM To: Histonet Subject: [Histonet] coverslipping bubbles I was wondering if anyone has tried a wetting agent in the distilled water prior to coverslipping with aqueous mounting media? If so what did you use and at what concentration. I just thought it might help with my constant bubble problem on immunos. Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Thu Dec 8 10:32:25 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Thu Dec 8 10:32:28 2005 Subject: [Histonet] fixation in 70% alcohol In-Reply-To: <43984B5B.9080903@cs.cmu.edu> Message-ID: <200512081632.jB8GWCN1001554@chip.viawest.net> Fawn, Working in calcified bone, I often use cold absolute methanol to fix, in our experience it best preserves the enzymes such as alk.phos. For osteoblasts and TRAP for osteoclasts as well as the fluorochorme labels which can sometimes be soluable in aqueous fixatives. We do not use 70%, we use absolute at 4dc overnight, after that it can be brought to rt. I do this for undecalcified tissue embedded in glycol methacrylate. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fawn Jones Sent: Thursday, December 08, 2005 8:04 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] fixation in 70% alcohol I just started working in a new lab and they use 70% Ethanol for fixation, however I do not know how long to keep the tissues in 70% to ensure adequate fixation. I am working with animal bones (mice, rats, sheep). Could anyone give me suggestions on fixation times? Thank you Fawn Jones _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ian.montgomery <@t> bio.gla.ac.uk Thu Dec 8 10:35:04 2005 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Thu Dec 8 10:35:16 2005 Subject: Fwd: Re: [Histonet] Tap Water in Staining Message-ID: <7.0.0.16.2.20051208163319.0360beb0@bio.gla.ac.uk> Benefit of living in Glasgow, the tap water. You can drink it. bathe in it, make whisky with it and blue sections. >Distilled or deionized ("pure") water nearly always has a pH >between 5 and 6 because of dissolved carbon dioxide (carbonic >acid) from the air. To change the colour of haemalum-stained >nuclei from red to blue the pH should be 7 or higher. Hard tap >water is neutral or slightly alkaline, and the dissolved calcium >bicarbonate can serve as a buffer. A drop of anything alkaline >will bring the pH of pure water >to a little over 7. I use limewater (saturated, that's about >0.3%, aqueous calcium hydroxide) because Ca(OH)2 is harmless and >cheap. > >John Kiernan >Anatomy, UWO >London, Canada >------------------------------------ >"Parker, Helayne" wrote: > > > > In using tap water for an H & E stain is there any documentation or > > testing that needs to be done to confirm that the tap water is ok for > > use. They have been using DI water here and there were complaints of > > slides being too pink from the Pathologist. I put the tap water into > > the regimen instead as I have always used tap for H & E staining -color > > is now fine - no problems. But I was asked by the other tech is that > > was acceptable. Having never been a supervisor I am not aware of any > > documentation needed. Can anyone advise. > > > > Helayne Parker, HT (A.S.C.P.) > > Histology Section Head > > Skaggs Community Health Center > > Branson, MO > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk From katherine-walters <@t> uiowa.edu Thu Dec 8 10:52:24 2005 From: katherine-walters <@t> uiowa.edu (Walters, Katherine S) Date: Thu Dec 8 10:52:30 2005 Subject: [Histonet] Tap Water in Staining Message-ID: Is that why they call it Scott's tap water? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ian Montgomery Sent: Thursday, December 08, 2005 10:35 AM To: histonet@lists.utsouthwestern.edu Subject: Fwd: Re: [Histonet] Tap Water in Staining Benefit of living in Glasgow, the tap water. You can drink it. bathe in it, make whisky with it and blue sections. >Distilled or deionized ("pure") water nearly always has a pH >between 5 and 6 because of dissolved carbon dioxide (carbonic >acid) from the air. To change the colour of haemalum-stained >nuclei from red to blue the pH should be 7 or higher. Hard tap >water is neutral or slightly alkaline, and the dissolved calcium >bicarbonate can serve as a buffer. A drop of anything alkaline >will bring the pH of pure water >to a little over 7. I use limewater (saturated, that's about >0.3%, aqueous calcium hydroxide) because Ca(OH)2 is harmless and >cheap. > >John Kiernan >Anatomy, UWO >London, Canada >------------------------------------ >"Parker, Helayne" wrote: > > > > In using tap water for an H & E stain is there any documentation or > > testing that needs to be done to confirm that the tap water is ok for > > use. They have been using DI water here and there were complaints of > > slides being too pink from the Pathologist. I put the tap water into > > the regimen instead as I have always used tap for H & E staining -color > > is now fine - no problems. But I was asked by the other tech is that > > was acceptable. Having never been a supervisor I am not aware of any > > documentation needed. Can anyone advise. > > > > Helayne Parker, HT (A.S.C.P.) > > Histology Section Head > > Skaggs Community Health Center > > Branson, MO > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Dr. Ian Montgomery, Histotechnology, IBLS Support Services, Graham Kerr Building, Institute of Biomedical & Life Sciences, University of Glasgow, Glasgow, G12 8QQ. Tel: 0141 339 8855 Office: 4652 Lab: 6644. Pager: 07623 975451 e-mail: ian.montgomery@bio.gla.ac.uk _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From tracey.couse <@t> ibb.gatech.edu Thu Dec 8 11:02:53 2005 From: tracey.couse <@t> ibb.gatech.edu (Tracey Couse) Date: Thu Dec 8 11:03:08 2005 Subject: [Histonet] Trying to locate Carlos Miller Message-ID: <4398673D.9020201@ibb.gatech.edu> Hello All, I am trying to locate Carlos Miller. Carlos, please contact me. If anyone has his current contact information, please forward to me. Thanks! -- Tracey Couse Laboratory Coordinator/Histologist Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.2291 From tracey.couse <@t> ibb.gatech.edu Thu Dec 8 11:15:46 2005 From: tracey.couse <@t> ibb.gatech.edu (Tracey Couse) Date: Thu Dec 8 11:16:04 2005 Subject: [Histonet] Histologist Position Available Message-ID: <43986A42.7020601@ibb.gatech.edu> Hello everyone, I will soon be leaving my current position as Laboratory Coordinator/Histologist here at Georgia Institute of Technology. I am assisting my supervisor in locating a replacement. If anyone is interested in a technically challenging as well as socially and academically stimulating position with the Gatech/Emory Center for the Engineering of Living Tissues, please read on. The lucky candidate will have the opportunity to work with a variety of personnel (graduate students, staff, and faculty) as well as sample types. The newly renovated (1 year old), fully equipped core lab currently utilizes frozen, paraffin, and plastic techniques. Below is a link to Gatech?s Human Resources website with the official posting. I have also included the formal job description below. If anyone is interested, please contact myself or supervisor with any questions or visit the link below. Sincerely, Tracey -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.2291 Steve Woodard Laboratory Manager/Safety Officer Petit Institute for Bioengineering and Bioscience Georgia Institute of Technology Email: steve.woodard@ibb.gatech.edu https://ea.ohr.gatech.edu/FullDescription.asp?jobid=CEW4987&type=3&typeofjob=ext&jobtitle=LABORATORY%20COORDINATOR Laboratory Coordinator Education: A Bachelor's Degree, preferably in Chemistry or Biochemistry or any equivalent combination of education and experience. Experience: Three years of job related experience is required; preferred computer experience to include Windows, Office 2000/PC. Preference for experience in frozen paraffin, and plastic histotechniques. Duties: Supervise the histology laboratory and support faculty and graduate student researchers. Instruct, perform experiments (e.g. immunohistochemistry); requires lab safety, compliance, and microscopy knowledge; ability to exchange information with academic/research faculty and graduate information. Position requires strong interpersonal communications skills for advising, exchanging information, training; responsibility for promoting an atmosphere of acceptance and respect among employees and customers. From victoria.spoon <@t> bassett.org Thu Dec 8 11:15:45 2005 From: victoria.spoon <@t> bassett.org (Spoon, Victoria) Date: Thu Dec 8 11:17:01 2005 Subject: [Histonet] surgical pathology reporting turn around times Message-ID: <052739589974CC44A7770DA22157C804099C9D@ex3.bassett.org> Is there a standard for turn around time reporting for surgical pathology? (CAP) What do other institutions have for standards? Thanks in advance. NOTICE OF CONFIDENTIALITY This electronic message, including attachments, is for the sole use of the named recipient and may contain confidential or privileged information protected by New York State, and Federal regulations. Any unauthorized review, use, disclosure, copying or distribution is strictly prohibited. If you are not the intended recipient or have received this communication in error please contact the sender or email.security@bassett.org and destroy all copies of the original message. Thank you. From gcallis <@t> montana.edu Thu Dec 8 11:37:16 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 8 11:37:35 2005 Subject: Bone for Re: [Histonet] fixation in 70% alcohol In-Reply-To: <001901c5fc13$0f787d60$690a4246@yourlk4rlmsu> References: <43984B5B.9080903@cs.cmu.edu> <001901c5fc13$0f787d60$690a4246@yourlk4rlmsu> Message-ID: <6.0.0.22.1.20051208102622.01b2daf8@gemini.msu.montana.edu> 70% alcohol has been a historically popular fixative for undecalcified bone destined for methyl methacrylate i.e.plastic embedding to help preserve water soluble fluorescent tetracycline labeling. So here are two questions Are you labeling the bones with tetracycline? Are you embedding in PMMA or paraffin? In general, 70% alcohol is a poor fixative for bone!!! Morphology of cells and soft tissues is bad compared to NBF fixation. If you planning to decalcify and embed in paraffin, it would be advisable to fix in neutral buffered formalin. In this case, alcohol slows down and can actually stop ionization of calcium until the bone rehydrates. If you are planning to PMMA embed, you will have superior morphology is you fix in 1. Neutral buffered formalin for general staining without tetracyline 2. Alcoholic formalin which preserves the tetracyline label, but gives you better morphology than 70%. One recommendation, Anatech has an excellent alcoholic formalin fixative if you don't want to mess with making it up inhouse. After fixation with NBF, you can store bone in 70% alcohol. If your bones are whole, fix rat, mice at least a week or more, and change the fixative - this also goes for NBF or alcoholic formalin. Sheep - cut slabs or open bone and fix a longer, it may be up to two weeks for whole big bones and maybe even longer if the bones are huge - have done sheep and know how big their bones really are!!!! You fix according to size of bone, less time for small, more for large and there is NO established time as it depends on the bone cortical versus mostly trabecular, removal of soft tissues, size of bone, age of animal, etc. If you open a bone after it has been in fixative and it is still reddish inside, you have not fixed long enough. Good luck At 09:18 AM 12/8/2005, you wrote: >70% ethanol is not considered a fixative. Ethanol fixes by dehydration, >so at a minimum you should be using 95% ethanol, and preferably >absolute. 70% can be used as a preservative after fixation. You have >unfixed tissue if they are put into it immediately after harvest. Since >it is not a fixative, the length of time is not really a factor. What is >most likely happening is that you then dehydrate with higher >concentrations of ethanol and the tissue is fixed at that time. Ethanol >alone is considered to be a very poor fixative, although it may be >necessary depending on your application. I suggest you either use a >proper fixative such as a formalin variant, preferably overnight, or put >your tissue directly into absolute ethanol for a few hours. > >Bryan Llewellyn > > >----- Original Message ----- From: "Fawn Jones" >To: >Sent: Thursday, December 08, 2005 7:03 AM >Subject: [Histonet] fixation in 70% alcohol > > >>I just started working in a new lab and they use 70% Ethanol for >>fixation, however I do not know how long to keep the tissues in 70% to >>ensure adequate fixation. I am working with animal bones (mice, rats, >>sheep). Could anyone give me suggestions on fixation times? >>Thank you >>Fawn Jones Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Dorothy.L.Webb <@t> HealthPartners.Com Thu Dec 8 13:19:50 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Thu Dec 8 13:20:02 2005 Subject: [Histonet] microwriters Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C4FE@hpes1.HealthPartners.int> Does anyone have a system in use that labels cassettes and slides and is hooked into your computer system, I believe they are called microwriters? They have available to the system a bar code reader. We are looking to go that route in 2006, but wondering of some feedback from users!! Also, how long do you use your recycled xylene? Do you have outdates or expiration dates on the recycled xylene? An inspector for JACHO asked the question, and we were not able to give a good answer! ________________________________________ 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 JWEEMS <@t> sjha.org Thu Dec 8 13:31:06 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Dec 8 13:31:14 2005 Subject: [Histonet] microwriters Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0130524A@sjhaexc02.sjha.org> We have Triple G LIS - and just yesterday completed interface with Sakura cassette and slide printers. It is amazing - bar code and all. Barcoding is not active yet, but will be with next version of our LIS. As far as recycled xylene - I will that to a better expert. To my knowledge it never expires! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Thursday, December 08, 2005 2:20 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] microwriters Does anyone have a system in use that labels cassettes and slides and is hooked into your computer system, I believe they are called microwriters? They have available to the system a bar code reader. We are looking to go that route in 2006, but wondering of some feedback from users!! Also, how long do you use your recycled xylene? Do you have outdates or expiration dates on the recycled xylene? An inspector for JACHO asked the question, and we were not able to give a good answer! ________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From JPaulB42 <@t> aol.com Thu Dec 8 14:52:25 2005 From: JPaulB42 <@t> aol.com (JPaulB42@aol.com) Date: Thu Dec 8 14:52:37 2005 Subject: [Histonet] formalin waste disposal Message-ID: <238.338efba.30c9f709@aol.com> Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL From kconway <@t> cmcc.ca Thu Dec 8 15:13:30 2005 From: kconway <@t> cmcc.ca (Kevin Conway) Date: Thu Dec 8 15:14:08 2005 Subject: [Histonet] re: BrdU on cultured cells Message-ID: Hi Steve, In addition to permeabilizing (we used .15% Triton X-100, no reason the saponin wouldn't also work), I added a denaturation step (x 10 min in ?2 N HCl (aq)- I will double-check the [HCl] and let you know for sure if you don't have a protocol. Just helps to keep the histones away :) Also, I'm not sure what antibody you're using, but we always had great success with the G3G4 Mab from DSHB. Good luck, Kevin -------------------------------------- Date: Wed, 7 Dec 2005 12:31:10 -0800 (PST) From: Steven Coakley Subject: [Histonet] Brdu staining fixed cultured cells. Good afternoon everyone, I'm working witha scientist who is attempting to stain previously fixed cultured cells in "wells" and cover slips with anti-Brdu. He has fixed the cells 1st in methanol, air dryed, then in 10%formalin/PBS and again air dryed. Our anti-BrdU/DNAase works great on tissue but not on the fixed/air dryed cultured cells. We tried a 30 minute 0.1% Saponin step to help permeablize the cells but w/o success. Any ideas. Steve --------------------------------- Kevin Conway, PhD Assistant Professor Department of Anatomy Canadian Memorial Chiropractic College 416-482-2340 x.258 kconway@cmcc.ca From jkiernan <@t> uwo.ca Thu Dec 8 22:50:48 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Thu Dec 8 22:51:00 2005 Subject: [Histonet] Re: Mast cells in spinal cord References: <200512082234.jB8MYOhq000138@itsa.ucsf.edu> Message-ID: <43990D28.2BACCB0A@uwo.ca> Dear Dr Cun, You need to do some more homework! You probably found my name because of two publications: Campbell, D. J. and Kiernan, J. A. 1966. Mast cells in the central nervous system. Nature 210, 756-757. Kiernan, J. A. 1976. A comparative survey of the mast cells of the mammalian brain. Journal of Anatomy 121, 303-311. These relate to mast cells in the normal CNS (which occur in only a few species, if you exclude the ones in connective tissue around blood vessels). In central nervous tissue, in contact with neurons and glial cells, mast cells occur only in the habenular nuclei and nearby dorsomedial thalamus, in some Insectivora and "lower" primates (tree-shrew, loris). In the 1990s Zhuang, Silver and others published a series of papers about mast cells in the medial habenular nucleus of birds - the place where they are most abundant in insectivores and prosimians. To the best of my knowledge, mast cells do not occur in the normal spinal cord of any vertebrate animal. Is your research with the normal or diseased spinal cord? Mast cells are abundant in the dura mater. All research with mast cells begins with a reading of Hans Selye's book "The Mast Cells" (1963). Have you consulted this book? It contains the answers to most of your questions. The fixation and staining problems had all been nailed down by 1963. For most species you are wasting your time trying to find mast cells in frozen/cryostat sections. The tryptase and chymase activities do not exist in all species. Rodents differ from humans and dogs. Discharged mast cell granules may hang around and be stainable as extracellular dots (rat), or they may quickly dissolve and contribute to anaphlaxis (dog, guinea-pig, some people). All this was known 40 years ago. John Kiernan Anatomy & Cell Biology London, Canada. ______________________________________ clcun@itsa.ucsf.edu wrote: > > Dear Dr. Kiernan, > > I am trying to study mast cells in the CNS of mice and your name has popped > up on several occasions. I have a few questions regarding proper prepping/ > staining of mast cells and would greatly appreciate your expertise on this > matter. > > I would like to locate mast cells in the spinal cord using several stains: > toludine blue, chloroacetate esterase, cKIT (ACK45) immunohistochemistry, > tryptase and chymase enzyme histochemistry. > > 1. What fixative is suitable for these stains? We normally perfuse our > animals with 4% paraformaldehyde and cryoprotect our tissues in 30% > sucrose, but we?ve heard that this may not work for some of these staining > protocols (especially, cKIT immuno and tryptase/chymase enzyme histo, > which some have recommended using frozen tissues). We would like to > minimize usage of different fixation methods, so it would be great if we could > get away with using paraformaldehyde, but only if it will give us good > preservation of mast cells. > > 2. What is the best way to embed our tissues? After fixation, we usually > cryoprotect with sucrose and block our tissues in OCT, but some people > have suggested paraffin embedded tissues. Unfortunately, our lab is not > equipped to do paraffin embedding. Would we be able to get good sections > with our cryoprotected OCT blocked tissues? > > 3. Which orientation would be best to see the distribution of mast cells > in the > spinal cord, longitudinal or cross-section? > > 4. Of the above stains that I?ve mentioned, which one do you think is best, in > terms of specificity for mast cells, ease, etc. > > Any comments and suggestions you can give would be greatly appreciated. > Thank you in advance for your time. > > Sincerely, > > Christine Cun > Department of Neurosurgery > University of California, San Francisco From godsgirlnow <@t> msn.com Fri Dec 9 09:18:39 2005 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Fri Dec 9 09:18:50 2005 Subject: [Histonet] cap ihc info Message-ID: ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa From pruegg <@t> ihctech.net Fri Dec 9 10:30:07 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Fri Dec 9 10:30:35 2005 Subject: [Histonet] cap ihc info In-Reply-To: Message-ID: <200512091630.jB9GU6ch068463@pro12.abac.com> This is an ongoing issue in IHC, when dealing with limited tissue how can we afford to use up sections for negative controls on each block. I haven't been updated on this for awhile, just wondering what is happening in the real world, are you including a negative (isotype matched reagent in place of the primary antibody) for each block even when it is a small biopsy specimen. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roxanne Soto Sent: Friday, December 09, 2005 8:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cap ihc info ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Fri Dec 9 10:32:44 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Fri Dec 9 10:33:12 2005 Subject: [Histonet] formalin waste disposal In-Reply-To: <238.338efba.30c9f709@aol.com> Message-ID: <200512091632.jB9GWh7t070031@pro12.abac.com> We always had to pour the formalin into separate disposal containers, then dispose of the tissue as biohazard. The tissue was picked up by someone and the chemical waste by another, they did not combine services. It has been a while since I looked into this, perhaps someone has met that service need by now. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JPaulB42@aol.com Sent: Thursday, December 08, 2005 1:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formalin waste disposal Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Fri Dec 9 10:47:25 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Dec 9 10:47:38 2005 Subject: [Histonet] formalin waste disposal Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305268@sjhaexc02.sjha.org> We have biohazard disposal on site and dispose of small containers in biohazard bags to which Tissue-Tek neutralizer has been added. We pour formalin off large specimens and neutralize with Tissue-Tek neutralizer. This is approved by CA EPA so we think it has to be safe. Happy Friday! j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of patsy ruegg Sent: Friday, December 09, 2005 11:33 AM To: JPaulB42@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin waste disposal We always had to pour the formalin into separate disposal containers, then dispose of the tissue as biohazard. The tissue was picked up by someone and the chemical waste by another, they did not combine services. It has been a while since I looked into this, perhaps someone has met that service need by now. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JPaulB42@aol.com Sent: Thursday, December 08, 2005 1:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formalin waste disposal Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From JWEEMS <@t> sjha.org Fri Dec 9 10:58:38 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Dec 9 10:58:50 2005 Subject: [Histonet] formalin waste disposal Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0130526A@sjhaexc02.sjha.org> And I should have said - dispose of neutralized formalin down the drain. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Friday, December 09, 2005 11:47 AM To: patsy ruegg; JPaulB42@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin waste disposal We have biohazard disposal on site and dispose of small containers in biohazard bags to which Tissue-Tek neutralizer has been added. We pour formalin off large specimens and neutralize with Tissue-Tek neutralizer. This is approved by CA EPA so we think it has to be safe. Happy Friday! j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of patsy ruegg Sent: Friday, December 09, 2005 11:33 AM To: JPaulB42@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin waste disposal We always had to pour the formalin into separate disposal containers, then dispose of the tissue as biohazard. The tissue was picked up by someone and the chemical waste by another, they did not combine services. It has been a while since I looked into this, perhaps someone has met that service need by now. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JPaulB42@aol.com Sent: Thursday, December 08, 2005 1:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formalin waste disposal Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From sa.drew <@t> hosp.wisc.edu Fri Dec 9 11:03:57 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Dec 9 11:04:09 2005 Subject: [Histonet] cap ihc info Message-ID: We have recently changed how we deal with the negative control situation. We run one negative control slide per specimen block, using our most aggressive pretreatment protocol, per panel of antibodies. We also document that within our positive control tissue there are negative tissue elements. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of patsy ruegg Sent: Friday, December 09, 2005 10:30 AM To: 'Roxanne Soto'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cap ihc info This is an ongoing issue in IHC, when dealing with limited tissue how can we afford to use up sections for negative controls on each block. I haven't been updated on this for awhile, just wondering what is happening in the real world, are you including a negative (isotype matched reagent in place of the primary antibody) for each block even when it is a small biopsy specimen. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roxanne Soto Sent: Friday, December 09, 2005 8:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cap ihc info ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa _______________________________________________ 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 settembr <@t> umdnj.edu Fri Dec 9 09:43:05 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Dec 9 11:19:31 2005 Subject: [Histonet] cap ihc info Message-ID: Roxanne, Most of your wondering is correct, such as using a negative control per block. Your volume will increase. In reference to "built in negative control" you should have that specifically mentioned in your procedure manual. The very best thing to do is to ask your specific question(s) to CAP directly. They have a e-mail address that I have use several times and they respond promptly and precisely. Here's the address: accred@cap.org Good Luck, Dana Settembre University Hospital - UMDNJ Newark, New Jersey >>> Roxanne Soto 12/9/2005 10:18:39 AM >>> ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Fri Dec 9 11:44:39 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Dec 9 11:46:43 2005 Subject: [Histonet] cap ihc info Message-ID: All, Just an Observation...Has anyone else ever wondered why we cannot charge for IHC's on multiple blocks that are considered the "same" sample: eg. A1, A2, A3, A4, A5 on a case where 3 IHC's are ordered on each block, only 3 are billable although we are really performing 15. Yet, when it comes to negative controls, one must be run on each of the blocks because they are seperate and distinct. The end result of this madness is that, if you count the negative control run on each of the blocks, although 20 IHC's are run for the above case, only 3 are billable. I find it strange that when it comes to being able to bill for a case like this, the blocks are too much the "same" to bill for all technical work done, but,when it comes to running negatives (& lets face it, negatives are largely procedural and not closely monitored by many pathologists), it is vital that one be run on each block because they are not the same, although it is all just one, big, happy specimen. The IHC lab is lucky enough to take it in the shorts on cases like this. Ain't Life Grand, Glen Dawson Milwaukee, WI From cfavara <@t> niaid.nih.gov Fri Dec 9 11:49:47 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Fri Dec 9 11:49:57 2005 Subject: [Histonet] cap ihc info Message-ID: Because like so many other things the people that made the rule have no idea what is going on! Have a good weekend! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Dawson, Glen [mailto:GDawson@dynacaremilwaukee.com] Sent: Friday, December 09, 2005 10:45 AM To: Histonet Subject: RE: [Histonet] cap ihc info All, Just an Observation...Has anyone else ever wondered why we cannot charge for IHC's on multiple blocks that are considered the "same" sample: eg. A1, A2, A3, A4, A5 on a case where 3 IHC's are ordered on each block, only 3 are billable although we are really performing 15. Yet, when it comes to negative controls, one must be run on each of the blocks because they are seperate and distinct. The end result of this madness is that, if you count the negative control run on each of the blocks, although 20 IHC's are run for the above case, only 3 are billable. I find it strange that when it comes to being able to bill for a case like this, the blocks are too much the "same" to bill for all technical work done, but,when it comes to running negatives (& lets face it, negatives are largely procedural and not closely monitored by many pathologists), it is vital that one be run on each block because they are not the same, although it is all just one, big, happy specimen. The IHC lab is lucky enough to take it in the shorts on cases like this. Ain't Life Grand, Glen Dawson Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tracey <@t> labcareer.com Fri Dec 9 12:24:48 2005 From: Tracey <@t> labcareer.com (Tracey Dunn) Date: Fri Dec 9 12:20:42 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 12 Message-ID: <26A7728EF768DA478F45954252293C204496F3@matrix.HCCI.local> Can't you call me on a break or something ? Tracey -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Friday, December 09, 2005 1:11 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, Issue 12 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. microwriters (Webb, Dorothy L) 2. RE: microwriters (Weems, Joyce) 3. formalin waste disposal (JPaulB42@aol.com) 4. re: BrdU on cultured cells (Kevin Conway) 5. Re: Mast cells in spinal cord (John Kiernan) 6. cap ihc info (Roxanne Soto) 7. RE: cap ihc info (patsy ruegg) 8. RE: formalin waste disposal (patsy ruegg) 9. RE: formalin waste disposal (Weems, Joyce) 10. RE: formalin waste disposal (Weems, Joyce) 11. RE: cap ihc info (Drew Sally A.) 12. Re: cap ihc info (Dana Settembre) 13. RE: cap ihc info (Dawson, Glen) 14. RE: cap ihc info (Favara, Cynthia (NIH/NIAID) [E]) ---------------------------------------------------------------------- Message: 1 Date: Thu, 08 Dec 2005 13:19:50 -0600 From: "Webb, Dorothy L" Subject: [Histonet] microwriters To: Histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C4FE@hpes1.HealthPartners.int> Content-Type: text/plain; charset="us-ascii" Does anyone have a system in use that labels cassettes and slides and is hooked into your computer system, I believe they are called microwriters? They have available to the system a bar code reader. We are looking to go that route in 2006, but wondering of some feedback from users!! Also, how long do you use your recycled xylene? Do you have outdates or expiration dates on the recycled xylene? An inspector for JACHO asked the question, and we were not able to give a good answer! ________________________________________ 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. ------------------------------ Message: 2 Date: Thu, 8 Dec 2005 14:31:06 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] microwriters To: "Webb, Dorothy L" , Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0130524A@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" We have Triple G LIS - and just yesterday completed interface with Sakura cassette and slide printers. It is amazing - bar code and all. Barcoding is not active yet, but will be with next version of our LIS. As far as recycled xylene - I will that to a better expert. To my knowledge it never expires! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Webb, Dorothy L Sent: Thursday, December 08, 2005 2:20 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] microwriters Does anyone have a system in use that labels cassettes and slides and is hooked into your computer system, I believe they are called microwriters? They have available to the system a bar code reader. We are looking to go that route in 2006, but wondering of some feedback from users!! Also, how long do you use your recycled xylene? Do you have outdates or expiration dates on the recycled xylene? An inspector for JACHO asked the question, and we were not able to give a good answer! ________________________________________ 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 Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------ Message: 3 Date: Thu, 8 Dec 2005 15:52:25 EST From: JPaulB42@aol.com Subject: [Histonet] formalin waste disposal To: Histonet@lists.utsouthwestern.edu Message-ID: <238.338efba.30c9f709@aol.com> Content-Type: text/plain; charset="US-ASCII" Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL ------------------------------ Message: 4 Date: Thu, 08 Dec 2005 16:13:30 -0500 From: "Kevin Conway" Subject: [Histonet] re: BrdU on cultured cells To: Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Steve, In addition to permeabilizing (we used .15% Triton X-100, no reason the saponin wouldn't also work), I added a denaturation step (x 10 min in ?2 N HCl (aq)- I will double-check the [HCl] and let you know for sure if you don't have a protocol. Just helps to keep the histones away :) Also, I'm not sure what antibody you're using, but we always had great success with the G3G4 Mab from DSHB. Good luck, Kevin -------------------------------------- Date: Wed, 7 Dec 2005 12:31:10 -0800 (PST) From: Steven Coakley Subject: [Histonet] Brdu staining fixed cultured cells. Good afternoon everyone, I'm working witha scientist who is attempting to stain previously fixed cultured cells in "wells" and cover slips with anti-Brdu. He has fixed the cells 1st in methanol, air dryed, then in 10%formalin/PBS and again air dryed. Our anti-BrdU/DNAase works great on tissue but not on the fixed/air dryed cultured cells. We tried a 30 minute 0.1% Saponin step to help permeablize the cells but w/o success. Any ideas. Steve --------------------------------- Kevin Conway, PhD Assistant Professor Department of Anatomy Canadian Memorial Chiropractic College 416-482-2340 x.258 kconway@cmcc.ca ------------------------------ Message: 5 Date: Thu, 08 Dec 2005 23:50:48 -0500 From: John Kiernan Subject: [Histonet] Re: Mast cells in spinal cord To: clcun@itsa.ucsf.edu Cc: Histonet Message-ID: <43990D28.2BACCB0A@uwo.ca> Content-Type: text/plain; charset=iso-8859-1 Dear Dr Cun, You need to do some more homework! You probably found my name because of two publications: Campbell, D. J. and Kiernan, J. A. 1966. Mast cells in the central nervous system. Nature 210, 756-757. Kiernan, J. A. 1976. A comparative survey of the mast cells of the mammalian brain. Journal of Anatomy 121, 303-311. These relate to mast cells in the normal CNS (which occur in only a few species, if you exclude the ones in connective tissue around blood vessels). In central nervous tissue, in contact with neurons and glial cells, mast cells occur only in the habenular nuclei and nearby dorsomedial thalamus, in some Insectivora and "lower" primates (tree-shrew, loris). In the 1990s Zhuang, Silver and others published a series of papers about mast cells in the medial habenular nucleus of birds - the place where they are most abundant in insectivores and prosimians. To the best of my knowledge, mast cells do not occur in the normal spinal cord of any vertebrate animal. Is your research with the normal or diseased spinal cord? Mast cells are abundant in the dura mater. All research with mast cells begins with a reading of Hans Selye's book "The Mast Cells" (1963). Have you consulted this book? It contains the answers to most of your questions. The fixation and staining problems had all been nailed down by 1963. For most species you are wasting your time trying to find mast cells in frozen/cryostat sections. The tryptase and chymase activities do not exist in all species. Rodents differ from humans and dogs. Discharged mast cell granules may hang around and be stainable as extracellular dots (rat), or they may quickly dissolve and contribute to anaphlaxis (dog, guinea-pig, some people). All this was known 40 years ago. John Kiernan Anatomy & Cell Biology London, Canada. ______________________________________ clcun@itsa.ucsf.edu wrote: > > Dear Dr. Kiernan, > > I am trying to study mast cells in the CNS of mice and your name has > popped up on several occasions. I have a few questions regarding > proper prepping/ staining of mast cells and would greatly appreciate > your expertise on this matter. > > I would like to locate mast cells in the spinal cord using several stains: > toludine blue, chloroacetate esterase, cKIT (ACK45) > immunohistochemistry, tryptase and chymase enzyme histochemistry. > > 1. What fixative is suitable for these stains? We normally perfuse > our animals with 4% paraformaldehyde and cryoprotect our tissues in > 30% sucrose, but we've heard that this may not work for some of these > staining protocols (especially, cKIT immuno and tryptase/chymase > enzyme histo, which some have recommended using frozen tissues). We > would like to minimize usage of different fixation methods, so it > would be great if we could get away with using paraformaldehyde, but > only if it will give us good preservation of mast cells. > > 2. What is the best way to embed our tissues? After fixation, we > usually cryoprotect with sucrose and block our tissues in OCT, but > some people have suggested paraffin embedded tissues. Unfortunately, > our lab is not equipped to do paraffin embedding. Would we be able to > get good sections with our cryoprotected OCT blocked tissues? > > 3. Which orientation would be best to see the distribution of mast > cells in the spinal cord, longitudinal or cross-section? > > 4. Of the above stains that I've mentioned, which one do you think is > best, in terms of specificity for mast cells, ease, etc. > > Any comments and suggestions you can give would be greatly appreciated. > Thank you in advance for your time. > > Sincerely, > > Christine Cun > Department of Neurosurgery > University of California, San Francisco ------------------------------ Message: 6 Date: Fri, 09 Dec 2005 10:18:39 -0500 From: "Roxanne Soto" Subject: [Histonet] cap ihc info To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa ------------------------------ Message: 7 Date: Fri, 9 Dec 2005 09:30:07 -0700 From: "patsy ruegg" Subject: RE: [Histonet] cap ihc info To: "'Roxanne Soto'" , Message-ID: <200512091630.jB9GU6ch068463@pro12.abac.com> Content-Type: text/plain; charset="US-ASCII" This is an ongoing issue in IHC, when dealing with limited tissue how can we afford to use up sections for negative controls on each block. I haven't been updated on this for awhile, just wondering what is happening in the real world, are you including a negative (isotype matched reagent in place of the primary antibody) for each block even when it is a small biopsy specimen. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roxanne Soto Sent: Friday, December 09, 2005 8:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cap ihc info ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Fri, 9 Dec 2005 09:32:44 -0700 From: "patsy ruegg" Subject: RE: [Histonet] formalin waste disposal To: , Message-ID: <200512091632.jB9GWh7t070031@pro12.abac.com> Content-Type: text/plain; charset="US-ASCII" We always had to pour the formalin into separate disposal containers, then dispose of the tissue as biohazard. The tissue was picked up by someone and the chemical waste by another, they did not combine services. It has been a while since I looked into this, perhaps someone has met that service need by now. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JPaulB42@aol.com Sent: Thursday, December 08, 2005 1:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formalin waste disposal Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 9 Date: Fri, 9 Dec 2005 11:47:25 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] formalin waste disposal To: "patsy ruegg" , , Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01305268@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" We have biohazard disposal on site and dispose of small containers in biohazard bags to which Tissue-Tek neutralizer has been added. We pour formalin off large specimens and neutralize with Tissue-Tek neutralizer. This is approved by CA EPA so we think it has to be safe. Happy Friday! j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of patsy ruegg Sent: Friday, December 09, 2005 11:33 AM To: JPaulB42@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin waste disposal We always had to pour the formalin into separate disposal containers, then dispose of the tissue as biohazard. The tissue was picked up by someone and the chemical waste by another, they did not combine services. It has been a while since I looked into this, perhaps someone has met that service need by now. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JPaulB42@aol.com Sent: Thursday, December 08, 2005 1:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formalin waste disposal Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------ Message: 10 Date: Fri, 9 Dec 2005 11:58:38 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] formalin waste disposal To: "Weems, Joyce" , "patsy ruegg" , , Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0130526A@sjhaexc02.sjha.org> Content-Type: text/plain; charset="utf-8" And I should have said - dispose of neutralized formalin down the drain. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Weems, Joyce Sent: Friday, December 09, 2005 11:47 AM To: patsy ruegg; JPaulB42@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin waste disposal We have biohazard disposal on site and dispose of small containers in biohazard bags to which Tissue-Tek neutralizer has been added. We pour formalin off large specimens and neutralize with Tissue-Tek neutralizer. This is approved by CA EPA so we think it has to be safe. Happy Friday! j:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of patsy ruegg Sent: Friday, December 09, 2005 11:33 AM To: JPaulB42@aol.com; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] formalin waste disposal We always had to pour the formalin into separate disposal containers, then dispose of the tissue as biohazard. The tissue was picked up by someone and the chemical waste by another, they did not combine services. It has been a while since I looked into this, perhaps someone has met that service need by now. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JPaulB42@aol.com Sent: Thursday, December 08, 2005 1:52 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] formalin waste disposal Dear Histonetters: I am looking for information on any company that picks up tissue specimens with formalin still on the specimen for disposal. We were recently reclassified by the EPA and need to make immediate changes in formalin disposal, alcohol disposal and xylene disposal. Any help would be appreciated. Jim Bradford Orlando Regional Medical Center Orlando, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------ Message: 11 Date: Fri, 9 Dec 2005 11:03:57 -0600 From: "Drew Sally A." Subject: RE: [Histonet] cap ihc info To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" We have recently changed how we deal with the negative control situation. We run one negative control slide per specimen block, using our most aggressive pretreatment protocol, per panel of antibodies. We also document that within our positive control tissue there are negative tissue elements. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of patsy ruegg Sent: Friday, December 09, 2005 10:30 AM To: 'Roxanne Soto'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] cap ihc info This is an ongoing issue in IHC, when dealing with limited tissue how can we afford to use up sections for negative controls on each block. I haven't been updated on this for awhile, just wondering what is happening in the real world, are you including a negative (isotype matched reagent in place of the primary antibody) for each block even when it is a small biopsy specimen. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roxanne Soto Sent: Friday, December 09, 2005 8:19 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] cap ihc info ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa _______________________________________________ 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: Fri, 09 Dec 2005 10:43:05 -0500 From: Dana Settembre Subject: Re: [Histonet] cap ihc info To: histonet@lists.utsouthwestern.edu, godsgirlnow@msn.com Message-ID: Content-Type: text/plain; charset=US-ASCII Roxanne, Most of your wondering is correct, such as using a negative control per block. Your volume will increase. In reference to "built in negative control" you should have that specifically mentioned in your procedure manual. The very best thing to do is to ask your specific question(s) to CAP directly. They have a e-mail address that I have use several times and they respond promptly and precisely. Here's the address: accred@cap.org Good Luck, Dana Settembre University Hospital - UMDNJ Newark, New Jersey >>> Roxanne Soto 12/9/2005 10:18:39 AM >>> ______________________________________________________________ >I have a question about a particular IHC that I do on a daily basis. I do a PIN-4 cocktail >stain on prostate cores. This cocktail contains p63, CK903, and P504S. My question is, do I >need to perform a negative control? >Right now we are not CAP certified, but we are looking to become certified. CLIA only >requires one negative per case, instead of one negative per block. We deal with very tiny >cores and I am currently following the CLIA guidelines in this matter. However, we are >looking to automate and work on CAP accreditation and this negative factor would greatly >increase our volume and therefor would mean that we would need to get a stainer to >accommodate the volume. >I know that whenever I do a CK903, I do not do a negative because the tissue has built in >controls. Is this now null and void because of the racemase? >Please advice. >Thank you >Roxanne Soto HT(ASCP) >Tampa _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 13 Date: Fri, 9 Dec 2005 11:44:39 -0600 From: "Dawson, Glen" Subject: RE: [Histonet] cap ihc info To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" All, Just an Observation...Has anyone else ever wondered why we cannot charge for IHC's on multiple blocks that are considered the "same" sample: eg. A1, A2, A3, A4, A5 on a case where 3 IHC's are ordered on each block, only 3 are billable although we are really performing 15. Yet, when it comes to negative controls, one must be run on each of the blocks because they are seperate and distinct. The end result of this madness is that, if you count the negative control run on each of the blocks, although 20 IHC's are run for the above case, only 3 are billable. I find it strange that when it comes to being able to bill for a case like this, the blocks are too much the "same" to bill for all technical work done, but,when it comes to running negatives (& lets face it, negatives are largely procedural and not closely monitored by many pathologists), it is vital that one be run on each block because they are not the same, although it is all just one, big, happy specimen. The IHC lab is lucky enough to take it in the shorts on cases like this. Ain't Life Grand, Glen Dawson Milwaukee, WI ------------------------------ Message: 14 Date: Fri, 9 Dec 2005 12:49:47 -0500 From: "Favara, Cynthia \(NIH/NIAID\) [E]" Subject: RE: [Histonet] cap ihc info To: "Dawson, Glen" , "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" Because like so many other things the people that made the rule have no idea what is going on! Have a good weekend! c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Dawson, Glen [mailto:GDawson@dynacaremilwaukee.com] Sent: Friday, December 09, 2005 10:45 AM To: Histonet Subject: RE: [Histonet] cap ihc info All, Just an Observation...Has anyone else ever wondered why we cannot charge for IHC's on multiple blocks that are considered the "same" sample: eg. A1, A2, A3, A4, A5 on a case where 3 IHC's are ordered on each block, only 3 are billable although we are really performing 15. Yet, when it comes to negative controls, one must be run on each of the blocks because they are seperate and distinct. The end result of this madness is that, if you count the negative control run on each of the blocks, although 20 IHC's are run for the above case, only 3 are billable. I find it strange that when it comes to being able to bill for a case like this, the blocks are too much the "same" to bill for all technical work done, but,when it comes to running negatives (& lets face it, negatives are largely procedural and not closely monitored by many pathologists), it is vital that one be run on each block because they are not the same, although it is all just one, big, happy specimen. The IHC lab is lucky enough to take it in the shorts on cases like this. Ain't Life Grand, Glen Dawson Milwaukee, WI _______________________________________________ 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 25, Issue 12 **************************************** From ASamra <@t> mrl.ubc.ca Fri Dec 9 13:06:35 2005 From: ASamra <@t> mrl.ubc.ca (Amrit Samra) Date: Fri Dec 9 13:07:25 2005 Subject: [Histonet] un-subscribe Message-ID: <4399653B0200008D0000185E@mail.mrl.ubc.ca> From BGapinski <@t> pathgroup.com Fri Dec 9 13:59:36 2005 From: BGapinski <@t> pathgroup.com (Bruce Gapinski) Date: Fri Dec 9 13:59:51 2005 Subject: [Histonet] microwave processing tips? Message-ID: Hey-now Histonians! We are going to start microwave tissue processing next year. (The Milestone manual processor RH-1 or 2 I guess) I'd sure like to hear about any experiences you may have had, and any tips you'd care to share. I am an old dog and this sure is a new trick for me (no xylene!?!). I confess this was NOT my idea, and I worry that I'm going to end up with lots of tissues that didn't process well because the pathologist cut specimens too thick. I (mis)understand that there is no way to reprocess after microwave processing. Is that true? If so why? Respectfully, Bruce (I am not a rep) Gapinski HT (ASCP) --------------------------------------------------------------------- Important Notice: This e-mail is intended for the use of the person to whom it is addressed and may contain information that is privileged and confidential. If you are not the intended recipient, any disclosure, copying, distribution, or use of the contents of this message is strictly prohibited. If you have received this e-mail in error, please destroy this message and contact the Security Officer at PathGroup, Inc. immediately at 615-221-4431. From HornHV <@t> archildrens.org Fri Dec 9 15:23:26 2005 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Fri Dec 9 15:23:44 2005 Subject: [Histonet] cap ihc info Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BDEF1@EMAIL.archildrens.org> Your right Glen and the same rules apply to special stains. Hazel Horn, HT/HTL (ASCP) Histology Supervisor Arkansas Children's Hospital 800 Marshall Mail Slot 820 Little Rock, AR 72202 phone- 501.364.4240 fax- 501.364.3912 visit us on the web at: www.archildrens.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Friday, December 09, 2005 11:45 AM To: Histonet Subject: RE: [Histonet] cap ihc info All, Just an Observation...Has anyone else ever wondered why we cannot charge for IHC's on multiple blocks that are considered the "same" sample: eg. A1, A2, A3, A4, A5 on a case where 3 IHC's are ordered on each block, only 3 are billable although we are really performing 15. Yet, when it comes to negative controls, one must be run on each of the blocks because they are seperate and distinct. The end result of this madness is that, if you count the negative control run on each of the blocks, although 20 IHC's are run for the above case, only 3 are billable. I find it strange that when it comes to being able to bill for a case like this, the blocks are too much the "same" to bill for all technical work done, but,when it comes to running negatives (& lets face it, negatives are largely procedural and not closely monitored by many pathologists), it is vital that one be run on each block because they are not the same, although it is all just one, big, happy specimen. The IHC lab is lucky enough to take it in the shorts on cases like this. Ain't Life Grand, Glen Dawson Milwaukee, WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From Eric <@t> ategra.com Fri Dec 9 13:56:39 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Fri Dec 9 19:28:32 2005 Subject: [Histonet] Openings for HistoTechs Bench, Supv, Mgrs(NH, OH, CA, MI, NM, OK, RI, MA, PA, VA, NY, IL, CO, GA) Message-ID: Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Research Histo Tech positions as well. Here are some of my Hottest jobs: 1. Virginia Opening for a Histotech(bench) No weekends, No Call, Top dollar 2. Florida Openings for Histotech Manager No weekends, No call 3. Ohio Opening for Histotechs(bench, NightShift and Dayshift) No Weekends, No Call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 4. Ohio Opening for CytoTech No Weekends, No Call, Top Dollar 5. Ohio(Dayton Area) HistoTech Manager No Weekends, No call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 6. Colorado (Greater Denver area) Opening for Several Bench Positions(Night Shift) No weekends, No call, Top Dollar 7. Georgia Openings for Histotech(Temp to Perm) No Weekends, No Call, Top Dollar, Excellent Benefits if you are interested, please call me today at 1-800-466-9919 ext 223 8. Pennsylvania(Philadelphia area) Openings for HistoTech(Bench) No Weekends, No Call, Top Dollar if you are interested, please call me today at 1-800-466-9919 ext 223 9. California( Southern) HistoTech Supervisor(2nd shift) Openings for Histotechs(Bench,2nd shift) Great Location, No weekends, No call, Top Dollar 10. California(Southern) Openings for HistoTechs(Bench) Great Location, No weekends, No call, Top Dollar 11. Pennsylvania(Multiple jobs in greater Pittsburgh area) HistoTech opening(Bench) if you are interested, please call me today at 1-800-466-9919 ext 223 12. Illinois(Multiple jobs in Greater Illinois area) Opening for Histotech(Bench) 13. Michigan(Multiple jobs in Greater Michigan area) Openings for Histotechs(Bench) if you are interested, please call me today at 1-800-466-9919 ext 223 14. Massachusetts (Several Opportunities in the Boston Area) Part time Histo Tech(Permanent) No weekends, No call 15. Virginia(Western Virginia) Opening for Bench Histotech No weekends, No call if you are interested, please call me today at 1-800-466-9919 ext 223 16. Florida(Greater Florida area) Openings for Bench Histotech No weekends, No call The clients are currently interviewing candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu ---------------------------------------------------------------- ---------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------------- From dpconsult <@t> earthlink.net Sat Dec 10 18:42:33 2005 From: dpconsult <@t> earthlink.net (Dick Paulson [Source Medical Products]) Date: Sat Dec 10 18:43:04 2005 Subject: [Histonet] microwave processing tips? Message-ID: Bruce, There are 2 books you should read to get a firm grasp on the state of microwave technology in the laboratory. ?Microwaves for the Art of Microscopy? by L.P. Kok and Mathilde E. Boon Coulomb Press Leyden, 2003. ?Microwave Technology for Light Microscopy and Ultrastructural Studies? by Anthony S-Y Leong, M.D. I submit to you that ?Microwaves for the Art of Microscopy? should be included in the Reference Library of any lab investigating or using microwave technology. It is a unique collaboration between a physicist and a pathologist. I have included below the full Table of Contents for both books to peak your interest. You can get these books at: Milestone Medical Products Phone (866) 995-5300 $115.00 - ?Microwaves for the Art of Microscopy? $ 55.00 - ?Microwave Technology for Light Microscopy and Ultrastructural Studies? Dick Paulson Source Medical Products dpconsult@earthlink.net ------ Microwaves for the Art of Microscopy by L. P. Kok, Biomedical Engineering, University of Groningen Mathilde E. Boon, Leiden Cytology and Pathology Laboratory, Leiden 370 Pages - 900 references Mathilde E. Boon is the senior pathologist at, and the director of, the Leiden Cytology and Pathology Laboratory. She was trained in the late sixties as a cytologist at Ann Arbor (Michigan), then as a pathologist at Leiden, The Netherlands. She has headed subsequently the Departments of Cytology at the Leiden University and at Delft. She has authored more than three hundred scientific papers on cytology, pathology, laboratory techniques, and microwaves. L. P. Kok is a theoretical physicist and a full professor in Theoretical Biomedical Engineering at the University of Groningen, The Netherlands. In that city he also obtained his Ph.D. degree, in 1969. He worked for extended periods in Italy, Israel, the US, and the USSR. He has authored more than two hundred and fifty publications, starting off in the field of quantum scattering theory of few-body systems. Books he (co)authored address subjects in quantum mechanics, stereology, and the use of microwaves in laboratory technique. Together they started to publish on the use of microwaves in the pathology laboratory some twenty years ago. This work led to four books on this subject, the most recent one being "Microwaves for the Art of Microscopy", appearing in 2003. With their background in physics and pathology they did more innovative work. In the nineties, they introduced neural-network scanning technology in cervical screening. This application is still very successful, until now some 700,000 patients were screened using this technique. In 1985, the pathologist Boon introduced microwaving and coagulant fixation in her laboratory. These unconventional methods are, in a fine-tuned manner, still used for all the incoming diagnostic material. They are the basics for the workshop at the NSH Convention. There, Dr Kok will give a presentation concerning the physics background of the use of microwaves in pathology. Dr Boon will present the practical side of the application of microwaves for fixation, (automated) histoprocessing, and other techniques. In addition, she will conduct the discussion of artifacts with clinical examples. Table of Contents Preface 1. Microwaves for beginners ? Introduction to light and heat ? Heat: convection, conduction, and radiation ? The electromagnetic spectrum ? The dual nature of microwaves ? New perspectives to the microscopist 2. Microwaves are electromagnetic waves ? Microwaves? For microscopy? ? Electromagnetic waves ? Electromagnetic spectrum ? Electromagnetic radiation ? Microwaves ? Terminology: (ir)radiation versus exposure and emission 3. Microwaves and society ? Origins of radar ? Applications of microwaves ? From radar, via cooking, to the laboratory ? Hazards of microwaves ? The 1982 ANSI standard and the 1988 IRPA standard ? The body as an antenna ? Hazards of microwave ovens 4. Interaction between microwaves and matter ? Introduction ? The optics of microwaves: reflection and refraction ? Concentration effects in the optics of microwaves ? Molecular picture of dielectric media in electric fields ? Macroscopic description of microwaves in media ? Dielectric data and depth of penetration of microwaves ? Depth of penetration into slightly conducting media ? Penetration into highly conducting media ? Antenna properties of exposed objects ? Standing waves and standing-wave phenomena ? ?The microwave effect? and ?The temperature effect?. What is temperature? ? Preferential stimulation; microwave stimulated diffusion ? Microwave-assisted chemistry and chemical-reaction rates ? Defrosting: runaway heating ? Conclusion: lessons from the food industry 5. From Domestic microwave ovens to laboratory microwave processors ? Introduction ? Operation and precautions ? The microwave oven as a resonant cavity. Hot spots ? Power-level control of microwave ovens ? Power and efficiency of microwave ovens ? Shielding and detection of unwanted microwaves ? Microwave-transparent materials, susceptors, and active packaging ? Domestic microwave oven or laboratory processor? A legal question 6. Microwave processors, temperature control, and robustness of procedures ? Temperature measurement in microwave environment ? Historical ways of measuring temperature in microwave cavities ? Infrared thermometry ? Variation of temperature of exposed objects ? A uniform microwave heating pattern is a fiction ? General characteristics of microwave processors ? The rise, usage, and fall of the water load 7. Transport properties ? Introduction ? Viscosity of liquids ? Diffusion in liquids ? Heat capacity ? Thermal conductivity and transport of heat ? Electric conductivity ? External versus internal heating: the beef experiment ? The power of vacuum and low-pressure histology ? High-altitude microscopy and superheating 8. History of the use of microwaves for microscopy ? Introduction ? The first two decades of microwave publications ? Microwave publications ordered by subject ? History of microwave fixation and stabilization ? History of microwave histoprocessing ? History of microwave-stimulated staining ? History of microwaves and immunostaining ? History of microwaves and enzyme incubations ? History of microwaves and ELISA ? History of microwaves and botany ? History of microwaves in entomology ? History of microwaves and antigen retrieval ? History of microwaves and electron microscopy ? Miscellany 9. Grossing The gross room ? Grossing large operation specimens ? Microwave hardening of large operation specimen ? Hardening of human brains ? Hardening of small human embryos ? Grossing small operation specimens ? Grossing biopsies ? Surgical margins of microwave-treated operation specimen ? From the grossing room to the pathology report ? In situ stabilization brains in live rats by L?berg and Torvik 10. Microwave-Stimulated fixation with formalin ? Introduction ? Formaldehyde fixation and microwave exposure ? DNA and RNA in formalin-fixed tissue Formaldehyde-glutaraldehyde mixture for histochemistry ? Microwave-stimulated fixation for in situ hybridization ? Microwave-stimulated formalin fixation of perfused laboratory animals ? Microwave exposure with low formalin concentrations for the detection of dopamine 11. Microwave stimulation of coagulant formalin-free fixatives ? Coagulant fixatives ? Ethyl alcohol ? polyethylene glycol fixatives: Kryofix and BoonFix ? Selecting a fixative for DNA and RNA preservation ? Experiences with Kryofix ? Comparing microwave coagulation with microwave-stimulated Kryofix fixation: the egg experiment ? Microwave-stimulated Kryofix fixation ? Flow of specimen in the Kryofix laboratory 12. Microwave stabilization of fresh tissue by microwave heating ? Introduction ? Microwave stabilization without postfixation ? Hybrid method: microwave stabilization with postfixation ? Microwave stabilization of lungs ? Perfusion of laboratory animals ? Combining microwave stabilization and paraffin embedding ? Combining microwave stabilization and frozen sectioning ? Comparison of different microwave stabilization and fixation methods ? Microwave stabilization of tissue and cells for immunocytology ? Microwave stabilization of myocard to delineate necrosis 13. Microwaves for paraffin histoprocessing ? Introduction ? The fixative factor ? Formalin as fixative for paraffin sections ? Coagulant fixatives for paraffin sections Reagents used for microwave histoprocessing ? Dehydration fluid ? The intermedium ? Paraffin wax Differences between microwave and conventional histoprocessing ? Vacuum histoprocessing in the Milestone LAVIS microwave processor ? Exploiting low boiling temperatures under vacuum ? Troubleshooting in the first-generation microwave vacuum histoprocessors Introduction of a vacuum-drying step in 1997 ? The Milestone MicroMED URM vacuum Histoprocessor ? The Milestone RHS-1 vacuum processor ? Vacuum histoprocessing using JFC solution in the Milestone processors ? Vacuum histoprocessing using ethyl alcohol/isopropanol solution in the Milestone processors ? Microwave histoprocessing in the Pelco processors ? The Tissue-Tek continuous rapid processor of Sakura ? The story of microwave histoprocessing using domestic ovens ? The story of the H2500 and the H2800 microwave processor ? The future of microwave histoprocessing and automation 14. Combining microwave and freeze techniques ? Introduction ? Combining cryostat-sectioning and microwave-stimulated fixation in surgical pathology ? Combining cryostat-sectioning and microwave-stimulated fixation for immunopathology ? Microwave-stimulated fixations of cryosections for histochemistry ? Microwave cryostat procedure for large specimen 15. Microwaves and staining ? Introduction ? Theory and practice of staining in the microwave processor ? Microwave Warthin-Starry method ? Microwave Southgate mucicarmine method ? Microwave Alcian-blue method ? Microwave Fontana-Masson method ? Microwave Grimelius silver method ? Microwave Grocott methenamine silver-nitrate method ? Microwave Jones periodic acid methenamine silver method ? Microwave Jones-Marres method ? Microwave Perls iron stain ? Microwave rhodamine method ? Microwave Bosma-Steiner method ? Microwave Elastica Van Gieson method ? Microwave Azan staining method ? Microwave Romanowsky-Giemsa method for plastic-embedding bone-marrow sections ? Microwave PAS staining method for glycol methacrylate sections ? Microwave Hanker-Giammara silver staining method of cytochemical-reation products ? Microwave AgNOR method ? Microwave staining method for nerve and muscle biopsies ? Microwave Nissl method ? Microwave Kl?ver-Barrera method ? Microwave Bodian silver method ? Microwave King silver method ? Microwave Rio-Hortega method ? Microwave H?ggqvist method for human brain 16. Microwaves for decalcification ? Introduction ? Decalcification in the Milestone processors ? Decalcification in the Pelco 3440 microwave processor ? Microwave-stimulated decalcification using nitric acid ? Microwave-stimulated decalcification using formic acid ? Microwave exposure for the study of bone canaliculi 17. Microwave exposure in immunostaining ? Introduction ? Temperature controlled microwave-enhanced incubations for immunostaining in the Milestone processors ? Immunostaining in the Pelco microwave processors 18. Microwaves and cell fixation for TEM and SEM ? Introduction ? Microwave-stimulated fixation of single cells ? Microwave stabilization of single cells ? Microwave-stimulated aldehyde fixation of cell monolayers ? Microwave-stimulated aldehyde fixation of cell pellets ? Comparison of microwave stabilization and fixation ? Cytochemistry and immunocytochemistry at EM level in microwaved cells ? Microwaving for immunoelectron microscopy of cultured cells ? Microwave stabilization for scanning electron microscopy ? Microwave-stimulated fixation of cells for scanning electron microscopy ? Microwave method for electron microscopy in aspiration cytology 19. Microwaves and tissue fixation for TEM and SEM ? Introduction ? Working with the Milestone REM Processor for EM ? Microwave-stimulated fixation for TEM using the Pelco 3440 ? Microwave procedure for TEM fixation with membrane protection ? Microwave-enhanced fixation for TEM of the parathyroid ? Microwave procedure for TEM fixation of brain slices ? Microwave stabilization for SEM ? Microwave-stimulated fixation for SEM ? Microwaving for electron microscopy in Boston 20. Microwave exposure in immunoelectron microscopy ? Introduction ? Temperature control in the immunogold technique ? Immunogold labeling of antigens on mesothelial cells using microwave exposure ? Lysosmal localization of acid phosphatase in prostate epithelium using microwave exposure ? Microwaving and preembedding in immunoelectron microscopy 21. Microwaves and embryology ? Introduction ? Combining microwave-stimulated formalin fixation and frozen sectioning ? Combining microwave stabilization and frozen sectioning ? Combining microwave stabilization and paraffin embedding ? Microwave-stimulated formalin fixation of embryos in fertilized fish eggs ? Microwave treatment of chimeric mouse embryos ? Microwave-stimulated Kryofix fixation of cuticulized embryos 22. Microwave exposure and epoxy-resin embedding for TEM ? Introduction ? Working with the Milestone REM Processor for EM embedding ? Working with the Pelco processors for EM embedding ? Microwave embedding for TEM using the Pelco processors ? Temperature control in the BioRad H2500 in the dehydrations steps ? Microwave embedding for TEM 23. Microwaves and botany ? Introduction ? The AgNOR story in botany ? Microwave-enhanced immunostainings in botanic sections ? Microwave exposure for rapid killing and fixing of plant tissue ? Microwave-stimulated glutaraldehyde and osmium tetroxide fixation of plant tissue ? Microwave-stimulated glutaraldehyde fixation of yeast cells ? Microwave-stimulated fixation for the preservation of soluble antigens ? Microwave-stimulated staining of plant virus inclusion bodies ? Microwave-stimulated staining of plastic sections ? Processing plant tissues for ultrastructural studies ? Processing plant tissue in the Pelco 3450 Processor 24. Microwaves and entomology ? Introduction ? Microwave exposure to extrude hidden cuticular parts for SEM 25. High-temperature heat treatment for immunhistochemistry and molecular techniques ? Introduction ? Chemical composition and pH of the retrieval solution ? Heat treatment in domestic microwave ovens ? The ER story ? The MiB-1 story ? The p53 story ? Working the Milestone T/T Mega for high-temperature heat treatment (retrieval) ? Microwave superheating in the Milestone Pressure Reactor ? Combining microwave heat treatment with signal amplification ? Multiple immunostainings: microwave heat treatment for blocking of cross-reactivity, false-positive staining and antigen retrieval ? Multiple immunoenzyme staining of paraffin tissue sections ? DNA and RNA retrieval in formalin-fixed tissue and sections by microwave heat treatment ? Microwave heating for RNA-ISH in formalin-fixed tissues ? Antigen retrieval for immunoelectron microscopy ? Antigen retrieval in plastic sections 26. Miscellany ? Introduction ? Mordanting in the microwave oven ? Bleaching melanin in the microwave oven ? Drying and attaching sections using the microwave oven ? Treating plastic and glass equipment (sterilizing?) ? Preparation of bacteriological-culture media ? Microwave-stimulated Ce-Pb conversion ? Cryoquenching and microwaving ? Treating cryosections in the microwave oven ? Microwave digestion for forensic pathology ? Destaining slides in the microwave oven ? Deparaffinizing sections in the microwave oven ? Various ? Microwaves and wine The formalin-free microwave laboratory ---------------------------------------------------------------------------- ------------------- Microwave Technology for Light Microscopy and Ultrastructural Studies by Anthony S-Y Leong, M.D. Dr. Anthony S-Y Leong is one of the world's best-known pioneers in the use of microwave technology for histopathology. This book is a compendium of the knowledge he has gathered in 20 years of activity. It describes the variety of purposes to which microwaves can be applied, providing detailed information on the techniques and procedures that he and his colleagues have developed. This book is an excellent primer for any pathologist or histologist who is considering the transition to microwaves. It is also good reading for anyone who has already made that transition and now wants to get the most out of their new technology. Table of Contents: 1. Introduction ? Effects of low dose microwave and radio frequency radiation on mammalian tissues ? Microwaves - physical properties ? Background to applications in histopathology 2. Tissue Fixation ? Microwave fixation of large specimens ? Microwave fixation of tisue blocks ? Motable differences in the properties of microwave -fixed and formalin-fixed sections ? Incorporation of microwave -fixation with conventional tissue processing ? Formaldehyde toxicity ? Principles of tissue fixation ? A classification of Microwave fixation ? Microwave -stimulated aqueous formaldehyde fixation ? Microwave fixation and microwave stimulated fixation of whole organs ? Microwaves for fixation in neurochemical analysis ? Optimal temperatures for Microwave fixation and Microwave stimulated fixation ? Protocol for Microwave fixation for light microscopy ? Are organisms destroyed during Microwave fixation? ? Microwave fixation for electron microscopy ? Ultrastructural enzyme and antigen preservation in Microwave -fixed tissues ? Protocol for Microwave stimulated fixation of specimens for electron microscopy ? Microwave stimulated fixation for accelerated processing of fine needle biopsy specimens for ultrastructural diagnosis ? Protocol for rapid Microwave stimulated fixation and processing of fine needle aspiration biopsies ? Microwave fixation for cytological preparations 3. Microwave Accelerated Demineralisation 4. Cryostat Sections 5. Histochemical and Immunohistological Staining ? Light microscopy ? Microwave -stimulated staining of reticulin in plastic sections with ammoniacal silver nitrate (Leong and Pulbrook, 1989) ? A new Microwave -stimulated stain for melanocytic lesions (Leong & Gillham, 1989) ? Electron microscopy ? Immunohistological staining ? General comments 6. Antigen Retrieval ? Introduction ? Microwave antigen retrieval ? Antigen retrieval for cytological preparations ? Protocol for immunolabelling of cytological preparations ? Antigen retrieval for electron microscopy ? Proposed mechanism of formalin fixation and antigen retrieval 7. Applications in Molecular Analyses 8. Rapid Tissue Processing ? Conventional Tissue Processing ? Microwave -Stimulated Tissue Processing ? Impact of Microwave -Stimulated Tissue Processing ? Protocol for Microwave -Stimulated Tissue Processing ? Rapid Processing for Electron Microscopy 9. Chemical and Industrial Applications 10. Conclusions From darkdaym <@t> mindspring.com Sun Dec 11 09:59:00 2005 From: darkdaym <@t> mindspring.com (Mark Ray) Date: Sun Dec 11 09:58:41 2005 Subject: [Histonet] Holiday Gift Message-ID: <439C4CC4.3090801@mindspring.com> Still searching for a suitable holiday gift for your Histopal or Histosignificantother? John Kiernan's _Histological and Histochemical Techniques: Theory and Practice_ is still available from most bookdealers. Many of us know it as an important basic text, and if you don't, you should. It's highly recommended by all authorities, so get it now before it goes out of print. I suggest that you refer to it only as ISBN 0750649364 when ordering. That will insure that you get the current Third Edition. Mark From bamur <@t> alaska.net Sun Dec 11 18:17:09 2005 From: bamur <@t> alaska.net (Barbara Murray) Date: Sun Dec 11 18:17:17 2005 Subject: [Histonet] Histologist position Message-ID: Greetings from Anchorage, Alaska, The Alaska Native Tribal Health Consortium has an opening for a fulltime Histologist. Please visit our website @ http://anthc.org . You may also call the Human Resources Dept. for more details. The phone number is: 907-729-1301. Barbara A. Murray, HT. (ASCP) Pathology Dept. The Alaska Native Medical Center Anchorage, Alaska 99508 (907-729-1804 From brucea <@t> unimelb.edu.au Sun Dec 11 22:27:33 2005 From: brucea <@t> unimelb.edu.au (Bruce Abaloz) Date: Sun Dec 11 22:27:56 2005 Subject: [Histonet] fixation in 70% alcohol In-Reply-To: <43984B5B.9080903@cs.cmu.edu> References: <43984B5B.9080903@cs.cmu.edu> Message-ID: Hi fawn, you will no doubt receive replies that 70%ETOH is not a fixative....we have been using it for over a decade now & it does a beautiful job. I discussed this @ a workshop in Toronto last year with Histology colleagues/Workshop on 'Fixation'. After initial 'gasps' (how could you use that) that it was my preference/choice of fixatives & gave us excellent results, another seven Histologists in the workshop 'admitted that they also use it in their labs......of course we still use 10% NBF, Bouins etc. when asked to fix in them specifically, but 90% of our tissues are fixed in 70% ETOH .......as this is a Research Lab. we have time on our side - minimum time for skin lesions etc. 24hrs. but today we also have the luxury of infiltrating under vacuum.... Cheers, Bruce in OZ -- BRUCE ABALOZ PH:61383446282 HISTOLOGIST FAX:61383447909 DEPT.of ZOOLOGY EMAIL: brucea@unimelb.edu.au THE UNIVERSITY Of MELBOURNE. VICTORIA. AUSTRALIA 3010 Nobody Can Make You Feel Inferior Without YOUR Permission - Eleanor Roosevelt DANCE LIKE NO-ONE'S WATCHING Q: What's a specimen? A: An Italian astronaut. ******************************************************************************** This email and any attachments may contain personal information or information that is otherwise privileged, confidential or otherwise protected from disclosure/subject of copyright. Any use, disclosure or copying of any part of it is prohibited. The University does not warrant that this email or any attachments are free from viruses or defects. Please check any attachments for viruses and defects before opening them. If this email is received in error please delete it and notify us by return email. The University does not necessarily share or endorse the views expressed in this email. From moolovescow <@t> yahoo.com.sg Sun Dec 11 23:39:25 2005 From: moolovescow <@t> yahoo.com.sg (Phua Shirley) Date: Sun Dec 11 23:39:36 2005 Subject: [Histonet] Pig's eyeballs Message-ID: <20051212053925.59109.qmail@web35210.mail.mud.yahoo.com> Hi All My lab will be working with pig's eyeballs soon and I would like to find out from anyone out there who have had experiences on it. Are there any special treatments dealing with the eyeballs, in terms of fixation, processing, sectioning and stainings? Your reply will be truely helpful/needful. Thanks, Shirley Phua Histopathology Laboratory Centre for Forensic Medicine Health Sciences Authority Singapore --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 1GB free storage! From vanann702 <@t> skmc.gov.ae Mon Dec 12 00:02:07 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sun Dec 11 23:59:11 2005 Subject: [Histonet] Pig's eyeballs Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D708@SKMCEMAIL.skmc.gov.ae> Yes indeed - my protocol was posted on this site a few years ago - you will find it in the archives. I did most of my 'trial runs' on pig eyes before getting it right. Feel free to contact me once you have tried it - if you hit any snags Good luck and have patience. Annieinarabia -----Original Message----- From: Phua Shirley [mailto:moolovescow@yahoo.com.sg] Sent: Monday, December 12, 2005 9:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pig's eyeballs Hi All My lab will be working with pig's eyeballs soon and I would like to find out from anyone out there who have had experiences on it. Are there any special treatments dealing with the eyeballs, in terms of fixation, processing, sectioning and stainings? Your reply will be truely helpful/needful. Thanks, Shirley Phua Histopathology Laboratory Centre for Forensic Medicine Health Sciences Authority Singapore --------------------------------- Do you Yahoo!? New and Improved Yahoo! Mail - 1GB free storage! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PatPatterson <@t> mhd.com Mon Dec 12 08:47:46 2005 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Mon Dec 12 08:47:58 2005 Subject: [Histonet] (no subject) Message-ID: <293C7C19EFF7D611AE1A0002A53F8114164ABF9B@omega.mhd.com> Hello to all HistoNet folks. Our lab is having inconsistent occurrences of "crispy" biopsies - especially Livers. They feel crispy during embedding and sectioning and the sections appear to have cracking - separating the tissue in unnatural directions. These are fixed in Carson Millonig formalin and then processed with 10% NBF, 70% alc, 95% alc, Abs alc, xylene to paraffin - total process about 2.5 hours. They are held in the 10% NBF at the beginning of the processing schedule and there is no heat on the retort until the paraffin steps. The biopsies are placed in "micro-cassettes" with no wrapping (these cassettes are not the micro-screen cassettes ones with metal like screening). We'd appreciate any ideas you might have - especially since our liver transplant service is increasing in volume as I type! Thanks! Pat Patterson Manager, AP Methodist Dallas Medical Center (214) 947-3538 *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From Tony.Crick <@t> rli.mbht.nhs.uk Mon Dec 12 08:51:35 2005 From: Tony.Crick <@t> rli.mbht.nhs.uk (Crick Tony) Date: Mon Dec 12 08:55:21 2005 Subject: [Histonet] (no subject) Message-ID: It sounds like processing issue being carried out too fast. Tony -----Original Message----- From: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: 12 December 2005 14:48 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] (no subject) Hello to all HistoNet folks. Our lab is having inconsistent occurrences of "crispy" biopsies - especially Livers. They feel crispy during embedding and sectioning and the sections appear to have cracking - separating the tissue in unnatural directions. These are fixed in Carson Millonig formalin and then processed with 10% NBF, 70% alc, 95% alc, Abs alc, xylene to paraffin - total process about 2.5 hours. They are held in the 10% NBF at the beginning of the processing schedule and there is no heat on the retort until the paraffin steps. The biopsies are placed in "micro-cassettes" with no wrapping (these cassettes are not the micro-screen cassettes ones with metal like screening). We'd appreciate any ideas you might have - especially since our liver transplant service is increasing in volume as I type! Thanks! Pat Patterson Manager, AP Methodist Dallas Medical Center (214) 947-3538 *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From Kemlo.Rogerson <@t> elht.nhs.uk Mon Dec 12 09:00:44 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Mon Dec 12 09:00:54 2005 Subject: [Histonet] (no subject) Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB237F3@elht-exch1.xelht.nhs.uk> But it then would not be 'crispy'. I have suggested poor fixation which would mean you are relying on the fixative properties of ethanol; which causes tissue to go hard. Rapidly processed tissue usually result in poor processing which is 'soft' not 'hard', IMHO. Good preliminary fixative will prevent a lot of problems subsequently. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Crick Tony (Morecambe Bay Hospitals NHS Trust) Sent: 12 December 2005 14:52 To: 'Patterson, Pat'; 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] (no subject) It sounds like processing issue being carried out too fast. Tony -----Original Message----- From: Patterson, Pat [mailto:PatPatterson@mhd.com] Sent: 12 December 2005 14:48 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] (no subject) Hello to all HistoNet folks. Our lab is having inconsistent occurrences of "crispy" biopsies - especially Livers. They feel crispy during embedding and sectioning and the sections appear to have cracking - separating the tissue in unnatural directions. These are fixed in Carson Millonig formalin and then processed with 10% NBF, 70% alc, 95% alc, Abs alc, xylene to paraffin - total process about 2.5 hours. They are held in the 10% NBF at the beginning of the processing schedule and there is no heat on the retort until the paraffin steps. The biopsies are placed in "micro-cassettes" with no wrapping (these cassettes are not the micro-screen cassettes ones with metal like screening). We'd appreciate any ideas you might have - especially since our liver transplant service is increasing in volume as I type! Thanks! Pat Patterson Manager, AP Methodist Dallas Medical Center (214) 947-3538 *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcox90 <@t> yahoo.com Mon Dec 12 09:48:31 2005 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Mon Dec 12 09:48:40 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: <20051212154831.56056.qmail@web52107.mail.yahoo.com> Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) From la.sebree <@t> hosp.wisc.edu Mon Dec 12 09:59:18 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Mon Dec 12 09:59:28 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: We do. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, December 12, 2005 9:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caron_fournier <@t> yahoo.ca Mon Dec 12 10:02:10 2005 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Mon Dec 12 10:02:18 2005 Subject: [Histonet] bone protocal Message-ID: <20051212160210.80757.qmail@web36307.mail.mud.yahoo.com> Hi All: I am new to a position where I am doing undecalcified bone sectioning and grinding. We have a project coming up that I need some imput on....we will be fixing and embedding sheep spine with implants but the implants are PMMA and I need to know if there is a protocal that I can follow that will embedd these in a resin hard enough to grind but that will not damage the existing PMMA of the implant. Can anyone help? Caron Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos From JWEEMS <@t> sjha.org Mon Dec 12 10:06:40 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Mon Dec 12 10:06:49 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: <83AACDB0810528418AA106F9AE9B7F7E0130528C@sjhaexc02.sjha.org> We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From Jackie.O'Connor <@t> abbott.com Mon Dec 12 10:08:24 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Dec 12 10:08:54 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: Check JCAHO - this is a safety issue. Anywhere there are hazardous chems, I personally would want an emergency shower. Been there, done that with formalin. "Sebree Linda A." Sent by: histonet-bounces@lists.utsouthwestern.edu 12/12/2005 09:59 AM To: "Jill Cox" , "Histonet@Lists. Utsouthwestern. Edu" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] Emergency showers in Lab We do. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, December 12, 2005 9:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mweirauch <@t> crittenton.com Mon Dec 12 11:16:33 2005 From: mweirauch <@t> crittenton.com (Maray Weirauch) Date: Mon Dec 12 11:24:18 2005 Subject: [Histonet] NSH Histopathology Productivity Task Force Message-ID: Is there anyone on histonet that was involved in the study published in The Journal of Histotechnology, Vol. 27, No 4, Dec 2004? We pulled the article to use for benchmarking information during a 'staffing evaluation' of our Histology Department by an outside agency. Our administration is now questioning the information listed in Table 1. It lists averages of personnel given 28,800 surgicals & 22,184 pap smears annually and 365 blocks & 649 slides daily. There are several categories of personnel included: pathologists, residents, secretaries, cytologists and pathologist assistants in addition to the histology positions. There is a line that reads 'Average FTEs for all the above positions - 11.30', but the total of the averages above this line comes to 42.7 (12.6 for strictly histo titles). My administration wants to interpret this as total employees for the pathology department. Does anyone know the way these numbers were meant to be interpreted? Thanks! Maray From john.mcginley <@t> colostate.edu Mon Dec 12 11:26:50 2005 From: john.mcginley <@t> colostate.edu (John McGinley) Date: Mon Dec 12 11:27:08 2005 Subject: [Histonet] IHCRG listserv Message-ID: <000701c5ff41$3cfc9330$23255281@CAS.ColoState.EDU> Hi, The following message is intended for NSH Immunohistochemistry Resource Group (IHCRG) subscribers: You may have noticed that the IHCRG listserv has been down for the past few days. The hard drive on the Linux server that runs our laboratory mail server and hosts the IHCRG list suffered a catastrophic failure last week. I was able to get the system up and running again in a crippled state. However, many files related to the IHCRG list were corrupted. I spent most of this weekend installing a new hard drive, reloading the operating system and reconfiguring files. I will be performing a fresh install of the mailman software this morning and I hope to have members re-subscribed to the IHCRG list by this afternoon. Members should see a new subscription message in their inbox along with a new password sometime later today. Keep in mind that since the files were corrupted I'm having to rebuild from scratch and that's why your passwords will be changing. All members will be re-subscribed with the individual email delivery option set. Digest and no delivery members will need to reset these options themselves by logging into the server with their password. The archives will need to wait until I have more time to re-integrate. Sorry for the inconvenience and thanks for your patience. Regards, John McGinley IHCRG Moderator From Jan.Minshew <@t> leica-microsystems.com Mon Dec 12 11:31:14 2005 From: Jan.Minshew <@t> leica-microsystems.com (Jan.Minshew@leica-microsystems.com) Date: Mon Dec 12 11:31:33 2005 Subject: [Histonet] Cleaning of microtomes and accessories Message-ID: Hello to all, Not long ago, I responded to the originator of a question about how to clean disposable blade holders on microtomes. Since there was a great deal of conversation afterwards, I decided to consult my colleagues at the factory and respond to the whole group. The following information is what Leica recommends, but the advice should be safe to use on any microtome. I hope that other manufacturers who monitor the Histonet will feel free to jump in with additional comments too. Microtomes should always be cleaned after use (don't forget to remove blades/knives before starting). Brush away any loose, easily removed debris and disposed of it according to the regulations in your facility. Debris that is stuck to a surface should be removed with a soft cloth or gauze and "elbow grease". Baby oil (or other light oils like mineral oil) work wonders for removing paraffin. Be sure to wipe well with a clean cloth to remove residues. Using xylene is not generally recommended for cleaning for a variety of reasons (i.e. absorbtion through the skin, potential damage to the housing and locking mechanisms on some models, etc.). If, occasionally, you wish to clean more thoroughly, place the blade holder on paper towels in a paraffin oven. After it has had time to heat, use the "elbow grease" method again. Be sure to follow with the lubricant recommended by the manufacturer. This is also a great way to clean block holders that have their jaw openings caked with paraffin. As a general rule, older model disposable blade holders should not be taken apart by individuals who have not been trained. Newer models are a little more forgiving, but it would still be wise to ask your sales or service representative for their recommendation for the model you are using. If you take the clamping plate off the disposable blade holder, treat it with extreme care. Damage to the clamping surface will cause sectioning difficulties and could require replacement. Lubricate according to the instructions received with the instrument. Use the lubricant recommended by the manufacturer on the areas recommended by the manufacturer. Do not exceed the recommended amount of lubricant. If a little is good, a lot is NOT better. Last, but not least, if you have soaked a block in decal solution, rinse it in water before sectioning. This will prolong the appearance and integrity of the accessories. Be sure to remove any water from rinsing or cooling with ice, too. These are just a few tips. Cleanliness and proper maintenance are extremely important for these precision instruments. The blade holder is one of the most important parts of the microtome and it has a great deal of influence on the quality of sections that can be obtained. Some blade holders are more delicate and finiky than others. If you have any questions about the ones in your lab, I'm sure the manufacturer would be happy to offer advice on the proper way to maintain them. Warm wishes to all and Happy Holidays (a little early), Jan Minshew HT(ASCP)HTL Marketing Manager Leica Microsystems, Inc. 2345 Waukegan Rd. Bannockburn, IL 60015 800.248.0123 x7015 (toll free) 847.405.7051 (direct) 847.405.7041 (fax) ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From asamra <@t> mrl.ubc.ca Mon Dec 12 11:36:54 2005 From: asamra <@t> mrl.ubc.ca (Amrit Samra) Date: Mon Dec 12 11:37:31 2005 Subject: [Histonet] un-subscribe Message-ID: <439D44B60200008D0000198D@mail.mrl.ubc.ca> un-subscribe please From pruegg <@t> ihctech.net Mon Dec 12 12:13:18 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Dec 12 12:13:11 2005 Subject: [Histonet] Emergency showers in Lab In-Reply-To: <83AACDB0810528418AA106F9AE9B7F7E0130528C@sjhaexc02.sjha.org> Message-ID: <200512121813.jBCID0N1019734@chip.viawest.net> We do. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Monday, December 12, 2005 9:07 AM To: Jill Cox; Histonet@Lists. Utsouthwestern. Edu Subject: RE: [Histonet] Emergency showers in Lab We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From pruegg <@t> ihctech.net Mon Dec 12 12:14:27 2005 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Mon Dec 12 12:14:16 2005 Subject: [Histonet] bone protocal In-Reply-To: <20051212160210.80757.qmail@web36307.mail.mud.yahoo.com> Message-ID: <200512121814.jBCIE8N1020089@chip.viawest.net> Why don't you embed in PMMA? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of caron fournier Sent: Monday, December 12, 2005 9:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone protocal Hi All: I am new to a position where I am doing undecalcified bone sectioning and grinding. We have a project coming up that I need some imput on....we will be fixing and embedding sheep spine with implants but the implants are PMMA and I need to know if there is a protocal that I can follow that will embedd these in a resin hard enough to grind but that will not damage the existing PMMA of the implant. Can anyone help? Caron Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Dec 12 12:27:03 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 12 12:27:14 2005 Subject: [Histonet] Emergency showers in Lab In-Reply-To: <20051212154831.56056.qmail@web52107.mail.yahoo.com> Message-ID: <20051212182703.93891.qmail@web61215.mail.yahoo.com> Jill: I had always have a shower in the lab, and that is what you want to have if handling chemicals (of any type) that could spill over you. This is why there are also showers in the Chemistry labs. It is a safety requirement in Florida. Rene J. Jill Cox wrote: Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From jladuc <@t> trudeauinstitute.org Mon Dec 12 12:51:26 2005 From: jladuc <@t> trudeauinstitute.org (Judith LaDuc) Date: Mon Dec 12 12:46:49 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: Hi Jill, You can find the info at www.osha.gov if you look through the formaldehyde standard. I can't remember it off the top of my head... 1410 - ? There are many other safety organizations out there. I have a link below that is using some ASNI information that may be helpful to you. A Google or other internet search will bring up tons of safety shower hits for you as well. I do remember that they should be able to be operated with one hand and stay on after activation. It is also important to flush them (eye washes) weekly if they are attached to plumbing to avoid stagnant solutions. I heard at a recent seminar that you may need to arrange for tepid (warm) water as well, in your showers, so that you don't add the shock of cold water to victims. I haven't seen documentation yet to verify this, so take it for what it is worth. It may have been a regional/state recommendation. The link I promised is: http://www.auburn.edu/administration/safety/standards.html Good luck! Judy LaDuc, HTL ASCP >>>Jill Cox 12/12/05 10:48 am >>> Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From japoteete <@t> saintfrancis.com Mon Dec 12 12:56:42 2005 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Mon Dec 12 12:57:01 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: We do,also. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Monday, December 12, 2005 12:13 PM To: 'Weems, Joyce'; 'Jill Cox'; 'Histonet@Lists. Utsouthwestern. Edu' Subject: RE: [Histonet] Emergency showers in Lab We do. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Monday, December 12, 2005 9:07 AM To: Jill Cox; Histonet@Lists. Utsouthwestern. Edu Subject: RE: [Histonet] Emergency showers in Lab We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From adafeldman <@t> anatechltdusa.com Mon Dec 12 12:57:22 2005 From: adafeldman <@t> anatechltdusa.com (Ada Feldman) Date: Mon Dec 12 12:57:39 2005 Subject: [Histonet] Emergency showers in Lab In-Reply-To: <20051212154831.56056.qmail@web52107.mail.yahoo.com> References: <20051212154831.56056.qmail@web52107.mail.yahoo.com> Message-ID: <876E5143-D75B-40AB-940A-41E4CF7E0186@anatechltdusa.com> Jill, In the Formaldehyde Standard 29 CFR 1910.1048 (i) (2) it states "If employees may become splashed with solutions containing 1% or greater formaldehyde, for example, because of equipment failure or improper work practices, the employer shall provide conveniently located quick drench showers and assure that affected employees use these facilities. Ada T. Feldman ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com On Dec 12, 2005, at 10:48 AM, Jill Cox wrote: > Hello Histonetters!! > Just wondering if anyone knows about regs for an emergency shower > in a Pathology lab? I can't find anything in CAP checklist or OSHA > on this. Is there somewhere else I can look? Who all has showers? > Thanks in advance, Jill > > > Jill Cox HT (ASCP) > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Ada T. Feldman ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com website: www.anatechltdusa.com From Jackie.O'Connor <@t> abbott.com Mon Dec 12 13:24:13 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Dec 12 13:24:41 2005 Subject: [Histonet] IHC Contractor Message-ID: A colleague of mine is looking for a contract lab to perform IHC with a quick TAT - any takers? Jackie From sjchtascp <@t> yahoo.com Mon Dec 12 15:42:55 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Mon Dec 12 15:43:03 2005 Subject: [Histonet] IHC on coverglass Message-ID: <20051212214255.99767.qmail@web90201.mail.scd.yahoo.com> Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From lldewe <@t> ucdavis.edu Mon Dec 12 15:46:19 2005 From: lldewe <@t> ucdavis.edu (Loralei Dewe) Date: Mon Dec 12 15:46:28 2005 Subject: [Histonet] IHC on coverglass In-Reply-To: <20051212214255.99767.qmail@web90201.mail.scd.yahoo.com> References: <20051212214255.99767.qmail@web90201.mail.scd.yahoo.com> Message-ID: Yep, adherent cells? Lorie On Mon, 12 Dec 2005, Steven Coakley wrote: > Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. > > Steve > > > --------------------------------- > Yahoo! Shopping > Find Great Deals on Holiday Gifts at Yahoo! Shopping > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From la.sebree <@t> hosp.wisc.edu Mon Dec 12 15:54:53 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Mon Dec 12 15:55:05 2005 Subject: [Histonet] IHC on coverglass Message-ID: Only under extreme duress! In our case, we mounted the coverslips on a regular slide and stained it on our automated instrument (Ventana). We weren't very successful but I think it was because these were bone marrow smears that sat around air-dried in a file for who knows how long. We weren't informed that they had not been fixed in any manner and we ended up losing most of the cells. In a best case scenario it might work a whole lot better. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Monday, December 12, 2005 3:43 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on coverglass Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbass <@t> bidmc.harvard.edu Mon Dec 12 16:02:23 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Mon Dec 12 16:02:32 2005 Subject: [Histonet] rat spinal cord longitudinal sections Message-ID: Hi Guys, I have a rat that was injected with a virus that produces a fluorescent protein. I would like to scan the entire spinal cord for the signal and thought the best way to do this was to section the spinal cord along the long axis. I don't know if I have the terminology right on this. I think this should be longitudinal sections. The histologists who is to section the tissue says that this is impractical. I know that cross sections (coronal?) are more common, but which method is better when we are simply screening for a visible signal? I am worried that I will have hundreds of cross sections to examine. If there is a published example of this anywhere, even a picture on a website I would love to see it. I have not been able to find anything that is helpful in deciding which route to go. Thanks for you help, Caroline Bass From PMonfils <@t> Lifespan.org Mon Dec 12 17:20:42 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Dec 12 17:20:57 2005 Subject: [Histonet] rat spinal cord longitudinal sections Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171762F@lsexch.lsmaster.lifespan.org> Sounds to me like a longitudinal section might be good for your specific purpose. Cross sections are better for studying the morphology of the cord, but if scanning the length of the cord is your objective, and you can sacrifice the specific advantages of cross sections, than that's the way to go. It is impractical to section the entire length of the uncut cord. But the cord could be cut into three or four lengths, mounted one above the other in a single block, and sectioned that way. It would be a fairly large block, but in my lab we frequently section larger ones. If you do go with cross sections, many of these could be mounted in a single block (15 should be no great problem - 3 rows of 5 each). The average clinical lab, trying to produce hundreds of slides a day on a deadline, may not have the time to devote to such special projects. But it is certainly doable. My lab, a core research facility, receives such special requests regularly. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Caroline Bass > Sent: Monday, December 12, 2005 2:02 PM > To: Histonet (E-mail) > Subject: [Histonet] rat spinal cord longitudinal sections > > Hi Guys, > > I have a rat that was injected with a virus that produces a > fluorescent protein. I would like to scan the entire spinal cord for > the signal and thought the best way to do this was to section the > spinal cord along the long axis. I don't know if I have the > terminology right on this. I think this should be longitudinal > sections. The histologists who is to section the tissue says that > this is impractical. I know that cross sections (coronal?) are more > common, but which method is better when we are simply screening for a > visible signal? I am worried that I will have hundreds of cross > sections to examine. > > If there is a published example of this anywhere, even a picture on a > website I would love to see it. I have not been able to find > anything that is helpful in deciding which route to go. > > Thanks for you help, > > Caroline Bass > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From grupo.crs <@t> gmail.com Mon Dec 12 18:39:11 2005 From: grupo.crs <@t> gmail.com (crs grupo) Date: Mon Dec 12 18:39:20 2005 Subject: [Histonet] DAB info disposabel/neutralization/tratment Message-ID: Hello! I'm doing a research about the DAB and his tratment/neutralization/disposabel... I need some information about DAB and what to do whit it after use. If someone can give me any information, I appreciated. From rjbuesa <@t> yahoo.com Tue Dec 13 06:51:28 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 13 06:51:37 2005 Subject: [Histonet] DAB info disposabel/neutralization/tratment In-Reply-To: Message-ID: <20051213125128.98084.qmail@web61216.mail.yahoo.com> Hello CRS Group: DAB disposal constitutes an everyday need and there are several approaches, the most common being to treat it with bleach until it decolorizes but this practice is not recommended because the final product of this treatment is not known. DAB can be treat in the following way: 1-Prepare a stock solution (A) of 0.2 M potassium permanganate (=31.6 g/L) and another stock sol. (B) of 2.0 M sulfuric acid (112mL conc./L). 2- Dilute the DAB solution up to a conc. that does not exceeds 0.9 mg/mL 3- To each 10 mL of the DAB sol. add 5 mL of sol. A + 5 mL of sol. B 4- Allow the mixture to react for at least 10 hours after which time it will be non-mutagenic. 5- Add ascorbic acid in powder until the mixture is decolorized. 6- Add sodium bicarbonate to neutralize the solution (test with pH paper). 7- Now you can discard the solution down the drain, provided that your local authorities give their approval, OR you can give your DAB to a waste management company (WMC) but always remembering that you, and not the WMC, will be responsible for the disposed product. In my laboratory we used to give our DAB to a WMC after we treated it with bleach. Hope this will help you! Rene J. crs grupo wrote: Hello! I'm doing a research about the DAB and his tratment/neutralization/disposabel... I need some information about DAB and what to do whit it after use. If someone can give me any information, I appreciated. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From vd38 <@t> georgetown.edu Tue Dec 13 09:42:46 2005 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Tue Dec 13 09:45:30 2005 Subject: [Histonet] anti-mouse PR primary Ab Message-ID: <439EEBF6.1030900@georgetown.edu> Hi Histonet, I need to stain FFPE mouse tissue for Progesterone Receptor. I have tried Santa Cruz rabbit polyclonal but it is not showing the type of staining that I would expect. The luminal epithelial cells of normal mouse uterus are negative while the lamina propria stains positive. My understanding is they should both be positive in normal uterus. Also, I can only get the lamina propria staining at 1:50 in the uterus but when I stain tumor tissue at 1:50 I get tremendous amounts of background (tissue not necrotic and I see very nice, clean staining when using Santa Cruz ER Ab at 1:1500) Can anyone recommend another product that has been successful. Many thanks, -Vernon Dailey From asmith <@t> mail.barry.edu Tue Dec 13 10:12:49 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Tue Dec 13 10:15:54 2005 Subject: [Histonet] Emergency showers in Lab Message-ID: <5D2189E74151CC42BEC02906BA8996322B9139@exchsrv01.barrynet.barry.edu> Forget the regs: think about your own skin! Sulfuric or nitric acid will do serious damage faster than you can undress. If you handle either, you need a shower. If you ever use an open flame, you need a shower. Showers not only extinguish burning clothing; they also cool the skin underneath, thus reducing the severity of the burn. You should also have an eyewash station next to the shower. Keep the water in the eyewash station clean by flushing it once a week. Even if you wear goggles, eyewash stations are handy for washing the face around the goggles after a splash. (Making colloidal iron solutions or opening a hot container of decloaking buffer involves a spatter risk.) My shower and eyewash are next to my fume hood, where I use my nasty stuff. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From Jackie.O'Connor <@t> abbott.com Tue Dec 13 10:57:01 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Tue Dec 13 10:57:30 2005 Subject: [Histonet] FFPE CD31 on mouse Message-ID: I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' From koellinr <@t> amgen.com Tue Dec 13 11:17:03 2005 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Tue Dec 13 11:19:24 2005 Subject: [Histonet] FFPE CD31 on mouse Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CCA28@wa-mb4-sea.amgen.com> Jackie, Not too long ago I posted on Histonet our CD31 procedure we've used for a couple years. On mouse tissue using the Pharmingen antibody. Works beautifully but need to enzyme retrieve and amplify signal. Should be in archive. Ray Raymond Koelling Pathology Research scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Tuesday, December 13, 2005 8:57 AM To: Histonet (E-mail) Subject: [Histonet] FFPE CD31 on mouse I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Dec 13 11:30:19 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Dec 13 11:30:40 2005 Subject: Protocols that have worked using BD PharminGen monoclonal Re: [Histonet] FFPE CD31 on mouse In-Reply-To: References: Message-ID: <6.0.0.22.1.20051213102205.01b288c8@gemini.msu.montana.edu> Jackie, Here are two messages copied from Histonet Archives using CD31 BD Pharm MEC 13.3, on FFPE murine tissues with success: 1. I have used a rat anti mouse CD31 from Pharmingen(BD Biosciences).Cat. #553370. I use a rabbit anti rat(mouse adsorbed)secondary-biotinylated,from Vector Labs. For retreival, use pepsin at 37C for 20-40 min. depending on fixation. I used the ABC detection from Vector with DAB. 2. If you are using the DAKO CD31 antibody - it will not react with mouse endothelium. Pharmigen CD31 (MEC13.3) works well on FFPE with Vectastain Elite- ABC followed by NEN TSA-kit. Both work well on FFPE mouse tissues. Barb Wright Jamie Erickson supplied this info to me using BD Pharmingen clone, MEC 13.3, 1.25 ug/ml concentration, using proteinase K digestion on FFPE tissue. There may be more messages in Histonet Archives for this antibody At 09:57 AM 12/13/2005, you wrote: >I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody >that USED TO work on FFPE murine tissue - but with no luck. >I've scoured the internet for two days looking for a viable replacement - >nada. >Does anyone have a method for murine CD31 in FFPE that works without the >Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but >it didn't work. Boo. > >Jackie O' >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From DDDeltour <@t> mar.med.navy.mil Tue Dec 13 11:30:38 2005 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Tue Dec 13 11:31:02 2005 Subject: [Histonet] Florida License Message-ID: <3F500F8B416C554EBB21FF16642F72E959CD8C@marxchg03.mar.med.navy.mil> Hello All, I am looking to get my Florida License but I am currently living in Virginia. Is there an online course that covers the required HIV class? Can anyone give me any advice how to get this process done without much pain :-) Thanks Douglas D. Deltour HT(ASCP) Histology Technician NMCP Portsmouth Va (757) 953-1525/1523 From gcallis <@t> montana.edu Tue Dec 13 11:34:11 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Dec 13 11:34:27 2005 Subject: [Histonet] Another Histonet Archive CD31 for murine FFPE tissue Message-ID: <6.0.0.22.1.20051213103225.01b07c10@gemini.msu.montana.edu> It seems most people use enzyme digestion with FFPE CD31 on mouse, rather than an antigen retrieval i.e. with buffers/HIER. Other message: I just completed a project usind CD31 on FFPE mouse tissue with good results. The supplier of the antibody is Research Diagnostics (973-584-7093; www.researchd.com). I used rat anti-mouse CD31, monoclonal; Catalog# RDI-mCD31abrt. They also suggested zinc fixation, but it worked on formalin fixation. I used DAKO's Prot K to open up the sights. Bob Meyer Pathology Core Facility Northwestern University Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From angela.mcnabola.b <@t> bayer.com Tue Dec 13 12:12:16 2005 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Tue Dec 13 12:12:03 2005 Subject: [Histonet] FFPE CD31 on mouse In-Reply-To: Message-ID: We received the new lot of CD31 from Santa Cruz, and it is working, though we are going to have to use it at a more concentrated diution than our current (1:750). We haven't had the time to see what the new concentration will be, but I'm guessing somewhere around 1:200 or so. If the CD31 you have is not working, Santa Cruz will replace it to try it for yourself. We got a couple of vials for free as replacements. Another alternative is CD34 (can give you some of the same information, depedning on your work). Abcam has a great rat polyclonal that works well in FFPE mouse tissues. Angela Angela McNabola, MS, HT(ASCP)SLS, QIHC Bayer Helathcare 400 Morgan Lane West Haven, CT 06516 203-812-5001 angela.mcnabola.b@bayer.com "Jackie M O'Connor" To: "Histonet (E-mail)" Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] FFPE CD31 on mouse western.edu 12/13/2005 11:57 AM I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From thoward <@t> unm.edu Tue Dec 13 12:23:39 2005 From: thoward <@t> unm.edu (Tamara Howard) Date: Tue Dec 13 12:23:47 2005 Subject: [Histonet] RE: IHC on coverslips Message-ID: Hi - we do this in our sleep! What do you need? Tamara |--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward@unm.edu |--------------------------------------------------| From rjbuesa <@t> yahoo.com Tue Dec 13 12:28:58 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Dec 13 12:29:07 2005 Subject: [Histonet] Florida License In-Reply-To: <3F500F8B416C554EBB21FF16642F72E959CD8C@marxchg03.mar.med.navy.mil> Message-ID: <20051213182858.46593.qmail@web61218.mail.yahoo.com> Douglas: Try contacting Jerry Santiago jerry.santiago@jax.ufl.edu president of the Florida HS that may be able to giude you. Rene J. DDDeltour@mar.med.navy.mil wrote: Hello All, I am looking to get my Florida License but I am currently living in Virginia. Is there an online course that covers the required HIV class? Can anyone give me any advice how to get this process done without much pain :-) Thanks Douglas D. Deltour HT(ASCP) Histology Technician NMCP Portsmouth Va (757) 953-1525/1523 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From angela.mcnabola.b <@t> bayer.com Tue Dec 13 12:35:01 2005 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Tue Dec 13 12:34:45 2005 Subject: [Histonet] FFPE CD31 on mouse In-Reply-To: <6.0.0.22.1.20051213112609.01b43948@gemini.msu.montana.edu> Message-ID: Gayle is right....it is a monclonal....I was thinking of something else...thank you! Gayle Callis edu> cc: Subject: Re: [Histonet] FFPE CD31 on mouse 12/13/2005 01:27 PM Isn't the ABCAM rat antiMouse CD34 a monoclonal, the only CD34 they have that is polyclonal is chicken anti CD34 - that cross reacts to human, mouse and rat? At 11:12 AM 12/13/2005, you wrote: >We received the new lot of CD31 from Santa Cruz, and it is working, though we >are going to have to use it at a more concentrated diution than our current >(1:750). > >We haven't had the time to see what the new concentration will be, but I'm >guessing somewhere around 1:200 or so. If the CD31 you have is not working, >Santa Cruz will replace it to try it for yourself. We got a couple of >vials for >free as replacements. > >Another alternative is CD34 (can give you some of the same information, >depedning on your work). Abcam has a great rat polyclonal that works well in >FFPE mouse tissues. > > >Angela > > >Angela McNabola, MS, HT(ASCP)SLS, QIHC >Bayer Helathcare >400 Morgan Lane >West Haven, CT 06516 >203-812-5001 >angela.mcnabola.b@bayer.com > > > > > > "Jackie M > O'Connor" > > To: > "Histonet (E-mail)" > Sent > by: cc: > > histonet-bounces@lists.utsouth Subject: > [Histonet] FFPE CD31 on mouse > western.edu > > > > > > 12/13/2005 11:57 > AM > > > > > > > >I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody >that USED TO work on FFPE murine tissue - but with no luck. >I've scoured the internet for two days looking for a viable replacement - >nada. >Does anyone have a method for murine CD31 in FFPE that works without the >Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but >it didn't work. Boo. > >Jackie O' >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From AFeatherstone <@t> KaleidaHealth.Org Tue Dec 13 12:39:38 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Tue Dec 13 12:39:55 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 16 Message-ID: <139141F8BAF4A642A945ECC528511AF0012A7B1E@kalmb02.kaleidahealth.org> We need a different protocol for Glucose Tolerance Test. Does anyone out there have one they would be willing to email me or fax? My fax # is 716 8591853. thanks in advance Annette Featherstone HT?MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, December 13, 2005 13:05 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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: Emergency showers in Lab (Patsy Ruegg) 2. RE: bone protocal (Patsy Ruegg) 3. Re: Emergency showers in Lab (Rene J Buesa) 4. Re: Emergency showers in Lab (Judith LaDuc) 5. RE: Emergency showers in Lab (Poteete, Jacquie A.) 6. Re: Emergency showers in Lab (Ada Feldman) 7. IHC Contractor (Jackie M O'Connor) 8. IHC on coverglass (Steven Coakley) 9. Re: IHC on coverglass (Loralei Dewe) 10. RE: IHC on coverglass (Sebree Linda A.) 11. rat spinal cord longitudinal sections (Caroline Bass) 12. RE: rat spinal cord longitudinal sections (Monfils, Paul) 13. DAB info disposabel/neutralization/tratment (crs grupo) 14. Re: DAB info disposabel/neutralization/tratment (Rene J Buesa) 15. anti-mouse PR primary Ab (Vernon Dailey) 16. RE: Emergency showers in Lab (Smith, Allen) 17. FFPE CD31 on mouse (Jackie M O'Connor) 18. RE: FFPE CD31 on mouse (Koelling, Ray) 19. Protocols that have worked using BD PharminGen monoclonal Re: [Histonet] FFPE CD31 on mouse (Gayle Callis) 20. Florida License (DDDeltour@mar.med.navy.mil) 21. Another Histonet Archive CD31 for murine FFPE tissue (Gayle Callis) ---------------------------------------------------------------------- Message: 1 Date: Mon, 12 Dec 2005 11:13:18 -0700 From: "Patsy Ruegg" Subject: RE: [Histonet] Emergency showers in Lab To: "'Weems, Joyce'" , "'Jill Cox'" , "'Histonet@Lists. Utsouthwestern. Edu'" Message-ID: <200512121813.jBCID0N1019734@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" We do. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Monday, December 12, 2005 9:07 AM To: Jill Cox; Histonet@Lists. Utsouthwestern. Edu Subject: RE: [Histonet] Emergency showers in Lab We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------ Message: 2 Date: Mon, 12 Dec 2005 11:14:27 -0700 From: "Patsy Ruegg" Subject: RE: [Histonet] bone protocal To: "'caron fournier'" , Message-ID: <200512121814.jBCIE8N1020089@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Why don't you embed in PMMA? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of caron fournier Sent: Monday, December 12, 2005 9:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone protocal Hi All: I am new to a position where I am doing undecalcified bone sectioning and grinding. We have a project coming up that I need some imput on....we will be fixing and embedding sheep spine with implants but the implants are PMMA and I need to know if there is a protocal that I can follow that will embedd these in a resin hard enough to grind but that will not damage the existing PMMA of the implant. Can anyone help? Caron Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 12 Dec 2005 10:27:03 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Emergency showers in Lab To: Jill Cox , "Histonet@Lists. Utsouthwestern. Edu" Message-ID: <20051212182703.93891.qmail@web61215.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Jill: I had always have a shower in the lab, and that is what you want to have if handling chemicals (of any type) that could spill over you. This is why there are also showers in the Chemistry labs. It is a safety requirement in Florida. Rene J. Jill Cox wrote: Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping ------------------------------ Message: 4 Date: Mon, 12 Dec 2005 13:51:26 -0500 From: "Judith LaDuc" Subject: Re: [Histonet] Emergency showers in Lab To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Jill, You can find the info at www.osha.gov if you look through the formaldehyde standard. I can't remember it off the top of my head... 1410 - ? There are many other safety organizations out there. I have a link below that is using some ASNI information that may be helpful to you. A Google or other internet search will bring up tons of safety shower hits for you as well. I do remember that they should be able to be operated with one hand and stay on after activation. It is also important to flush them (eye washes) weekly if they are attached to plumbing to avoid stagnant solutions. I heard at a recent seminar that you may need to arrange for tepid (warm) water as well, in your showers, so that you don't add the shock of cold water to victims. I haven't seen documentation yet to verify this, so take it for what it is worth. It may have been a regional/state recommendation. The link I promised is: http://www.auburn.edu/administration/safety/standards.html Good luck! Judy LaDuc, HTL ASCP >>>Jill Cox 12/12/05 10:48 am >>> Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 12 Dec 2005 12:56:42 -0600 From: "Poteete, Jacquie A." Subject: RE: [Histonet] Emergency showers in Lab To: 'Patsy Ruegg' , "'Weems, Joyce'" , 'Jill Cox' , "'Histonet@Lists. Utsouthwestern. Edu'" Message-ID: Content-Type: text/plain We do,also. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Monday, December 12, 2005 12:13 PM To: 'Weems, Joyce'; 'Jill Cox'; 'Histonet@Lists. Utsouthwestern. Edu' Subject: RE: [Histonet] Emergency showers in Lab We do. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Monday, December 12, 2005 9:07 AM To: Jill Cox; Histonet@Lists. Utsouthwestern. Edu Subject: RE: [Histonet] Emergency showers in Lab We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp ------------------------------ Message: 6 Date: Mon, 12 Dec 2005 13:57:22 -0500 From: Ada Feldman Subject: Re: [Histonet] Emergency showers in Lab To: Jill Cox Cc: "Histonet@Lists. Utsouthwestern. Edu" Message-ID: <876E5143-D75B-40AB-940A-41E4CF7E0186@anatechltdusa.com> Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Jill, In the Formaldehyde Standard 29 CFR 1910.1048 (i) (2) it states "If employees may become splashed with solutions containing 1% or greater formaldehyde, for example, because of equipment failure or improper work practices, the employer shall provide conveniently located quick drench showers and assure that affected employees use these facilities. Ada T. Feldman ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com On Dec 12, 2005, at 10:48 AM, Jill Cox wrote: > Hello Histonetters!! > Just wondering if anyone knows about regs for an emergency shower > in a Pathology lab? I can't find anything in CAP checklist or OSHA > on this. Is there somewhere else I can look? Who all has showers? > Thanks in advance, Jill > > > Jill Cox HT (ASCP) > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Ada T. Feldman ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com website: www.anatechltdusa.com ------------------------------ Message: 7 Date: Mon, 12 Dec 2005 13:24:13 -0600 From: "Jackie M O'Connor" Subject: [Histonet] IHC Contractor To: "Histonet@Lists. Utsouthwestern. Edu" Message-ID: Content-Type: text/plain; charset="us-ascii" A colleague of mine is looking for a contract lab to perform IHC with a quick TAT - any takers? Jackie ------------------------------ Message: 8 Date: Mon, 12 Dec 2005 13:42:55 -0800 (PST) From: Steven Coakley Subject: [Histonet] IHC on coverglass To: Histonet@lists.utsouthwestern.edu Message-ID: <20051212214255.99767.qmail@web90201.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping ------------------------------ Message: 9 Date: Mon, 12 Dec 2005 13:46:19 -0800 (PST) From: Loralei Dewe Subject: Re: [Histonet] IHC on coverglass To: Steven Coakley Cc: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII Yep, adherent cells? Lorie On Mon, 12 Dec 2005, Steven Coakley wrote: > Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. > > Steve > > > --------------------------------- > Yahoo! Shopping > Find Great Deals on Holiday Gifts at Yahoo! Shopping > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 10 Date: Mon, 12 Dec 2005 15:54:53 -0600 From: "Sebree Linda A." Subject: RE: [Histonet] IHC on coverglass To: "Steven Coakley" , Message-ID: Content-Type: text/plain; charset="us-ascii" Only under extreme duress! In our case, we mounted the coverslips on a regular slide and stained it on our automated instrument (Ventana). We weren't very successful but I think it was because these were bone marrow smears that sat around air-dried in a file for who knows how long. We weren't informed that they had not been fixed in any manner and we ended up losing most of the cells. In a best case scenario it might work a whole lot better. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Monday, December 12, 2005 3:43 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on coverglass Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Mon, 12 Dec 2005 17:02:23 -0500 From: Caroline Bass Subject: [Histonet] rat spinal cord longitudinal sections To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Hi Guys, I have a rat that was injected with a virus that produces a fluorescent protein. I would like to scan the entire spinal cord for the signal and thought the best way to do this was to section the spinal cord along the long axis. I don't know if I have the terminology right on this. I think this should be longitudinal sections. The histologists who is to section the tissue says that this is impractical. I know that cross sections (coronal?) are more common, but which method is better when we are simply screening for a visible signal? I am worried that I will have hundreds of cross sections to examine. If there is a published example of this anywhere, even a picture on a website I would love to see it. I have not been able to find anything that is helpful in deciding which route to go. Thanks for you help, Caroline Bass ------------------------------ Message: 12 Date: Mon, 12 Dec 2005 18:20:42 -0500 From: "Monfils, Paul" Subject: RE: [Histonet] rat spinal cord longitudinal sections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171762F@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" Sounds to me like a longitudinal section might be good for your specific purpose. Cross sections are better for studying the morphology of the cord, but if scanning the length of the cord is your objective, and you can sacrifice the specific advantages of cross sections, than that's the way to go. It is impractical to section the entire length of the uncut cord. But the cord could be cut into three or four lengths, mounted one above the other in a single block, and sectioned that way. It would be a fairly large block, but in my lab we frequently section larger ones. If you do go with cross sections, many of these could be mounted in a single block (15 should be no great problem - 3 rows of 5 each). The average clinical lab, trying to produce hundreds of slides a day on a deadline, may not have the time to devote to such special projects. But it is certainly doable. My lab, a core research facility, receives such special requests regularly. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Caroline Bass > Sent: Monday, December 12, 2005 2:02 PM > To: Histonet (E-mail) > Subject: [Histonet] rat spinal cord longitudinal sections > > Hi Guys, > > I have a rat that was injected with a virus that produces a > fluorescent protein. I would like to scan the entire spinal cord for > the signal and thought the best way to do this was to section the > spinal cord along the long axis. I don't know if I have the > terminology right on this. I think this should be longitudinal > sections. The histologists who is to section the tissue says that > this is impractical. I know that cross sections (coronal?) are more > common, but which method is better when we are simply screening for a > visible signal? I am worried that I will have hundreds of cross > sections to examine. > > If there is a published example of this anywhere, even a picture on a > website I would love to see it. I have not been able to find > anything that is helpful in deciding which route to go. > > Thanks for you help, > > Caroline Bass > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 13 Date: Tue, 13 Dec 2005 00:39:11 +0000 From: crs grupo Subject: [Histonet] DAB info disposabel/neutralization/tratment To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hello! I'm doing a research about the DAB and his tratment/neutralization/disposabel... I need some information about DAB and what to do whit it after use. If someone can give me any information, I appreciated. ------------------------------ Message: 14 Date: Tue, 13 Dec 2005 04:51:28 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] DAB info disposabel/neutralization/tratment To: crs grupo , Histonet@lists.utsouthwestern.edu Message-ID: <20051213125128.98084.qmail@web61216.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello CRS Group: DAB disposal constitutes an everyday need and there are several approaches, the most common being to treat it with bleach until it decolorizes but this practice is not recommended because the final product of this treatment is not known. DAB can be treat in the following way: 1-Prepare a stock solution (A) of 0.2 M potassium permanganate (=31.6 g/L) and another stock sol. (B) of 2.0 M sulfuric acid (112mL conc./L). 2- Dilute the DAB solution up to a conc. that does not exceeds 0.9 mg/mL 3- To each 10 mL of the DAB sol. add 5 mL of sol. A + 5 mL of sol. B 4- Allow the mixture to react for at least 10 hours after which time it will be non-mutagenic. 5- Add ascorbic acid in powder until the mixture is decolorized. 6- Add sodium bicarbonate to neutralize the solution (test with pH paper). 7- Now you can discard the solution down the drain, provided that your local authorities give their approval, OR you can give your DAB to a waste management company (WMC) but always remembering that you, and not the WMC, will be responsible for the disposed product. In my laboratory we used to give our DAB to a WMC after we treated it with bleach. Hope this will help you! Rene J. crs grupo wrote: Hello! I'm doing a research about the DAB and his tratment/neutralization/disposabel... I need some information about DAB and what to do whit it after use. If someone can give me any information, I appreciated. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping ------------------------------ Message: 15 Date: Tue, 13 Dec 2005 10:42:46 -0500 From: Vernon Dailey Subject: [Histonet] anti-mouse PR primary Ab To: Histonet Message-ID: <439EEBF6.1030900@georgetown.edu> Content-Type: text/plain; charset="iso-8859-1" Hi Histonet, I need to stain FFPE mouse tissue for Progesterone Receptor. I have tried Santa Cruz rabbit polyclonal but it is not showing the type of staining that I would expect. The luminal epithelial cells of normal mouse uterus are negative while the lamina propria stains positive. My understanding is they should both be positive in normal uterus. Also, I can only get the lamina propria staining at 1:50 in the uterus but when I stain tumor tissue at 1:50 I get tremendous amounts of background (tissue not necrotic and I see very nice, clean staining when using Santa Cruz ER Ab at 1:1500) Can anyone recommend another product that has been successful. Many thanks, -Vernon Dailey ------------------------------ Message: 16 Date: Tue, 13 Dec 2005 11:12:49 -0500 From: "Smith, Allen" Subject: RE: [Histonet] Emergency showers in Lab To: "Jill Cox" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5D2189E74151CC42BEC02906BA8996322B9139@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Forget the regs: think about your own skin! Sulfuric or nitric acid will do serious damage faster than you can undress. If you handle either, you need a shower. If you ever use an open flame, you need a shower. Showers not only extinguish burning clothing; they also cool the skin underneath, thus reducing the severity of the burn. You should also have an eyewash station next to the shower. Keep the water in the eyewash station clean by flushing it once a week. Even if you wear goggles, eyewash stations are handy for washing the face around the goggles after a splash. (Making colloidal iron solutions or opening a hot container of decloaking buffer involves a spatter risk.) My shower and eyewash are next to my fume hood, where I use my nasty stuff. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 17 Date: Tue, 13 Dec 2005 10:57:01 -0600 From: "Jackie M O'Connor" Subject: [Histonet] FFPE CD31 on mouse To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="us-ascii" I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' ------------------------------ Message: 18 Date: Tue, 13 Dec 2005 09:17:03 -0800 From: "Koelling, Ray" Subject: RE: [Histonet] FFPE CD31 on mouse To: "'Jackie M O'Connor'" , "Histonet (E-mail)" Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CCA28@wa-mb4-sea.amgen.com> Content-Type: text/plain; charset="iso-8859-1" Jackie, Not too long ago I posted on Histonet our CD31 procedure we've used for a couple years. On mouse tissue using the Pharmingen antibody. Works beautifully but need to enzyme retrieve and amplify signal. Should be in archive. Ray Raymond Koelling Pathology Research scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Tuesday, December 13, 2005 8:57 AM To: Histonet (E-mail) Subject: [Histonet] FFPE CD31 on mouse I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 13 Dec 2005 10:30:19 -0700 From: Gayle Callis Subject: Protocols that have worked using BD PharminGen monoclonal Re: [Histonet] FFPE CD31 on mouse To: "Jackie M O'Connor" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051213102205.01b288c8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Jackie, Here are two messages copied from Histonet Archives using CD31 BD Pharm MEC 13.3, on FFPE murine tissues with success: 1. I have used a rat anti mouse CD31 from Pharmingen(BD Biosciences).Cat. #553370. I use a rabbit anti rat(mouse adsorbed)secondary-biotinylated,from Vector Labs. For retreival, use pepsin at 37C for 20-40 min. depending on fixation. I used the ABC detection from Vector with DAB. 2. If you are using the DAKO CD31 antibody - it will not react with mouse endothelium. Pharmigen CD31 (MEC13.3) works well on FFPE with Vectastain Elite- ABC followed by NEN TSA-kit. Both work well on FFPE mouse tissues. Barb Wright Jamie Erickson supplied this info to me using BD Pharmingen clone, MEC 13.3, 1.25 ug/ml concentration, using proteinase K digestion on FFPE tissue. There may be more messages in Histonet Archives for this antibody At 09:57 AM 12/13/2005, you wrote: >I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody >that USED TO work on FFPE murine tissue - but with no luck. >I've scoured the internet for two days looking for a viable replacement - >nada. >Does anyone have a method for murine CD31 in FFPE that works without the >Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but >it didn't work. Boo. > >Jackie O' >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 20 Date: Tue, 13 Dec 2005 12:30:38 -0500 From: DDDeltour@mar.med.navy.mil Subject: [Histonet] Florida License To: histonet@lists.utsouthwestern.edu Message-ID: <3F500F8B416C554EBB21FF16642F72E959CD8C@marxchg03.mar.med.navy.mil> Content-Type: text/plain Hello All, I am looking to get my Florida License but I am currently living in Virginia. Is there an online course that covers the required HIV class? Can anyone give me any advice how to get this process done without much pain :-) Thanks Douglas D. Deltour HT(ASCP) Histology Technician NMCP Portsmouth Va (757) 953-1525/1523 ------------------------------ Message: 21 Date: Tue, 13 Dec 2005 10:34:11 -0700 From: Gayle Callis Subject: [Histonet] Another Histonet Archive CD31 for murine FFPE tissue To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051213103225.01b07c10@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed It seems most people use enzyme digestion with FFPE CD31 on mouse, rather than an antigen retrieval i.e. with buffers/HIER. Other message: I just completed a project usind CD31 on FFPE mouse tissue with good results. The supplier of the antibody is Research Diagnostics (973-584-7093; www.researchd.com). I used rat anti-mouse CD31, monoclonal; Catalog# RDI-mCD31abrt. They also suggested zinc fixation, but it worked on formalin fixation. I used DAKO's Prot K to open up the sights. Bob Meyer Pathology Core Facility Northwestern University Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 25, Issue 16 **************************************** CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. 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From jlinda <@t> ces.clemson.edu Tue Dec 13 13:01:43 2005 From: jlinda <@t> ces.clemson.edu (Linda Jenkins) Date: Tue Dec 13 13:01:34 2005 Subject: [Histonet] Embedding PMMA implants Message-ID: <5.2.1.1.2.20051213134125.01ed5b40@mailhost.ces.clemson.edu> Caron, There are numerous plastics to embed hard tissue in BUT, if your implant is PMMA, you definitely DO NOT want to embed in PMMA as it will dissolve most of your implant. I have had very good results with Technovit 7200 which is a light curing resin and is sold by the Exakt company. There are many pre-made kits on the market to embed hard tissue. Since you are just starting out, you might want to use a pre-made kit(removes a lot of the "head-banging" that goes along with hard tissue). I would also urge you to do a pilot study of your implant material. In other words, take a sample of your implant material and embed it in a variety of monomers and see what happens. Also, bear in mind, that an implant will usually maintain more of its integrity when it is implanted and then processed. Good Luck, Linda Linda Jenkins, HT Clemson University Dept. of Bioengineering Clemson, SC 29634-0905 864.656.5553 http://www.ces.clemson.edu/bio/research/histo.htm From LJohns53 <@t> CNTUS.JNJ.COM Tue Dec 13 13:13:48 2005 From: LJohns53 <@t> CNTUS.JNJ.COM (Johns, Laura [CNTUS]) Date: Tue Dec 13 13:14:17 2005 Subject: [Histonet] protocol for fixation of rat eyes Message-ID: <70F83FE9F65318468A612768E7043F8903753F34@cntusmaexs9.na.jnj.com> Does anyone have a good protocol for fixation and processing of rodent eyes using Pen-Fix? We will be embedding in paraffin. I saw a few emails in the archive from Lynn Gardner saying she had a good protocol for this, but I am not sure where she is now. Thanks! Laura Johns Laura M. Johns, Ph.D. Centocor, Inc. 145 King of Prussia Road (Mail Stop # R-4-2) Radnor, PA 19087 Phone: 610-240-8284 .... Fax: 610-240-8150.... Email: ljohns53@cntus.jnj.com > Confidentiality Notice: This message is intended for the individual(s) or > entity to which it is addressed and may contain information that is > privileged, confidential and exempt from disclosure under applicable law. > If the reader of this message is not the intended recipient, he/she is > 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 by telephone and delete > the e-mail from your system immediately. Thank you. > > From bliven.laura <@t> marshfieldclinic.org Tue Dec 13 13:52:14 2005 From: bliven.laura <@t> marshfieldclinic.org (bliven.laura@marshfieldclinic.org) Date: Tue Dec 13 13:52:24 2005 Subject: [Histonet] Factor XIIIa Antibody Message-ID: <16f8d01c6001e$b6b6ccd0$8e0110ac@mfldclinframe.org> Looking for info on a Factor XIIIa antibody that is IVD labeled. I've tried clone AC-1A1, which seems to give me some false positive nuclear staining in the dermis (even without antigen retrieval). This staining isn't unhear of or a big problem, but I would like to see if anything else is available before I commit to this antibody. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 From cbass <@t> bidmc.harvard.edu Tue Dec 13 14:06:32 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Tue Dec 13 14:06:43 2005 Subject: [Histonet] temperature for sectioning fixed liver Message-ID: <29F4D60A-99AC-4966-B945-1C900D3AE15C@bidmc.harvard.edu> Hello, I am going to section 4% formaldehyde fixed liver blocks on a cryostat this afternoon. It is the first time that I have sectioned this particular tissue in a cryostat. Could someone recommend a good starting temperature for specimen and chamber? I have a recommendation of -17 for frozen tissue, but is this good for fixed as well? I am going to try 10 um sections initially. Thanks, Caroline Bass From kl2215 <@t> columbia.edu Tue Dec 13 14:06:45 2005 From: kl2215 <@t> columbia.edu (Kim Lopez) Date: Tue Dec 13 14:06:54 2005 Subject: [Histonet] combined FISH/IHC protocol Message-ID: <1134504405.439f29d571a3b@cubmail.cc.columbia.edu> Hi all, I wanna do combined FISH and IHC on frozen rat brain using a rat y-chromosome probe and Ki67. I've already found one paper that has done something similar on mouse brains using a mouse y-chromosome probe and anti-GFP. It seemed to work well for them and I'm going to try and replicate using their protocol (Donadoni C, J Histochem & Cytochem, 2004). I was just wondering if anybody else has any experience with this. I will be very grateful for any tips or advice. Kim From la.sebree <@t> hosp.wisc.edu Tue Dec 13 14:09:03 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Tue Dec 13 14:09:13 2005 Subject: [Histonet] Factor XIIIa Antibody Message-ID: Laura, We use Biocare's polyclonal FXIIIa, an IVD antibody, with good results. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of bliven.laura@marshfieldclinic.org Sent: Tuesday, December 13, 2005 1:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Factor XIIIa Antibody Looking for info on a Factor XIIIa antibody that is IVD labeled. I've tried clone AC-1A1, which seems to give me some false positive nuclear staining in the dermis (even without antigen retrieval). This staining isn't unhear of or a big problem, but I would like to see if anything else is available before I commit to this antibody. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sa.drew <@t> hosp.wisc.edu Tue Dec 13 14:14:37 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Tue Dec 13 14:19:42 2005 Subject: [Histonet] Factor XIIIa Antibody Message-ID: We are using a polyclonal F13a from BioCare with no complaints from our dermpath people... Pretreatment is with protease. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of bliven.laura@marshfieldclinic.org Sent: Tuesday, December 13, 2005 1:52 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Factor XIIIa Antibody Looking for info on a Factor XIIIa antibody that is IVD labeled. I've tried clone AC-1A1, which seems to give me some false positive nuclear staining in the dermis (even without antigen retrieval). This staining isn't unhear of or a big problem, but I would like to see if anything else is available before I commit to this antibody. Thanks, Laura Bliven Histology Lab/IHC Marshfield Laboratories 1000 N. Oak Ave. Marshfield, WI 54449 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Dec 13 14:35:44 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Dec 13 14:37:22 2005 Subject: [Histonet] protocol for fixation of rat eyes In-Reply-To: <70F83FE9F65318468A612768E7043F8903753F34@cntusmaexs9.na.jn j.com> References: <70F83FE9F65318468A612768E7043F8903753F34@cntusmaexs9.na.jnj.com> Message-ID: <6.0.0.22.1.20051213133356.01b3c1d8@gemini.msu.montana.edu> Lynn wrote this up for the NSH VIR Animal Processing Manual. You can purchase the manual from NSH office. Her protocol also included how to section the eye after embedding. Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From lanbergld <@t> vcu.edu Tue Dec 13 19:18:21 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Tue Dec 13 19:18:30 2005 Subject: [Histonet] Cuting speed and sem-thin plastic sectioning Message-ID: Hello, I am a beginning microtomist - most part - in a Biochemistry laborato retinas using plastic (Spurr's Formula).&nbs MT-1000 ultramicrotome. My sectioning, staining and developed very well in the 4 months I have done this. I have little thing, though, that seems to be impeding progress: Speed of cut Currently, the only way I get quality sec mm/sec. Which is almost the slowest setting on t lucky, I may get three blocks done per hour; 20 section slides per block. This is slow output, from what I am told. I have experimented cutting these on a speed as high This greatly increases my output, obviously, but after staining-destaining the sections appear corrugated. But they are beautiful I am un cordurouy-like plastic (hard formulation self-made glass knives, can anybody possibly cut on the higher speeds, while mai section? Thank You & Larry Lanberg Student Worker IST Dept. Virginia Commonwealth University (804) From lhotaks <@t> mcmaster.ca Tue Dec 13 19:21:45 2005 From: lhotaks <@t> mcmaster.ca (Sarka Lhotak) Date: Tue Dec 13 19:21:55 2005 Subject: [Histonet] Re: IHC on coverslips Message-ID: Hi Netters, IHC on coverslips is easily done by performing the whole procedure, including fixation and permeabilization with Triton X, in a 6 well plate. Label the lid of the plate to know which slip is in which well. Rinse by filling the well with buffer, then aspirate. Before incubations, I aspirate as much liquid as possible from around the coverslip with a glass Pasteur pipette. This will limit spreading of your precious antibody into the well. Then I carefully place 5 drops of the antibody (50 ul), in the middle and four corners of the coverslip. I use this technique mostly to do immunofluorescence, but I have also done peroxidase/ AEC reactions, with great results. The coverslip is removed at the very end for "coverslipping" with a slide. The coverslip is cells up all the way, just make sure you coverslip it cells down. Hope this helps. Please, contact me for more details. Sarka Lhotak, PhD McMaster University Hamilton, Ontario, Canada From JEllin <@t> yumaregional.org Tue Dec 13 19:29:04 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Tue Dec 13 19:29:34 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Message-ID: Hello everyone I have a real problem here, We have just accquired a tissue proccesor (VIP 5) and found out the hard way like everything in Histology, there is a problem with the proccessing. Our pathologist are claiming that the small biopsies are coming out cooked, after reviewing the slide myself with the pathologist it is not a cooked looked but rather a glassy look and bubbly. Later found out that the latch of the tissue proccessor is not tight enough and is was allowing moisture in and there was a lot of condensation. I figured out what was wrong but here is the kicker. My Pathologist is wanting to take one of the small biopsies that was proccessed and regress the tissue and then reproccess the small biopsy by hand instead of throwing it back on the tissue proccessor. The biopsy is about .1 mm very tiny. I need all the help that I can get!!!!!! So please fellow histo-netters share the knowledge with me. Jesus Ellin Yuma Regional Medical Center From lanbergld <@t> vcu.edu Tue Dec 13 21:04:24 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Tue Dec 13 21:04:33 2005 Subject: [Histonet] Re: cutting speed and semi-thin plastic sectioning Message-ID: Thank you very much Dr. / Ms. Marcum. That makes sense. I ag the quality is A-#1 priority, so 0.3 mm/sec is the magic number fo me. I do know that my ultra-microtome is extremely sensiti distant vibrations, anywhere in the lab. In fact, I hold my bre (not exhale at all) when the block slowly makes its way through the bla some to b Thanks again. From vanann702 <@t> skmc.gov.ae Tue Dec 13 22:10:01 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Tue Dec 13 22:07:03 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E02F6D70E@SKMCEMAIL.skmc.gov.ae> What kind of tissue? Was it 'dried out' before placing in fixative? What is your processing protocol In my humble opinion - reprocessing this small bx will just make it worse I have used VIP5's for the past 10 years and they are wonderful processors. Its usually the times of the processing protocol that are the 'devil' - OR (I have found this to be a HUUUGE factor - how long the bx lies in 'fresh air' before it gets popped into formalin (beyond your control). This term 'cooked' is loosely applied by Paths. I have only one VIP and use the same programme overnight - for small gastrics, mastectomies, lymph nodes, liver and renal cores - a whole variety of tissue. The 'latch' of the VIP cannot be 'loose' - the retort has a safety mechanism - it wont process if it 'leaks' - perhaps the rubber gasket was not snug in its housing (whoever last used/cleaned machine should check) We often blame the equipment - when it is the operators fault Cars are not to blame for accidents - drivers are!!! Annieinarabie -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Wednesday, December 14, 2005 5:29 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Hello everyone I have a real problem here, We have just accquired a tissue proccesor (VIP 5) and found out the hard way like everything in Histology, there is a problem with the proccessing. Our pathologist are claiming that the small biopsies are coming out cooked, after reviewing the slide myself with the pathologist it is not a cooked looked but rather a glassy look and bubbly. Later found out that the latch of the tissue proccessor is not tight enough and is was allowing moisture in and there was a lot of condensation. I figured out what was wrong but here is the kicker. My Pathologist is wanting to take one of the small biopsies that was proccessed and regress the tissue and then reproccess the small biopsy by hand instead of throwing it back on the tissue proccessor. The biopsy is about .1 mm very tiny. I need all the help that I can get!!!!!! So please fellow histo-netters share the knowledge with me. Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AFeatherstone <@t> KaleidaHealth.Org Wed Dec 14 05:13:43 2005 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Wed Dec 14 05:14:09 2005 Subject: [Histonet] GLUCOSE TOLERANCE TEST Message-ID: <139141F8BAF4A642A945ECC528511AF0012A7B21@kalmb02.kaleidahealth.org> Does anyone out there have a protocol for a glucose tolerance test? I know this isn't histo but my resident needs a new protocol. Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Tuesday, December 13, 2005 13:05 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 25, 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: Emergency showers in Lab (Patsy Ruegg) 2. RE: bone protocal (Patsy Ruegg) 3. Re: Emergency showers in Lab (Rene J Buesa) 4. Re: Emergency showers in Lab (Judith LaDuc) 5. RE: Emergency showers in Lab (Poteete, Jacquie A.) 6. Re: Emergency showers in Lab (Ada Feldman) 7. IHC Contractor (Jackie M O'Connor) 8. IHC on coverglass (Steven Coakley) 9. Re: IHC on coverglass (Loralei Dewe) 10. RE: IHC on coverglass (Sebree Linda A.) 11. rat spinal cord longitudinal sections (Caroline Bass) 12. RE: rat spinal cord longitudinal sections (Monfils, Paul) 13. DAB info disposabel/neutralization/tratment (crs grupo) 14. Re: DAB info disposabel/neutralization/tratment (Rene J Buesa) 15. anti-mouse PR primary Ab (Vernon Dailey) 16. RE: Emergency showers in Lab (Smith, Allen) 17. FFPE CD31 on mouse (Jackie M O'Connor) 18. RE: FFPE CD31 on mouse (Koelling, Ray) 19. Protocols that have worked using BD PharminGen monoclonal Re: [Histonet] FFPE CD31 on mouse (Gayle Callis) 20. Florida License (DDDeltour@mar.med.navy.mil) 21. Another Histonet Archive CD31 for murine FFPE tissue (Gayle Callis) ---------------------------------------------------------------------- Message: 1 Date: Mon, 12 Dec 2005 11:13:18 -0700 From: "Patsy Ruegg" Subject: RE: [Histonet] Emergency showers in Lab To: "'Weems, Joyce'" , "'Jill Cox'" , "'Histonet@Lists. Utsouthwestern. Edu'" Message-ID: <200512121813.jBCID0N1019734@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" We do. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Monday, December 12, 2005 9:07 AM To: Jill Cox; Histonet@Lists. Utsouthwestern. Edu Subject: RE: [Histonet] Emergency showers in Lab We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. ------------------------------ Message: 2 Date: Mon, 12 Dec 2005 11:14:27 -0700 From: "Patsy Ruegg" Subject: RE: [Histonet] bone protocal To: "'caron fournier'" , Message-ID: <200512121814.jBCIE8N1020089@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Why don't you embed in PMMA? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of caron fournier Sent: Monday, December 12, 2005 9:02 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone protocal Hi All: I am new to a position where I am doing undecalcified bone sectioning and grinding. We have a project coming up that I need some imput on....we will be fixing and embedding sheep spine with implants but the implants are PMMA and I need to know if there is a protocal that I can follow that will embedd these in a resin hard enough to grind but that will not damage the existing PMMA of the implant. Can anyone help? Caron Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. --------------------------------- Find your next car at Yahoo! Canada Autos _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 3 Date: Mon, 12 Dec 2005 10:27:03 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] Emergency showers in Lab To: Jill Cox , "Histonet@Lists. Utsouthwestern. Edu" Message-ID: <20051212182703.93891.qmail@web61215.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Jill: I had always have a shower in the lab, and that is what you want to have if handling chemicals (of any type) that could spill over you. This is why there are also showers in the Chemistry labs. It is a safety requirement in Florida. Rene J. Jill Cox wrote: Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping ------------------------------ Message: 4 Date: Mon, 12 Dec 2005 13:51:26 -0500 From: "Judith LaDuc" Subject: Re: [Histonet] Emergency showers in Lab To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Hi Jill, You can find the info at www.osha.gov if you look through the formaldehyde standard. I can't remember it off the top of my head... 1410 - ? There are many other safety organizations out there. I have a link below that is using some ASNI information that may be helpful to you. A Google or other internet search will bring up tons of safety shower hits for you as well. I do remember that they should be able to be operated with one hand and stay on after activation. It is also important to flush them (eye washes) weekly if they are attached to plumbing to avoid stagnant solutions. I heard at a recent seminar that you may need to arrange for tepid (warm) water as well, in your showers, so that you don't add the shock of cold water to victims. I haven't seen documentation yet to verify this, so take it for what it is worth. It may have been a regional/state recommendation. The link I promised is: http://www.auburn.edu/administration/safety/standards.html Good luck! Judy LaDuc, HTL ASCP >>>Jill Cox 12/12/05 10:48 am >>> Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 12 Dec 2005 12:56:42 -0600 From: "Poteete, Jacquie A." Subject: RE: [Histonet] Emergency showers in Lab To: 'Patsy Ruegg' , "'Weems, Joyce'" , 'Jill Cox' , "'Histonet@Lists. Utsouthwestern. Edu'" Message-ID: Content-Type: text/plain We do,also. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK -----Original Message----- From: Patsy Ruegg [mailto:pruegg@ihctech.net] Sent: Monday, December 12, 2005 12:13 PM To: 'Weems, Joyce'; 'Jill Cox'; 'Histonet@Lists. Utsouthwestern. Edu' Subject: RE: [Histonet] Emergency showers in Lab We do. Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Monday, December 12, 2005 9:07 AM To: Jill Cox; Histonet@Lists. Utsouthwestern. Edu Subject: RE: [Histonet] Emergency showers in Lab We do. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp ------------------------------ Message: 6 Date: Mon, 12 Dec 2005 13:57:22 -0500 From: Ada Feldman Subject: Re: [Histonet] Emergency showers in Lab To: Jill Cox Cc: "Histonet@Lists. Utsouthwestern. Edu" Message-ID: <876E5143-D75B-40AB-940A-41E4CF7E0186@anatechltdusa.com> Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Jill, In the Formaldehyde Standard 29 CFR 1910.1048 (i) (2) it states "If employees may become splashed with solutions containing 1% or greater formaldehyde, for example, because of equipment failure or improper work practices, the employer shall provide conveniently located quick drench showers and assure that affected employees use these facilities. Ada T. Feldman ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com On Dec 12, 2005, at 10:48 AM, Jill Cox wrote: > Hello Histonetters!! > Just wondering if anyone knows about regs for an emergency shower > in a Pathology lab? I can't find anything in CAP checklist or OSHA > on this. Is there somewhere else I can look? Who all has showers? > Thanks in advance, Jill > > > Jill Cox HT (ASCP) > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet Ada T. Feldman ANATECH LTD. 1020 Harts Lake Road Battle Creek, MI 49015 Phone: 800.262.8324 Fax: 269.964.8084 email: adafeldman@anatechltdusa.com website: www.anatechltdusa.com ------------------------------ Message: 7 Date: Mon, 12 Dec 2005 13:24:13 -0600 From: "Jackie M O'Connor" Subject: [Histonet] IHC Contractor To: "Histonet@Lists. Utsouthwestern. Edu" Message-ID: Content-Type: text/plain; charset="us-ascii" A colleague of mine is looking for a contract lab to perform IHC with a quick TAT - any takers? Jackie ------------------------------ Message: 8 Date: Mon, 12 Dec 2005 13:42:55 -0800 (PST) From: Steven Coakley Subject: [Histonet] IHC on coverglass To: Histonet@lists.utsouthwestern.edu Message-ID: <20051212214255.99767.qmail@web90201.mail.scd.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping ------------------------------ Message: 9 Date: Mon, 12 Dec 2005 13:46:19 -0800 (PST) From: Loralei Dewe Subject: Re: [Histonet] IHC on coverglass To: Steven Coakley Cc: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: TEXT/PLAIN; charset=US-ASCII Yep, adherent cells? Lorie On Mon, 12 Dec 2005, Steven Coakley wrote: > Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. > > Steve > > > --------------------------------- > Yahoo! Shopping > Find Great Deals on Holiday Gifts at Yahoo! Shopping > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > ------------------------------ Message: 10 Date: Mon, 12 Dec 2005 15:54:53 -0600 From: "Sebree Linda A." Subject: RE: [Histonet] IHC on coverglass To: "Steven Coakley" , Message-ID: Content-Type: text/plain; charset="us-ascii" Only under extreme duress! In our case, we mounted the coverslips on a regular slide and stained it on our automated instrument (Ventana). We weren't very successful but I think it was because these were bone marrow smears that sat around air-dried in a file for who knows how long. We weren't informed that they had not been fixed in any manner and we ended up losing most of the cells. In a best case scenario it might work a whole lot better. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Monday, December 12, 2005 3:43 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC on coverglass Does anyone out there do IHC on coverglass. Appears to be a "tad" tricky. Steve --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Mon, 12 Dec 2005 17:02:23 -0500 From: Caroline Bass Subject: [Histonet] rat spinal cord longitudinal sections To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed Hi Guys, I have a rat that was injected with a virus that produces a fluorescent protein. I would like to scan the entire spinal cord for the signal and thought the best way to do this was to section the spinal cord along the long axis. I don't know if I have the terminology right on this. I think this should be longitudinal sections. The histologists who is to section the tissue says that this is impractical. I know that cross sections (coronal?) are more common, but which method is better when we are simply screening for a visible signal? I am worried that I will have hundreds of cross sections to examine. If there is a published example of this anywhere, even a picture on a website I would love to see it. I have not been able to find anything that is helpful in deciding which route to go. Thanks for you help, Caroline Bass ------------------------------ Message: 12 Date: Mon, 12 Dec 2005 18:20:42 -0500 From: "Monfils, Paul" Subject: RE: [Histonet] rat spinal cord longitudinal sections To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE0171762F@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" Sounds to me like a longitudinal section might be good for your specific purpose. Cross sections are better for studying the morphology of the cord, but if scanning the length of the cord is your objective, and you can sacrifice the specific advantages of cross sections, than that's the way to go. It is impractical to section the entire length of the uncut cord. But the cord could be cut into three or four lengths, mounted one above the other in a single block, and sectioned that way. It would be a fairly large block, but in my lab we frequently section larger ones. If you do go with cross sections, many of these could be mounted in a single block (15 should be no great problem - 3 rows of 5 each). The average clinical lab, trying to produce hundreds of slides a day on a deadline, may not have the time to devote to such special projects. But it is certainly doable. My lab, a core research facility, receives such special requests regularly. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Caroline Bass > Sent: Monday, December 12, 2005 2:02 PM > To: Histonet (E-mail) > Subject: [Histonet] rat spinal cord longitudinal sections > > Hi Guys, > > I have a rat that was injected with a virus that produces a > fluorescent protein. I would like to scan the entire spinal cord for > the signal and thought the best way to do this was to section the > spinal cord along the long axis. I don't know if I have the > terminology right on this. I think this should be longitudinal > sections. The histologists who is to section the tissue says that > this is impractical. I know that cross sections (coronal?) are more > common, but which method is better when we are simply screening for a > visible signal? I am worried that I will have hundreds of cross > sections to examine. > > If there is a published example of this anywhere, even a picture on a > website I would love to see it. I have not been able to find > anything that is helpful in deciding which route to go. > > Thanks for you help, > > Caroline Bass > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 13 Date: Tue, 13 Dec 2005 00:39:11 +0000 From: crs grupo Subject: [Histonet] DAB info disposabel/neutralization/tratment To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hello! I'm doing a research about the DAB and his tratment/neutralization/disposabel... I need some information about DAB and what to do whit it after use. If someone can give me any information, I appreciated. ------------------------------ Message: 14 Date: Tue, 13 Dec 2005 04:51:28 -0800 (PST) From: Rene J Buesa Subject: Re: [Histonet] DAB info disposabel/neutralization/tratment To: crs grupo , Histonet@lists.utsouthwestern.edu Message-ID: <20051213125128.98084.qmail@web61216.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello CRS Group: DAB disposal constitutes an everyday need and there are several approaches, the most common being to treat it with bleach until it decolorizes but this practice is not recommended because the final product of this treatment is not known. DAB can be treat in the following way: 1-Prepare a stock solution (A) of 0.2 M potassium permanganate (=31.6 g/L) and another stock sol. (B) of 2.0 M sulfuric acid (112mL conc./L). 2- Dilute the DAB solution up to a conc. that does not exceeds 0.9 mg/mL 3- To each 10 mL of the DAB sol. add 5 mL of sol. A + 5 mL of sol. B 4- Allow the mixture to react for at least 10 hours after which time it will be non-mutagenic. 5- Add ascorbic acid in powder until the mixture is decolorized. 6- Add sodium bicarbonate to neutralize the solution (test with pH paper). 7- Now you can discard the solution down the drain, provided that your local authorities give their approval, OR you can give your DAB to a waste management company (WMC) but always remembering that you, and not the WMC, will be responsible for the disposed product. In my laboratory we used to give our DAB to a WMC after we treated it with bleach. Hope this will help you! Rene J. crs grupo wrote: Hello! I'm doing a research about the DAB and his tratment/neutralization/disposabel... I need some information about DAB and what to do whit it after use. If someone can give me any information, I appreciated. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping ------------------------------ Message: 15 Date: Tue, 13 Dec 2005 10:42:46 -0500 From: Vernon Dailey Subject: [Histonet] anti-mouse PR primary Ab To: Histonet Message-ID: <439EEBF6.1030900@georgetown.edu> Content-Type: text/plain; charset="iso-8859-1" Hi Histonet, I need to stain FFPE mouse tissue for Progesterone Receptor. I have tried Santa Cruz rabbit polyclonal but it is not showing the type of staining that I would expect. The luminal epithelial cells of normal mouse uterus are negative while the lamina propria stains positive. My understanding is they should both be positive in normal uterus. Also, I can only get the lamina propria staining at 1:50 in the uterus but when I stain tumor tissue at 1:50 I get tremendous amounts of background (tissue not necrotic and I see very nice, clean staining when using Santa Cruz ER Ab at 1:1500) Can anyone recommend another product that has been successful. Many thanks, -Vernon Dailey ------------------------------ Message: 16 Date: Tue, 13 Dec 2005 11:12:49 -0500 From: "Smith, Allen" Subject: RE: [Histonet] Emergency showers in Lab To: "Jill Cox" Cc: histonet@lists.utsouthwestern.edu Message-ID: <5D2189E74151CC42BEC02906BA8996322B9139@exchsrv01.barrynet.barry.edu> Content-Type: text/plain; charset="us-ascii" Forget the regs: think about your own skin! Sulfuric or nitric acid will do serious damage faster than you can undress. If you handle either, you need a shower. If you ever use an open flame, you need a shower. Showers not only extinguish burning clothing; they also cool the skin underneath, thus reducing the severity of the burn. You should also have an eyewash station next to the shower. Keep the water in the eyewash station clean by flushing it once a week. Even if you wear goggles, eyewash stations are handy for washing the face around the goggles after a splash. (Making colloidal iron solutions or opening a hot container of decloaking buffer involves a spatter risk.) My shower and eyewash are next to my fume hood, where I use my nasty stuff. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jill Cox Sent: Monday, December 12, 2005 10:49 AM To: Histonet@Lists. Utsouthwestern. Edu Subject: [Histonet] Emergency showers in Lab Hello Histonetters!! Just wondering if anyone knows about regs for an emergency shower in a Pathology lab? I can't find anything in CAP checklist or OSHA on this. Is there somewhere else I can look? Who all has showers? Thanks in advance, Jill Jill Cox HT (ASCP) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) ------------------------------ Message: 17 Date: Tue, 13 Dec 2005 10:57:01 -0600 From: "Jackie M O'Connor" Subject: [Histonet] FFPE CD31 on mouse To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="us-ascii" I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' ------------------------------ Message: 18 Date: Tue, 13 Dec 2005 09:17:03 -0800 From: "Koelling, Ray" Subject: RE: [Histonet] FFPE CD31 on mouse To: "'Jackie M O'Connor'" , "Histonet (E-mail)" Message-ID: <16834C6DFFA6004C88DE4507FB8AE544018CCA28@wa-mb4-sea.amgen.com> Content-Type: text/plain; charset="iso-8859-1" Jackie, Not too long ago I posted on Histonet our CD31 procedure we've used for a couple years. On mouse tissue using the Pharmingen antibody. Works beautifully but need to enzyme retrieve and amplify signal. Should be in archive. Ray Raymond Koelling Pathology Research scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jackie M O'Connor Sent: Tuesday, December 13, 2005 8:57 AM To: Histonet (E-mail) Subject: [Histonet] FFPE CD31 on mouse I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody that USED TO work on FFPE murine tissue - but with no luck. I've scoured the internet for two days looking for a viable replacement - nada. Does anyone have a method for murine CD31 in FFPE that works without the Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but it didn't work. Boo. Jackie O' _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 13 Dec 2005 10:30:19 -0700 From: Gayle Callis Subject: Protocols that have worked using BD PharminGen monoclonal Re: [Histonet] FFPE CD31 on mouse To: "Jackie M O'Connor" , Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051213102205.01b288c8@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Jackie, Here are two messages copied from Histonet Archives using CD31 BD Pharm MEC 13.3, on FFPE murine tissues with success: 1. I have used a rat anti mouse CD31 from Pharmingen(BD Biosciences).Cat. #553370. I use a rabbit anti rat(mouse adsorbed)secondary-biotinylated,from Vector Labs. For retreival, use pepsin at 37C for 20-40 min. depending on fixation. I used the ABC detection from Vector with DAB. 2. If you are using the DAKO CD31 antibody - it will not react with mouse endothelium. Pharmigen CD31 (MEC13.3) works well on FFPE with Vectastain Elite- ABC followed by NEN TSA-kit. Both work well on FFPE mouse tissues. Barb Wright Jamie Erickson supplied this info to me using BD Pharmingen clone, MEC 13.3, 1.25 ug/ml concentration, using proteinase K digestion on FFPE tissue. There may be more messages in Histonet Archives for this antibody At 09:57 AM 12/13/2005, you wrote: >I've been waiting for Santa Cruz to grow a new goat for the CD31 antibody >that USED TO work on FFPE murine tissue - but with no luck. >I've scoured the internet for two days looking for a viable replacement - >nada. >Does anyone have a method for murine CD31 in FFPE that works without the >Santa Cruz antibody? I've tried the Pharmingen anti-mouse with AR - but >it didn't work. Boo. > >Jackie O' >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) ------------------------------ Message: 20 Date: Tue, 13 Dec 2005 12:30:38 -0500 From: DDDeltour@mar.med.navy.mil Subject: [Histonet] Florida License To: histonet@lists.utsouthwestern.edu Message-ID: <3F500F8B416C554EBB21FF16642F72E959CD8C@marxchg03.mar.med.navy.mil> Content-Type: text/plain Hello All, I am looking to get my Florida License but I am currently living in Virginia. Is there an online course that covers the required HIV class? Can anyone give me any advice how to get this process done without much pain :-) Thanks Douglas D. Deltour HT(ASCP) Histology Technician NMCP Portsmouth Va (757) 953-1525/1523 ------------------------------ Message: 21 Date: Tue, 13 Dec 2005 10:34:11 -0700 From: Gayle Callis Subject: [Histonet] Another Histonet Archive CD31 for murine FFPE tissue To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20051213103225.01b07c10@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed It seems most people use enzyme digestion with FFPE CD31 on mouse, rather than an antigen retrieval i.e. with buffers/HIER. Other message: I just completed a project usind CD31 on FFPE mouse tissue with good results. The supplier of the antibody is Research Diagnostics (973-584-7093; www.researchd.com). I used rat anti-mouse CD31, monoclonal; Catalog# RDI-mCD31abrt. They also suggested zinc fixation, but it worked on formalin fixation. I used DAKO's Prot K to open up the sights. Bob Meyer Pathology Core Facility Northwestern University Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 25, Issue 16 **************************************** CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to it are confidential and intended solely for the use of the individual or entity to whom they are addressed. 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From jtrynak <@t> hotmail.com Wed Dec 14 05:48:45 2005 From: jtrynak <@t> hotmail.com (John Rynak) Date: Wed Dec 14 05:48:56 2005 Subject: [Histonet] Wanted - Consulting MD - Pathologist/Hematologist , Boston Area Message-ID: Consulting MD - Pathologist/Hematologist LMS has developed a cell separation technology that captures fetal nucleated red cells in maternal blood samples. These cells are then used for prenatal genetic screening, as an eventual alternative to amniocentesis. We need a consultant who can enumerate (count) these cells when fixed on slides and comment on their morphology. The cells will have a background of other nucleated cells (WBCs) and a some RBCs. We are generating slides faster than our staff can evaluate and have a backlog of ~75 sides and expect to have 10-15 additional slides per week for evaluation by the consultant. Each slide takes ~1 hr to evaluate. Ideally, the person could work at LMS so that we can learn from their expertise, but this is not essential after the initial slides have been read here. We also will have digital images of the slides with the coordinates of the cells of interest in a software file (Bioview system). This feature allows the Bioview system to find them quickly for evaluation. If interested please forward your CV to [1]hr@livingmicrosystems.com References 1. mailto:hr@livingmicrosystems.com From JEllin <@t> yumaregional.org Wed Dec 14 06:58:27 2005 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Wed Dec 14 06:58:58 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Message-ID: What kind of tissue? The tissue is a gastric biopsy Was it 'dried out' before placing in fixative? No, it is brought over in Formalin and filtered and placed again in formalin. the only time were it would be dryied would be in removal and we have no control over this, but I even doubt that. What is your processing protocol The protocol is has been used for about 25 years and we have not seen this but only 3 times. We used another VIP that was about 14 years old and it is a work horse. The problems before came from someone switching reagents around, and we cured that, the other was we check our alchols with a hydrometer and they were all fine, we even checked our xylene and nothing so we changed the entire machine and it cured it. We have had this proccessor for only a week and when you go and run it it sounds if there is a leak. I found it to have an excessive amount of condensation on the lid. But the question I have is , CAN we reproccess the biopsy Manually of line and if so HOW?????? I agree that we are going to do more harm than good , but my pathologist is insisting that there is a way, So that is why I am asking can we do it, and if so how??? OR better yet if we can not someone give me reasons for why we can not do this. What they are seeing is a glassy look, sort of like if you are caring water. We think it is do to the fact that when you seal the proccessor or Latch it down that we are still getting moisture within the chamber. We can hear a leek, of air escaping the chamber that it is not tight enough. In my humble opinion - reprocessing this small bx will just make it worse. I totally agree!!!!!!!!! Anyone please Help on this one. Jesus Ellin Yuma Regional Medical Center From cgfields <@t> lexhealth.org Wed Dec 14 07:20:36 2005 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Wed Dec 14 07:18:03 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Message-ID: We had a problem with a biopsy being left in the cleaning cycle of the processor and found the next day. It looked like a rock. We put it in Ruffer's Solution. 5% Sodium Carbonate Sol. 0.6 gm Sodium Carbonate to 42 ml. H2O 18 ml. Absolute Ethanol Mix Sod. Carb. Solution and add 18 ml Alcohol mixture Place tissue in solution for 8 hours or longer depending on the size of the tissue. Then proceed to processing tissue through alcohols. The alcohols will refix the tissue. Finish processing and embedding in paraffin. We left the tissue 8-12 hours washed it and reprocessed it on the VIP. It was not totally perfect but readable. I found this procedure on histonet@pathology.swmed.edu. Gayle Callis posted this and she was at Montana State University, Bozeman, MT, 404-994-4705. Good luck, Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 -----Original Message----- From: Jesus Ellin [mailto:JEllin@yumaregional.org] Sent: Tuesday, December 13, 2005 8:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Hello everyone I have a real problem here, We have just accquired a tissue proccesor (VIP 5) and found out the hard way like everything in Histology, there is a problem with the proccessing. Our pathologist are claiming that the small biopsies are coming out cooked, after reviewing the slide myself with the pathologist it is not a cooked looked but rather a glassy look and bubbly. Later found out that the latch of the tissue proccessor is not tight enough and is was allowing moisture in and there was a lot of condensation. I figured out what was wrong but here is the kicker. My Pathologist is wanting to take one of the small biopsies that was proccessed and regress the tissue and then reproccess the small biopsy by hand instead of throwing it back on the tissue proccessor. The biopsy is about .1 mm very tiny. I need all the help that I can get!!!!!! So please fellow histo-netters share the knowledge with me. Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ This email and any files transmitted with it may contain PRIVILEGED or CONFIDENTIAL information and may be read or used only by the intended recipient. If you are not the intended recipient of the email or any of its attachments, please be advised that you have received this email in error and that any use, dissemination, distribution, forwarding, printing, or copying of this email or any attached files is strictly prohibited. If you have received this email in error, please immediately purge it and all attachments and notify the sender by reply email or contact the sender at the number listed above if one is provided. From tracey.couse <@t> ibb.gatech.edu Wed Dec 14 07:37:44 2005 From: tracey.couse <@t> ibb.gatech.edu (Tracey Couse) Date: Wed Dec 14 07:37:54 2005 Subject: [Histonet] re: rat spinal cord longitudinal sections Message-ID: <43A02028.2030701@ibb.gatech.edu> Carolyn, I agree with you. I would section longitudinally so as to limit the section number and time analyzing, especially if this is preliminary work and you are simply screening. I work with graduates students who currently perform similar studies in the sense that they are looking for a fluorescent signal in rat/mouse spinal cord and many of them section longitudinally. Good luck! Tracey -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.2291 From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Caroline Bass > Sent: Monday, December 12, 2005 2:02 PM > To: Histonet (E-mail) > Subject: [Histonet] rat spinal cord longitudinal sections > > Hi Guys, > > I have a rat that was injected with a virus that produces a > fluorescent protein. I would like to scan the entire spinal cord for > the signal and thought the best way to do this was to section the > spinal cord along the long axis. I don't know if I have the > terminology right on this. I think this should be longitudinal > sections. The histologists who is to section the tissue says that > this is impractical. I know that cross sections (coronal?) are more > common, but which method is better when we are simply screening for a > visible signal? I am worried that I will have hundreds of cross > sections to examine. > > If there is a published example of this anywhere, even a picture on a > website I would love to see it. I have not been able to find > anything that is helpful in deciding which route to go. > > Thanks for you help, > > Caroline Bass From germckeon <@t> excite.com Wed Dec 14 08:34:34 2005 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Wed Dec 14 08:34:49 2005 Subject: [Histonet] Myelin in spinal cord Message-ID: <20051214143434.9FB6BB58E6@xprdmxin.myway.com> Hello, I have a problem and am in need of some advice. I am working on spinal cord of zebrafish and will be staining for myelin in fine detail. I have been strongly advised not to use paraffin embedding and so am currently doing gelatin embedded sectioning on a cryostat. Unfortunately this has led to tissue loss due to poor adherence to the slide. I tried superfrost plus and superfrost gold slides with no change. gelatin coating the slides greatly improved this but staining clarity and specifity is very poor. I have tried varying times and even stains (Kultschitsky's verses Weil's) but no improvement. Does anyone have any suggestions on how to improve this or what other other techniques (including possibly other stains) I could try? Thanking you, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From ROrr <@t> enh.org Wed Dec 14 08:44:12 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Dec 14 08:44:23 2005 Subject: [Histonet] IHC controls for Cytology Message-ID: Hi everyone, We have very recently begun to destain pap stained smears as well as unstained cytology smears, and run IHC. Because my Docs are happy with the results, I anticipate receiving more of these types of smears to run. I only have FFPE tissue in stock to use as control, so I give them the slides with the disclaimer that the control has been treated differently than the smear. This seems to be acceptable as long as they can detect some positive stained cells (TTF, PanCk, WT-1 Mart-1 for example). But if the smear is negative, how can I be sure it's a true negative? My antibodies aren't really titered for smears.... I would appreciate some comments from anyone who is running a standardized protocol for smears. Do you have a set of smears you can use as controls? I have access to our molecular lab and there is potential to make my own cell cultured controls, what do you think? Many thanks, Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From wstover <@t> vet.uga.edu Wed Dec 14 08:48:18 2005 From: wstover <@t> vet.uga.edu (Wanda Stover) Date: Wed Dec 14 08:48:26 2005 Subject: [Histonet] Antibody Message-ID: <43A030B2.10506@vet.uga.edu> Does anyone know where I can order a Sarcocystis Neuroma Immunohistochemistry antibody? I have looked all over. Thanks WS \UGA Vet Med From germckeon <@t> excite.com Wed Dec 14 08:49:25 2005 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Wed Dec 14 08:49:33 2005 Subject: [Histonet] Celloidin Embedding Message-ID: <20051214144925.12002109F64@xprdmailfe1.nwk.excite.com> Hello I wonder could I also ask would anyone have a protocol for celloidin embedding? Many thanks in advance for any help, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From cwscouten <@t> myneurolab.com Wed Dec 14 08:52:33 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Wed Dec 14 08:53:17 2005 Subject: [Histonet] Picture Needed Message-ID: <5784D843593D874C93E9BADCB87342AB9169DE@tpiserver03.Coretech-holdings.com> Can any one send a 10 or 20 x picture of classic "swiss cheese" freezing artifact to histonet.org for posting? There ought to be one there, and I have need for comparison ? Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com From la.sebree <@t> hosp.wisc.edu Wed Dec 14 09:08:23 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Wed Dec 14 09:08:39 2005 Subject: [Histonet] IHC controls for Cytology Message-ID: Hi Becky, When we were doing IHC on cytospins and ThinPreps, we used frozen tissue sections that had been acetone fixed the same way as the cytology preps. We were able to optimize our antibodies using the frozen sections and the protocols developed always worked for the cytology preps as well. We figured frozens were a much closer preparation methodology than paraffin sections and our pathologists were happy with the compromise we came up with. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Wednesday, December 14, 2005 8:44 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC controls for Cytology Hi everyone, We have very recently begun to destain pap stained smears as well as unstained cytology smears, and run IHC. Because my Docs are happy with the results, I anticipate receiving more of these types of smears to run. I only have FFPE tissue in stock to use as control, so I give them the slides with the disclaimer that the control has been treated differently than the smear. This seems to be acceptable as long as they can detect some positive stained cells (TTF, PanCk, WT-1 Mart-1 for example). But if the smear is negative, how can I be sure it's a true negative? My antibodies aren't really titered for smears.... I would appreciate some comments from anyone who is running a standardized protocol for smears. Do you have a set of smears you can use as controls? I have access to our molecular lab and there is potential to make my own cell cultured controls, what do you think? Many thanks, Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 14 09:14:07 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Wed Dec 14 09:17:55 2005 Subject: [Histonet] IHC controls for Cytology Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EB23813@elht-exch1.xelht.nhs.uk> Aren't controls supposed to be treated exactly the same as the test? So unless you treat them exactly the same you will never know if you have a false negative and I'm not bright enough to know if you could have false positives too. Hi everyone, We have very recently begun to destain pap stained smears as well as unstained cytology smears, and run IHC. Because my Docs are happy with the results, I anticipate receiving more of these types of smears to run. I only have FFPE tissue in stock to use as control, so I give them the slides with the disclaimer that the control has been treated differently than the smear. This seems to be acceptable as long as they can detect some positive stained cells (TTF, PanCk, WT-1 Mart-1 for example). But if the smear is negative, how can I be sure it's a true negative? My antibodies aren't really titered for smears.... I would appreciate some comments from anyone who is running a standardized protocol for smears. Do you have a set of smears you can use as controls? I have access to our molecular lab and there is potential to make my own cell cultured controls, what do you think? Many thanks, Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Kemlo.Rogerson <@t> elht.nhs.uk Wed Dec 14 09:19:45 2005 From: Kemlo.Rogerson <@t> elht.nhs.uk (Rogerson Kemlo (ELHT) Pathology) Date: Wed Dec 14 09:20:09 2005 Subject: [Histonet] Celloidin Embedding Message-ID: <53A22BBB01D6184EBF7A4DC18A021E9EBBC1C6@elht-exch1.xelht.nhs.uk> Page 92 of 'Histopathologic Technic and Practical Histochemistry' RD Lillie and Harold M Fullmer, McGraw-Hill Inc 1976. ------------------------------------------------------------------------ ---- Hello I wonder could I also ask would anyone have a protocol for celloidin embedding? Many thanks in advance for any help, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jgemmanual <@t> wlgore.com Wed Dec 14 09:24:17 2005 From: jgemmanual <@t> wlgore.com (Jeannie G Emmanuel) Date: Wed Dec 14 09:24:35 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! In-Reply-To: Message-ID: Jesus: I agree with Anne Van Binsbergen re the VIP5. I quote her: "The 'latch' of the VIP cannot be 'loose' - the retort has a safety mechanism - it wont process if it 'leaks' - perhaps the rubber gasket was not snug in its housing (whoever last used/cleaned machine should check)". I just bought one and it works great! "Jesus Ellin" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/13/2005 06:29 PM To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! Hello everyone I have a real problem here, We have just accquired a tissue proccesor (VIP 5) and found out the hard way like everything in Histology, there is a problem with the proccessing. Our pathologist are claiming that the small biopsies are coming out cooked, after reviewing the slide myself with the pathologist it is not a cooked looked but rather a glassy look and bubbly. Later found out that the latch of the tissue proccessor is not tight enough and is was allowing moisture in and there was a lot of condensation. I figured out what was wrong but here is the kicker. My Pathologist is wanting to take one of the small biopsies that was proccessed and regress the tissue and then reproccess the small biopsy by hand instead of throwing it back on the tissue proccessor. The biopsy is about .1 mm very tiny. I need all the help that I can get!!!!!! So please fellow histo-netters share the knowledge with me. Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Dec 14 09:59:29 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Dec 14 10:02:06 2005 Subject: [Histonet] Re: Myelin in spinal cord In-Reply-To: <20051214143434.9FB6BB58E6@xprdmxin.myway.com> References: <20051214143434.9FB6BB58E6@xprdmxin.myway.com> Message-ID: <6.0.0.22.1.20051214085322.01b70db8@gemini.msu.montana.edu> I'm curious as to why paraffin was not advised? Reasons? We had terrible luck with frozen sections and prefer to do paraffin sections for myelin - one can either do a Kluver Barrera luxol fast blue or you can do John Kiernans solochrome cyanin (gives you black myelin instead of blue). There are many new myelin staining technics out there, some are fluorescent and a just published method in Dec issue of J Histochemistry Cytochemistry. Check out this journal for other myelin staining methods too. Molecular Probes has another fluorescent myelin staining kit - go to their website to check on what they offer. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From mcauliff <@t> umdnj.edu Wed Dec 14 10:02:18 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Dec 14 10:05:14 2005 Subject: [Histonet] GLUCOSE TOLERANCE TEST In-Reply-To: <139141F8BAF4A642A945ECC528511AF0012A7B21@kalmb02.kaleidahealth.org> References: <139141F8BAF4A642A945ECC528511AF0012A7B21@kalmb02.kaleidahealth.org> Message-ID: <43A0420A.8070302@umdnj.edu> Hi Annette: If your Resident, and the hospital he/she works for, does not want to get sued for malpractice a trip to the clinical lab or the library is in order. Think about it, the Resident is willing to rely on heresay to treat a patient?? Geoff Featherstone, Annette wrote: >Does anyone out there have a protocol for a glucose tolerance test? I know >this isn't histo but my resident needs a new protocol. >Annette Featherstone HT/MLT > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >histonet-request@lists.utsouthwestern.edu >Sent: Tuesday, December 13, 2005 13:05 >To: histonet@lists.utsouthwestern.edu >Subject: Histonet Digest, Vol 25, Issue 16 > > > > <>_______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > -- > ********************************************** > Geoff McAuliffe, Ph.D. > Neuroscience and Cell Biology > Robert Wood Johnson Medical School > 675 Hoes Lane, Piscataway, NJ 08854 > voice: (732)-235-4583; fax: -4029 > mcauliff@umdnj.edu > ********************************************** From Dorothy.L.Webb <@t> HealthPartners.Com Wed Dec 14 10:17:25 2005 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Wed Dec 14 10:17:35 2005 Subject: [Histonet] basics Message-ID: <0E394B648E5284478A6CCB78E5AFDA2718C51A@hpes1.HealthPartners.int> Out of curiosity over a recent discussion I had, what is everyone using for a control slide for your daily H&E check and how long do you dry your slides before staining and at what temperature? ( not interested in microwave drying, thanks!) ________________________________________ 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 rjbuesa <@t> yahoo.com Wed Dec 14 10:17:35 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 14 10:17:45 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! In-Reply-To: Message-ID: <20051214161735.28204.qmail@web61213.mail.yahoo.com> Hi Jes?s: If you have a new VIP5 that you think has a problem, try to check it out by the Sakura people so you can be sure it works properly. I don't think that the latch does not close properly (which you will have to excuse me for saying this) because VIP5 has a safety mechanism that will not allow to operate with a loose latch. Even if this is the case I don't think that some moisture (just a fraction of the air that may have entered the chamber) can influence a biopsy immersed in almost 4 liters of reagents. The problem most likely was due to the processing protocol and the ratio it has between the dehydtaring and the (antemedium+infiltrating) times which for small biopsies should be small. Finally I don't think that you will considerably improve the quality of the sections you have now by reprocessing that very small biopsy. In truth, there is not such a thing and a sucessesful "refixing" or "reprocessing". I would not jeopardize such a small biopsy with a "reprocessing" it by hand or otherwise. This is my opinion and I just hope it will help you. Rene J. Jesus Ellin wrote: Hello everyone I have a real problem here, We have just accquired a tissue proccesor (VIP 5) and found out the hard way like everything in Histology, there is a problem with the proccessing. Our pathologist are claiming that the small biopsies are coming out cooked, after reviewing the slide myself with the pathologist it is not a cooked looked but rather a glassy look and bubbly. Later found out that the latch of the tissue proccessor is not tight enough and is was allowing moisture in and there was a lot of condensation. I figured out what was wrong but here is the kicker. My Pathologist is wanting to take one of the small biopsies that was proccessed and regress the tissue and then reproccess the small biopsy by hand instead of throwing it back on the tissue proccessor. The biopsy is about .1 mm very tiny. I need all the help that I can get!!!!!! So please fellow histo-netters share the knowledge with me. Jesus Ellin Yuma Regional Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From cindy38017 <@t> aol.com Wed Dec 14 10:32:09 2005 From: cindy38017 <@t> aol.com (cindy38017@aol.com) Date: Wed Dec 14 10:32:29 2005 Subject: [Histonet] Needed-Lab to perform PCR testing for TB on a paraffin block Message-ID: <8C7CEE31056B48F-1E10-36424@mblk-d14.sysops.aol.com> A co-worker and I have been searching for a lab that performs PCR testing for TB on a paraffin block and we haven't had any success. Does anyone out there have an address for a lab that performs this particular test..... From Barry.R.Rittman <@t> uth.tmc.edu Wed Dec 14 10:38:33 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Dec 14 10:38:43 2005 Subject: [Histonet] Celloidin Embedding Message-ID: Gerald Are you sure that you want to embed in celloidin and do you mean celloidin or low viscosity nitrocellulose. Had a lot of experience in the past with these embedding media. Please contact me to discuss your application. Thanks Barry 713 - 500 -4134 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rogerson Kemlo (ELHT) Pathology Sent: Wednesday, December 14, 2005 9:20 AM To: germckeon@excite.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Celloidin Embedding Page 92 of 'Histopathologic Technic and Practical Histochemistry' RD Lillie and Harold M Fullmer, McGraw-Hill Inc 1976. ------------------------------------------------------------------------ ---- Hello I wonder could I also ask would anyone have a protocol for celloidin embedding? Many thanks in advance for any help, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! _______________________________________________ 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 Jackie.O'Connor <@t> abbott.com Wed Dec 14 10:39:39 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Dec 14 10:40:12 2005 Subject: [Histonet] Re: IHC vendor Message-ID: Thanks to everyone who contacted me regarding IHC vendors. I've forwarded the information I've received to the Investigator who will be in touch. Thanks again! Jackie From rjbuesa <@t> yahoo.com Wed Dec 14 10:45:08 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 14 10:45:26 2005 Subject: [Histonet] Myelin in spinal cord In-Reply-To: <20051214143434.9FB6BB58E6@xprdmxin.myway.com> Message-ID: <20051214164508.39672.qmail@web61220.mail.yahoo.com> Hi Gerald: Many years ago I used to work with lizard and frog spinal cords; I used paraffin embedding and there was not a problem. Besides the Luxol Fast Blue-Cresyl Etch Violet there are nomerous other well recognized techniques. I personally have used (and like) Lillie's and Olivecrona's method. I would advise you to change to paraffin embedding. Hope this will help you! Ren? J. "germckeon@excite.com" wrote: Hello, I have a problem and am in need of some advice. I am working on spinal cord of zebrafish and will be staining for myelin in fine detail. I have been strongly advised not to use paraffin embedding and so am currently doing gelatin embedded sectioning on a cryostat. Unfortunately this has led to tissue loss due to poor adherence to the slide. I tried superfrost plus and superfrost gold slides with no change. gelatin coating the slides greatly improved this but staining clarity and specifity is very poor. I have tried varying times and even stains (Kultschitsky's verses Weil's) but no improvement. Does anyone have any suggestions on how to improve this or what other other techniques (including possibly other stains) I could try? Thanking you, Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From MCKEELA <@t> UCMAIL.UC.EDU Wed Dec 14 11:02:53 2005 From: MCKEELA <@t> UCMAIL.UC.EDU (McKee, Lynn (mckeela)) Date: Wed Dec 14 11:03:07 2005 Subject: [Histonet] unsubscribe Message-ID: <9871F0C990DF9B4681F1EB312A4D02F80C6DECCC@ucmail8.ad.uc.edu> From Heather.A.Harper <@t> pcola.med.navy.mil Wed Dec 14 11:05:11 2005 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Wed Dec 14 11:05:38 2005 Subject: [Histonet] Looking for a comfortable grossing chair, ergonomic Message-ID: <807FE48C5A7CC940B973B58D32E701431C0CF79A@nhpens-exch1.pcola.med.navy.mil> Hi everybody, We currently have a pathology chair by MOPEC and we are looking for a new chair and I have been searching but haven't found anything except for the same chair by MOPEC. If you have any links, or names of companies, that make a good comfortable chair for grossing, please let me know. Thank you in advance. Heather A. Harper Supervisor of Histology Naval Hospital of Pensacola, FL From lanbergld <@t> vcu.edu Wed Dec 14 11:15:05 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Wed Dec 14 11:15:17 2005 Subject: [Histonet] Cuting speed and sem-thin plastic sectioning Message-ID: Thanks for the reply. I make my own glass knives (part of the pro tocol for this lab) and I do attach a little boat on the back so that they 990 nm. < I also made a mistake in my initial post, I see. Its a - not 1000. As I said, when I cut on the higher speeds the after staining and under 400x mag - will appear as corrugated. St riped. I am wonder if i try to go back to the 'soft' Spurr mix. LDL From MDiCarlo <@t> KaleidaHealth.Org Wed Dec 14 11:28:34 2005 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Wed Dec 14 11:28:50 2005 Subject: [Histonet] tissue processor Message-ID: <139141F8BAF4A642A945ECC528511AF001F5ED57@kalmb02.kaleidahealth.org> Histonetters/ Vendors, I am in need of a small tissue processor to mostly process large human bones but I need one that will process longer than 24 hours. Is there one out there that you can program to do this? Thanks for any information you have. Peggy DiCarlo HT (ASCP) Orthopedics Bone Lab Buffalo General Hospital 100 High St. Buffalo, NY 14203 716-859-1293 _____ Upgrade Your Email - Click here! CONFIDENTIALITY NOTICE: This email transmission and any documents, files, or previous e-mail messages attached to 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, or a person responsible for delivering it to the intended recipient, you are hereby notified that any further review, disclosure, copying, dissemination, distribution, or use of any of the information contained in or attached to this e-mail transmission is strictly prohibited. If you have received this message in error, please notify the sender immediately by e-mail, discard any paper copies, and delete all electronic files of the message. If you are unable to contact the sender or you are not sure as to whether you are the intended recipient, please e-mail ISTSEC@KaleidaHealth.org or call (716) 859-7777. From jkiernan <@t> uwo.ca Wed Dec 14 11:32:20 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Dec 14 11:32:15 2005 Subject: [Histonet] Myelin in spinal cord References: <20051214143434.9FB6BB58E6@xprdmxin.myway.com> Message-ID: <43A05724.E1361332@uwo.ca> Ordinary myelin stains work well on paraffin sections of formaldehyde-fixed tissue. Are you doing some additional method that works only with cryostat sections? Another factor may be age. If the fishes are very young, they may not yet have myelin sheaths in their central nervous systems. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ "germckeon@excite.com" wrote: > > Hello, > > I have a problem and am in need of some advice. > > I am working on spinal cord of zebrafish and will be staining for > > myelin in fine detail. I have been strongly advised not to use paraffin embedding and so am currently doing gelatin embedded sectioning on a cryostat. Unfortunately this has led to tissue loss due to poor adherence to the slide. I tried superfrost plus and superfrost gold slides with no change. gelatin coating the slides > > greatly improved this but staining clarity and specifity is very poor. I have tried varying times and even stains (Kultschitsky's verses Weil's) but no improvement. > > Does anyone have any suggestions on how to improve this or what other other techniques (including possibly other stains) I could try? > > Thanking you, > > Gerald McKeon > From GDawson <@t> dynacaremilwaukee.com Wed Dec 14 12:25:59 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Wed Dec 14 12:27:59 2005 Subject: [Histonet] IHC controls for Cytology Message-ID: FFPE tissue sections are not appropriate controls for smears. Kemlo is correct, the control tissue/slide must be processed/stained exactly like the test slide in order for it to be valid. I do IHC on smears/PAPs occasionally when absolutely nothing else is available, but I am sure to tell the ordering pathologist that I do not have an appropriate control to run. I routinely do a rapid melanoma procedure on smears so I have a bank of known positive melanoma smears in my freezer, but that is the only one. It is unrealistic to believe that an IHC lab could have appropriate controls for any situaion that may come up, but I can also see how a pathologist needs to use whatever is available in an emergency situation. I believe that many of them will gleen what they can from these smears but not put them in their report since an appropriate control is not available. Then, as with so many other situations, the IHC lab takes one for the team when it comes time to bill. Interesting thread, Glen Dawson IHC Manager Milwaukee, WI -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Wednesday, December 14, 2005 9:14 AM To: Orr, Rebecca; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC controls for Cytology Aren't controls supposed to be treated exactly the same as the test? So unless you treat them exactly the same you will never know if you have a false negative and I'm not bright enough to know if you could have false positives too. Hi everyone, We have very recently begun to destain pap stained smears as well as unstained cytology smears, and run IHC. Because my Docs are happy with the results, I anticipate receiving more of these types of smears to run. I only have FFPE tissue in stock to use as control, so I give them the slides with the disclaimer that the control has been treated differently than the smear. This seems to be acceptable as long as they can detect some positive stained cells (TTF, PanCk, WT-1 Mart-1 for example). But if the smear is negative, how can I be sure it's a true negative? My antibodies aren't really titered for smears.... I would appreciate some comments from anyone who is running a standardized protocol for smears. Do you have a set of smears you can use as controls? I have access to our molecular lab and there is potential to make my own cell cultured controls, what do you think? Many thanks, Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ 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 jqb7 <@t> cdc.gov Wed Dec 14 12:22:08 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Wed Dec 14 12:40:04 2005 Subject: [Histonet] coverslipping stations Message-ID: Hi everyone: Hope you are all ready for the holidays. I need some help, please. Are there any manual coverslipping stations on the market today that do not release xylene fumes into the air? Vendors are welcome to reply as well as techs. Thanks, Jeanine Bartlett, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 (404) 639-3590 From tmmrosla <@t> healtheast.org Wed Dec 14 13:52:23 2005 From: tmmrosla <@t> healtheast.org (Mrosla, Tina M) Date: Wed Dec 14 13:53:28 2005 Subject: [Histonet] non-specific staining IHC Message-ID: <22E6C4C32AFB3E44B7056C7EC3E1F2040531269E@HECLUSTER.HEALTHEAST.LOC> Hello, Our lab has recently noticed non-specific staining on our negative antibodies (mouse and rabbit) since we changed our protocols to comply with CAP guidelines. We apply our harshest pre-treatments to our negative controls, including cell conditioner, protease, blocker, and amplification kits. We have a Ventana BenchMark and use their products. I have never experienced background staining before, so I need some advice. Is this a common problem and can it be fixed? Thank you! Tina Mrosla St. Joseph's Hospital St. Paul, MN The information included in this e-mail message, including any attachments, is intended only for the person or organization to which it is addressed. This e-mail message may contain information that is privileged or confidential. If you receive this e-mail message and are not the intended recipient or responsible for delivering the message to the intended recipient, you may not use, disseminate, distribute or copy the information included in this e-mail and any attachments. If you received this e-mail message by mistake, please reply by e-mail and destroy all copies of this message and any attachments. Thank you. From cbass <@t> bidmc.harvard.edu Wed Dec 14 14:16:23 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Wed Dec 14 14:16:31 2005 Subject: [Histonet] DAPI vs. Hoescht Message-ID: <494AC97F-36FF-456E-ABC5-3010687A606F@bidmc.harvard.edu> Hello, I would like to stain my sections with something to highlight the nucleus. I have heard that both DAPI and Hoescht are good. Is there are reason to use one over the other? I am looking for something very quick, easy and inexpensive. I know that Molecular Probes makes an anti-fade with DAPI, but this is rather expensive and I am worried about not being able to optimize the conditions. I am mainly interested in hearing info on the best formulation to use, a reliable company to purchase from, and any simple protocol you may have on hand. I am staining liver sections, 10 micron, NBF fixed, cryoprotected. Thanks, Caroline Bass From gcallis <@t> montana.edu Wed Dec 14 14:44:55 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Dec 14 14:45:15 2005 Subject: [Histonet] DAPI vs. Hoescht In-Reply-To: <494AC97F-36FF-456E-ABC5-3010687A606F@bidmc.harvard.edu> References: <494AC97F-36FF-456E-ABC5-3010687A606F@bidmc.harvard.edu> Message-ID: <6.0.0.22.1.20051214134046.01b47b08@gemini.msu.montana.edu> You can try Vectashield hard set with DAPI, and not find it so pricey. It also has antifade properties. One thing is to have the correct filter on the microscope for DAPI, and it takes a special laser for confocal. I would vote for the DAPI, it is right in the mounting media and is done in an instant when you mount the coverslip, no optimization is needed. It stains the DNA without problems, and extremely easy. If you have autofluorescence, look into Molecular Probes Image iT signal enhancer. We love the Molecular Probes Prolong Gold antifade with DAPI, hard set - and ignore the expense in favor of excellent results. At 01:16 PM 12/14/2005, you wrote: >Hello, > >I would like to stain my sections with something to highlight the >nucleus. I have heard that both DAPI and Hoescht are good. Is there >are reason to use one over the other? I am looking for something >very quick, easy and inexpensive. I know that Molecular Probes makes >an anti-fade with DAPI, but this is rather expensive and I am worried >about not being able to optimize the conditions. > >I am mainly interested in hearing info on the best formulation to >use, a reliable company to purchase from, and any simple protocol you >may have on hand. > >I am staining liver sections, 10 micron, NBF fixed, cryoprotected. > >Thanks, > >Caroline Bass > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From lanbergld <@t> vcu.edu Wed Dec 14 15:14:13 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Wed Dec 14 15:14:22 2005 Subject: [Histonet] Cutting speed and semi-thin plastic sectioning Message-ID: I'll look these pads up. Who knows, maybe I can convince my P.I. purchase one. Also, if you or anyone else can recommend a book or public appreciated also. I doing this, but the few books really dated and dealt more with stai Thanks for all your kind From lanbergld <@t> vcu.edu Wed Dec 14 17:44:09 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Wed Dec 14 17:44:19 2005 Subject: [Histonet] Cutting speed and semi-thin plastic sectioning Message-ID: Thank you again. The sectioning I'm doing, right now, i microscopy. Reason: Examining the width of a nuclear layer (pho toreceptor nucleii). Maybe that's just microtomy. Eventually I will be prog I think the uniq knows I don't get paid theoretically I could be doi certainly let you know that my mak as you mentioned to do, provides a benef this tomorrow PM. From JWEEMS <@t> sjha.org Wed Dec 14 17:47:19 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Dec 14 17:47:27 2005 Subject: [Histonet] Billing Cytology Special Stains Message-ID: <83AACDB0810528418AA106F9AE9B7F7E013052F9@sjhaexc02.sjha.org> For those of you in Cytology, do you bill a GMS for the ThinPrep as well as the cell block when you only have one specimen? Thanks in advance for your help and Happy Holiday! Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From lillinda70 <@t> aol.com Wed Dec 14 18:03:03 2005 From: lillinda70 <@t> aol.com (lillinda70@aol.com) Date: Wed Dec 14 18:03:17 2005 Subject: [Histonet] histonet-unsubscribe Message-ID: <8C7CF220D782C0E-12AC-2CD6@mblk-d31.sysops.aol.com> From IKirbis <@t> onko-i.si Thu Dec 15 03:32:14 2005 From: IKirbis <@t> onko-i.si (=?ISO-8859-1?Q?Kirbis_Srebotnik_Irena?=) Date: Thu Dec 15 03:32:29 2005 Subject: [Histonet] IHC controls for Cytology Message-ID: I agree with Glen, that control slide should be processed in the same manner as the test slide and this should be a basic rule especially for cytology samples. It's not an easy job to adopt, introduce and organize immuno for cytology samples giving consistent, interpretable and reliable results but it can be done and it is worth to do it. in average for last 10 yrs, we have 3000 immuno reactions per year, 40-60% are QA/QC reactions (positive, negative controls, testings for new markers...) majority of reactions were performed on cytospins prepared from FNAB and effusions, we use home made cell medium for this purpose which allows short term storage of this samples, this cell suspensions can be also used for other diagnostic methods - flow cytometry immunophenotyping,immunocytochemistry, DNA measurements... control slides are prepared in the same way as diagnostic samples from highly cellular diagnostic FNAB samples, effusions, cell cultures, FNAB of resected tumors. immunocytochemistry is done on: - Papanicolaou stained cytospin without destaining - cytoplasmic antigens, not for nuclear and membrane antigens (very convenient for long term storage of positive controls) - cytospins immediately fixed and stored in methanol (up to 1 month) for all antigens including Ki67, TTF, ER, CD's (not possible to prepare positive controls for long term storage for all antigens!) best regards Irena Srebotnik Kirbis, MSc Institute of Oncology Dept. of Cytopathology Zaloska 2 1000 Ljubljana Slovenia phone +386 15879 705 fax +38615879400 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dawson, Glen Sent: Wednesday, December 14, 2005 7:26 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC controls for Cytology FFPE tissue sections are not appropriate controls for smears. Kemlo is correct, the control tissue/slide must be processed/stained exactly like the test slide in order for it to be valid. I do IHC on smears/PAPs occasionally when absolutely nothing else is available, but I am sure to tell the ordering pathologist that I do not have an appropriate control to run. I routinely do a rapid melanoma procedure on smears so I have a bank of known positive melanoma smears in my freezer, but that is the only one. It is unrealistic to believe that an IHC lab could have appropriate controls for any situaion that may come up, but I can also see how a pathologist needs to use whatever is available in an emergency situation. I believe that many of them will gleen what they can from these smears but not put them in their report since an appropriate control is not available. Then, as with so many other situations, the IHC lab takes one for the team when it comes time to bill. Interesting thread, Glen Dawson IHC Manager Milwaukee, WI -----Original Message----- From: Rogerson Kemlo (ELHT) Pathology [mailto:Kemlo.Rogerson@elht.nhs.uk] Sent: Wednesday, December 14, 2005 9:14 AM To: Orr, Rebecca; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] IHC controls for Cytology Aren't controls supposed to be treated exactly the same as the test? So unless you treat them exactly the same you will never know if you have a false negative and I'm not bright enough to know if you could have false positives too. Hi everyone, We have very recently begun to destain pap stained smears as well as unstained cytology smears, and run IHC. Because my Docs are happy with the results, I anticipate receiving more of these types of smears to run. I only have FFPE tissue in stock to use as control, so I give them the slides with the disclaimer that the control has been treated differently than the smear. This seems to be acceptable as long as they can detect some positive stained cells (TTF, PanCk, WT-1 Mart-1 for example). But if the smear is negative, how can I be sure it's a true negative? My antibodies aren't really titered for smears.... I would appreciate some comments from anyone who is running a standardized protocol for smears. Do you have a set of smears you can use as controls? I have access to our molecular lab and there is potential to make my own cell cultured controls, what do you think? Many thanks, Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ 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 wasielewski.reinhard.von <@t> mh-hannover.de Thu Dec 15 06:26:14 2005 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Thu Dec 15 06:26:29 2005 Subject: [Histonet] CD 4, CD8 and C5a in mouse FFPE In-Reply-To: Message-ID: <43A16EF6.13544.297C5B95@localhost> Dear histonetters, this is the second try to get valueable information on how to stain these epitopes in fixed mouse tissue. CD 4 in mouse tissue, FFPE CD 8 in mouse tissue, FFPE C5a in mouse tissue, FFPE Maybe I missed the answers to my first question or something with my server went wrong. Could you give me informations about clones and protocols that work reliably ? Many thanks in advance Reinhard von Wasielewski PD Dr. med. Reinhard von Wasielewski From sharon.willman <@t> bms.com Thu Dec 15 06:35:48 2005 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Thu Dec 15 06:38:10 2005 Subject: [Histonet] Histonet-5-(p-dimethylamino-benthylamino-benzylidine) Message-ID: <43A16324.9070104@bms.com> Hi, We are wanting to purchase 5-(p-dimethylaminobenzylidine) for rhodanine for a copper stain. Does anyone know a vendor that supplies this? Any help would be most appreciated. Thanks, Sharon From akbitting <@t> geisinger.edu Thu Dec 15 07:13:21 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Thu Dec 15 07:14:06 2005 Subject: [Histonet] Acetylcholinesterase special stain controls Message-ID: What is everyone using for ACE controls, other than the obvious positive case blocks? IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From katri <@t> cogeco.ca Thu Dec 15 07:54:06 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Thu Dec 15 07:54:17 2005 Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! References: Message-ID: <00f901c6017f$03d56970$6a9a9618@Katri> Jesus, If the biopsy is unreadable, there is nothing to lose to follow Carole's suggestion. Obviously you'll have to melt the paraffin block and the specimen first, remove the wax with xylene and bring it through to a low cocentration alcohol (50%) before putting it in the sodium carbonate solution. Good luck! Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Carole Fields" To: "'Jesus Ellin'" Cc: Sent: Wednesday, December 14, 2005 8:20 AM Subject: RE: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! > We had a problem with a biopsy being left in the cleaning cycle of the > processor and found the next day. It looked like a rock. We put it in > Ruffer's Solution. > > 5% Sodium Carbonate Sol. > > 0.6 gm Sodium Carbonate to 42 ml. H2O > 18 ml. Absolute Ethanol > > Mix Sod. Carb. Solution and add 18 ml Alcohol mixture > > Place tissue in solution for 8 hours or longer depending on the size of > the > tissue. Then proceed to processing tissue through alcohols. The alcohols > will refix the tissue. Finish processing and embedding in paraffin. > > We left the tissue 8-12 hours washed it and reprocessed it on the VIP. It > was not totally perfect but readable. I found this procedure on > histonet@pathology.swmed.edu. Gayle Callis posted this and she was at > Montana State University, Bozeman, MT, 404-994-4705. > > Good luck, > Carole Fields, HT,ASCP > Pathology Supervisor > Lexington Medical Center > 2720 Sunset Blvd. > W. Columbia, SC 29169 > > > > > > -----Original Message----- > From: Jesus Ellin [mailto:JEllin@yumaregional.org] > Sent: Tuesday, December 13, 2005 8:29 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue Proccessing HELP!!!!!!!!!!!!!! > > > Hello everyone I have a real problem here, We have just accquired a tissue > proccesor (VIP 5) and found out the hard way like everything in Histology, > there is a problem with the proccessing. Our pathologist are claiming > that > the small biopsies are coming out cooked, after reviewing the slide > myself > with the pathologist it is not a cooked looked but rather a glassy look > and > bubbly. Later found out that the latch of the tissue proccessor is not > tight enough and is was allowing moisture in and there was a lot of > condensation. I figured out what was wrong but here is the kicker. > > My Pathologist is wanting to take one of the small biopsies that was > proccessed and regress the tissue and then reproccess the small biopsy by > hand instead of throwing it back on the tissue proccessor. The biopsy is > about .1 mm very tiny. I need all the help that I can get!!!!!! So > please > fellow histo-netters share the knowledge with me. > > > Jesus Ellin > Yuma Regional Medical Center > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________ > This email and any files transmitted with it may contain PRIVILEGED or > CONFIDENTIAL information and may be read or used only by the intended > recipient. If you are not the intended recipient of the email or any of > its > attachments, please be advised that you have received this email in error > and that any use, dissemination, distribution, forwarding, printing, or > copying of this email or any attached files is strictly prohibited. If you > have received this email in error, please immediately purge it and all > attachments and notify the sender by reply email or contact the sender at > the number listed above if one is provided. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dpahisto <@t> yahoo.com Thu Dec 15 07:59:12 2005 From: dpahisto <@t> yahoo.com (Cindy DuBois) Date: Thu Dec 15 07:59:20 2005 Subject: [Histonet] Re: Basics Message-ID: <20051215135912.34240.qmail@web33405.mail.mud.yahoo.com> I have a set of "blocks of all different types of tissue" ie. uterus, cervix, skin, tonsils, colon, just to name a few. I pick out one or two each day and they are cut at the end of the run(by a different tech each day) and stained with the last run. Everyday, we use a different tissue and I try to cover all the basics within the week. We dry our slides on a warming plate @60C for 10 minutes, then place in 60C oven for 20 minutes. I worked at another place that didn't have a warming plate, so we let the slides air dry for 20 minutes then put them in oven for 20 minutes. Cindy DuBois Integrated Pathology Stockton, CA --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From klapka <@t> uni-duesseldorf.de Thu Dec 15 08:07:16 2005 From: klapka <@t> uni-duesseldorf.de (klapka) Date: Thu Dec 15 08:06:41 2005 Subject: [Histonet] Double fluorescence labelling of paraffin sections Message-ID: <43A17894.8000701@uni-duesseldorf.de> Hello, does anyone have any suggestions how to eliminate background fluorescence of paraffin slices? I tried the autofluorescence eliminator, but that wasnt enough. Maybe the deparaffinization is not well enough with Rotihistol? Thanks, Nicole Klapka -- Nicole Klapka PhD Labor f?r Molekulare Neurobiologie TVA, Geb. 22.22 Heinrich-Heine Universit?t D?sseldorf Universit?tsstr. 1 40225 D?sseldorf Tel: ++49 211 81 14437 From JKOBLER <@t> PARTNERS.ORG Thu Dec 15 09:23:21 2005 From: JKOBLER <@t> PARTNERS.ORG (Kobler, James) Date: Thu Dec 15 09:23:32 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 13 Message-ID: Hello Histonet, I have a question regarding registration of paraffin sections with in vivo optical sections. We are doing a study of vocal fold pathology where we need to correlate images acquired with a new optical coherence tomography (OCT) device and images from histological sections. The OCT imaging will be performed on excised specimens (e.g. partial or total laryngectomy specimens) in the operating room via a small catheter that is pressed against the vocal fold tissue. 5mm long OCT sections are acquired that are perpendicular to the tissue surface with an effective depth of about 2 mm. We would like to mark the tissue in a reliable way so that the OCT sections can be aligned with subsequent paraffin sections. India ink works well for tattooing the tissue because it shows up nicely in the OCT sections. We can place 2 small tattoos about 5 mm apart and then take an OCT section that shows these marks at both ends. The problem is maintaining the 3D shape of the specimen through the fixation/embedding process and then orienting the block so that a few sections catch both tattoo marks. The paraffin sections would be cut perpendicular to the surface of the tissue and thus parallel to the tattoo tracks. Routine H&E staining will be used. I would appreciate your ideas/experience regarding tissue processing so that we can best align the 2 types of sections. Thanks very much! Jim Kobler, Department of Surgery, Mass General Hospital, jkobler@partners.org From jkiernan <@t> uwo.ca Thu Dec 15 10:14:07 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Thu Dec 15 10:14:16 2005 Subject: [Histonet] Histonet-5-(p-dimethylamino-benthylamino-benzylidine) References: <43A16324.9070104@bms.com> Message-ID: <43A1964F.86282DA3@uwo.ca> The reagent is 5-(4-dimethylaminobenzylidene)rhodanine. It's in the Sigma catalogue (D-1801) and I'm sure other vendors will also have it. The name may also be written as p-dimethylaminobenzylidenerhodanine. Rhodanine is a different chemical, which wouldn't work (and isn't in the Sigma catalogue). It is wrong to call the histochemical method for copper a "rhodanine stain," even though this phrase is widely used informally and is occasionally seen in print. (Dyes and fluorochromes with "rhodamine" in their names are completely unrelated despite differing by only one letter. ) John Kiernan Anatomy, UWO London, Canada ------------------------ Sharon E Willman wrote: > > Hi, > We are wanting to purchase 5-(p-dimethylaminobenzylidine) for rhodanine > for a copper stain. Does anyone know a vendor that supplies this? Any > help would be most appreciated. > > Thanks, > Sharon > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Dec 15 10:17:33 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 15 10:17:45 2005 Subject: [Histonet] Double fluorescence labelling of paraffin sections In-Reply-To: <43A17894.8000701@uni-duesseldorf.de> References: <43A17894.8000701@uni-duesseldorf.de> Message-ID: <6.0.0.22.1.20051215091644.01b4f900@gemini.msu.montana.edu> Did you try the Molecular Probes autofluorescence blocking reagent? Image-iT FX signal enhancer? At 07:07 AM 12/15/2005, you wrote: >Hello, > >does anyone have any suggestions how to eliminate background fluorescence >of paraffin slices? I tried the autofluorescence eliminator, but that >wasnt enough. >Maybe the deparaffinization is not well enough with Rotihistol? >Thanks, >Nicole Klapka > >-- >Nicole Klapka PhD >Labor f?r Molekulare Neurobiologie >TVA, Geb. 22.22 >Heinrich-Heine Universit?t D?sseldorf >Universit?tsstr. 1 >40225 D?sseldorf >Tel: ++49 211 81 14437 > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From tp2 <@t> medicine.wisc.edu Thu Dec 15 10:33:10 2005 From: tp2 <@t> medicine.wisc.edu (Thomas Pier) Date: Thu Dec 15 10:34:02 2005 Subject: [Histonet] F4/80, IL-12 flourescent Message-ID: Hello, I hope that everyone is doing well. My current project requires me to stain frozen mouse tumors for F4/80 and IL-12. I was wondering if anyone out there has a good protocol for flourescent staining of either of these antibodies. The antibodies that were provided for me are for flow or westerns. They have not given me good results (not that that's a shocker). Buying IHC specific antibodies is probably going to be the next step, so if anybody knows of some good ones please let me know. Thanks alot. Happy Holidays. Tom Pier From choltonzenner <@t> earthlink.net Wed Dec 7 09:19:54 2005 From: choltonzenner <@t> earthlink.net (Claire Holton-Zenner) Date: Thu Dec 15 12:00:22 2005 Subject: [Histonet] Histology Job Opportunity Message-ID: <18334A665430264780053F077275738E0DA905@exchange.unipathllc.corp> On behalf of Bill Chauvin, HT (ASCP) Technical Manager of Histology at UniPath Pathology Laboratory, we are actively searching for experienced Histotechnicians, Histotechnologists with production experience to join our team at our new state of the art laboratory located in Denver, Colorado. We have full-time night and early morning shifts available. We offer competitive pay with a complete benefits package including education assistance, 401(k), Profit Sharing and relocation assistance. Contact Bill Chauvin at 303-512-2243 or email resume to cholton@unipathllc.com. Resumes also accepted via fax at: 303-512-2259. For more information visit our website at www.unipathllc.com. Claire Holton-Zenner, PHR Manager of Human Resources 303.512.2234 - office 303.512.2259 - fax Comments: The information contained in this transmission is intended solely for the addressee(s) named above and is privileged and/or confidential. If the reader of this message is not the intended recipient or the person responsible to deliver it to the intended recipient, you are prohibited from reading or disclosing the information contained in this transmission. Any examination, use, dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by telephone for instructions. From LewisS <@t> pediatrics.ohio-state.edu Fri Dec 9 09:18:34 2005 From: LewisS <@t> pediatrics.ohio-state.edu (Lewis, Sarah) Date: Thu Dec 15 12:00:24 2005 Subject: [Histonet] SDH protocol Message-ID: <714B9F12B4E18C4C843B66E8E190F2AD447631@res2k3ms01.CRII.ORG> Fellow Neuromuscular Labs!! I am in need of a Great SDH protocol. I have been struggling with this stain for a long time. I purchase new NBT and it works pretty well for a few weeks. Then is starts fading into nothing within 1month. I use a protocol of on the WashU website (St. Louis MO). It works better than any other I have tried... but still very pale, However my pathologist said that the SDH's in the past have always been more blue. Mine come out purple/ pink. I have also used the protocol from past histologist in this lab with no luck. I must be missing some small technique that isn't written in the protocol. Please help!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net From lwuerges <@t> rochester.rr.com Mon Dec 12 18:25:00 2005 From: lwuerges <@t> rochester.rr.com (lwuerges) Date: Thu Dec 15 12:00:25 2005 Subject: [Histonet] negative controls with IHC Message-ID: <000801c5ff7b$a823b5a0$566acd45@rochester.rr.com> I have a question regarding negative controls for a manual immunoperoxidase procedure. Do you use a neg control for each antibody used with each block? or one neg control per block regardless of how many antibodies for that block. The latter is what I was told, but the first seems more likely on account of each antibody may be treated differently with some needing antigen retrieval--obviously the contol should receive identical treatment. What is the usual neg control protocol? Thank you. Lisa Wuerges From Clarke.Harding <@t> samc.com Tue Dec 13 09:29:57 2005 From: Clarke.Harding <@t> samc.com (Clarke Harding) Date: Thu Dec 15 12:00:27 2005 Subject: [Histonet] Histo Supervisor position available in central California Message-ID: St Agnes Medical Center is a 400+ bed hospital located in Fresno, California. Our histology dept processes over 20,000 surgicals per year and offers a full complement of special stains, including an extensive library of immunohistochemical stains. We are in the process of installing a Sakura rapid tissue processor, which we anticipate will be on-line by the end of this month, and will soon offer same-day turnaround. Fresno is located in the center of California. There are abundant recreational opportunities in the nearby (1 hr drive) Sierra Nevada mountains, including Yosemite National Park. The Pacific Ocean is a 2.5 hr drive; San Francisco and Los Angeles are 3 hrs away. We need an energetic, experienced individual to lead and grow our histo lab. Interested individuals may contact me by phone or email. Clarke T. Harding, III, M.D. Dept. of Pathology St. Agnes Medical Center 1303 E. Herndon Ave. Fresno, CA 93720 (559) 450 - 3130 (voice) (559) 450 - 2035 (fax) hardcl@samc.com -------------------------------------------------------------- Confidentiality Note: This e-mail is intended only for the person or entity to which it is addressed, and may contain information that is privileged, confidential, or otherwise protected from disclosure. Dissemination, distribution, or copying of this e-mail or the information herein by anyone other than the intended recipient(s) is prohibited. If you have received this e-mail in error, please notify the sender by reply e-mail and destroy the original message and all copies. -------------------------------------------------------------- From Jennifer.Morrison <@t> DrexelMed.edu Thu Dec 15 11:33:46 2005 From: Jennifer.Morrison <@t> DrexelMed.edu (Morrison, Jennifer M) Date: Thu Dec 15 12:00:28 2005 Subject: [Histonet] Job Opportunity in Philadelphia for Histotechnician!!!!!!!!!!!!!!! Message-ID: <22E4FD336D99624A8D0CA6C506CE07C90105FE2E@NCB-SV-003.DUCOM.edu> Histotechnician: Drexel University College of Medicine, Department of Pathology is currently seeking a full-time histotechnician with at least one-year experience to perform the following duties: embedding, microtomy, special staining, and the ability for accurate production of slides. Must be computer literate, and possess excellent organizational/communication skills. HT certification or eligibility required. We offer competitive salary and benefits package including tuition reimbursement. Please fax resume to Jennifer Morrison at 215-893-5534 and apply online at www.drexelmedjobs.com. From PIXLEYSK <@t> UCMAIL.UC.EDU Thu Dec 15 12:30:40 2005 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Thu Dec 15 12:30:46 2005 Subject: [Histonet] RE: DAP vs Hoescht Message-ID: Dear Caroline: We have found that both are very easy to use, but the problem with Hoechst is that it CANNOT be used with an aqueous mounting medium. Both can be purchased as the relatively cheap solutions (i.e., DAPI: Mol. Probes D-1306; Hoechst 33258: Sigma catalog # B2883). Both can be diluted in an aqueous buffer (PB or PBS) and then you incubate your sections for 5 mins, then rinse and coverslip. But DAPI can be used with any aqueous mounting medium, like Gelvatol or Fluoromount, etc., while with Hoechst, you have to dehydrate, go to xylene or its equivalent and mount with a standard mounting medium like Permount or Depex. Hope this helps. Sarah Pixley From Vfierke <@t> SNBLUSA.com Thu Dec 15 13:06:55 2005 From: Vfierke <@t> SNBLUSA.com (Vaughn Fierke) Date: Thu Dec 15 13:10:29 2005 Subject: [Histonet] adjusting pH for formalin Message-ID: <3CB1E2E5EC8E9C48A06C8E0967EB82B20E5CB6@MAIL01.snblusa.com> Anyone with an idea to bring up the pH of 10% formalin to 7.0. Our recycler tests 5 gallon batches at 6.2 and the paths would like everything at 7.0. One source says add 1% sodium phosphate dibasic; others say use SP dibasic saturated solution. Another source says that using 1% sodium hydroxide is ok and won't alter fixation. Any consistent suggestions? From rjbuesa <@t> yahoo.com Thu Dec 15 14:10:05 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 15 14:10:15 2005 Subject: [Histonet] negative controls with IHC In-Reply-To: <000801c5ff7b$a823b5a0$566acd45@rochester.rr.com> Message-ID: <20051215201005.94608.qmail@web61216.mail.yahoo.com> Hi Lisa: I used to run 1 negative control per block. The rationale was that you want to find out how the tissue itself reacts to those "non-antibody" components of the procedure. What I also did was to run 1 negative with "non-immune rabbit IgG" and another for mouse IgG (when I had polyclonal and monoclonal antibodies in the same run). Hope this will help you! Ren? J. lwuerges wrote: I have a question regarding negative controls for a manual immunoperoxidase procedure. Do you use a neg control for each antibody used with each block? or one neg control per block regardless of how many antibodies for that block. The latter is what I was told, but the first seems more likely on account of each antibody may be treated differently with some needing antigen retrieval--obviously the contol should receive identical treatment. What is the usual neg control protocol? Thank you. Lisa Wuerges _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From dellav <@t> musc.edu Thu Dec 15 14:08:53 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Thu Dec 15 14:10:31 2005 Subject: [Histonet] histology teaching position available Message-ID: Come join the excitement at the MUSC Medical Center in Charleston, South Carolina. Explore employment opportunities in our state of the art facility as you explore our city. Visitors travel to Charleston each year to walk down the cobblestone streets, relive its history and enjoy the beaches. Experienced instructor wanted for newly accredited, 12 month, post-baccalaureate certificate Histotechnology Program (HTL). Details about our program may be found at www.musc.edu/histoprogram The successful candidate will possess a bachelor of science degree in biology, medical technology or other life science and at least five years of clinical histology experience with a strong emphasis in special stains and immunohistochemistry. A Master's degree in science, education or healthcare administration or equivalent is preferred. Certification in histotechnology, with the American Society of Clinical Pathologists (ASCP) at the technologist level (HTL) is required. Certification at the technician level may be substituted with ten or more years of clinical histology experience that include emphasis in the areas of specialization indicated above. Prior teaching experience is strongly preferred. Applicants will be required to conduct a formal presentation to the selection committee at interview. Interested applicants are invited to contact Vinnie Della Speranza at dellav@musc.edu Discover the excellent salaries, competitive benefits, tuition reimbursement and flexible schedules that only a true leader in the health care field can provide. From rjbuesa <@t> yahoo.com Thu Dec 15 14:18:51 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 15 14:19:00 2005 Subject: [Histonet] adjusting pH for formalin In-Reply-To: <3CB1E2E5EC8E9C48A06C8E0967EB82B20E5CB6@MAIL01.snblusa.com> Message-ID: <20051215201851.9211.qmail@web61212.mail.yahoo.com> Any strong base (like sodium hydroxide) could be used. The reaction will increase the pH value and sodium will end as a sodium salt with no effects on fixation. Another solution would be to use phosphate salts to reconstitute the "classical" 10% NBF (neutral buffered formalin). Ren? J. Vaughn Fierke wrote: Anyone with an idea to bring up the pH of 10% formalin to 7.0. Our recycler tests 5 gallon batches at 6.2 and the paths would like everything at 7.0. One source says add 1% sodium phosphate dibasic; others say use SP dibasic saturated solution. Another source says that using 1% sodium hydroxide is ok and won't alter fixation. Any consistent suggestions? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From jfish <@t> gladstone.ucsf.edu Thu Dec 15 14:27:01 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Thu Dec 15 14:27:15 2005 Subject: [Histonet] (no subject) Message-ID: <000001c601b5$e7dc9860$8903010a@JFISH> Hi histonetters, I have a question for you about muscle processing for paraffin embedding. Starting from fixation to embedding, can you tell me the best and most successful way to do this? I have processed leg muscle (gastroc and soleus) after initial overnight fixation in 3% paraformaldehyde using the following schedule on our Excelsior: Alcohols 1 through 6 45 minutes each Xylenes substitute 1 through 3 1 hour each 1st Paraffin @ 62 degrees 1.5 hours 2nd Paraffin 1.5 hours 3rd Paraffin 2 hours I know the paraffin sounds long, but we usually process fatty tissue and brain that seems to require a longer infiltration time. What do you think, am I missing the mark? Our muscle seems to have a little more space surrounding the bundles than the investigator had at their old institute. I really appreciate any help you can give. Take care, Jo Dee Fish Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-2567 Fax: (415) 355-0230 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From TJJ <@t> Stowers-Institute.org Thu Dec 15 14:33:05 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Dec 15 14:33:26 2005 Subject: [Histonet] RE: Double fluorescence labelling of paraffin sections Message-ID: Nicole, Some authors have had some success using Sudan Black B to "quench" autofluorescence. The Journal of Histochemistry and cytochemistry has some articles on this if you do a PubMed search, you can find the references (sorry, I'm having a busy day, so I can't provide them at the moment). Our imaging department also offers the availability of using spectral imaging techniques. If you're fortunate enough to have good (read: expensive) equipment, you can possibly use imaging to subtract the autofluorescence. Most of those types of techniques require microscopes hooked up to computers to provide these techniques. You may also have some luck switching to a different fluorophore that won't show autofluorescence in the tissues as easily as the FITC or green channels do. Good luck! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From Vickroy.Jim <@t> mhsil.com Thu Dec 15 14:45:43 2005 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Thu Dec 15 14:46:35 2005 Subject: [Histonet] USE OF AGAR AS A MWTHOD TO ORIENT SMALL SPECIMENS Message-ID: I have a new pathology assistant that want's to sue some agar to help orient her small specimens so that the orientation is done at the grossing station and not the embedding center. Does anybody have experience with this and if you do can you send me a sample procedure. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 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 Vickroy.Jim <@t> mhsil.com Thu Dec 15 14:50:00 2005 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Thu Dec 15 14:51:18 2005 Subject: [Histonet] method of using agar for an embedding tool Message-ID: Has anyone used agar at the embedding stations to help orient small specimens? I am told by my new pathology assistant that this would prevent the embedders from having to reorient them at the embedding center? If you have could someone send me a procedure? Please disregard the spelling errors in the last email. Obviously spell check should have been used. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 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 DeBrosse_Beatrice <@t> Allergan.com Thu Dec 15 14:53:55 2005 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Thu Dec 15 14:54:36 2005 Subject: [Histonet] SDH protocol Message-ID: Sarah, Give this procedure a try. It worked very well for us when I worked at Children's Hospital in Cincinnati. We purchased the substrate from SIGMA and used it, until it was gone. The reaction should be a deep blue. Hope it works for you! Sincerely, Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lewis, Sarah Sent: Friday, December 09, 2005 7:19 AM To: HISTONET (E-mail) Subject: [Histonet] SDH protocol Fellow Neuromuscular Labs!! I am in need of a Great SDH protocol. I have been struggling with this stain for a long time. I purchase new NBT and it works pretty well for a few weeks. Then is starts fading into nothing within 1month. I use a protocol of on the WashU website (St. Louis MO). It works better than any other I have tried... but still very pale, However my pathologist said that the SDH's in the past have always been more blue. Mine come out purple/ pink. I have also used the protocol from past histologist in this lab with no luck. I must be missing some small technique that isn't written in the protocol. Please help!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DeBrosse_Beatrice <@t> Allergan.com Thu Dec 15 14:59:12 2005 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Thu Dec 15 14:59:43 2005 Subject: [Histonet] SDH protocol Message-ID: I guess the attachment didn't come through..........Here is the procedure: SUCCINIC ACID DEHYDROGENASE (SDH) Solutions: Prepare this substrate solution at least on e hour before incubating and preheat in 37?C. 0.1M Sodium succinate 15ml 0.1M Phosphate buffer pH 7.6 15ml Nitro Blue Tetrazolium (refrigerated) 15mg Method: 1. Frozen sections are cut 6 ? 8 microns and stay in the cryostat until ready to Incubate. 2. Place slides in coplin jar and incubate for 30 minutes at 37?C. 3. Rinse slides with distilled water and coverslip from water with Aqua mount. Ref. Histochemistry, A.G. Parse, page 910 Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Lewis, Sarah Sent: Friday, December 09, 2005 7:19 AM To: HISTONET (E-mail) Subject: [Histonet] SDH protocol Fellow Neuromuscular Labs!! I am in need of a Great SDH protocol. I have been struggling with this stain for a long time. I purchase new NBT and it works pretty well for a few weeks. Then is starts fading into nothing within 1month. I use a protocol of on the WashU website (St. Louis MO). It works better than any other I have tried... but still very pale, However my pathologist said that the SDH's in the past have always been more blue. Mine come out purple/ pink. I have also used the protocol from past histologist in this lab with no luck. I must be missing some small technique that isn't written in the protocol. Please help!!! Sarah E. Lewis Histotechnician Childrens Research Center for Gene Therapy 700 Childrens Dr Rm WA3112 Columbus OH 43205 (614)-722-2204 LewisS@ccri.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Dec 15 15:10:11 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Dec 15 15:10:39 2005 Subject: [Histonet] USE OF AGAR AS A MWTHOD TO ORIENT SMALL SPECIMENS Message-ID: My personal experience is that if the sample is badly oriented in the agar, you have no recourse to fix it. I don't like using agar. A former lab used it for endocervical currettings, but the currettings were suspended in the agar - no way to get to them unless you serial sectioned the whole lump of agar. Leave the orientation to the histotechs. "Vickroy, Jim" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/15/2005 02:45 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] USE OF AGAR AS A MWTHOD TO ORIENT SMALL SPECIMENS I have a new pathology assistant that want's to sue some agar to help orient her small specimens so that the orientation is done at the grossing station and not the embedding center. Does anybody have experience with this and if you do can you send me a sample procedure. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 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 PMonfils <@t> Lifespan.org Thu Dec 15 15:11:26 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Dec 15 15:11:35 2005 Subject: [Histonet] adjusting pH for formalin Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717633@lsexch.lsmaster.lifespan.org> You recycle formalin? I have heard of recycling solvents but how is formalin recycled? Is it simply filtered? Seems like that would allow a lot of dissolved contaminants to remain. Is it distilled? Seems like that would remove formaldehyde. Also, how does one deal with the fact that formaldehyde is removed from the solution whenever anything is fixed in it, so that used 10% formalin is no longer 10%? Do they add new formaldehyde? Would appreciate some information on this. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Vaughn Fierke > Sent: Thursday, December 15, 2005 11:06 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] adjusting pH for formalin > > > > Anyone with an idea to bring up the pH of 10% formalin to 7.0. Our > recycler tests 5 gallon batches at 6.2 and the paths would like > everything at 7.0. One source says add 1% sodium phosphate dibasic; > others say use SP dibasic saturated solution. Another source says that > using 1% sodium hydroxide is ok and won't alter fixation. Any consistent > suggestions? > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From jamie.erickson <@t> abbott.com Thu Dec 15 15:13:29 2005 From: jamie.erickson <@t> abbott.com (Jamie E Erickson) Date: Thu Dec 15 15:30:25 2005 Subject: [Histonet] Re: CD4 and CD8 mouse FFPE tissues Message-ID: HI Reinhard, I have tried many and I repeat many times to get CD4 and CD8 to work in FFPE tissues mostly with the purified monoclonals antibody from BD biosciences and have failed. I have been doing mouse IHC for 12 years and have come to the conclusion after many try's and conversations with others in the field (Gayle Callis and others) that for CD4 and CD8 just doesn't work for mouse FFPE IHC. We do research and I'm going to start doing more frozen mouse work. Frozens are faster and you don't have to beat your head against the wall to think of more antigen retrieval to try. This may not be everyone's experience but for me I've had enough FFPE CD4/CD8 and others.. That's my opinion for what it worth. _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com From gcallis <@t> montana.edu Thu Dec 15 15:36:47 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 15 15:37:07 2005 Subject: [Histonet] USE OF AGAR AS A MWTHOD TO ORIENT SMALL SPECIMENS In-Reply-To: References: Message-ID: <6.0.0.22.1.20051215142759.01b28e18@gemini.msu.montana.edu> Jackie makes an excellent point. In addition to letting the histotechs orient small samples at embedding, provide your techs with good magnifiers to see the samples - they come attached to embedding centers which tend to get in the way at times so we prefer to wear magnifying glasses instead. Safety glasses are available with bifocal magnifiers built right into lenses, and at different magnifications and you simply wear them OVER your prescription glasses. There are flip up magnifiers that attach to prescription glasses, but if safety glasses are required, the flip models are probably inappropriate for safety regulations. WE have 1.5 and 3X and love them. At 02:10 PM 12/15/2005, you wrote: >My personal experience is that if the sample is badly oriented in the >agar, you have no recourse to fix it. >I don't like using agar. A former lab used it for endocervical >currettings, but the currettings were suspended in the agar - no way to >get to them unless you serial sectioned the whole lump of agar. Leave the >orientation to the histotechs. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From anh2006 <@t> med.cornell.edu Thu Dec 15 15:58:46 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Thu Dec 15 15:58:59 2005 Subject: [Histonet] RE: DAP vs Hoescht In-Reply-To: References: Message-ID: Dear Sarah, I must respectfully disagree. I use Hoechst 33342 diluted to 0.1ug/ml for 10 minutes on a daily basis with various aqueous mountants including Pro-Long Gold (I agree with Gayle that it is a good one), Slow Fade, Vectashield, Fluormount-G, Gelmount etc etc. Works like a charm, never a problem, gorgeous counterstain Sincerely, Andrea At 1:30 PM -0500 12/15/05, Pixley, Sarah (pixleysk) wrote: >Dear Caroline: > >We have found that both are very easy to use, but the problem with >Hoechst is that it CANNOT be used with an aqueous mounting medium. Both >can be purchased as the relatively cheap solutions (i.e., DAPI: Mol. >Probes D-1306; Hoechst 33258: Sigma catalog # B2883). Both can be >diluted in an aqueous buffer (PB or PBS) and then you incubate your >sections for 5 mins, then rinse and coverslip. But DAPI can be used with >any aqueous mounting medium, like Gelvatol or Fluoromount, etc., while >with Hoechst, you have to dehydrate, go to xylene or its equivalent and >mount with a standard mounting medium like Permount or Depex. >Hope this helps. >Sarah Pixley > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From AFoshey <@t> chw.org Thu Dec 15 16:23:00 2005 From: AFoshey <@t> chw.org (Foshey, Annette) Date: Thu Dec 15 16:23:13 2005 Subject: [Histonet] Powerpath Users - Cassette labelers that interface with Powerpath. Message-ID: <9E6D52F532809247BDA1783680E92C56586867@CHWEXC.chwi.chswi.org> We are evaluating the Shurmark TBS cassette labeler, the IPC Leica, and the Thermo Shandon Microwriter to interface with Powerpath. We don't have enough time to demo the units. Please give me your comments on the units both pro and con. Thank you, Annette Foshey Team Leader in Histology Children's Hospital of Wisconsin P.O. Box 1997, M.S. 701 Milwaukee, WI 53201 (414) 266-2524 ************************** Children's Hospital and Health System recognizes that unencrypted e-mail is insecure and does not guarantee confidentiality. The confidentiality of replies to this message cannot be guaranteed unless the replies are encrypted. From jfish <@t> gladstone.ucsf.edu Thu Dec 15 16:24:19 2005 From: jfish <@t> gladstone.ucsf.edu (Jo Dee Fish) Date: Thu Dec 15 16:24:31 2005 Subject: [Histonet] Paraffin processing of muscle Message-ID: <000d01c601c6$4a983800$8903010a@JFISH> Hi histonetters, Sorry, I forgot to put anything in the subject with my first post, so I am resubmitting it with a subject line! I have a question for you about muscle processing for paraffin embedding. Starting from fixation to embedding, can you tell me the best and most successful way to do this? I have processed leg muscle (gastroc and soleus) after initial overnight fixation in 3% paraformaldehyde using the following schedule on our Excelsior: Alcohols 1 through 6 45 minutes each Xylenes substitute 1 through 3 1 hour each 1st Paraffin @ 62 degrees 1.5 hours 2nd Paraffin 1.5 hours 3rd Paraffin 2 hours I know the paraffin sounds long, but we usually process fatty tissue and brain that seems to require a longer infiltration time. What do you think, am I missing the mark? Our muscle seems to have a little more space surrounding the bundles than the investigator had at their old institute. I really appreciate any help you can give. Take care, Jo Dee Fish Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease Telephone: (415) 734-2567 Fax: (415) 355-0230 E-mail: jfish@gladstone.ucsf.edu Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158 From Malcolm.McCallum <@t> tamut.edu Thu Dec 15 18:40:20 2005 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Thu Dec 15 18:43:47 2005 Subject: [Histonet] FW: Anti- Endnagered Specis Bill Introduced into Senate Message-ID: From: PARC on behalf of Allen Salzberg Sent: Thu 12/15/2005 2:30 PM To: PARC@LISTSERV.UGA.EDU Subject: Anti- Endnagered Specis Bill Introduced into Senate Senator Crapo (R-ID) Introduces Bill to Undermine Endangered Species Act Summary from HerpDigest editor - Senator Crapo is sttempting to have his version of Rep. Pombo's, endangered species bill passed by the Senate. Pombo's bill passed the House, it is a bill that every environmental organization agrees only purpose is to eviscerate the esa. So does the Senator's, which is basically the same bill. The Senator is attempting to do this by avoiding any real debate on his bill, and so possible changes as a result of the debate. Debate that would occcur in the Fisheries, Wildlife, and Water Subcommittee, under the leadership of Sen. Lincoln Chafee (R-RI). The committee is already considering ESA reauthorization of its own, but has committed to developing such legislation only after gathering adequate information, and hearing from agencies, experts and stakeholders. The Senator is obviously scared of that gathering of this informaton. So the introduction of the Crapo bill is clearly a rush to purposely sidestep that deliberative process. Links to text of bill and an overview of the bill is enclosed in the following press release supplied by the Center for Biological Diversity. WASHINGTON, DC-Thursday, December 15, 2005-Senator Mike Crapo (R-ID) introduced a bill today aimed at undermining protections for endangered species. The Senate bill, S.2110, cynically titled the "Collaboration and Recovery of Endangered Species Act," would create arbitrary roadblocks to protecting endangered species and habitat, greatly increase delays and political manipulation in conservation decisions, and cuts federal oversight of projects that threaten endangered species. (Overview of the Crapo bill follows below. The text of the bill is available at www.biologicaldiversity.org) The Crapo bill pays lip service to encouraging landowners to conserve endangered species on private land, and idea long supported by conservation organizations. However, the Crapo bill focuses on giving large tax breaks to large-scale land developers and eliminating habitat protections rather than encouraging or enabling conservation on private land. "Senator Crapo's proposal alone would be terrible for endangered species conservation," said Melissa Waage, legislative advocate for the Center for Biological Diversity. "But the bill introduced today is part of an even bigger plan to gut the Endangered Species Act by teaming up with Rep. Pombo to adopt the worst provisions of Pombo's House bill behind closed doors." On September 29, the House passed HR 3824 by Rep. Pombo (R-CA)-a bill that would repeal entire sections of the Endangered Species Act. (A detailed analysis and the text of the Pombo bill are available at www.biologicaldiversity.org.) ESA bills that pass the Senate this year would be referred to a conference committee to be merged with the Pombo bill from the House. The two leaders of such a conference committee would be Rep. Pombo and Senator Inhofe (R-OK), who has an environmental voting score of 0 according to the League of Conservation Voters. Crapo told E&E TV on October 6: "I think the House [Pombo] bill is a very good bill and although we may not be able to get the necessary 60 votes for every part of the House bill, and I don't know that yet, that doesn't, that wouldn't change my support for the whole bill as is. I mean it's a good bill [the Pombo bill], but my objective here is to make sure that we get a bill that has as much of those reforms that the House [Pombo bill] has and maybe even some more, that we can get consensus on, through the Senate." Crapo has an environmental voting score of 0 according to the League of Conservation Voters. "Crapo has sponsored a poorly written bill with the worst intentions and terrible implications for wildlife," said Kieran Suckling, policy director of the Center for Biological Diversity. "The Endangered Species Act is the safety net for America's imperiled plants and animals. This bill rips down endangered species protections and creates road blocks to endangered species recovery." The Fisheries, Wildlife, and Water Subcommittee, under the leadership of Sen. Lincoln Chafee (R-RI), is considering ESA reauthorization of its own, but has committed to developing such legislation only after gathering adequate information, and hearing from agencies, experts and stakeholders. The introduction of the Crapo bill today appears to be a rush to purposely sidestep that deliberative process. The Endangered Species Act protects 1,300 of America's most endangered plants and animals. Originally created in 1973, it has a saved over 99% of these species from extinction including the Bald Eagle, Grizzly Bear, Gray Wolf, Sea Otter, and Grizzly Bear. Overview of Crapo bill follows. Overview of S.2110, the "Collaboration and Recovery of Endangered Species Act" Introduced by Senator Crapo (R-ID) Thursday, December 15, 2005 Creates a Habitat Conservation Shell Game (page 36-41) The Crapo bill would create an ill-defined conservation bank for developers to buy and sell credits for destroying endangered species habitat. This would allow developers to extensively destroy the habitat for one species in exchange for habitat credits for another species or by purchasing habitat that could never be developed in the first place. There are numerous ways that this would lead to a great loss of habitat and no gain or improvement of any habitat at all. Delays Protections for Species and Habitat (pages 10-14) The Crapo bill would delay for three years the listing of imperiled species as threatened and endangered. Current law requires a final listing rule within 12 months of a proposed listing; the Crapo bill would extend that delay to 3 years. The Crapo bill would delay critical habitat designation until three years after the adoption of a recovery plan. Current law requires critical habitat designated immediately after listing; the Crapo bill would extend that delay to 3 years after adoption of a recovery plan. The Crapo bill sets no deadline for recovery plans. Undermines Recovery Plans (pages 21-28) The Crapo bill would create a new convoluted recovery planning process that allows industry to rewrite and overrule the decisions of wildlife experts. A newly created "executive committee" made up of industry interests would make final edits and revisions to the recovery plan developed by scientists and agency biologists. Furthermore, the Crapo bill explicitly makes recovery plans "non-binding and advisory." Creates Roadblocks to Listing Endangered Species (pages 16-18) The Crapo bill would create an ambiguous priority system for listing endangered species that includes industry interests. Current law requires endangered species listings to be based solely on the biological needs of the species. Eliminates Federal Oversight of Endangered Species (page 15) The Crapo bill would require Fish and Wildlife Service to provide a "provisional permit" for any project on private property (except for "ground clearing") if there is no recovery plan in place. The permit would remain in effect until a habitat conservation plan is approved. This would allow activities like mining and logging in endangered species habitat to proceed indefinitely with no federal oversight. Restricts Wildlife Agencies from Improving Conservation Agreements (pages 50-53) The Crapo bill would take "No Surprises"-a highly controversial administrative regulation-and make it law. The Fish and Wildlife Service would be unable to update or revoke a permit (HCP) that authorizes harm to an endangered species, even if new information indicates that the original plan was inadequate and even if it is causing the extinction of the species. Pays Off Developers to Not Violate the Law (page 56) The Crapo bill would create tax breaks to compensate private landowners for conservation work done on private property. However, the Crapo bill fails to limit these tax breaks to landowners who engage in active conservation-the creation or enhancement of endangered species habitat. Therefore, land developers who are required to set aside some portion of their land from development would also be eligible for these tax breaks. That is, instead of paying private landowners to create new habitat, the Crapo bill would primarily be paying developers to comply with the law, creating no new habitat. ******* PRESS RELEASE Center for Biological Diversity December 15, 2005 Contact: Melissa Waage mwaage@biologicaldiversity.org 202-736-5760 or Brian Nowicki bnowicki@biologicaldiversity.org 520-623-5252 x311 Allen Salzberg HerpDigest.org Subscribe now To The Only E-zine That Reports on The Latest News on Herpetological Conservation and Science (Two issues a week). www.herpdigest.org HerpArts.com Gifts for Herp Lovers: Reptile and Amphibian Jewelry, Art, Toys for Adults And Kids, CDs, Candles and More www.herparts.com * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Partners in Amphibian and Reptile Conservation www.parcplace.org * Replies to this message will go only to the original sender unless the default option is changed. * Need to unsubscribe? 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Click on the following web link: http://listserv.uga.edu From Tony_Reilly <@t> health.qld.gov.au Thu Dec 15 20:23:48 2005 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Thu Dec 15 21:15:17 2005 Subject: [Histonet] method of using agar for an embedding tool Message-ID: Hi Jim We have employed this technique on occasions when the clinician has sent endoscopic biopsies with 10 or more pieces, which makes it very difficult for the histologist to embed them all on edge before the wax solidifies. We get our micro dept to provide us with neutral agar in 5ml aliquots because the continual heating and cooling of the agar affects its viability and therefore the smaller aliquots means less wastage. 1. Melt the agar in microwave or waterbath. 2.Pippette onto a glass slide. The agar will spread but the viscosity prevents it from becoming too thin. 3. As agar cools orientate your tissue as desired. 4.Allow to cool on the bench or in refrigerator. be careful if using the refrigerator that the specimen is not forgotten. 5.When solid lift from slide using an old microtome bade or razor blade. 6.Wrap in paper( whatever you use) place into cassette, place casette in formalin. the formalin will help to harden the agar even further. 7. Process overnight as the agar will not process on a short cycle. Note it is important that the tissue is well fixed prior to placing into the agar or else you may get some heat artifact. as I said we only use this for those specimens where normal embedding is made difficult by the number of pieces of tissue in the biopsy. I would not recommend it as a routine method. regards Tony Reilly Chief Scientist Anatomical Pathology QHPS-Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au >>> "Vickroy, Jim" 12/16/05 6:50 am >>> Has anyone used agar at the embedding stations to help orient small specimens? I am told by my new pathology assistant that this would prevent the embedders from having to reorient them at the embedding center? If you have could someone send me a procedure? Please disregard the spelling errors in the last email. Obviously spell check should have been used. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 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 ***************************************************************** 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. 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Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. **************************************************************** From MVaughan4 <@t> ucok.edu Thu Dec 15 22:05:15 2005 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Thu Dec 15 22:05:44 2005 Subject: [Histonet] DAPI vs Hoechst Message-ID: I have used both DAPI and Hoechst, they excitation/emission (360/470). If you buy the DAPI chemi stain with it at the same time as the secondary antibody, and should use it at a very low conc. (about 1:10,000, I think), so your st Mel From arunams <@t> interchange.ubc.ca Fri Dec 16 01:36:17 2005 From: arunams <@t> interchange.ubc.ca (arunams) Date: Fri Dec 16 01:36:28 2005 Subject: [Histonet] Histology Technologist job opening in Vancouver, BC Canada Message-ID: <26942746.1134718577857.JavaMail.myubc2@portal9.itservices.ubc.ca> Wax-it Histology Services Inc. Wax-it Histology Services is a fast growing Vancouver based contract research organization that is providing histology related solutions to academic institutes, hospitals, biotechnology and pharmaceutical industries. We are specialized in both animal and plant tissue related solutions. Wax-it Histology Services has an opening for a histotechnologist to assist in day-to-day operations of the lab. Major Responsibilities The successful incumbent will perform routine histology methods such as grossing tissue, processing, sectioning, cryo processing and sectioning, immunohistochemistry and special histological stains. The position also involves the evaluation and development of new technologies and includes routine QC testing. Emphasis will be placed on the maintenance of neat, accurate and precise documentation of all work performed. Other research and general lab duties will be performed as required. Requirements The successful candidate will have at least a BSc and 2-4 years relevant work experience, with basic knowledge of standard laboratory procedures including lab safety. They should have strong organizational, multi-tasking and time management abilities. Further to that the candidate will have good problem solving skills and posses well-developed communication skills. The candidate must also be competent and self motivated and be able to integrate into a fast growing and dynamic company with the ability to work independently or collaboratively in a team environment. Your resume must clearly demonstrate that you meet all of the above qualifications and preferences. We thank all applicants for their interest, but only those candidates short-listed will be contacted for an interview. How to Apply: Please send your resume, cover letter and three referees with salary expectations to Mr. Kenneth To Address: Wax-it Histology Services 310-2386 East Mall Vancouver, BC, V6T 1Z3 E-mail: kto@waxitinc.com From sharon.willman <@t> bms.com Fri Dec 16 08:25:17 2005 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Fri Dec 16 08:25:57 2005 Subject: [Histonet] 10% formalin Fixation time for Non-Human Primate Message-ID: <43A2CE4D.3040305@bms.com> Hi, I am trying to find the fixation time in 10% NBF before the decal process (13% formic acid) for non-human primate, canine, and rodent nasal cavities. I would appreciate any help someone could offer. Thanks, Sharon Willman From sa.drew <@t> hosp.wisc.edu Fri Dec 16 09:12:49 2005 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Dec 16 09:13:00 2005 Subject: [Histonet] Leu-enkephalin Message-ID: We have a pathologist who would like to send some slides somewhere for leu-enkephalin and met-enkephalin immunostaining. Is anyone aware of a reference lab that does these two? Thank you! Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 From stamptrain <@t> yahoo.com Fri Dec 16 09:53:18 2005 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Fri Dec 16 09:53:29 2005 Subject: [Histonet] 10% formalin Fixation time for Non-Human Primate In-Reply-To: <43A2CE4D.3040305@bms.com> Message-ID: <20051216155318.43511.qmail@web50304.mail.yahoo.com> While penetration is usually complete by 24 hours, fixation in formalin (formaldehyde) is not considered to be complete until about 48 to 72 hours. We have found that 48hours is the minimum time we can use to get optimum preservation. Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals, Inc. Rigdefield, CT --- Sharon E Willman wrote: > Hi, > I am trying to find the fixation time in 10% NBF > before the decal > process (13% formic acid) > for non-human primate, canine, and rodent nasal > cavities. I would > appreciate any help someone could offer. > Thanks, > Sharon Willman > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From LINDA.MARGRAF <@t> childrens.com Fri Dec 16 10:24:06 2005 From: LINDA.MARGRAF <@t> childrens.com (LINDA MARGRAF) Date: Fri Dec 16 10:24:35 2005 Subject: [Histonet] job opportunity Message-ID: Here is a message Tim asked me to post for him. Please contact him not me. Thanks and happy holidays. Linda M (Histonet administrator) BioGenex, a technology leader in molecular pathology, provides Total Solutions for complete automation of cell and tissue testing. BioGenex is the inventor of Antigen Retrieval and offers a broad range of reagents, detection kits, antibodies, probes, special stains, tissue microarrays, automated staining and imaging systems to streamline and standardize cellular and molecular diagnostics. Because of our growth in 2005 we need to expand our application specialist group. We are looking for a couple of professional histotechs in the western area of the country, CA, OR, WA, AZ, NV, CO, TX to provide onsite support in the areas of stain optimizations, sales support and training to accounts in the western region. We offer a competitive salary, bonus, car allowance and stock options. If you are interested please call or email Tim Nicholson, Western Region Manager at 904-616-6117 or t.nicholson@biogenex.com. Thanks for your Tim Nicholson Western Region Manager BioGenex Cell 904-616-6117 office 904-247-1737 fax 904-247-9758 From sbreeden <@t> nmda.nmsu.edu Fri Dec 16 11:14:56 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Fri Dec 16 11:15:05 2005 Subject: [Histonet] Salary Surveys? Message-ID: Are there any current histo/technician/logist salary surveys posted anywhere? The most comprehensive I can locate is the Advance magazine's November 2005 issue; I am certain other recent surveys are/were done by (perhaps) ASCP or NSH although I cannot quickly locate any. Appreciate any directions you can provide. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From gcallis <@t> montana.edu Fri Dec 16 11:54:16 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Dec 16 11:55:43 2005 Subject: [Histonet] 10% formalin Fixation time for Non-Human Primate In-Reply-To: <43A2CE4D.3040305@bms.com> References: <43A2CE4D.3040305@bms.com> Message-ID: <6.0.0.22.1.20051216104427.01b53d38@gemini.msu.montana.edu> The bigger the specimen, the longer you need to fix - a whole canine nose? This may take two weeks or more. If you cut it into slabs? Maybe up to a week depending on thickness of slabs. I guess the main question is what do you do to the sample before you start fixation - reduce in size? Cut into slabs? If you cut open a bone or slab it after it has been in fixative and see it is still pink, go back to NBF - it isn't fixed. Be sure to change the fixative once or twice during fixation of large bones, suspend in cheesecloth in NBF. You might want to get rid of any air pockets in nasal passages by applying a vacuum briefly (you will see bubbles come off) This is to merely pull NBF into the vacant air spaces so fixative comes in contact with all surfaces efficiently. Vacuum really doesn't affect speed of fixation as that is a chemical reaction. Rat and Mouse whole heads with eye, tongue and lower jaw disarticulated, skin with ears dissected off skull, and brain removed - we like to fix a week and change the NBF once - use a large volume. If you leave the brain in skull, or are not dissecting away cheek muscles, eyes, etc, - you may have to go 10 days to 2 weeks. At 07:25 AM 12/16/2005, you wrote: >Hi, >I am trying to find the fixation time in 10% NBF before the decal process >(13% formic acid) >for non-human primate, canine, and rodent nasal cavities. I would >appreciate any help someone could offer. >Thanks, >Sharon Willman > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From MVaughan4 <@t> ucok.edu Fri Dec 16 12:26:06 2005 From: MVaughan4 <@t> ucok.edu (MVaughan4@ucok.edu) Date: Fri Dec 16 12:26:29 2005 Subject: [Histonet] Agar embedding protocol In-Reply-To: Message-ID: Jim, Bio-Rad agar 3% in water, boil to melt, let cool to about 60 degrees, quickly pour into a cryoembedding mold and orient sample before the agar gels. Once the agar hardens you can cut out the sample surrounded by agar. Mounting works best using a dissecting scope if you have a small sample. You might be able to use low-melt agar if your sample is sensitive to heat. The only problem I have noticed is that sometimes there is nonspecific background staining of the tissue or loss of antigen, probably if the agar is too hot. You might test on an unimportant sample first. Mel Melville B. Vaughan, Ph. D. Assistant Professor Department of Biology University of Central Oklahoma Edmond, OK 73034 From eshields <@t> bhset.org Fri Dec 16 12:39:48 2005 From: eshields <@t> bhset.org (E Sharon Shields) Date: Fri Dec 16 12:40:17 2005 Subject: [Histonet] c-myc Message-ID: Histonetters, Is any one doing c-myc protein in the Ventana Benchmark? If so would you please share your protocol with me? Thank you and Merry Christmas to all, Sharon Baptist Hospital of E TN Knoxville, TN 865 549-4351 From dennijc <@t> vetmed.auburn.edu Fri Dec 16 12:50:51 2005 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Fri Dec 16 12:51:42 2005 Subject: [Histonet] 10% formalin Fixation time for Non-Human Primate In-Reply-To: <43A2CE4D.3040305@bms.com> References: <43A2CE4D.3040305@bms.com> Message-ID: Sharon What parts of that skull region are you interested in looking at? Are you planning to dissect out specific tissues/structures or just plunk heads into buckets of fixative (I've done that, too)? John Carroll Dennis Anatomy, Physiology, and Pharmacology 109 Greene Hall Auburn University, AL 36849 On Fri, 16 Dec 2005, Sharon E Willman wrote: > Hi, > I am trying to find the fixation time in 10% NBF before the decal process > (13% formic acid) > for non-human primate, canine, and rodent nasal cavities. I would appreciate > any help someone could offer. > Thanks, > Sharon Willman > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jamie.erickson <@t> abbott.com Fri Dec 16 13:19:56 2005 From: jamie.erickson <@t> abbott.com (Jamie E Erickson) Date: Fri Dec 16 13:20:10 2005 Subject: [Histonet] Mouse Autoradiography of fixed frozen tissue Message-ID: Hello everyone, I was hoping that I can get some assistance on a project I'm going to do. An Investigator here would like to look at radiolabeled mouse Sm. intestines for there specific radiolabeled protein. My problem is that I don't want to use our only processor to process radioactive material ( I 125 or H3, not sure yet) My other histology colleges may not like me so much if I did that. So I plan on fixing the material in 10% NBF (24-48 hours), washing in water (Blot) then cryopreserving them (LN2 Isopentane). This way I can Cryosection and decontaminate the less used cryostat. Next I wash in water and lastly dipping slides in Photographic Emulsion and wait, and wait....My question is, has anyone done this that could help me out with the finer details and will this be OK? I've done this with paraffin samples dip wait 2,4,7,14, 21, etc days for a signal, but not with frozens.. If anyone has done this and could help me out I'd really appreciate it.. Thanks Jamie _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com From la.sebree <@t> hosp.wisc.edu Fri Dec 16 13:53:15 2005 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Dec 16 13:53:25 2005 Subject: [Histonet] Mouse Autoradiography of fixed frozen tissue Message-ID: Its been a long time Jamie, but when I did autoradiography on frozen rabbit heart, I simply cut the frozen sections in a dark room, air-dried and dipped in emulsion. Then I sealed them in a light-tight box to develop. Linda A. Sebree University of Wisconsin Hospital & Clinics IHC/ISH Clinical & Research Laboratory DM223-VA 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jamie E Erickson Sent: Friday, December 16, 2005 1:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mouse Autoradiography of fixed frozen tissue Hello everyone, I was hoping that I can get some assistance on a project I'm going to do. An Investigator here would like to look at radiolabeled mouse Sm. intestines for there specific radiolabeled protein. My problem is that I don't want to use our only processor to process radioactive material ( I 125 or H3, not sure yet) My other histology colleges may not like me so much if I did that. So I plan on fixing the material in 10% NBF (24-48 hours), washing in water (Blot) then cryopreserving them (LN2 Isopentane). This way I can Cryosection and decontaminate the less used cryostat. Next I wash in water and lastly dipping slides in Photographic Emulsion and wait, and wait....My question is, has anyone done this that could help me out with the finer details and will this be OK? I've done this with paraffin samples dip wait 2,4,7,14, 21, etc days for a signal, but not with frozens.. If anyone has done this and could help me out I'd really appreciate it.. Thanks Jamie _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PBugelsk <@t> CNTUS.JNJ.COM Fri Dec 16 14:12:29 2005 From: PBugelsk <@t> CNTUS.JNJ.COM (Bugelski, Peter [CNTUS]) Date: Fri Dec 16 14:12:43 2005 Subject: [Histonet] Mouse Autoradiography of fixed frozen tissue Message-ID: <7BF70FA941B9AE4783EAAF733762F1B50B3A488E@CNTUSMAEXS3.na.jnj.com> Why not process to paraffin "by hand"? Process through xylene at room temp and then transfer to paraffin in an oven overnight. Do everything in scintillation vials. This way you can keep the volumes low and check for leaching radioactivity by either gamma or scintillation counting. I've done this many times. There should be no difference in the exposure time required. The only steps that need to be in the dark are the melting of the emulsion, the dipping, the exposur, the development and photographic fixing. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jamie E Erickson Sent: Friday, December 16, 2005 2:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Mouse Autoradiography of fixed frozen tissue Hello everyone, I was hoping that I can get some assistance on a project I'm going to do. An Investigator here would like to look at radiolabeled mouse Sm. intestines for there specific radiolabeled protein. My problem is that I don't want to use our only processor to process radioactive material ( I 125 or H3, not sure yet) My other histology colleges may not like me so much if I did that. So I plan on fixing the material in 10% NBF (24-48 hours), washing in water (Blot) then cryopreserving them (LN2 Isopentane). This way I can Cryosection and decontaminate the less used cryostat. Next I wash in water and lastly dipping slides in Photographic Emulsion and wait, and wait....My question is, has anyone done this that could help me out with the finer details and will this be OK? I've done this with paraffin samples dip wait 2,4,7,14, 21, etc days for a signal, but not with frozens.. If anyone has done this and could help me out I'd really appreciate it.. Thanks Jamie _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Sat Dec 17 04:09:02 2005 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Dec 17 04:09:07 2005 Subject: AW: [Histonet] c-myc In-Reply-To: Message-ID: We used c-myc by Novocastra; 1:100, mild CC1, 8 min Protease, ink.temp. 37, ink.time 28 min. I don't know, if it was really successfull, because recently we took this antibody out of our program. Gudrun Lang -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von E Sharon Shields Gesendet: Freitag, 16. Dezember 2005 19:40 An: histonet@lists.utsouthwestern.edu Betreff: [Histonet] c-myc Histonetters, Is any one doing c-myc protein in the Ventana Benchmark? If so would you please share your protocol with me? Thank you and Merry Christmas to all, Sharon Baptist Hospital of E TN Knoxville, TN 865 549-4351 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Sat Dec 17 05:08:52 2005 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Sat Dec 17 05:11:08 2005 Subject: [Histonet] method of using agar for an embedding tool Message-ID: <3C83687E8F6AE04792E361ABE2D385B841807B@cht-mail2-2k.xchristie.nhs.uk> There was a report in the Journal of Med Lab Tech, in the 70s that described use of agar to embedd small bits, Cut the end off plastic syringes to reveal the whole bore. Clip or otherwise mount the syringe upright and pull back to leave say 3mm above the plunger. 2% agar, meleted in a microwave for 20 secs. Pipette in agar Orient tissue Allow to cool syringe depressed and block come out Ready to go in a cassette Bob's your uncle If you would like the reference then RSVP David Christie Hospital Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vickroy, Jim Sent: 15 December 2005 20:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] method of using agar for an embedding tool Has anyone used agar at the embedding stations to help orient small specimens? I am told by my new pathology assistant that this would prevent the embedders from having to reorient them at the embedding center? If you have could someone send me a procedure? Please disregard the spelling errors in the last email. Obviously spell check should have been used. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 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 ****************************************************************************************************************** This e-mail and any files transmitted with it are confidential and solely for the use of the intended recipient. If you have received this e-mail in error you should not disseminate, distribute or copy it. Please notify the sender immediately and delete this e-mail from your system. ******************************************************************************************************************* From lbeaudet <@t> ctcsurge.com Sat Dec 17 12:14:33 2005 From: lbeaudet <@t> ctcsurge.com (Laura Beaudet) Date: Sat Dec 17 12:18:34 2005 Subject: [Histonet] automated response Message-ID: <10512171314.AA05864@mail1.ctcsurge.com> I will be out of the office on Monday, December 19, 2005. If the matter is urgent, please call Computer Trust Corporation at 617-557-9264 for immediate assistance. From pruegg <@t> ihctech.net Sat Dec 17 13:02:12 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Sat Dec 17 13:02:27 2005 Subject: [Histonet] method of using agar for an embedding tool In-Reply-To: <3C83687E8F6AE04792E361ABE2D385B841807B@cht-mail2-2k.xchristie.nhs.uk> Message-ID: <200512171902.jBHJ22rH083020@pro12.abac.com> I use a product for Richard Allan called Histogel to do this. I tried making my own agar but it was a hassle, this gel comes in tubes, you put the tube in the microwave (I put the tube in a beaker of water), heat it til it is liquid, I then use a transfer pipette to pull some out to fill a cryomold, let it sit at RT or even on a ice pack, it sets up slowly allowing you to orient the sample. The gel with the oriented sample can then be processed maintaining the orientation. I use this to embed mouse bile ducts on end, don't know what I would do without it. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Edmondson David (RBV) NHS Christie Tr Sent: Saturday, December 17, 2005 4:09 AM To: Vickroy, Jim Cc: Histonet (E-mail 2) Subject: RE: [Histonet] method of using agar for an embedding tool There was a report in the Journal of Med Lab Tech, in the 70s that described use of agar to embedd small bits, Cut the end off plastic syringes to reveal the whole bore. Clip or otherwise mount the syringe upright and pull back to leave say 3mm above the plunger. 2% agar, meleted in a microwave for 20 secs. Pipette in agar Orient tissue Allow to cool syringe depressed and block come out Ready to go in a cassette Bob's your uncle If you would like the reference then RSVP David Christie Hospital Manchester UK -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vickroy, Jim Sent: 15 December 2005 20:50 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] method of using agar for an embedding tool Has anyone used agar at the embedding stations to help orient small specimens? I am told by my new pathology assistant that this would prevent the embedders from having to reorient them at the embedding center? If you have could someone send me a procedure? Please disregard the spelling errors in the last email. Obviously spell check should have been used. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 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 **************************************************************************** ************************************** This e-mail and any files transmitted with it are confidential and solely for the use of the intended recipient. If you have received this e-mail in error you should not disseminate, distribute or copy it. Please notify the sender immediately and delete this e-mail from your system. **************************************************************************** *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Sat Dec 17 14:09:32 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Sat Dec 17 14:09:50 2005 Subject: [Histonet] Re: CD4 and CD8 mouse FFPE tissues In-Reply-To: Message-ID: <200512172009.jBHK9NNo003913@pro12.abac.com> Jamie, I agree with you on this. I gave up trying to do cd4 and cd8 on ffpe mouse tissue and do it only as frozens now. I am one that can't even get them to work on human ffpe tissues, people have told me their methods but when I try I fail even with human tissues with these t cell subsets. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jamie E Erickson Sent: Thursday, December 15, 2005 2:13 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: CD4 and CD8 mouse FFPE tissues HI Reinhard, I have tried many and I repeat many times to get CD4 and CD8 to work in FFPE tissues mostly with the purified monoclonals antibody from BD biosciences and have failed. I have been doing mouse IHC for 12 years and have come to the conclusion after many try's and conversations with others in the field (Gayle Callis and others) that for CD4 and CD8 just doesn't work for mouse FFPE IHC. We do research and I'm going to start doing more frozen mouse work. Frozens are faster and you don't have to beat your head against the wall to think of more antigen retrieval to try. This may not be everyone's experience but for me I've had enough FFPE CD4/CD8 and others.. That's my opinion for what it worth. _______________________________ Jamie Erickson Sr. Research Associate Department: DSMP Abbott Bioresearch Center 100 Research Drive Worcester, MA 01605-4341 508-688-3134 FAX: 508-793-4895 e-mail: jamie.erickson@abbott.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From wasielewski.reinhard.von <@t> mh-hannover.de Sun Dec 18 13:49:21 2005 From: wasielewski.reinhard.von <@t> mh-hannover.de (wasielewski.reinhard.von@mh-hannover.de) Date: Sun Dec 18 13:49:32 2005 Subject: [Histonet] CD4 and CD8 in mouse / human FFPE Message-ID: <43A5CB51.20138.3A8527E7@localhost> Hi Patsy, we do a lot of CD4 and CD8 in human FFPE tissues, even in BM after decalification. It works very good and is daily routine in our lab. If you need advice, give me a mail. We now got some hints how both markers could work in mouse tissue, too. If we succeed, we'll let you know. best regards Reinhard. PD Dr. med. Reinhard von Wasielewski From laurie.reilly <@t> jcu.edu.au Sun Dec 18 17:09:49 2005 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Sun Dec 18 17:10:30 2005 Subject: [Histonet] CD4 and CD8 in mouse / human FFPE In-Reply-To: <43A5CB51.20138.3A8527E7@localhost> Message-ID: <5.1.0.14.0.20051219090417.00c4a238@mail.jcu.edu.au> Reinhard, Could I please have a copy of your protocol, we have had lots of trouble trying to stain with CD4 and CD8 in bovine tissues. Any advice you can give would be most welcome, particularly about antigen retrieval. Hope everyone has a happy Christmas, Regards, Laurie. At 08:49 PM 18/12/2005 +0100, you wrote: >Hi Patsy, >we do a lot of CD4 and CD8 in human FFPE tissues, even in BM after >decalification. >It works very good and is daily routine in our lab. If you need advice, >give me a mail. >We now got some hints how both markers could work in mouse tissue, too. If we >succeed, we'll let you know. > >best regards >Reinhard. > > > >PD Dr. med. Reinhard von Wasielewski > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 School of Veterinary & Biomedical Science Fax 07 4779 1526 James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From luc136677 <@t> aol.com Mon Dec 19 12:25:47 2005 From: luc136677 <@t> aol.com (luc136677@aol.com) Date: Mon Dec 19 12:26:03 2005 Subject: [Histonet] preservation of unstained 20 um sections on slides for later DNA research Message-ID: <8C7D2E0C43B0AA3-193C-9E19@MBLK-M42.sysops.aol.com> Can anyone recommend ways of preserving paraffin embedded 20 um prostate tissue sections, unstained, on the slides? Thanks, J From settembr <@t> umdnj.edu Mon Dec 19 12:38:57 2005 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Mon Dec 19 12:40:32 2005 Subject: [Histonet] negative controls with IHC Message-ID: Lisa, Your best bet is to contact CAP directly with your question(s). They have an e-mail address that I have used several times. They answer promptly and precisely. The address is: www.accred@cap.org Good Luck, Dana Settembre University Hospital - UMDNJ Newark, NJ >>> lwuerges 12/12/2005 7:25:00 PM >>> I have a question regarding negative controls for a manual immunoperoxidase procedure. Do you use a neg control for each antibody used with each block? or one neg control per block regardless of how many antibodies for that block. The latter is what I was told, but the first seems more likely on account of each antibody may be treated differently with some needing antigen retrieval--obviously the contol should receive identical treatment. What is the usual neg control protocol? Thank you. Lisa Wuerges _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Dec 19 12:51:44 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Dec 19 12:51:52 2005 Subject: [Histonet] preservation of unstained 20 um sections on slides for later DNA research In-Reply-To: <8C7D2E0C43B0AA3-193C-9E19@MBLK-M42.sysops.aol.com> Message-ID: <20051219185144.34422.qmail@web61219.mail.yahoo.com> Try to isolate the slides from air as much as possible. In small plastic mailers inside a sealed Kapak pouch in the refrigerator would be good. Try to obtain the 2 following publications: Lab.Invest., 14 June/04 and Am.Soc.Invest.Pathol., vol 161, pp 1961-1972 (2002) Ren? J. luc136677@aol.com wrote: Can anyone recommend ways of preserving paraffin embedded 20 um prostate tissue sections, unstained, on the slides? Thanks, J _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From cbass <@t> bidmc.harvard.edu Mon Dec 19 12:51:42 2005 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Mon Dec 19 12:51:55 2005 Subject: [Histonet] sealing slides with diluted permount Message-ID: <47CD9724-9E0E-4CBA-8CA2-F42500FEE307@bidmc.harvard.edu> Hello, I have some slides of GFP expressing tissue. I am using the prolong gold mounting media from Molecular Probes to mount the coverslips but now I have to seal them. I have read that the best way to do this is with diluted permount. Unfortunately I have never sealed slides before and am having a hard time visualizing the process. What is a good starting dilution for the permount? I was thinking 1:10 with xylenes to start, but this is just a guess. How exactly do you apply the permount? Can you just pipet it around the edges or do you need to brush it on in some manner? Does it have to be flush with the coverslip or can you have some buildup? Any advice would be appreciated. Thanks, Caroline Bass From hfedor <@t> jhmi.edu Mon Dec 19 12:52:02 2005 From: hfedor <@t> jhmi.edu (Helen Fedor) Date: Mon Dec 19 12:52:21 2005 Subject: [Histonet] preservation of unstained 20 um sections on slides for later DNA research Message-ID: We have had pretty good success with drying them overnight, vertically, then putting them face to face, wrapping them tightly in parafilm and then putting them into a ziploc and expelling the air. And then store at -20. We have only done this on 4 to 5 micron sections, and have them stored for up to 3 years and they retain their antigenicity fairly well. Helen Helen L. Fedor B.S. Johns Hopkins University Pathology Department 600 N Wolfe St Marburg Room 406 Baltimore MD 21287 email: hfedor@jhmi.edu Phone: 410 614-1660 Pager: 410 283-3419 WARNING: E-mail sent over the Internet is not secure. Information sent by e-mail may not remain confidential. DISCLAIMER: This e-mail is intended only for the individual to whom it is addressed. It may be used only in accordance with applicable laws. If you received this e-mail by mistake, notify the sender and destroy the e-mail. From Malcolm.McCallum <@t> tamut.edu Mon Dec 19 13:01:20 2005 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Mon Dec 19 13:03:18 2005 Subject: [Histonet] sealing slides with diluted permount Message-ID: The research I do has involved simply mounting specimens on slides with permount. With reference to sealing, I would appreciate any comments on this technique. What are some methods and how does this help with longivity of the slide? Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Caroline Bass Sent: Mon 12/19/2005 12:51 PM To: Histonet (E-mail) Subject: [Histonet] sealing slides with diluted permount Hello, I have some slides of GFP expressing tissue. I am using the prolong gold mounting media from Molecular Probes to mount the coverslips but now I have to seal them. I have read that the best way to do this is with diluted permount. Unfortunately I have never sealed slides before and am having a hard time visualizing the process. What is a good starting dilution for the permount? I was thinking 1:10 with xylenes to start, but this is just a guess. How exactly do you apply the permount? Can you just pipet it around the edges or do you need to brush it on in some manner? Does it have to be flush with the coverslip or can you have some buildup? Any advice would be appreciated. Thanks, Caroline Bass _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Dec 19 13:04:46 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Dec 19 13:05:01 2005 Subject: [Histonet] preservation of unstained 20 um sections on slides for later DNA research In-Reply-To: <8C7D2E0C43B0AA3-193C-9E19@MBLK-M42.sysops.aol.com> References: <8C7D2E0C43B0AA3-193C-9E19@MBLK-M42.sysops.aol.com> Message-ID: <6.0.0.22.1.20051219120244.01b54e98@gemini.msu.montana.edu> You can store paraffin sections in a 100 capacity slide box with lid, in a cool, dry place. We have stored unstained paraffin sections for years this way many time. At 11:25 AM 12/19/2005, you wrote: >Can anyone recommend ways of preserving paraffin embedded 20 um prostate >tissue sections, unstained, on the slides? >Thanks, >J >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From Malcolm.McCallum <@t> tamut.edu Mon Dec 19 13:04:57 2005 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Mon Dec 19 13:05:42 2005 Subject: [Histonet] sealing slides with diluted permount Message-ID: I found this. http://www.clontech.com/clontech/techinfo/faqs/LivingColors/fixation.shtml Q What is the best method of sealing coverslips? A We recommend sealing coverslips with molten agarose or rubber cement. We do not recommend using nail polish, because GFP is very sensitive to some nail polishes. However, when using ProLong Antifade from Molecular Probes (www.probes.com ) no further sealing is necessary. Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Caroline Bass Sent: Mon 12/19/2005 12:51 PM To: Histonet (E-mail) Subject: [Histonet] sealing slides with diluted permount Hello, I have some slides of GFP expressing tissue. I am using the prolong gold mounting media from Molecular Probes to mount the coverslips but now I have to seal them. I have read that the best way to do this is with diluted permount. Unfortunately I have never sealed slides before and am having a hard time visualizing the process. What is a good starting dilution for the permount? I was thinking 1:10 with xylenes to start, but this is just a guess. How exactly do you apply the permount? Can you just pipet it around the edges or do you need to brush it on in some manner? Does it have to be flush with the coverslip or can you have some buildup? Any advice would be appreciated. Thanks, Caroline Bass _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Dec 19 13:15:36 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Dec 19 13:15:47 2005 Subject: [Histonet] Re:sealing slides with diluted permount In-Reply-To: <47CD9724-9E0E-4CBA-8CA2-F42500FEE307@bidmc.harvard.edu> References: <47CD9724-9E0E-4CBA-8CA2-F42500FEE307@bidmc.harvard.edu> Message-ID: <6.0.0.22.1.20051219120516.01b569e8@gemini.msu.montana.edu> We buy cheap clear finger nail polish and dump it out!!! Rinse out the little polish bottle and brush with acetone and let it dry. Put Permount in the bottle, add some xylene or toluene (if toluene based). When you buy fingernail polish, look at the type that is for base or top coat, tip it so it flows and look at consistency or thickness of this polish. This is the thickness you want, I don't bother with proportions, as you can use thick goo if you want - thinner is easier to apply and it dries almost instantly. To apply the diluted Permount with the little polish brush to edges of coverslip, generally a put a little over the edge to see a good seal. If you pipette it, you will have a mess going everywhere. You can also use an artists brush like we use for sectioning, and wash it with solvent afterwards. However the fingernail polish brush is ideal, stiff, short bristles that you can control application of media which is now stored in the little polish bottle. Reusing the fingernail polish bottle is ideal. I generally do two edges over the glass slide, and not bother with edges of coverglass on outside edges of slide, if you have 24 x 24 coverglass, you have to be careful not to get media on bottom of slide. Some people might like a 22 x 22 coverglass or 22 x 30, or 40 so you can seal ALL edges. At 11:51 AM 12/19/2005, you wrote: >Hello, > >I have some slides of GFP expressing tissue. I am using the prolong >gold mounting media from Molecular Probes to mount the coverslips but >now I have to seal them. I have read that the best way to do this is >with diluted permount. Unfortunately I have never sealed slides >before and am having a hard time visualizing the process. > >What is a good starting dilution for the permount? I was thinking >1:10 with xylenes to start, but this is just a guess. How exactly do >you apply the permount? Can you just pipet it around the edges or do >you need to brush it on in some manner? Does it have to be flush >with the coverslip or can you have some buildup? > >Any advice would be appreciated. > >Thanks, > >Caroline Bass > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Dec 19 13:25:06 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Dec 19 13:25:15 2005 Subject: Clontech and Molecular Probes recommendations for RE: [Histonet] sealing slides with diluted permount In-Reply-To: References: Message-ID: <6.0.0.22.1.20051219121648.01b62630@gemini.msu.montana.edu> Malcolm, We found agarose and rubber cement to be terribly messy, and since had conversations with Clontechs tech services on using diluted mounting media. The same for Molecular Probes as their technical services one on one did recommend sealing the coverslip with Prolong Gold anitfade hard set - it does retract when dry, as we learned the hard way. They were also recommending the messy way. Another reason for sealing is to keep the coverslip from sliding all over the place with potential for ruining a stained section (had it happen). The only reason you can't use nail polish (per a Science publication on GFP) is that is contains isopropyl alcohol, and that is what ruins the GFP since the alcohol leaches into the aqueous mounting medias. That is why we now use toluene or xylene based mounting medias since these two solvents are not miscible with water. At 12:04 PM 12/19/2005, you wrote: >I found this. >http://www.clontech.com/clontech/techinfo/faqs/LivingColors/fixation.shtml > >Q What is the best method of sealing coverslips? > >A We recommend sealing coverslips with molten agarose or rubber cement. We >do not recommend using nail polish, because GFP is very sensitive to some >nail polishes. However, when using ProLong Antifade from Molecular Probes >(www.probes.com ) no further >sealing is necessary. > > >Malcolm L. McCallum >Assistant Professor >Department of Biological Sciences >Texas A&M University Texarkana >2600 Robison Rd. >Texarkana, TX 75501 >O: 1-903-233-3134 >H: 1-903-791-3843 >Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html > > >________________________________ > >From: histonet-bounces@lists.utsouthwestern.edu on behalf of Caroline Bass >Sent: Mon 12/19/2005 12:51 PM >To: Histonet (E-mail) >Subject: [Histonet] sealing slides with diluted permount > > > >Hello, > >I have some slides of GFP expressing tissue. I am using the prolong >gold mounting media from Molecular Probes to mount the coverslips but >now I have to seal them. I have read that the best way to do this is >with diluted permount. Unfortunately I have never sealed slides >before and am having a hard time visualizing the process. > >What is a good starting dilution for the permount? I was thinking >1:10 with xylenes to start, but this is just a guess. How exactly do >you apply the permount? Can you just pipet it around the edges or do >you need to brush it on in some manner? Does it have to be flush >with the coverslip or can you have some buildup? > >Any advice would be appreciated. > >Thanks, > >Caroline Bass > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From andrae <@t> u.washington.edu Mon Dec 19 13:29:36 2005 From: andrae <@t> u.washington.edu (A. Erickson) Date: Mon Dec 19 13:29:46 2005 Subject: [Histonet] Rat eyes in paraffin Message-ID: Hi all, I have a problem when blocking the rat eyes in paraffin. The eye tend to colapse and what I need is the round eye. My paraffin procedure is fix the eyes for 15 minute in 70%, 80%, 95%, 100%, mix 100%+Toluene, 2X Toluene. Then 30 minutein mix Toluene+Paraffin, Paraffin, Paraffin+vaccum. Most of the eyes were colaps. Is anyone has another protocol to prevent the colapsing of the eyes? Thank you so much. Regards, Dayat From plott <@t> uab.edu Mon Dec 19 14:21:07 2005 From: plott <@t> uab.edu (Patricia F Lott) Date: Mon Dec 19 14:21:15 2005 Subject: [Histonet] MMA sections coming off! Message-ID: Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! From PMonfils <@t> Lifespan.org Mon Dec 19 14:33:59 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Dec 19 14:34:14 2005 Subject: [Histonet] Rat eyes in paraffin Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717634@lsexch.lsmaster.lifespan.org> Assuming that you will be sectioning the eyes in the sagittal plane, try making a small incision on one lateral side of the eyeball with a #11 scalpel blade. Immerse the specimen in a petri dish or other shallow container before making the incision, and don't hold the specimen with forceps when making the incision, Otherwise the pressure of the forceps will collapse the specimen, forcing fluid out through the incision. This will allow freer flow of solvents into and out of the sphere, and will thereby avoid the buildup of differential pressure that occurs when one solvent diffuses through the wall faster than the other, which is what causes the collapse. Of course, if you are sectioning in a plane other than sagittal, just make the incision on a part of the eyeball that won't appear in the actual section. If the collapse occurs at a specific point in the process, such as just after the switch from 100% alcohol into toluene, then make the incision just before that switch, in this example while the specimens are in 100% alcohol. The hardening effect of the dehydration will make it easier to cut the incision. It is more difficult to cut the fresh, unfixed specimen because of the softness of the tissue. I notice you do not use formalin, and I assume there is a reason why you don't. However, the hardening effect of formalin also helps protect against collapse as well as facilitating the cutting of an incision. This method works well on various kinds of whole, hollow specimens such as gallbladders or urinary bladders, swim bladders of fish, cysts of various kinds, etc. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of A. > Erickson > Sent: Monday, December 19, 2005 11:29 AM > To: HistoNet Server > Subject: [Histonet] Rat eyes in paraffin > > Hi all, > I have a problem when blocking the rat eyes in paraffin. The eye tend to > colapse and what I need is the round eye. > > My paraffin procedure is fix the eyes for 15 minute in 70%, 80%, 95%, > 100%, mix 100%+Toluene, 2X Toluene. Then 30 minutein mix > Toluene+Paraffin, Paraffin, Paraffin+vaccum. > > Most of the eyes were colaps. Is anyone has another protocol to prevent > the colapsing of the eyes? > > Thank you so much. > Regards, > Dayat > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From portera <@t> msu.edu Mon Dec 19 14:51:47 2005 From: portera <@t> msu.edu (Amy Porter) Date: Mon Dec 19 14:52:50 2005 Subject: [Histonet] Agar embedding protocol References: Message-ID: <000501c604de$07314c80$8e7a0923@HistoJJ> Mel - I know that this is an older posting, but I am just now getting through my bunches of histonet save items. Are you using the agar for paraffin embedding or for frozens? I missed the original query into what you are using this for. Our laboratory is trying to embed one or multiple segments of 6-8 week old mouse mesenteric veins for cross sections! We are having a heck of a time with these and I am trying to find a method which will withstand automated processing and paraffing embedding. If you or anyone else out there would have any suggestions that would be great. Thanks in advance for your responses and suggestions. ----- Original Message ----- From: To: Sent: Friday, December 16, 2005 1:26 PM Subject: [Histonet] Agar embedding protocol > Jim, > Bio-Rad agar 3% in water, boil to melt, let cool to about 60 degrees, > quickly pour into a cryoembedding mold and orient sample before the agar > gels. > Once the agar hardens you can cut out the sample surrounded by agar. > Mounting works best using a dissecting scope if you have a small sample. > You might be able to use low-melt agar if your sample is sensitive to > heat. The only problem I have noticed is that sometimes there is > nonspecific background staining of the tissue or loss of antigen, probably > if the agar is too hot. You might test on an unimportant sample first. > Mel > Melville B. Vaughan, Ph. D. > Assistant Professor > Department of Biology > University of Central Oklahoma > Edmond, OK 73034 > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From cfavara <@t> niaid.nih.gov Mon Dec 19 15:03:12 2005 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Mon Dec 19 15:03:20 2005 Subject: [Histonet] protocols Message-ID: Does any one have a template that weds NAACLS protocols with automated immunohistochemistry? I am in the process of writing SOP's and need all the help I can get. Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From Jackie.O'Connor <@t> abbott.com Mon Dec 19 15:07:02 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Dec 19 15:07:31 2005 Subject: [Histonet] Agar embedding protocol Message-ID: Have you tried Sally Schlessinger's "double embedding" method? You embed multiple veins (or nerves or gut or anything long) longitudinally in a shallow amount of paraffinin a mold, then when the paraffin has hardened, you can cross section the longitudinal veins, and re-embed them at a 90 degree angle. She wrote an article somewhere about this, about 10 years ago, I think. It works great. "Amy Porter" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/19/2005 02:51 PM Please respond to Amy Porter To: , cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: Re: [Histonet] Agar embedding protocol Mel - I know that this is an older posting, but I am just now getting through my bunches of histonet save items. Are you using the agar for paraffin embedding or for frozens? I missed the original query into what you are using this for. Our laboratory is trying to embed one or multiple segments of 6-8 week old mouse mesenteric veins for cross sections! We are having a heck of a time with these and I am trying to find a method which will withstand automated processing and paraffing embedding. If you or anyone else out there would have any suggestions that would be great. Thanks in advance for your responses and suggestions. ----- Original Message ----- From: To: Sent: Friday, December 16, 2005 1:26 PM Subject: [Histonet] Agar embedding protocol > Jim, > Bio-Rad agar 3% in water, boil to melt, let cool to about 60 degrees, > quickly pour into a cryoembedding mold and orient sample before the agar > gels. > Once the agar hardens you can cut out the sample surrounded by agar. > Mounting works best using a dissecting scope if you have a small sample. > You might be able to use low-melt agar if your sample is sensitive to > heat. The only problem I have noticed is that sometimes there is > nonspecific background staining of the tissue or loss of antigen, probably > if the agar is too hot. You might test on an unimportant sample first. > Mel > Melville B. Vaughan, Ph. D. > Assistant Professor > Department of Biology > University of Central Oklahoma > Edmond, OK 73034 > _______________________________________________ > 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 Emily.Wiesner <@t> medecine.unige.ch Mon Dec 19 15:08:48 2005 From: Emily.Wiesner <@t> medecine.unige.ch (Emily.Wiesner@medecine.unige.ch) Date: Mon Dec 19 15:08:57 2005 Subject: [Histonet] fluorescence immunohistochemistry Message-ID: <1135026528.43a72160c086f@webmail.medecine.unige.ch> Hi All, I am new to fluorescence immunohistochemistry and I was wondering if anyone can let me know the simple steps involved in performing this technique. I know the dilutions required for the primary and secondary antibodies, I just need to known what solutions are used for washes and the steps in between! Thanks in advance, Emily From Luis.Chiriboga <@t> med.nyu.edu Mon Dec 19 16:37:28 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Mon Dec 19 16:34:32 2005 Subject: [Histonet] Ab Info Request Message-ID: Hi everyone Any source/supplier recommendations for anti-CD3, CD20, CD68, CD34/Factor VIII for use in FF-PE rabbit tissue. Any protocol info would also be greatly appreciated. Thanks in advance & Happy Holidays to all Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 From marjoh3 <@t> telus.net Mon Dec 19 19:36:03 2005 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Mon Dec 19 19:36:10 2005 Subject: [Histonet] Parts For Old Ultra-Technicon Tissue Processor Message-ID: <002f01c60505$bd2c99f0$6501a8c0@VALUED20606295> Hi Histonetters, I would like to refurbish an old Ultra-Technicon tissue processor. Can anyone please refer me to some Histology Lab./company that still may have one of these old tissue processor available for parts? Specifically, I require the motor-driven stirrers to circulate the oil in the processing pots and paraffin baths. Thanks in advance to any replies. Happy Holidays! Marilyn Johnson Alberta Agriculture Edmonton, Alberta, Canada From anh2006 <@t> med.cornell.edu Mon Dec 19 19:37:49 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Mon Dec 19 19:38:00 2005 Subject: [Histonet] anti-human VEGF antibody Message-ID: What's the current consensus on the best antibody to use for anti-human VEGF immunostaining on human FFPE sections (frozens as well if possible). Thanks so much!! Andrea -- From katri <@t> cogeco.ca Mon Dec 19 20:10:05 2005 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Mon Dec 19 20:10:16 2005 Subject: [Histonet] anti-human VEGF antibody References: Message-ID: <00ba01c6050a$7e68d850$6a9a9618@Katri> Andrea, I cannot say this is the best anti-VEGF antibody, but I am doing a research project on FFPE human brain tissue and using placenta as a control with following antibody: monoclonal mouse anti human VEGF, clone VG1 from ID Labs (www.idlabs.com), dilution 1:100, HIER in citrate buffer pH 6.0. Detection system is from Zymed, Histostain Plus Kit.. Glioblastoma stains strongly with this antibody. I have not used this with frozen sections. Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Andrea T. Hooper" To: "Histonet" Sent: Monday, December 19, 2005 8:37 PM Subject: [Histonet] anti-human VEGF antibody > What's the current consensus on the best antibody to use for anti-human > VEGF immunostaining on human FFPE sections (frozens as well if possible). > > Thanks so much!! > Andrea > -- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jason.PALMER <@t> svhm.org.au Mon Dec 19 20:55:35 2005 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Mon Dec 19 20:56:03 2005 Subject: [Histonet] Dako streptavidin / FITC for immuno's Message-ID: Dear all, Just wondering if anybody has used Dako streptavidin / FITC (F0422) in an immunohisto or immunocytochemistry run, and roughly what dilution you found to be best? We have very limited material to work with and so can't really do a titration to find the best dilution for ourselves (which is what we would normally do). It is to be used with a Stro-1 primary and biotinylated goat anti mouse secondary on cultured cells. The spec sheet has a suggested dilution range for flow cytometry only. Much obliged if anyone can help. Cheers! Jason Jason Palmer Bernard O'Brien Institute of Microsurgery 42 Fitzroy St, Fitzroy Victoria 3065 Australia tel +61 3 9288 4018 fax +61 3 9416 0926 email: palmerj@svhm.org.au Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From holling <@t> med-in.uni-saarland.de Tue Dec 20 01:54:21 2005 From: holling <@t> med-in.uni-saarland.de (holling@med-in.uni-saarland.de) Date: Tue Dec 20 01:54:33 2005 Subject: [Histonet] MMA sections coming off! In-Reply-To: References: Message-ID: <2612.134.96.44.146.1135065261.squirrel@134.96.44.146> Hello, I had have the same problems with the "+" and the "ULTRA +" slides. I tried the CHROME ALUM ADHESIVE and it worked much better. I don?t know Haupt?s adhesive, maybe it?s the same. Nicole Hollinger > Help! We are losing our thin plastic sections! We have cut PMMA > plastic sections at 5 microns and put them on "+" slides, and the > sections fall off either during de-plasticizing or during staining! We > have tried: Haupt's adhesive, longer drying times, "+" slides, plus > slides and Haupt's together, etc....no luck! Any suggestions would be > welcome! > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From mauger <@t> email.chop.edu Tue Dec 20 08:39:43 2005 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Tue Dec 20 08:40:51 2005 Subject: [Histonet] Re:Sections coming off! Message-ID: Hi Patricia, Are you using slides from Erie? We buy ours through Fisher, and have been having trouble keeping sections on + slides as well. It is as if they have no charge at all. We think the lot may be bad. The company claims to have no complaints. Anyone else having similar problem? Jo Mauger >>> "Patricia F Lott" 12/19/05 3:21 PM >>> Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mauger <@t> email.chop.edu Tue Dec 20 08:41:50 2005 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Tue Dec 20 08:42:40 2005 Subject: [Histonet] MMA sections coming off! Message-ID: I should also say we are doing regular paraffin blocks for routine histology and Immuno. Jo >>> "Patricia F Lott" 12/19/05 3:21 PM >>> Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Tue Dec 20 09:52:56 2005 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Tue Dec 20 09:53:12 2005 Subject: Fwd: [Histonet] Parts For Old Ultra-Technicon Tissue Processor Message-ID: <6.0.1.1.0.20051220094618.01a704b8@mailhost.vetmed.auburn.edu> Hello, the last company I had contact with that services Autotechnicon Ultra's was: G. & G. Instrument Corp., 466 Saw Mill River Road, Ardsley, NY 10502. Phone: 1-914-693-6000 or 1-800-882-2288, Fax: 1-914-693-6738. However, it's been approximately 1 year since I last contacted them. Best wishes, Atoska >From: "Marilyn Johnson" >To: >Date: Mon, 19 Dec 2005 18:36:03 -0700 >X-Mailer: Microsoft Outlook Express 6.00.2800.1437 >X-Scan-Signature: 0a4a30e5e11c556c77da169e50c2508a >X-Content-Filtered-By: Mailman/MimeDel 2.1.5 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >List-Id: For the exchange of information pertaining to histotechnology and > related fields >List-Unsubscribe: >, > > >List-Archive: >List-Post: >List-Help: >List-Subscribe: , > >Sender: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 2fb82c3b59105628058b6e8043468fa4 >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Parts For Old Ultra-Technicon Tissue Processor >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=5.5 tests=none autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >X-spam: ESP<-83>=RBL:<-140> RDNS:<0> SHA:<57> UHA:<0> SLS:<0> BAYES:<0> > SenderID:<0> URL Substring Dictionary (TRU8):<0> Spam > Dictionary (TRU8):<0> NigeriaScam Dictionary (TRU8):<0> HTML > Dictionary (TRU8):<0> Porn Dictionary (TRU8):<0> Embed HTML > Dictionary (TRU8):<0> Obscenities Dictionary (TRU8):<0> URL > Dictionary (TRU8):<0> CAN-SPAM Compliance Dictionary (TRU8):<0> > >Hi Histonetters, >I would like to refurbish an old Ultra-Technicon tissue processor. >Can anyone please refer me to some Histology Lab./company that still may >have one of these old tissue processor available for parts? >Specifically, I require the motor-driven stirrers to circulate the oil >in the processing pots and paraffin baths. >Thanks in advance to any replies. >Happy Holidays! > >Marilyn Johnson >Alberta Agriculture >Edmonton, Alberta, Canada > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Malcolm.McCallum <@t> tamut.edu Tue Dec 20 12:11:01 2005 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Tue Dec 20 12:11:15 2005 Subject: [Histonet] online journals Message-ID: Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html From Karen.Heckford <@t> CHW.edu Tue Dec 20 12:17:21 2005 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Tue Dec 20 12:16:53 2005 Subject: [Histonet] Purple blue haze Message-ID: Dear Histonetters, Has anyone had any problems with a purple- blue haze on their H&E using Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I am still getting the haze. I have changed nothing, it just appeared a couple of weeks ago. Any help would be greatly appreciated. Happy Holidays, Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center Histology Department 450 Stanyan St. San Francisco, Ca. 94117 1-415-668-1000 ext. 6167 email: kheckfor@chw.edu From jkiernan <@t> uwo.ca Tue Dec 20 12:42:34 2005 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Dec 20 12:42:25 2005 Subject: [Histonet] online journals References: Message-ID: <43A8509A.9B855C2A@uwo.ca> You raise some interesting points. Clearly you're polling a wide range of scientists; please send out a summary when you've analysed the results. Here's my two-centsworth. Paying to read the full text doesn't apply if your institution subscribes to the journal (or more usually to a large batch from the publisher). Since most people who want the full text are in institutions with libraries (or with employers such as drug companies that will pay for individual articles), avoiding page charges is the way to go. Why should the researcher and author pay the publisher? It should be the other way round. Prestigious journals commonly do charge authors, but anyone publishing in such a journal is sure to have a big enough research grant to cover the fees, which are small compared with salaries and items for the lab. Some journals published by learned societies do not charge authors anything, even for coloured pictures if these are needed to make a point. Three examples that come to mind are the Journal of Anatomy, the Journal of Histochemistry and Cytochemistry, and Biotechnic & Histochemistry. I'm sure there are many others. All three that I mentioned come free with membership in the society. The cost works out at about $5 per issue (print, and access online). I've not heard of the option of the author paying and the reader not paying. It's hard to see that working; even non-profit societies cannot run their journals at a loss. Libraries have to pay much more than individual members. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Malcolm McCallum wrote: > > Hi > Which of the below three options do you think is best regarding a journal with both print and online versions???? > > 1) Journal is open access, the publisher will charge page charges to authors. > 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. > 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. > > In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. > > Thanks for the feedback, its actually very important! > > Malcolm L. McCallum > Assistant Professor > Department of Biological Sciences > Texas A&M University Texarkana > 2600 Robison Rd. > Texarkana, TX 75501 > O: 1-903-233-3134 > H: 1-903-791-3843 > Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Shannon.Bryce <@t> UTSouthwestern.edu Tue Dec 20 13:01:17 2005 From: Shannon.Bryce <@t> UTSouthwestern.edu (Shannon Bryce) Date: Tue Dec 20 13:01:38 2005 Subject: [Histonet] Re:Sections coming off! In-Reply-To: References: Message-ID: <43A8009D0200009900000BC0@swnw124.swmed.edu> We get our slides which are the + coated slides from Fisher. Our MMA sections are also falling off here lately so I went to another department and borrowed a box of theirs that they bought through VWR and have had no problems so I would guess it's a bad lot.....anyone have lot numbers? Thanks, Shannon Bryce UT Southwestern Medical Center >>> "Joanne Mauger" 12/20/05 8:39 AM >>> Hi Patricia, Are you using slides from Erie? We buy ours through Fisher, and have been having trouble keeping sections on + slides as well. It is as if they have no charge at all. We think the lot may be bad. The company claims to have no complaints. Anyone else having similar problem? Jo Mauger >>> "Patricia F Lott" 12/19/05 3:21 PM >>> Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! _______________________________________________ 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 tracey.couse <@t> ibb.gatech.edu Tue Dec 20 14:15:42 2005 From: tracey.couse <@t> ibb.gatech.edu (Tracey Couse) Date: Tue Dec 20 14:15:51 2005 Subject: [Histonet] MMA sections coming off! Message-ID: <43A8666E.2030005@ibb.gatech.edu> Patricia, Try manually coating plain glass slides with poly-l-lysine, chrome/gelatin, or glycerin/gelatin. We use all of these for our thin plastic sections. I also suggest setioning a micron or two thinner (cut at 3 or 4um). A thinner section will reduce the surface area to volume ratio and may promote better section adherance. You may need to dry the sldies using a slide press and dry longer or with higher temperatures. Also, try searching the Histonet archives for alternative slide coating suggestions. Good luck! -- Tracey Couse Laboratory Coordinator Georgia Tech/Emory Center for the Engineering of Living Tissues Georgia Institute of Technology IBB, Room 1123 Office: 404.385.2611 Lab: 404-385-6735 Fax: 404.894.229 Date: Mon, 19 Dec 2005 14:21:07 -0600 From: "Patricia F Lott" Subject: [Histonet] MMA sections coming off! To: Message-ID: Content-Type: text/plain; charset="us-ascii" Help! We are losing our thin plastic sections! We have cut PMMA plastic sections at 5 microns and put them on "+" slides, and the sections fall off either during de-plasticizing or during staining! We have tried: Haupt's adhesive, longer drying times, "+" slides, plus slides and Haupt's together, etc....no luck! Any suggestions would be welcome! 1 From dharclerode <@t> cytoritx.com Tue Dec 20 15:06:17 2005 From: dharclerode <@t> cytoritx.com (Donna Harclerode) Date: Tue Dec 20 15:05:26 2005 Subject: [Histonet] Re 17. fluorescence immunohistochemistry Message-ID: <3DE0F644E093DF4BAE80C254176696A505B869@mp-mailserver.macropore.com> Message: 17 Date: Mon, 19 Dec 2005 22:08:48 +0100 From: Emily.Wiesner@medecine.unige.ch Subject: [Histonet] fluorescence immunohistochemistry To: histonet@lists.utsouthwestern.edu Message-ID: <1135026528.43a72160c086f@webmail.medecine.unige.ch> Content-Type: text/plain; charset=ISO-8859-1 Hi All, I am new to fluorescence immunohistochemistry and I was wondering if anyone can let me know the simple steps involved in performing this technique. I know the dilutions required for the primary and secondary antibodies, I just need to known what solutions are used for washes and the steps in between! Thanks in advance, Emily Dear Emily I do very simple IHC using fluorescent secondaries from Jackson Immunoresearch and purified primary abs. Depending on what you are doing, there can be more steps added. I usually am staining sections on a slide or cells on chamber slides or in a plate. For the unfixed frozen sections on slides I normally would fix 5 min in 75% acetone 25% ethanol and then right into PBS- do not allow the fixed slides to dry. The only change would be if the antibody prefers formalin fixation to acetone/ alcohol (thanks Gayle- I love this fixative for almost everything). I load them into a Sequenza(tm) rack (Thermo Electron, formerly Shandon) or you could put them in a humid chamber of some sort, wiping excess buffer before adding antibody or blocking solutions. OPTIONAL I have recently started using Image-iT(tm) FX signal enhancer, I36933, $80 Invitrogen for auto fluorescence (not sure it helps yet, but sure does not hurt) 30 minutes. I would not use this on routinely- my sections have infracted areas that have been a problem in the past) After blocking, I rinse 1 time in the Sequenza rack (3 x 5 minutes if you are doing them by hand) Add 100ul of the antibody diluted in Dako antibody diluent S0809- usually 1:100- 1:1k (dilutions are antibody dependant, but I have found in fluorescence too high a concentration is not as big a problem as with HRP formats) If you are using a humid chamber be sure to cover all the tissue. I have incubated slides at Room temperature for 1-2 hours, but now am staining overnight in the fridge (not so good in clinical situations) After primary incubation, I rinse 1 time in the Sequenza rack (3 x 5 minutes if you are doing them by hand) Next is Jackson secondary at a 1:100 dilution (I use Cy3 conjugates for single color and FITC and RITC for dual color. All my secondaries are minimal cross (mouse to rat), but if all your tissue is human that is not necessary. 30 minutes I add DAPI to my secondaries to give a blue nuclear stain so I can tell what I am looking at and localize the stain. Rinse again 3 times in PBS and coverslip with your choice of aqueous media. For media that never gets hard, I prefer Immunomount (Thermo Electron, formerly Shandon) or Aqua Mount (Lerner from VWR) same thing different label or the Prolonge Mounting media P36930 for $101 that will harden. I do not like the hard mount media from Vector- I got many air bubbles forming day after the sldes were coverslipped. For cells on plates or chamber slides I usually fix with 10% NBF or 4% PFA (same concentration of fixative, 4%PFA will contain no methanol) for 5-30 minutes. CD makers can be very hard to stain with NBF but with a short fix, I have been pretty luck so far. I rinse 3 x 10 minutes with PBS I have not used the blocking reagents on cells as I have not seen any autofluorescence in our cardiac myocytes, neurons or stem cells. The rest of the procedure is the same except in a 6 well plate you need a lot of antibody to cover the plate (about 600u). I try to use a rocker if it is available to help to be sure of good coverage. I do not usually cover my slides or plates, but if your room is very bright or your signal very week, it would be a good idea to protect the secondaries from light. I like to use isotype controls (negative controls) or rabbit IgGs for much of my work, but in a pinch I use just the antibody diluent to be sure the stain I see, is a result of the primary antibody and not the secondary binding to some part of the cell. In cell assays I always want to stain a known positive and a known negative cell if at all possible in addition to the isotype controls. If you are making your own diluent (to me Dako diluent is great and I have used it successfully for over 10 years in many applications) I use 0.3% Triton, 5 % Normal serum from the species that your secondary is made in - (all my secondaries are made in donkey so I would use donkey serum) in PBS. I have used donkey secondaries when possible because when I tested them years ago, they were a bit cleaner on the CD markers I was using. Dako diluent must have a similar make up, but it lasts a year in the fridge and it permeablizes intact cells very nicely. One big thing when purchasing fluorophores, make sure the filters on your scope can see the secondaries you are purchasing. I found out at my current job after staining with Cy3 and RITC that the reason I could not get any stain was they had a Texas red filter not the Cy3, RITC filter. Ideally if you are doing multicolor you want narrow band filters so you can separate the colors, with broad pass filters the colors blur together and it is hard to separate the results. Good Luck! Donna Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 322 dharclerode@cytoritx.com From algranth <@t> u.arizona.edu Tue Dec 20 15:19:47 2005 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Tue Dec 20 15:19:59 2005 Subject: [Histonet] Purple blue haze In-Reply-To: Message-ID: <4.3.2.7.2.20051220140523.00ce4520@algranth.inbox.email.arizona.edu> Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From laurie.colbert <@t> huntingtonhospital.com Tue Dec 20 17:12:17 2005 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Tue Dec 20 17:12:30 2005 Subject: [Histonet] Purple blue haze Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628006307C76@EXCHANGE1.huntingtonhospital.com> Your discussion rang a bell with me, and I've been meaning to call Richard Allan. We use their Type 9 paraffin, and for the last 3 or 4 months it also has been "blowing apart on the waterbath." We lowered the temp on the waterbaths, and this seems to have helped. Our housekeeping personnel have even noticed a difference in the paraffin (scraping it off the floor). They say it is stickier lately. I think something has been changed. Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Tuesday, December 20, 2005 1:20 PM To: Histonet (E-mail) Subject: Re: [Histonet] Purple blue haze Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> ihctech.net Tue Dec 20 19:56:17 2005 From: pruegg <@t> ihctech.net (pruegg@ihctech.net) Date: Tue Dec 20 19:56:31 2005 Subject: [Histonet] anti-human VEGF antibody In-Reply-To: <00ba01c6050a$7e68d850$6a9a9618@Katri> References: <00ba01c6050a$7e68d850$6a9a9618@Katri> Message-ID: <43434.207.200.116.10.1135130177.squirrel@207.200.116.10> I am using mouse anti-vegf from Santa Cruz with good results on human and baboon ffpe tissues, it is not easy, but I do HIER in a water bath in high ph buffer at 85dc for 3 hrs., followed by overnight incubation of the primary 1:200, detected with mouse labeled polymer (I use Envision from DAKO). Patsy > Andrea, > I cannot say this is the best anti-VEGF antibody, but I am doing a > research > project on FFPE human brain tissue and using placenta as a control with > following antibody: > monoclonal mouse anti human VEGF, clone VG1 from ID Labs (www.idlabs.com), > dilution 1:100, HIER in citrate buffer pH 6.0. Detection system is from > Zymed, Histostain Plus Kit.. > Glioblastoma stains strongly with this antibody. I have not used this with > frozen sections. > > Katri > > Katri Tuomala > Hamilton, Ontario, Canada > ----- Original Message ----- > From: "Andrea T. Hooper" > To: "Histonet" > Sent: Monday, December 19, 2005 8:37 PM > Subject: [Histonet] anti-human VEGF antibody > > >> What's the current consensus on the best antibody to use for anti-human >> VEGF immunostaining on human FFPE sections (frozens as well if >> possible). >> >> Thanks so much!! >> Andrea >> -- >> >> _______________________________________________ >> 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 Wed Dec 21 01:10:48 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Dec 21 01:10:29 2005 Subject: [Histonet] 17. fluorescence immunohistochemistry References: <3DE0F644E093DF4BAE80C254176696A505B869@mp-mailserver.macropore.com> Message-ID: <43A8FFF8.D988E7C6@uwo.ca> When I was a student in Birmingham, England (that's the Birmingham with the silent h), the immunology and serology lecturer told us: "water is unphysiological and beer is expensive, so we use saline." He must have been a good lecturer because I can still recall that apophthegm 40 years on when afar and asunder! John Kiernan London, Ontario. ------------------------------- > Hi All, > I am new to fluorescence immunohistochemistry and I was wondering if > anyone can let me know the simple steps involved in performing this > technique. I know the dilutions required for the primary and secondary > antibodies, I just need to known what solutions are used for washes and > the steps in between! > Thanks in advance, From Jacqueline.Malam <@t> rli.mbht.nhs.uk Wed Dec 21 06:00:58 2005 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Wed Dec 21 06:05:47 2005 Subject: [Histonet] Re: non-specific staining IHC Message-ID: We use the Benchmark XT but make our own prep kits up for neg controls (poly and mono) and are not experiencing any more non-sp staining than we normally do which is minimal. Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From nancy.troiano <@t> yale.edu Wed Dec 21 06:20:53 2005 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Wed Dec 21 06:20:56 2005 Subject: [Histonet] MMA sections falling off Message-ID: <5.2.1.1.2.20051221071248.00c2c038@email.med.yale.edu> We use Chrome-alum gel coated slides and prepare them as follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled water (solution A) while gently heating solution. Dissolve 4 g Chromium Potassium Sulfate (Fisher #C337-500) in distilled water (solution B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with 19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in this solution. Allow to dry vertically, then put in hot oven overnight (60 deg. C). To adhere sections to slides we place the 5 micron MMA sections on slide, spread with 70% ethanol, cover with a piece of plastic and stack slides then clamp with an office clamp. Put slides in 37deg C oven for two nights or for one night in 60 deg C oven (don't do the hot oven if you are subsequently staining for enzymes). For immuno we mount our 5 micron plastic sections on hybridization slides from Scientific Device Laboratory, Cat. #063 and place in hot oven 60 deg C overnite. Hope this helps! From sharon.willman <@t> bms.com Wed Dec 21 07:18:32 2005 From: sharon.willman <@t> bms.com (Sharon E Willman) Date: Wed Dec 21 07:19:24 2005 Subject: [Histonet] Modified Davidson's Rat Eyes and Testes Fixation Times Message-ID: <43A95628.7060609@bms.com> Hi, I was needing input on how long you fix rat eyes and testes in Modified Davidson's. Any suggestions would be most appreciated. Thanks in advance, Sharon Willman From rjr6 <@t> psu.edu Wed Dec 21 07:25:26 2005 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Wed Dec 21 07:25:40 2005 Subject: [Histonet] Embedding Centers Message-ID: My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta From akbitting <@t> geisinger.edu Wed Dec 21 07:49:01 2005 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Wed Dec 21 07:49:29 2005 Subject: [Histonet] Glass coverslippers Message-ID: Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From jqb7 <@t> cdc.gov Wed Dec 21 07:56:05 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Wed Dec 21 07:56:44 2005 Subject: [Histonet] Glass coverslippers Message-ID: We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Wed Dec 21 08:12:16 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 21 08:12:25 2005 Subject: [Histonet] Embedding Centers In-Reply-To: Message-ID: <20051221141216.20482.qmail@web61215.mail.yahoo.com> Hi Roberta: I would suggest you to call Sakura-Finetek. They have a new embedding center that has all the advantages of the old Tissue-Tek, along with the Sakura reliability. Ask for a "demo" instrument (they usually accomodate that type of request). Ren? J. Roberta Horner wrote: My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Jackie.O'Connor <@t> abbott.com Wed Dec 21 08:25:29 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Dec 21 08:28:36 2005 Subject: [Histonet] San Antonio Histology connection Message-ID: Could someone in the San Antonio area contact me directly? Thanks. Jackie O' From vanann702 <@t> skmc.gov.ae Wed Dec 21 08:20:37 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Wed Dec 21 08:45:16 2005 Subject: [Histonet] Embedding Centers Message-ID: <7E070A7B959A9F42BE732545E5CF621094ACBA@SKMCEMAIL.skmc.gov.ae> Im a fan of the Sakura embedding system - it's ergonomically designed and VERY user friendly Can be 'left handed' or 'right handed' Cassettes left/right Moulds left/right Cold plate left/right Forceps left/right Nothing more annoying (being a lefty) to be forced to work like a non-lefty!! Everything works independently of everything else - ie - micro adjustments can be made to temp in almost any area to your requirements Nice broad anti burn protection to rest hands/fingers while embedding And if you have a VIP5 the basket fits snugly into the holding area - left or right as you prefer of course!! Like I said - I am THE fan of Sakura And I just KNOW that some of you are smiling out there - you know who you are!!! Annieinarabia -----Original Message----- From: Roberta Horner [mailto:rjr6@psu.edu] Sent: Wednesday, December 21, 2005 5:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Embedding Centers My embedding center just died. Last time our maintenance looked at it they couldn't get the parts (its about 17 years old). I need to get information quick on which ones you like and are good. So far I have quotes from Surgipath and on the Leica but I don't know anything about them. Any comments please? Roberta _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Wed Dec 21 08:24:12 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Wed Dec 21 08:45:18 2005 Subject: [Histonet] Glass coverslippers Message-ID: <7E070A7B959A9F42BE732545E5CF6210948C5F@SKMCEMAIL.skmc.gov.ae> Try the Sakura Fast, efficient, small 'footprint' - will fit into a small space Had a Leica once - was so very patient - 2 exchanges (new machines) later - still hated it Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 5:56 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Dec 21 09:14:11 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Dec 21 09:14:24 2005 Subject: [Histonet] Re: MMA sections falling off In-Reply-To: <5.2.1.1.2.20051221071248.00c2c038@email.med.yale.edu> References: <5.2.1.1.2.20051221071248.00c2c038@email.med.yale.edu> Message-ID: <6.0.0.22.1.20051221081017.01b39c08@gemini.msu.montana.edu> To increase the holding power, the gelatin can be 275 bloom, a larger gelatin molecule made from pig collagen. This subbing solution suggested by Nancy is tried and true, also works for the nasty decalcified bone sections, generally larger ones. However, if the subbing solution causes too much blue background staining from hematoxylin, one can dip the subbed slides in NBF a couple of times, rinse well, dry and store in cool, dry place. The NBF crosslinks the gelatin a bit and reduces blue background. At 05:20 AM 12/21/2005, you wrote: >We use Chrome-alum gel coated slides and prepare them as >follows: dissolve 9 g gelatin(Fisher #G8-500) per 1000 ml distilled >water (solution A) while gently heating solution. Dissolve 4 g Chromium >Potassium Sulfate (Fisher #C337-500) in distilled water (solution >B). Prepare gelatin solution for slide dip: Mix 500 ml Solution a with >19.25 ml solution B. Heat to 70-75 Deg. C, dip slides 2 -4 minutes in >this solution. Allow to dry vertically, then put in hot oven overnight >(60 deg. C). To adhere sections to slides we place the 5 micron MMA >sections on slide, spread with 70% ethanol, cover with a piece of plastic >and stack slides then clamp with an office clamp. Put slides in 37deg C >oven for two nights or for one night in 60 deg C oven (don't do the hot >oven if you are subsequently staining for enzymes). For immuno we mount >our 5 micron plastic sections on hybridization slides from Scientific >Device Laboratory, Cat. #063 and place in hot oven 60 deg C >overnite. Hope this helps! > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From pmarcum <@t> vet.upenn.edu Wed Dec 21 09:14:58 2005 From: pmarcum <@t> vet.upenn.edu (Pamela Marcum) Date: Wed Dec 21 09:15:15 2005 Subject: [Histonet] Embedding Centers In-Reply-To: References: Message-ID: <6.1.1.1.2.20051221101125.01997318@mail.vet.upenn.edu> At 08:25 AM 12/21/2005, Roberta Horner wrote: >My embedding center just died. Last time our maintenance looked at it >they couldn't get the parts (its about 17 years old). I need to get >information quick on which ones you like and are good. So far I have >quotes from Surgipath and on the Leica but I don't know anything about >them. Any comments please? >Roberta >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Hi, I will be totally different as i am buying the Thermo Shandon Embedding Center. I had it in for demo and it had all the features the others have for timing and pre-sets as well as some independent temperature ranges I likes. Also the cold plate is seperate so I can have it on either side or in another area if I want for special projects. Good Luck, I think they all work well it just depends for me on features and price and Shandon also had the right price. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu From SAllen <@t> exchange.hsc.mb.ca Wed Dec 21 09:17:14 2005 From: SAllen <@t> exchange.hsc.mb.ca (Sharon Allen) Date: Wed Dec 21 09:18:25 2005 Subject: [Histonet] decal rapid Message-ID: <36FE435D6D2F1D489174B22362A961B66B830B@hscxntmx0006.hsc.mb.ca> Hi, Does anyone know about or what supplier "Decal Rapid" by "Pathtech" can be purchased from? We had used it a few years ago but have no idea where we got it from. I tried searching the internet & came up with Pathtech in Australia but would like a supplier a little closer. Thanks in advance Sharon sallen@hsc.mb.ca This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original. From dsantana <@t> pmaonline.com Wed Dec 21 09:27:06 2005 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Wed Dec 21 09:22:30 2005 Subject: [Histonet] FW: artifact Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB04D7E282@MAILPMA> > -----Original Message----- > From: Santana, Diane > Sent: Tuesday, December 20, 2005 10:20 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: artifact > > I am having a problem with artifact on some of my GI bx's. Actually a > paper published by Anatech "The Innovator", summer2004 shows this same > artifact on the front page. They believe it is a fixation problem, and or > retrieval by the endo Drs. My pathologist rules both these ideas out. I > tend to agree with him, at lease on the fixation. Our bx's go into the > processor around 5 pm and the bx run starts around 2 am. > It is a loss of nuclear detail. This artifact can be on a small section of > a bx. Even if there is 5 bx's in one cassette maybe 1 will have this > artifact where the others look great. I thought at first it was a clearing > problem or nuclear bubbling I changed times in clear rite and oven. It did > not eliminate the problem. My pathologist has seen this by other labs > also, but very seldom. For us it could be 1 or 2 slides a day. Sometimes > not at all. Weekend or weekdays. No consistently. The best way I can > describe it is that it looks like a smudge. > Our bx run is this; > > 10% NBF 30 min ea x2 > 80% Alcohol 10mins > 95% alcohol 10 mins ea x2 > 100% alcohol 17 mins ea x3 > Clear rite 22 mins ea x2 > Paraffin 15 mins ea x3 > > I would appreciate any feedback on this. > Diane Santana > Pentucket Medical Assoc. > Haverhill, Mass. From sjchtascp <@t> yahoo.com Wed Dec 21 09:22:41 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Dec 21 09:22:52 2005 Subject: [Histonet] antibody search Message-ID: <20051221152241.11885.qmail@web90204.mail.scd.yahoo.com> Gooday everyone, I'd like to get a few recommendations for anti-insulin and anti-glucagon antibodies for FFPE mouse tissue. If anyone has used a specific companies antibodies that work well please let me know. I'm still learning this research IHC stuff so I can use all the help I can "muster" Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Luis.Chiriboga <@t> med.nyu.edu Wed Dec 21 08:54:28 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Wed Dec 21 09:44:29 2005 Subject: [Histonet] Mouse liver tissue processing Message-ID: Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! From histosci <@t> shentel.net Wed Dec 21 10:02:33 2005 From: histosci <@t> shentel.net (HSRL) Date: Wed Dec 21 10:02:53 2005 Subject: [Histonet] Mouse liver tissue processing In-Reply-To: Message-ID: <000c01c60647$f47a7ab0$0200a8c0@HSRLMAIN> Luis, Have a tried keeping them on a WET ice for 45 minutes to an hour? If you have tried it and it doesn't work, it is a processing problem that you must rectify. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Wednesday, December 21, 2005 9:54 AM To: Histonet Subject: [Histonet] Mouse liver tissue processing Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jengirl1014 <@t> yahoo.com Wed Dec 21 10:32:05 2005 From: jengirl1014 <@t> yahoo.com (Jennifer Sipes) Date: Wed Dec 21 10:32:15 2005 Subject: [Histonet] Re: Mouse Liver Message-ID: <20051221163205.22628.qmail@web60612.mail.yahoo.com> Luis, I agree. Try a long soak in cold water. It sounds like the tissue needs to be rehydrated a bit. It's way to overdry and you need to let the people who do your processing know that otherwise they'll do it to other people. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From mary.ann.deathridge <@t> Vanderbilt.Edu Wed Dec 21 10:40:46 2005 From: mary.ann.deathridge <@t> Vanderbilt.Edu (Deathridge, Mary Ann) Date: Wed Dec 21 10:40:56 2005 Subject: [Histonet] unsubscribe Message-ID: <59342A85CAA485429AF89AAA42FFD9DF36B87D@mailbe10.mc.vanderbilt.edu> Thanks MaryAnn Deathridge, HT (ASCP) Supervisor, Histopathology TVC 4532 3-7012 From anh2006 <@t> med.cornell.edu Wed Dec 21 10:47:04 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Dec 21 10:47:18 2005 Subject: [Histonet] Mouse liver tissue processing In-Reply-To: References: Message-ID: Hi Luis! Are these livers being processed with a human processing protocol? If so you need to ask the facility to use an abbreviated protocol especially for mouse tissue. Do you know the details of the processing protocol they have been using? I find mouse livers are notoriously "crunchy" especially when overprocessed. Even when using the special mouse tissue processing protocol I still sometimes find the need to do a nice icy water bath soak for a good 10-20 minutes on some blocks to get good sections. Good luck! Andrea >Hi everyone >We are having a real serious problem getting good sections from mouse >livers. The tissue curls and shreds out of the block as soon as it comes in >contact with the knife, end up with a section that has the exact outline of >the tissue but no tissue. The tissue consistency could best be described as >"dry?" Interestingly, it is only the normal livers that have this problem. >We have tried sectioning thick, thin, no soak, cold soak......but out of >ideas. Anyone have any suggestions? Unfortunately we have no control over >the tissue processing, but perhaps can convince to change. >Thanks Luis > >Happy Holidays to all!! >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- From Karen.Heckford <@t> CHW.edu Wed Dec 21 11:00:19 2005 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Wed Dec 21 11:04:04 2005 Subject: [Histonet] Purple-blue haze conclusion Message-ID: I would like to thank everyone for their input. I will keep everything in mind for the future. I did up my rinse time after the Hematoxylin 7211 and also the Clarifier(Clarifier 1). This seemed to do the trick. Call me crazy but it seems when the weather starts getting colder the hematoxylin does not like it. Perhaps it can only get happy when the weather is warm. Again, thanks to everyone for your help, Happy Holidays and Happy New Year, Karen Heckford HT (ASCP) CE Lead Histology Technician St. Mary's Medical Center Histology Department 450 Stanyan St. San Francisco, Ca. 94117 1-415-668-1000 ext. 6167 email: kheckfor@chw.edu From c.ingles <@t> hosp.wisc.edu Wed Dec 21 11:08:52 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Wed Dec 21 11:13:27 2005 Subject: [Histonet] Mouse liver tissue processing References: <000c01c60647$f47a7ab0$0200a8c0@HSRLMAIN> Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE86@uwhis-xchng2.hosp.wisc.edu> Try a 5-10% aqueous ammonium hydroxide. We used to put this in a small sandwich size tupperware container on top of our ice trays and soaked the blocks in them after facing for about 10-15 minutes before taking sections. They cut great after that, especially small GI and liver bx's that were processed with the longer protocols with our big tissues. You may have to resoak after taking a few sections, but there you go. I usually gave a quick swish in regular cold water to get the excess amm. hydroxide off the block before sectioning. Claire Ingles UW Mohs Clinic Madison WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of HSRL Sent: Wed 12/21/2005 10:02 AM To: 'Luis Chiriboga'; histonet@pathology.swmed.edu Subject: RE: [Histonet] Mouse liver tissue processing Luis, Have a tried keeping them on a WET ice for 45 minutes to an hour? If you have tried it and it doesn't work, it is a processing problem that you must rectify. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Luis Chiriboga Sent: Wednesday, December 21, 2005 9:54 AM To: Histonet Subject: [Histonet] Mouse liver tissue processing Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From krat18 <@t> aol.com Wed Dec 21 11:51:59 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Wed Dec 21 11:52:17 2005 Subject: [Histonet] Embedding Centers In-Reply-To: <6.1.1.1.2.20051221101125.01997318@mail.vet.upenn.edu> References: <6.1.1.1.2.20051221101125.01997318@mail.vet.upenn.edu> Message-ID: <8C7D46E60053F94-A00-1903@mblk-d42.sysops.aol.com> We've had lots of problems with our Microm in a little over a year, compressors mainly --- we've gone through two! So we would not recommend this company. Karen Raterman St. Louis, MO krat18@aol.com -----Original Message----- From: Pamela Marcum To: Roberta Horner ; histonet@lists.utsouthwestern.edu Sent: Wed, 21 Dec 2005 10:14:58 -0500 Subject: Re: [Histonet] Embedding Centers At 08:25 AM 12/21/2005, Roberta Horner wrote: >My embedding center just died. Last time our maintenance looked at it >they couldn't get the parts (its about 17 years old). I need to get >information quick on which ones you like and are good. So far I have >quotes from Surgipath and on the Leica but I don't know anything about >them. Any comments please? >Roberta >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Hi, I will be totally different as i am buying the Thermo Shandon Embedding Center. I had it in for demo and it had all the features the others have for timing and pre-sets as well as some independent temperature ranges I likes. Also the cold plate is seperate so I can have it on either side or in another area if I want for special projects. Good Luck, I think they all work well it just depends for me on features and price and Shandon also had the right price. Best Regards, Pamela A Marcum Manager, Histology Special Procedures University of Pennsylvania School of Veterinary Medicine R.S. Reynolds Jr. CORL New Bolton Center 382 West Street Road Kennett Square, PA 19348 Phone - 610-925-6278 Fax - 610-925-8120 E-mail - pmarcum@vet.upenn.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From krat18 <@t> aol.com Wed Dec 21 12:10:20 2005 From: krat18 <@t> aol.com (krat18@aol.com) Date: Wed Dec 21 12:10:38 2005 Subject: [Histonet] Purple blue haze In-Reply-To: <0BE6ADFAE4E7E04496BF21ABD346628006307C76@EXCHANGE1.huntingtonhospital.com> References: <0BE6ADFAE4E7E04496BF21ABD346628006307C76@EXCHANGE1.huntingtonhospital.com> Message-ID: <8C7D470F0B3F444-18F8-315D@FWM-D18.sysops.aol.com> Funny about this paraffin problem --- we were using Surgipath paraffin, and we noticed that the paraffin looked "dirty" with some kind of sediment floating in it. So we switched to Richard Allan Type 9, and we've had no problems with it! We've been happy with the Richard Allan so far. Karen Raterman St. Mary's Health Center St. Louis, MO krat18@aol.com -----Original Message----- From: Laurie Colbert To: Andrea Grantham ; Histonet (E-mail) Sent: Tue, 20 Dec 2005 15:12:17 -0800 Subject: RE: [Histonet] Purple blue haze Your discussion rang a bell with me, and I've been meaning to call Richard Allan. We use their Type 9 paraffin, and for the last 3 or 4 months it also has been "blowing apart on the waterbath." We lowered the temp on the waterbaths, and this seems to have helped. Our housekeeping personnel have even noticed a difference in the paraffin (scraping it off the floor). They say it is stickier lately. I think something has been changed. Laurie Colbert Huntington Hospital Pasadena, CA -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Tuesday, December 20, 2005 1:20 PM To: Histonet (E-mail) Subject: Re: [Histonet] Purple blue haze Karen, Yes I have recently noticed the "haze" of the H&E stained slides. I have been using this hematoxylin for years and this is the first time that I have had a problem with it. Like you I have filtered it but it hasn't seemed to help. I also have been using their Type 6 paraffin and in the last few months have noticed that it blows apart on the waterbath. I have lowered the temp. but it didn't help -> 35-39 degrees C. A different lot number that they sent to me was only marginally better. A couple of weeks ago one of my clients brought a project consisting of blocks that had been processed and embedded here over 4-5 years. The paraffin throughout was RAS Type 6. The blocks from the first 4 years cut fine and laid out on the waterbath fine but when I got to recently embedded blocks they started the rapid expanding and blowing apart when they were placed on the waterbath. A histotech in CA told me they have been having the same problem in their lab. Hmmmmm.....could they have changed the formula? Maybe this is so with the Hematoxylin as well. I'm trying different paraffins looking for one comparable to the Type 6 only more stable on the waterbath. I really don't want to do this with Hematoxylin because my clients love my staining but I hate ugly slides. Andi At 11:17 AM 12/20/2005 -0700, Heckford, Karen - SMMC-SF wrote: >Dear Histonetters, >Has anyone had any problems with a purple- blue haze on their H&E using >Richard Allen Scientific Hematoxylin 7211. I am currently filtering it. I >am still getting the haze. I have changed nothing, it just appeared a >couple of weeks ago. Any help would be greatly appreciated. > >Happy Holidays, > > >Karen Heckford HT (ASCP) CE >Lead Histology Technician >St. Mary's Medical Center >Histology Department >450 Stanyan St. >San Francisco, Ca. 94117 >1-415-668-1000 ext. 6167 >email: kheckfor@chw.edu > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Dec 21 12:12:32 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Dec 21 12:12:46 2005 Subject: [Histonet] Mouse Liver In-Reply-To: <20051221163205.22628.qmail@web60612.mail.yahoo.com> References: <20051221163205.22628.qmail@web60612.mail.yahoo.com> Message-ID: <6.0.0.22.1.20051221110652.01b5ad68@gemini.msu.montana.edu> One can also try a very short warm water soak (even room temperature water) followed by cold water to recool the block. Warm water penetrates a tidge easier than really cold water. Becareful when sectioning the block - the soaked portion is only a few micrometers deep and you may only get a few sections. If your tissue protrude from the block face after a soak, beware, that is not good either - Re-address processing, it sounds as though there is too much dehydration, clearing, or too long in paraffins one or all at the same time and if they add heat to processing in dehydrants and clearing agents, don't do that - it only dries out mouse tissues even more since they are lean little critters, not a fatty as human tissues. At 09:32 AM 12/21/2005, you wrote: >Luis, > > I agree. Try a long soak in cold water. It sounds like the tissue > needs to be rehydrated a bit. It's way to overdry and you need to let > the people who do your processing know that otherwise they'll do it to > other people. Gayle Callis Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From BMolinari <@t> heart.thi.tmc.edu Wed Dec 21 12:54:58 2005 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Dec 21 12:59:26 2005 Subject: [Histonet] Glass coverslippers Message-ID: I have the Sakura and love it. I do not know if it is faster than the Hacker model because I have never used it. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. 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 Angela Bitting Sent: Wednesday, December 21, 2005 7:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ROrr <@t> enh.org Wed Dec 21 13:27:13 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Dec 21 13:27:23 2005 Subject: [Histonet] Hematoxylin Haze Message-ID: No crazy name calling here, Karen! I'm glad things are working better for you and for your comment on the weather change...I was going to ask you if you were using a tap water rinse and if that tap water could be running at a different temperature now. Maybe the "Cold" is running more cold or even more warm? Who knows what water plants have to do to compensate for quality water that ends up in our faucets. If you are running a DI water, I would ask if that filter might need changed, or is it coming from a commercial tap before it gets filtered through the dionization process.... But when I read Andi's reply, I thought it might be something else, as well. I have learned that when I know I haven't changed a "thing" and my results go wacko, to always look at the water. Cheers! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From escott8 <@t> houston.rr.com Wed Dec 21 14:05:19 2005 From: escott8 <@t> houston.rr.com (Edward Scott) Date: Wed Dec 21 14:05:09 2005 Subject: [Histonet] PI Reports Message-ID: <001601c60669$dea1db80$6400a8c0@thescotts> Hello to all in histoland. I would like to know what type of PI reports are being done by histo supervisors and managers. I am currently doing turnaround time for frozen sections and immuno send outs. We dont do our own immuno. I also do specimen discrepancy as it pertains to to requisitions that we receive. For example, we us Copath and at the end o f the day I pull out a specimen discrepancy log that has documentation for things like no procedure date, no clinical history, no patient history, no location, no attending physicians name. If anyone would like to share some of the reports that they do I would be greatly appreciative. I am looking to make some changes. Merry Christmas to all. Allison Scott Histology Supervisor LBJ Hospital Houston , Texas From bills <@t> icpmr.wsahs.nsw.gov.au Wed Dec 21 14:25:38 2005 From: bills <@t> icpmr.wsahs.nsw.gov.au (Bill Sinai) Date: Wed Dec 21 14:27:23 2005 Subject: [Histonet] FW: artifact In-Reply-To: <4C96AA62BADFD81180CB0002A5AD67FB04D7E282@wsahs.nsw.gov.au> Message-ID: <001701c6066c$b4eeeeb0$796a080a@wsahs.nsw.gov.au> Diane, You have described the classic signs of the biopsy drying before being placed in the fixative. The endoscopists we deal with usually have no problems, however, one had a new nurse assisting and as he gave her the biopsy she placed them on a piece of dry paper. The first biospy taken always showed the artefact the rest OK. A similar problem happens in curettings where the clinician scapes, each time the curette is used, onto a piece of gauze, the outside epithelium always looks hazy. We solved the problem by asking that they place the biopsied material into a small volume of saline or onto saline damped paper or gauze and we have had no problems since. Talk to whoever assists in the procedure and explain what is happening and they are usually willing to cooperate. Seasons greetings Bill Sinai Chief Hospital Scientist Tissue Pathology ICPMR Westmead Hospital Westmead NSW 2145 Phone 02 9845 7774 Fax 02 9687 2330 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Santana, Diane Sent: Thursday, 22 December 2005 2:27 AM To: 'Histonet@lists.utsouthwestern.edu' Subject: [Histonet] FW: artifact > -----Original Message----- > From: Santana, Diane > Sent: Tuesday, December 20, 2005 10:20 AM > To: 'histonet@lists.utsouthwestern.edu' > Subject: artifact > > I am having a problem with artifact on some of my GI bx's. Actually a > paper published by Anatech "The Innovator", summer2004 shows this same > artifact on the front page. They believe it is a fixation problem, and > or retrieval by the endo Drs. My pathologist rules both these ideas > out. I tend to agree with him, at lease on the fixation. Our bx's go > into the processor around 5 pm and the bx run starts around 2 am. > It is a loss of nuclear detail. This artifact can be on a small > section of a bx. Even if there is 5 bx's in one cassette maybe 1 will > have this artifact where the others look great. I thought at first it > was a clearing problem or nuclear bubbling I changed times in clear > rite and oven. It did not eliminate the problem. My pathologist has > seen this by other labs also, but very seldom. For us it could be 1 or > 2 slides a day. Sometimes not at all. Weekend or weekdays. No > consistently. The best way I can describe it is that it looks like a smudge. > Our bx run is this; > > 10% NBF 30 min ea x2 > 80% Alcohol 10mins > 95% alcohol 10 mins ea x2 > 100% alcohol 17 mins ea x3 > Clear rite 22 mins ea x2 > Paraffin 15 mins ea x3 > > I would appreciate any feedback on this. > Diane Santana > Pentucket Medical Assoc. > Haverhill, Mass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________________________ This electronic message and any attachments may be confidential. If you are not the intended recipient of this message would you please delete the message and any attachments and advise the sender. Western Sydney Area Health Services (WSAHS) uses virus scanning software but excludes any liability for viruses contained in any email or attachment. This email may contain privileged and confidential information intended only for the use of the addressees named above. If you are not the intended recipient of this email, you are hereby notified that any use, dissemination, distribution, or reproduction of this email is prohibited. If you have received this email in error, please notify WSAHS immediately. Any views expressed in this email are those of the individual sender except where the sender expressly and with authority states them to be the views of WSAHS. From victor <@t> pathology.washington.edu Wed Dec 21 14:29:25 2005 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Wed Dec 21 14:29:35 2005 Subject: [Histonet] PI Reports In-Reply-To: <001601c60669$dea1db80$6400a8c0@thescotts> References: <001601c60669$dea1db80$6400a8c0@thescotts> Message-ID: <43A9BB25.9030903@pathology.washington.edu> Allison, We are using PowerPath and for frozen sections, we have fields setup to enter the received time and completed time. During our recent CAP inspection we were asked about our TAT. I ran a quick report and our average came in under 20 min. so we were OK. We also have a section for specimen adequacy which is similiar to your specimen discrepancy log. I don't have a clue as to how often it is run. We have a variety of reports for management, such as TAT statistics, overdue cases, etc. Victor Edward Scott wrote: >Hello to all in histoland. I would like to know what type of PI reports are >being done by histo supervisors and managers. I am currently doing >turnaround time for frozen sections and immuno send outs. We dont do our >own immuno. I also do specimen discrepancy as it pertains to to >requisitions that we receive. For example, we us Copath and at the end o f >the day I pull out a specimen discrepancy log that has documentation for >things like no procedure date, no clinical history, no patient history, no >location, no attending physicians name. If anyone would like to share some >of the reports that they do I would be greatly appreciative. I am looking >to make some changes. Merry Christmas to all. > >Allison Scott >Histology Supervisor >LBJ Hospital >Houston , Texas >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From sjchtascp <@t> yahoo.com Wed Dec 21 14:33:35 2005 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Dec 21 14:33:43 2005 Subject: [Histonet] pancreas special stains Message-ID: <20051221203335.25623.qmail@web90201.mail.scd.yahoo.com> Has anyone had any experiance with any of the special stains for pancreatic alpha, beta and delta cells. Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From benoit.delatour <@t> ibaic.u-psud.fr Wed Dec 21 14:59:48 2005 From: benoit.delatour <@t> ibaic.u-psud.fr (Delatour =?iso-8859-1?Q?Beno=EEt?=) Date: Wed Dec 21 15:00:26 2005 Subject: [Histonet] Harris Hematoxylin troubleshooting Message-ID: <6.0.1.1.2.20051221215837.02ff8650@pop3.club-internet.fr> Dear all, I recently tried to do a H&E stain on mice brain frozen sections using a protocol I sucessfully used with paraffin sections. Staining is very bad with frozen sections, showing awful uneven labeling (i have just posted a PDF file on http://www.histonet.org/ with some horrible pics ; the filename is HE-pics.pdf). Here are some details: -Brains were fixed in buffered formaldehyde 10%, then cryoprotected with DMSO-glycerol overnight and cut on a sliding microtome (40?m free floatting sections). Sections were then mounted on Superfrost+ glass slides, dehydrated overnight at 40?C and then processed for H&E staining. -These sections can be perfectly stained using thionin, cresyl violet, Vector nuclear fast red etc... They can also be processed with success for IHC, histochemistry etc... -My protocol is very (too?) simple: rehydratation in tap water - staining in Harris H. (filtered) for 2-4 min - tap water - acidic alcohol ... -My H&E attemps made use of the Harris hemtoxylin from (c)Roth (see http://www.fr.carlroth.com/catalogue/catalogue.do;jsessionid=D750394EC8EA27E19AB00DEE85E744BA?id=6284&favOid=000000000002812b00060023&act=showBookmark&lang=en&catId=FR) -The problem does not come from dewaxing as... brains were not embedded in paraffin! -The problem does not seem to come from to short rehydratation as 20 min under tap water before hematein does not change anything. -I have mounted sections on glass slides using gelatin-PBS or distilled water : does not make any change. -I have increased "incubation" times in the Harris solution (from 2 to 15 min): always uneven stain. All suggestions/comments are welcome. Thanks! Dr. B. Delatour _______________________________ B. Delatour Laboratoire de Neurobiologie de l'Apprentissage, de la M?moire & de la Communication, NAMC, CNRS UMR 8620, B?t 446 Universit? Paris-Sud 91405 Orsay Cedex, FRANCE Email benoit.delatour@ibaic.u-psud.fr Web http://www.namc.u-psud.fr From rjbuesa <@t> yahoo.com Wed Dec 21 15:06:41 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 21 15:06:49 2005 Subject: [Histonet] pancreas special stains In-Reply-To: <20051221203335.25623.qmail@web90201.mail.scd.yahoo.com> Message-ID: <20051221210641.37871.qmail@web61223.mail.yahoo.com> Steve: I strongly recommend you to use the Mallory-Aaan classical trichromic stain widely used by Alexander Alexandrovich Maximow, it will differentiate α, ? and delta cells. Also Bowie's procedure will stain α cells blue; ? cells purple and delta cells red. I have used both procedures with very good results (with pancreas fixed in Zenker's though!). If you don't have the procedure, please let me know. Ren? J. Steven Coakley wrote: Has anyone had any experiance with any of the special stains for pancreatic alpha, beta and delta cells. Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From kbowden <@t> ucsd.edu Wed Dec 21 15:09:01 2005 From: kbowden <@t> ucsd.edu (Bowden) Date: Wed Dec 21 15:09:16 2005 Subject: [Histonet] unsubscribe In-Reply-To: <59342A85CAA485429AF89AAA42FFD9DF36B87D@mailbe10.mc.vanderbilt.edu> References: <59342A85CAA485429AF89AAA42FFD9DF36B87D@mailbe10.mc.vanderbilt.edu> Message-ID: <43A9C46D.4040108@ucsd.edu> > > From bamoe <@t> gundluth.org Wed Dec 21 15:52:53 2005 From: bamoe <@t> gundluth.org (bamoe@gundluth.org) Date: Wed Dec 21 15:53:03 2005 Subject: [Histonet] ER quantitative controls Message-ID: Hello all - For those labs reporting ER results as quantitative (1+, 2+, 3+) --- two questions: 1) Does your control tissue show representation of the 3 different grades - meaning that there is tissue staining as 1+, another piece staining as 2+, and a third piece showing 3+ staining? Or do you simply show a positive reaction? 2) If you do show representation of 1+, 2+, 3+ could you please share your process of obtaining control tissue? Thank you for any comments! Barb Moe Gundersen Lutheran Medical Center La Crosse WI From rjbuesa <@t> yahoo.com Wed Dec 21 16:15:55 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 21 16:16:03 2005 Subject: [Histonet] ER quantitative controls In-Reply-To: Message-ID: <20051221221555.56073.qmail@web61222.mail.yahoo.com> Hello Barb: We used the Dako HercepTest and there are positive controls with it (they are paraffin embedded cultured cells pearls). Since there are only few slides/test kit, we used to batch our tests and use 1 control slide/run of several cases. Sometimes we had + cases to be used as controls but a case has a particular positivity condition, and the slides provided by Dako contain the different + values in the same slide. Perhaps you could contact them and find out if they can sell you + slides. Ren? J. bamoe@gundluth.org wrote: Hello all - For those labs reporting ER results as quantitative (1+, 2+, 3+) --- two questions: 1) Does your control tissue show representation of the 3 different grades - meaning that there is tissue staining as 1+, another piece staining as 2+, and a third piece showing 3+ staining? Or do you simply show a positive reaction? 2) If you do show representation of 1+, 2+, 3+ could you please share your process of obtaining control tissue? Thank you for any comments! Barb Moe Gundersen Lutheran Medical Center La Crosse WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From rjbuesa <@t> yahoo.com Wed Dec 21 16:21:52 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Dec 21 16:22:02 2005 Subject: [Histonet] ER quantitative controls In-Reply-To: Message-ID: <20051221222152.60979.qmail@web61220.mail.yahoo.com> Hello all: My mistake! I was referring to the Her-2-Neu test, and Barb's question was about ER! Regarding ER we never reported it as positive grades; just as + or - Our pathologists considered it irrelevant due to many variables that could affect the + degree of the reaction. My excuses to Barb and all! Ren? J. bamoe@gundluth.org wrote: Hello all - For those labs reporting ER results as quantitative (1+, 2+, 3+) --- two questions: 1) Does your control tissue show representation of the 3 different grades - meaning that there is tissue staining as 1+, another piece staining as 2+, and a third piece showing 3+ staining? Or do you simply show a positive reaction? 2) If you do show representation of 1+, 2+, 3+ could you please share your process of obtaining control tissue? Thank you for any comments! Barb Moe Gundersen Lutheran Medical Center La Crosse WI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From mtitford <@t> aol.com Wed Dec 21 16:42:25 2005 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Wed Dec 21 16:42:37 2005 Subject: [Histonet] Holiday greetings + thanks! Message-ID: <8C7D496F30257FA-914-30A9@mblk-d45.sysops.aol.com> Seasons greetings to everyone from the heart of Dixie! and thanks to Drs Linda Margraf and Herb Hagler and others (and the University of Texas Southwestern Medical School) for hosting the "Histonet". I find it very rewarding! Mike Titford USA Pathology Mobile AL USA From lanbergld <@t> vcu.edu Wed Dec 21 17:55:40 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Wed Dec 21 17:55:51 2005 Subject: [Histonet] Q: Drop of Ammonia to Avoid Wrinkling Message-ID: A Happy Holiday Season to All! When I (archived)&n plastic 10% ammonia wa message is familiar able clarify my understanding After I release the sections onto& slide, I add one small drop of ammon usual. That's the way I interpreted what I rea right? Thanks. &nbs ps. I make 990 nm sections of murine retina, the eye is embedd Spurr's Formula and I use toluidene blue stain. From anh2006 <@t> med.cornell.edu Wed Dec 21 18:40:32 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Dec 21 18:40:46 2005 Subject: [Histonet] CY2/CY3 vs Alexa488/546-555 Message-ID: Has anyone done a SIDE-BY-SIDE comparison of Cy2 or Cy3 as compared with Alexa 488 or Alexa 546/555? I am ideally looking for experimental results not anecdotal information if possible .... I know we have all heard that Alexas are the best and this is certainly true as compared to FITC. But are they better compared to CY2 and CY3 from Jackson Labs? I ask b/c I love Jackson Labs secondaries and they don't sell Alexa conjugates :( I use Alexa Streptavidin from Molecular Probes but am not such a fan of their secondaries for TISSUE staining (they give me some strange artifacts sometimes although they work great for cell culture work). I want to keep ordering Jackson ImmunoResearch quality secondaries - they are just SO SPECIFIC for mouse work but they only sell CY2 and CY3 (which have worked for me thus far). Decisions, decisions .... Comments, opinions, suggestions? THANKS!! Andrea -- From bharatm <@t> glenmarkpharma.com Wed Dec 21 22:28:18 2005 From: bharatm <@t> glenmarkpharma.com (Bharat Mani) Date: Wed Dec 21 22:30:57 2005 Subject: FW: [Histonet] pancreas special stains Message-ID: <2E1FBE2AC0130748B591FD48AC7056E301B3706D@e2k1mhp.glenamrk.com> Bharat Mani Email: bharatm@glenmarkpharma.com -----Original Message----- From: Bharat Mani Sent: Thursday, December 22, 2005 9:57 AM To: 'Rene J Buesa'; 'histonet-bounces@lists.utsouthwestern.edu '; 'sjchtascp@yahoo.com' Subject: RE: [Histonet] pancreas special stains Dear Rene & Steve We have tried the following technique - http://home.primus.com.au/royellis/ST/PANCREAS.htm and observed satisfactory results. Rene, would be glad if you could share the procedures you have tried... Thanks & Regards Bharat Mani Email: bharatm@glenmarkpharma.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Thursday, December 22, 2005 2:37 AM To: Steven Coakley; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] pancreas special stains Steve: I strongly recommend you to use the Mallory-Aaan classical trichromic stain widely used by Alexander Alexandrovich Maximow, it will differentiate α, ? and delta cells. Also Bowie's procedure will stain α cells blue; ? cells purple and delta cells red. I have used both procedures with very good results (with pancreas fixed in Zenker's though!). If you don't have the procedure, please let me know. Ren? J. Steven Coakley wrote: Has anyone had any experiance with any of the special stains for pancreatic alpha, beta and delta cells. Steve __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet DISCLAIMER: The information contained in this email communication is intended only for the personal and confidential use of the designated recipient named above. This message may be an attorney-client communication, and as such is privileged and confidential. If the reader of this message is not the intended recipient, you are hereby notified that you have received this communication in error, and that any review, dissemination, distribution, or copying of the message is strictly prohibited. If you have received this transmission in error, please destroy this transmission and notify Glenmark Pharmaceuticals Limited immediately by telephone and/or send an email to supportho@glenmarkpharma.com From dubya <@t> swbell.net Thu Dec 22 08:47:35 2005 From: dubya <@t> swbell.net (Warren Hammond) Date: Thu Dec 22 08:47:43 2005 Subject: [Histonet] Technicon Parts Message-ID: <20051222144735.79871.qmail@web81107.mail.mud.yahoo.com> Hello, Someone requested parts info for Technicon instruments. The current manufacturer is G&G Instrument and their phone number is 1-800-882-2288 or email to sales@datacut.com. They also have complete units. Merry Christmas Warren Hammond Microscope & Microtome Service Company 2033 Military Parkway Mesquite, Texas 74149 Ph 972-289-9430 Fax 972-289-4488 warren@mandmservicecompany.com From igor.nasonkin <@t> jhmi.edu Thu Dec 22 08:57:31 2005 From: igor.nasonkin <@t> jhmi.edu (IGOR NASONKIN) Date: Thu Dec 22 08:57:41 2005 Subject: [Histonet] CY2/CY3 vs Alexa488/546-555 Message-ID: Andrea, i use both Alexa 546/488 and Cy2/3. The only crucial difference i found so far is that Cy2 does not work with Vectashield mounting medium, while A's naturally do. The choice may also depend on the filter setup of your fluorescent scope. Dr. Igor O. Nasonkin Ph.D. Postdoctoral Fellow Pathology/Neuropathology Johns Hopkins University Ross Research Bldg Rm 563 720 Rutland Ave Baltimore MD 21205 tel: 617-388-4104 Igor.Nasonkin@jhmi.edu ----- Original Message ----- From: "Andrea T. Hooper" Date: Wednesday, December 21, 2005 7:40 pm Subject: [Histonet] CY2/CY3 vs Alexa488/546-555 > Has anyone done a SIDE-BY-SIDE comparison of Cy2 or Cy3 as compared > with Alexa 488 or Alexa 546/555? I am ideally looking for > experimental results not anecdotal information if possible .... I > know we have all heard that Alexas are the best and this is > certainly > true as compared to FITC. But are they better compared to CY2 and > CY3 > from Jackson La bs? > > I ask b/c I love Jackson Labs secondaries and they don't sell Alexa > conjugates :( I use Alexa Streptavidin from Molecular Probes but am > not such a fan of their secondaries for TISSUE staining (they give > me > some strange artifacts sometimes although they work great for cell > culture work). I want to keep ordering Jackson ImmunoResearch > quality > secondaries - they are just SO SPECIFIC for mouse work but they > only > sell CY2 and CY3 (which have worked for me thus far). > > Decisions, decisions .... Comments, opinions, suggestions? > > THANKS!! > Andrea > -- > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Luis.Chiriboga <@t> med.nyu.edu Thu Dec 22 09:09:51 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Thu Dec 22 09:08:53 2005 Subject: [Histonet] Responses to mouse liver tissue processing In-Reply-To: Message-ID: everyone suggested changing the processing schedule (I would if I had could) + permutations on cold water soaking to get good sections. Thanks to everyone who responded. Peace & Happy Holidays to all Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Luis Chiriboga Sent: Wednesday, December 21, 2005 9:54 AM To: Histonet Subject: [Histonet] Mouse liver tissue processing Hi everyone We are having a real serious problem getting good sections from mouse livers. The tissue curls and shreds out of the block as soon as it comes in contact with the knife, end up with a section that has the exact outline of the tissue but no tissue. The tissue consistency could best be described as "dry?" Interestingly, it is only the normal livers that have this problem. We have tried sectioning thick, thin, no soak, cold soak......but out of ideas. Anyone have any suggestions? Unfortunately we have no control over the tissue processing, but perhaps can convince to change. Thanks Luis Happy Holidays to all!! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pathrm35 <@t> adelphia.net Thu Dec 22 10:07:23 2005 From: pathrm35 <@t> adelphia.net (pathrm35@adelphia.net) Date: Thu Dec 22 10:07:31 2005 Subject: [Histonet] SE Florida position Message-ID: <14533509.1135267643033.JavaMail.root@web14.mail.adelphia.net> Fellow Tech's, We are currently seeking a candidate for a tech position in our newly built dermpath lab in SE Florida. This position requires HT and/or HTL (ASCP) certification. A State of Florida license as a Histology Technologist (prefered) or Technician. Duties include: grossing ( CLIA requirements apply), IHC and Special Stains (we have a new Biogenex stainer with great results!!), embedding and sectioning of derm specimens. We are located in a professional atmosphere and have year round sunshine!! Ron Martin, BS, HT(ASCP)HTL,QIHC fax 561-721-1249 772-579-9350 From gcallis <@t> montana.edu Thu Dec 22 10:11:09 2005 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Dec 22 10:11:28 2005 Subject: [Histonet] Re: CY2/CY3 vs Alexa488/546-555 In-Reply-To: References: Message-ID: <6.0.0.22.1.20051222090222.01b50a18@gemini.msu.montana.edu> To get around the problem of Alexa conjugated secondaries you don't like from Molecular Probes, buy your favorite secondary from Jackson, and conjugate it yourself with an Alexa dye conjugation kit from Molecular Probes. You will get the best of both (of your favorite) worlds. The kit conjugates three antibodies, and is a very simple to use. This may be the only way to guarantee the quality you desire and still have your favorite Alexas available. We have experienced similar problems as you with MP Alexa conjugated secondaries. However, last week's IFA using a highly adsorbed goat antiRabbit-Alexa 488 at 0.5ug/ml with a rabbit polyclonal for a bacteria in frozen mouse lung sections gave spectacular results - a pleasant surprise for me and one PI. At 05:40 PM 12/21/2005, you wrote: >Has anyone done a SIDE-BY-SIDE comparison of Cy2 or Cy3 as compared with >Alexa 488 or Alexa 546/555? I am ideally looking for experimental results >not anecdotal information if possible .... I know we have all heard that >Alexas are the best and this is certainly true as compared to FITC. But >are they better compared to CY2 and CY3 from Jackson Labs? > >I ask b/c I love Jackson Labs secondaries and they don't sell Alexa >conjugates :( I use Alexa Streptavidin from Molecular Probes but am not >such a fan of their secondaries for TISSUE staining (they give me some >strange artifacts sometimes although they work great for cell culture >work). I want to keep ordering Jackson ImmunoResearch quality >secondaries - they are just SO SPECIFIC for mouse work but they only sell >CY2 and CY3 (which have worked for me thus far). > >Decisions, decisions .... Comments, opinions, suggestions? > >THANKS!! >Andrea >-- Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From PMonfils <@t> Lifespan.org Thu Dec 22 10:58:22 2005 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Thu Dec 22 10:58:36 2005 Subject: [Histonet] Histological Holiday Greetings Message-ID: <09C945920A6B654199F7A58A1D7D1FDE01717635@lsexch.lsmaster.lifespan.org> Thought I would share with you the microslide greeting I distributed to our staff this year ... http://members.cox.net/paulcyp/DSC01125 http://members.cox.net/paulcyp/DSC01127 (staining technique - Basic Fuchsin and Fast Green) Since a lot of people are not too comfortable with the idea of animal organs hanging on their tree, I used baloney as the tissue type (still animal organs of course, but in a more palatable form). Merry Christmas. From jeffmilz <@t> yahoo.com Thu Dec 22 11:20:05 2005 From: jeffmilz <@t> yahoo.com (jeff milz) Date: Thu Dec 22 11:20:15 2005 Subject: [Histonet] unsubscribe In-Reply-To: <43A9C46D.4040108@ucsd.edu> Message-ID: <20051222172005.88108.qmail@web30006.mail.mud.yahoo.com> Bowden wrote: > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From nellisk <@t> mail.nih.gov Thu Dec 22 12:23:26 2005 From: nellisk <@t> mail.nih.gov (Nellis, Kevin Lee (NIH/NCI) [E]) Date: Thu Dec 22 12:23:37 2005 Subject: [Histonet] NOW RECRUITING Supervisory Histology Technician Message-ID: Please pass this information to qualified candidates. The National Cancer Institute is seeking a Supervisory Histology Technician, GS-0616-11/12. The salary range for this position is $52,468 to $81,747. Relocation expenses and recruitment bonus may be paid. The Laboratory of Pathology (LP) within the National Cancer Institute, Center for Cancer Research provides expertise and diagnostic services in the field of anatomic pathology for all of the categorical institutes of the National Institutes of Health and Clinical Center patients; collaborates with research staff in investigations which involve the tissue and study of human pathological materials, provides instruction in the discipline of surgical pathology to residents; participates in teaching and interdepartmental conferences with staff; provides consultant services to NIH clinical staff and the oncology and general medical community including pathologists throughout the country and conducts independent applied research in surgical pathology. Within the LP, the Histology Laboratory provides technical support for Surgical Pathology, Autopsy, and Cytopathology services. The successful candidate will oversee and perform a full range of clinical histopathology techniques, maintains regulatory requirements, assures successful accreditation, and improves the quality of clinical patient care. He or she will analyze technical and organizational issues to make process improvements, priorities, and schedules that integrate with multiple sections within a department. Applicants should apply online through USA Jobs located at http://www.usajobs.gov/ See job announcement # NCI-05-105257 The closing date for applications is January 06, 2006. Be sure to read application instructions carefully and provide all information including statements of Knowledge Skills and Abilities (KSAs). The DHHS and NIH are Equal Opportunity Employers. Kevin L. Nellis, MS, MT(ASCP) Clinical Laboratory Manager Laboratory of Pathology CCR/NCI/NIH/HHS (301) 594-9532 http://home.ncifcrf.gov/ccr/lop/Clinical/default.asp From benoit.delatour <@t> ibaic.u-psud.fr Thu Dec 22 12:34:55 2005 From: benoit.delatour <@t> ibaic.u-psud.fr (Delatour =?iso-8859-1?Q?Beno=EEt?=) Date: Thu Dec 22 12:35:34 2005 Subject: [Histonet] Harris Hematoxylin troubleshooting (photos) Message-ID: <6.0.1.1.2.20051222193211.0302c820@pop3.club-internet.fr> Dear all, As histonet.org seems to be the wrong solution, here is a new link http://b_delat.club.fr/bd_stuff/HE-pics.pdf to download my H&E pics (related to the message I sent Yesterday). B. _______________________________ B. Delatour Laboratoire de Neurobiologie de l'Apprentissage, de la M?moire & de la Communication, NAMC, CNRS UMR 8620, B?t 446 Universit? Paris-Sud 91405 Orsay Cedex, FRANCE Email benoit.delatour@ibaic.u-psud.fr Web http://www.namc.u-psud.fr From Emilien.Gantelet <@t> unil.ch Thu Dec 22 12:52:31 2005 From: Emilien.Gantelet <@t> unil.ch (Emilien.Gantelet@unil.ch) Date: Thu Dec 22 12:52:40 2005 Subject: [Histonet] Toluidine blue with urea Message-ID: Hello histonetters, I am trying to perform direct counting of motor neurons in mice spinal cord. The fixative does not penetrate correctly in the heart of grey matter, so my usual recipe of 1% Toluidine Blue + 1% borax in distilled water (which is soooo reliable in other cases) just appears to be inefficient : there is strong persistance of a pale haloin the center of my sections (embedded in epoxy). That's why I'm interested today in adding urea to this recipe. I started with 1% urea. I got perfect results on 2 microns thick sections, but the contrast is far too weak on 1 micron thick sections. Do you think of anything I could try, to increase this contrast? Or is the 1 micron thickness totally unsufficient to show correctly Nissl with toluidine blue? Thank you all for your suggestions! Merry Christmas and happy new 2006 year!!!! Emilien Gantelet --------------------------------------------------------------------- Emilien Gantelet, assistant-doctorant D?partement de Biologie Cellulaire et de Morphologie (DBCM) Universit? de Lausanne Rue du Bugnon, 9 1005 Lausanne Switzerland From sbreeden <@t> nmda.nmsu.edu Thu Dec 22 13:59:18 2005 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Thu Dec 22 13:59:27 2005 Subject: [Histonet] Happy or Merry Message-ID: As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From Jackie.O'Connor <@t> abbott.com Thu Dec 22 14:09:44 2005 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Dec 22 14:10:13 2005 Subject: [Histonet] Happy or Merry Message-ID: To try to be perfectly politically correct - - Merry Christmakwanzukah to everyone! Let us not forget Happy Festivus for the Rest of us! Jackie O' "Breeden, Sara" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/22/2005 01:59 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Happy or Merry As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From c.ingles <@t> hosp.wisc.edu Thu Dec 22 15:25:18 2005 From: c.ingles <@t> hosp.wisc.edu (Ingles Claire) Date: Thu Dec 22 15:28:39 2005 Subject: [Histonet] Happy or Merry References: Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3025ACE89@uwhis-xchng2.hosp.wisc.edu> Hey, isn't Ramadan during the holiday season this year too? Don't forget Winter Solstice either! :) Happy Sacred time of the year everyone! Claire Ingles ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jackie M O'Connor Sent: Thu 12/22/2005 2:09 PM To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Happy or Merry To try to be perfectly politically correct - - Merry Christmakwanzukah to everyone! Let us not forget Happy Festivus for the Rest of us! Jackie O' "Breeden, Sara" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/22/2005 01:59 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Happy or Merry As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ 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 Thu Dec 22 15:50:45 2005 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Dec 22 15:50:49 2005 Subject: [Histonet] Happy or Merry Message-ID: Hey Ramadan has finished but ...... political correctness is being carried to ridiculous extremes throughout the country. I prefer to call the holidays by their given names. Calling this Christmas holiday is by no means a reflection on other religions. Let us respect all religions and cultures at this time of year and hope that we carry this philosophy throughout the new year. A very merry Christmas to y'all and a big thank you to Histonet for yet another excellent year. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Thursday, December 22, 2005 3:25 PM Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Hey, isn't Ramadan during the holiday season this year too? Don't forget Winter Solstice either! :) Happy Sacred time of the year everyone! Claire Ingles ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jackie M O'Connor Sent: Thu 12/22/2005 2:09 PM To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Happy or Merry To try to be perfectly politically correct - - Merry Christmakwanzukah to everyone! Let us not forget Happy Festivus for the Rest of us! Jackie O' "Breeden, Sara" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/22/2005 01:59 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Happy or Merry As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ 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 jseaton <@t> wlgore.com Thu Dec 22 17:02:49 2005 From: jseaton <@t> wlgore.com (Janella Seaton) Date: Thu Dec 22 17:05:26 2005 Subject: [Histonet] Janella Seaton/WLGORE is out of the office. Message-ID: I will be out of the office starting 12/22/2005 and will not return until 12/27/2005. I will respond to your message when I return. From vazquezr <@t> ohsu.edu Thu Dec 22 17:26:08 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Thu Dec 22 18:11:47 2005 Subject: [Histonet] Happy or Merry Message-ID: WOW!!! Couldn't of said it better myself...thanks and merry Christmas to all... Robyn From Eric <@t> ategra.com Thu Dec 22 12:16:05 2005 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Thu Dec 22 18:13:57 2005 Subject: [Histonet] Immediate Job Opportunities For HistoTechs (perm and contract) Message-ID: Histonetters - I have temporary travel assignments and permanent HistoTech openings available for you right now throughout the Southeast and the rest of the country! Right now, I have a temeporary assignment in Florida which does require a Florida license. If you are currently available, please contact me ASAP at (800) 466-9919, ext. 223. Our clients are now hiring and need to fill these positions ASAP. Thanks for your time/attention... Eric Dye (800) 466-9919, ext. 223 PS - If you know anyone else who is currently looking, I would appreciate it if you could email me there phone number/email address. (Your friend would also appreciate it too!) PSS - If YOU are interested in any of these opportunities, please call me ASAP @ (800) 466 9919 x223. To speed things up, please send me a copy of your resume, (if you haven't already done so) - There is absolutely N O C O S T to you whatsoever! Our services are entirely paid for by the hiring companies. Eric Dye (800) 466-9919, ext. 223 --------------------------------------------------------- Note: this message is intended for: Histonetters at histonet@lists.utsouthwestern.edu --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd Winter Park, FL 32792 (800)466-9919 ext 231 FAX: (407) 671-6075 eric@ategra.com To Learn More About Ategra: http://www.ategra.com -------------------------------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again -------------------------------------------------------------------------------------- From CrochiereSteve <@t> aol.com Thu Dec 22 20:51:15 2005 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Thu Dec 22 20:51:29 2005 Subject: [Histonet] mitochondrial peroxidase Message-ID: <1a2.42369acc.30dcc023@aol.com> Does anyone know of a blocking reagent to eliminate endogenous mitochondrial peroxidase? I have a tumor case that is staining positive (nonspecifically) for everything and the pathologist seem s to think it's caused by what he's calling "Mitochondrial peroxidase" in the tumor cells. Any ideas how to block this? Have a merry Christmas, Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 From petepath <@t> yahoo.com Fri Dec 23 06:53:22 2005 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Fri Dec 23 06:53:29 2005 Subject: [Histonet] Histological Holiday Greetings Message-ID: <20051223125322.31778.qmail@web30401.mail.mud.yahoo.com> Your ornaments are beautiful Paul. Happy and healthy holiday to you and all of our histonet colleagues. And thanks to you and all the wizards for sharing all of their tricks. stephen From Luis.Chiriboga <@t> med.nyu.edu Fri Dec 23 07:25:09 2005 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Dec 23 07:22:11 2005 Subject: [Histonet] Happy or Merry In-Reply-To: Message-ID: I agree....... but just to be safe.........have a happy Saturday Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Rittman, Barry R Sent: Thursday, December 22, 2005 4:51 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Hey Ramadan has finished but ...... political correctness is being carried to ridiculous extremes throughout the country. I prefer to call the holidays by their given names. Calling this Christmas holiday is by no means a reflection on other religions. Let us respect all religions and cultures at this time of year and hope that we carry this philosophy throughout the new year. A very merry Christmas to y'all and a big thank you to Histonet for yet another excellent year. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Thursday, December 22, 2005 3:25 PM Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Hey, isn't Ramadan during the holiday season this year too? Don't forget Winter Solstice either! :) Happy Sacred time of the year everyone! Claire Ingles ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jackie M O'Connor Sent: Thu 12/22/2005 2:09 PM To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Happy or Merry To try to be perfectly politically correct - - Merry Christmakwanzukah to everyone! Let us not forget Happy Festivus for the Rest of us! Jackie O' "Breeden, Sara" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/22/2005 01:59 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Happy or Merry As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ 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 GDawson <@t> dynacaremilwaukee.com Fri Dec 23 08:09:56 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Dec 23 08:12:07 2005 Subject: [Histonet] Happy or Merry Message-ID: All, Sorry to be a Grinch, but this topic can be gone over at nauseum by clicking on the FOX news channel so it need not be the main topic on today's histonet. Bah Humbug, Glen Dawson Milwaukee, WI -----Original Message----- From: Rittman, Barry R [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Thursday, December 22, 2005 3:51 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Hey Ramadan has finished but ...... political correctness is being carried to ridiculous extremes throughout the country. I prefer to call the holidays by their given names. Calling this Christmas holiday is by no means a reflection on other religions. Let us respect all religions and cultures at this time of year and hope that we carry this philosophy throughout the new year. A very merry Christmas to y'all and a big thank you to Histonet for yet another excellent year. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Ingles Claire Sent: Thursday, December 22, 2005 3:25 PM Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Hey, isn't Ramadan during the holiday season this year too? Don't forget Winter Solstice either! :) Happy Sacred time of the year everyone! Claire Ingles ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jackie M O'Connor Sent: Thu 12/22/2005 2:09 PM To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Happy or Merry To try to be perfectly politically correct - - Merry Christmakwanzukah to everyone! Let us not forget Happy Festivus for the Rest of us! Jackie O' "Breeden, Sara" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/22/2005 01:59 PM To: cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Happy or Merry As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ 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 vazquezr <@t> ohsu.edu Fri Dec 23 08:44:08 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Fri Dec 23 08:44:11 2005 Subject: [Histonet] Happy or Merry Message-ID: Who said it is the main topic, it is simply holiday cheer amongst histonetters. Why ruin it for us with your comments? Robyn From rjbuesa <@t> yahoo.com Fri Dec 23 08:59:28 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Dec 23 08:59:36 2005 Subject: [Histonet] mitochondrial peroxidase In-Reply-To: <1a2.42369acc.30dcc023@aol.com> Message-ID: <20051223145928.49896.qmail@web61225.mail.yahoo.com> Hi Steven: That term of "mitochondrial peroxidase" is somewhat confusing for me. Peroxidase exists in the cells (as you well know) to allow to use oxygen delivered by the oxygenated hemoglobin. As far as I remember respiration is a mitochondrial processin all eukaryotes, therefore peroxidase should always be present in the mitochondria. If you can quench peroxidase, if you can "spent" that enzime with the hydrogen peroxide step of the IHC procedure, you should be able to block the peroxidase no matter where it is present, including (or even limited to) the mitochondria. If the hydrogen peroxide you use is enough to block peroxidase from other sections or cases and not in this particular case, perhaps what you could do is to double the blocking time from 5 to 10 minutes to see if you can eliminate the problem. Do you have this same problem in the negative control also? If it is also in the negative control this unspecific staining has nothing to do with the primary antibody. I hope this will help you! Ren? J. CrochiereSteve@aol.com wrote: Does anyone know of a blocking reagent to eliminate endogenous mitochondrial peroxidase? I have a tumor case that is staining positive (nonspecifically) for everything and the pathologist seem s to think it's caused by what he's calling "Mitochondrial peroxidase" in the tumor cells. Any ideas how to block this? Have a merry Christmas, Steven M. Crochiere, HT(ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center Springfield, MA 01104 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! for Good - Make a difference this year. From GDawson <@t> dynacaremilwaukee.com Fri Dec 23 09:17:37 2005 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Dec 23 09:20:19 2005 Subject: [Histonet] Happy or Merry Message-ID: You're right. Sorry...it's already been a rough morning. Comment away. Glen D. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: Friday, December 23, 2005 8:44 AM To: GDawson@dynacaremilwaukee.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Who said it is the main topic, it is simply holiday cheer amongst histonetters. Why ruin it for us with your comments? Robyn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From donna <@t> milestonemed.com Fri Dec 23 10:14:34 2005 From: donna <@t> milestonemed.com (Donna Willis) Date: Fri Dec 23 11:03:33 2005 Subject: [Histonet] Happy or Merry In-Reply-To: Message-ID: <1135357405_3123@osiris.serverlogistics.com> Glen, Hope you day gets better and you have time to enjoy the holidays. Merry Christmas and Happy New Year Everyone, Donna Willis, HT/HTL(ASCP) Milestone Medical North American Application Manager Gulf Coast Sales Rep -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Friday, December 23, 2005 9:18 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry You're right. Sorry...it's already been a rough morning. Comment away. Glen D. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: Friday, December 23, 2005 8:44 AM To: GDawson@dynacaremilwaukee.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Who said it is the main topic, it is simply holiday cheer amongst histonetters. Why ruin it for us with your comments? Robyn _______________________________________________ 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 lbeaudet <@t> ctcsurge.com Fri Dec 23 12:01:21 2005 From: lbeaudet <@t> ctcsurge.com (Laura Beaudet) Date: Fri Dec 23 12:05:21 2005 Subject: [Histonet] automated response Message-ID: <10512231301.AA07433@mail1.ctcsurge.com> I will be out of the office from Friday afternoon, December 23, 2005 through Tuesday, December 27, 2005. If the matter is urgent, please call Computer Trust Corporation at 617-557-9264 for immediate assistance. From mucram11 <@t> comcast.net Fri Dec 23 14:11:56 2005 From: mucram11 <@t> comcast.net (Pam Marcum) Date: Fri Dec 23 14:12:07 2005 Subject: [Histonet] Peloris Processor Message-ID: <122320052011.27285.43AC5A0C0001D35200006A952207020953CECE030E9D0C9A03@comcast.net> Hi All, I know it is just before the holiday however, I need some input please. Does anyone have the Peloris processor? How is it working out for you? I have a PI who thinks it would be a great idea for his research and he has some money to spent. I think it is burning a hole in his pocket and he heard about microwave processing and had to have one. If you want to answer offline directly to me that is fine. I just don't enough about this to say yes or no and he wants to put it in my lab. I don't really need it as we do bone both non and decalcified. Please help with anything you know. Pam Marcum From scoop <@t> mail.nih.gov Fri Dec 23 20:49:29 2005 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Dec 23 20:49:42 2005 Subject: [Histonet] Happy or Merry Message-ID: Dear Histonetters, I just want to wish all the Histonetters a very happy holiday and thank everyone for all the help they've given to me, a relatively ignorant non-histologist who occasionally attempts to do some IHC and histology. In particular, thanks for attempting to answer and not making fun of my stupider question. Most of the histotechs and other histonetters I've met online and in person seem to be among those people who do what they do primarily to help and make a difference for others and you all deserve to be recognized at this time of year! Thanks and have a great holiday! Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-8417 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From e.kaplan <@t> uaeu.ac.ae Fri Dec 23 21:43:12 2005 From: e.kaplan <@t> uaeu.ac.ae (Evelyn Kaplan) Date: Fri Dec 23 21:41:52 2005 Subject: [Histonet] Happy or Merry In-Reply-To: Message-ID: <000001c6083c$2a521440$cb67a00a@aa.uaeu.ac.ae> To all Histonetters out there (and I know there will be some of you in work today, Christmas Eve), a safe and healthy Christmas and New Year. Evelyn Kaplan B.Sc. MLT Coordinator, Department of Medical Education, Faculty of Medicine & Health Sciences, PO Box 17666, United Arab Emirates University, Al Ain, UAE. Tel: 00 9713 7137216 (O) 00 971 0504717457 (Mobile) 00 9713 7672167 (Fax) e.kaplan@uaeu.ac.ae -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Thursday, December 22, 2005 11:59 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Happy or Merry As I sign off to begin the holidays, I wish for each of you non-explosive paraffin, antibodies that work and whirled peas. Merry Christmas to all and a successful 2006! Read you next year. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Fri Dec 23 22:43:54 2005 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Dec 23 22:40:35 2005 Subject: [Histonet] Glass coverslippers Message-ID: <7E070A7B959A9F42BE732545E5CF621094ACBD@SKMCEMAIL.skmc.gov.ae> Yes And before that we had a CV 5000 - it was taken back to its manufacturer for 'repair', got it back - same problem, replaced with new CV 5030 - which also developed problems. That was taken away too....hated both of them with a passion The slips were 'dropped', glass shards all over the place... slides snapped - was without a slipper for almost 4 months!!! Perhaps we were just unlucky - im not sure - some labs swear by them. Im just feeling a 'once bitten, twice shy' kind of feeling Merry Christmas everyone Think of us - working a normal day over here in Abu Dhabi Greetings Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 8:01 PM To: Anne Van Binsbergen Subject: RE: [Histonet] Glass coverslippers Anne: Was the Leica you had the CV5030? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne Van Binsbergen Sent: Wednesday, December 21, 2005 9:24 AM To: Bartlett, Jeanine; Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers Try the Sakura Fast, efficient, small 'footprint' - will fit into a small space Had a Leica once - was so very patient - 2 exchanges (new machines) later - still hated it Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 5:56 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ 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 Sat Dec 24 00:34:14 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sat Dec 24 00:34:23 2005 Subject: [Histonet] Histological Holiday Greetings References: <20051223125322.31778.qmail@web30401.mail.mud.yahoo.com> Message-ID: <43ACEBE6.5F877A81@uwo.ca> For which holy day do you refer? "Stephen Peters M.D." wrote: > > Your ornaments are beautiful Paul. Happy and healthy holiday to you and all of > our histonet colleagues. And thanks to you and all the wizards for sharing all of > their tricks. > > stephen > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Sat Dec 24 00:35:27 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Sat Dec 24 00:35:38 2005 Subject: [Histonet] Happy or Merry Christmas. References: Message-ID: <43ACEC2F.185B48F9@uwo.ca> Why holiday cheer rather than Christmas cheer? Christmas is one of many holy days. Back in the 1950s a holiday in the northern hemisphere was a week or two by the sea in the summer. The word holiday had already changed its meaning long ago. Vile people are trying to impose "holidays" (always plural, but why?) as a replacement for "Christmas." Is this an attempt to remove the original reason for having a few days off work at the end of December? We don't have to use the vile terminology. We get our few days off because of the Christian Christmas. If the vile ones have their way, 25th December will become just another working day for everyone. A happy Christmas to all (and a merry one, within reason for all non-drivers). John Kiernan Anatomy, UWO London, Canada ___________________________________________________ Robyn Vazquez wrote: > > Who said it is the main topic, it is simply holiday cheer amongst histonetters. Why ruin it for us with your comments? > > Robyn > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From e.kaplan <@t> uaeu.ac.ae Sat Dec 24 01:00:11 2005 From: e.kaplan <@t> uaeu.ac.ae (Evelyn Kaplan) Date: Sat Dec 24 00:58:51 2005 Subject: [Histonet] Happy or Merry Christmas. In-Reply-To: <43ACEC2F.185B48F9@uwo.ca> Message-ID: <000001c60857$af0587b0$680dfea9@aa.uaeu.ac.ae> John, I wholeheartedly agree. I live in an Islamic country where we are allowed to celebrate Christmas in its true meaning. Nobody says 'Happy Holidays' to us; always Merry Christmas. All over the UAE, there is Christmas music, and are Christmas lights and decorations. I find it unbelievable that in the UK folks are being told by our politically-correct busybodies we aren't allowed to say merry Christmas anymore for fear of offending non-Christians. Jesus, Mary and Joseph are mentioned in the Koran and Jesus is regarded as a prophet. So what's the problem? Here in my department we have Muslims, Christians and Hindus all working together and all wishing one another well on their chosen holidays without fear or even a flicker of a problem. Wishing you all a very merry Christmas, Evelyn Evelyn Kaplan C Sc C Biol MSc FIBMS DMLM FRMS MBiol MNSH B.Sc. MLT Coordinator, Department of Medical Education, Faculty of Medicine & Health Sciences, PO Box 17666, United Arab Emirates University, Al Ain, UAE. Tel: 00 9713 7137216 (O) 00 971 0504717457 (Mobile) 00 9713 7672167 (Fax) e.kaplan@uaeu.ac.ae -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Saturday, December 24, 2005 10:35 AM To: Robyn Vazquez Cc: GDawson@dynacaremilwaukee.com; histonet@lists.utsouthwestern.edu Subject: [Histonet] Happy or Merry Christmas. Why holiday cheer rather than Christmas cheer? Christmas is one of many holy days. Back in the 1950s a holiday in the northern hemisphere was a week or two by the sea in the summer. The word holiday had already changed its meaning long ago. Vile people are trying to impose "holidays" (always plural, but why?) as a replacement for "Christmas." Is this an attempt to remove the original reason for having a few days off work at the end of December? We don't have to use the vile terminology. We get our few days off because of the Christian Christmas. If the vile ones have their way, 25th December will become just another working day for everyone. A happy Christmas to all (and a merry one, within reason for all non-drivers). John Kiernan Anatomy, UWO London, Canada ___________________________________________________ Robyn Vazquez wrote: > > Who said it is the main topic, it is simply holiday cheer amongst > histonetters. Why ruin it for us with your comments? > > Robyn > > _______________________________________________ > 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 llewllew <@t> shaw.ca Sat Dec 24 01:33:10 2005 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Sat Dec 24 01:35:06 2005 Subject: [Histonet] Happy or Merry Christmas. References: <43ACEC2F.185B48F9@uwo.ca> Message-ID: <000a01c6085c$4ac53750$690a4246@yourlk4rlmsu> Strange isn't it? The message was, as far as I recall, "Peace on earth, goodwill toward men" What a pity some can't show it! Snide remarks because someone expresses a wish for happiness within some other tradition than Christmas? That is nonsense, and completely unchristian. Good wishes should be accepted for what they are - a spiritual wish or blessing. As a committed atheist I accept them as such, so thank you all. Happy Holidays - Christmas, Yule, Saturnalia, Solstice, Hanukkah and a myriad of other midwinter celebrations ancient and modern. Bryan Llewellyn From JWEEMS <@t> sjha.org Sat Dec 24 06:48:25 2005 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Sat Dec 24 06:53:25 2005 Subject: [Histonet] Happy or Merry Christmas. Message-ID: <83AACDB0810528418AA106F9AE9B7F7E018251EC@sjhaexc02.sjha.org> I think the problem in our country is our legal system, which has become so twisted. The field needs professionals like us to help straighten it out! While trying to equalize things for some - the system completely undoes what was more equal in the first place. And then the journalists stir the pot and there goes the runaway train. But in the midst - there are people still trying to spread love and peace to all and that is my wish for all of you. Joyce Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From jqb7 <@t> cdc.gov Sat Dec 24 09:47:17 2005 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Sat Dec 24 09:46:53 2005 Subject: [Histonet] Glass coverslippers Message-ID: I can't blame you for that. We have had much better luck. We have 2 of the CV 5000's. One had a pump problem but they replaced the unit and that was that. We use one for resin mountant and one for aqueous for our IHC slides. People just have to be neat and not drop mountant on the walking beam...if they are careful of that then there are no problems. We had a demo CV 5030 for about 3 months and everyone in the group almost cried when it left because it was so maintenance free. No pump! At any rate, I hope you find one you are happy with. As soon as out budget is official we are getting the CV 5030. It's on the top of our lab's "wish list". -----Original Message----- From: Anne Van Binsbergen [mailto:vanann702@skmc.gov.ae] Sent: Fri 12/23/2005 11:43 PM To: Bartlett, Jeanine Cc: histonet-bounces@lists.utsouthwestern.edu; histonet@pathology.swmed.edu Subject: RE: [Histonet] Glass coverslippers Yes And before that we had a CV 5000 - it was taken back to its manufacturer for 'repair', got it back - same problem, replaced with new CV 5030 - which also developed problems. That was taken away too....hated both of them with a passion The slips were 'dropped', glass shards all over the place... slides snapped - was without a slipper for almost 4 months!!! Perhaps we were just unlucky - im not sure - some labs swear by them. Im just feeling a 'once bitten, twice shy' kind of feeling Merry Christmas everyone Think of us - working a normal day over here in Abu Dhabi Greetings Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 8:01 PM To: Anne Van Binsbergen Subject: RE: [Histonet] Glass coverslippers Anne: Was the Leica you had the CV5030? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne Van Binsbergen Sent: Wednesday, December 21, 2005 9:24 AM To: Bartlett, Jeanine; Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers Try the Sakura Fast, efficient, small 'footprint' - will fit into a small space Had a Leica once - was so very patient - 2 exchanges (new machines) later - still hated it Annieinarabia -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, December 21, 2005 5:56 PM To: Angela Bitting; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Glass coverslippers We recently demo'd the new one from Leica and it was great! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Wednesday, December 21, 2005 8:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Glass coverslippers Can anyone recommend a automated glass coverslipper? I've used Hacker in the past. Any that are faster, better? Thanks. IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ 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 cindipqr <@t> wowway.com Sat Dec 24 10:02:37 2005 From: cindipqr <@t> wowway.com (cindipqr@wowway.com) Date: Sat Dec 24 10:04:00 2005 Subject: [Histonet] Happy or Merry Christmas. In-Reply-To: <000a01c6085c$4ac53750$690a4246@yourlk4rlmsu> References: <43ACEC2F.185B48F9@uwo.ca> <000a01c6085c$4ac53750$690a4246@yourlk4rlmsu> Message-ID: <20051224155735.M93624@wowway.com> "Peace on earth, good will towards men" was announced by the angels at Jesus' birth. It means peace on earth good will towards men from God, to those who accept Him. Not man to man. However, you are right the Bible also says Christians are supposed to treat each other very well, thus showing others that they are Christians. Merry Christmas all! ---------- Original Message ----------- From: Bryan Llewellyn To: Histonet Sent: Fri, 23 Dec 2005 23:33:10 -0800 Subject: Re: [Histonet] Happy or Merry Christmas. > Strange isn't it? The message was, as far as I recall, "Peace on > earth, goodwill toward men" What a pity some can't show it! Snide > remarks because someone expresses a wish for happiness within some > other tradition than Christmas? That is nonsense, and completely > unchristian. Good wishes should be accepted for what they are - a > spiritual wish or blessing. As a committed atheist I accept them as > such, so thank you all. > > Happy Holidays - Christmas, Yule, Saturnalia, Solstice, Hanukkah and > a myriad of other midwinter celebrations ancient and modern. > > Bryan Llewellyn > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------- End of Original Message ------- From pruegg <@t> ihctech.net Sun Dec 25 11:56:56 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Sun Dec 25 11:57:02 2005 Subject: [Histonet] Happy or Merry In-Reply-To: <1135357405_3123@osiris.serverlogistics.com> Message-ID: <200512251756.jBPHuc0D014356@pro12.abac.com> Personally I prefer to hear and say "happy holiday season", I am not interested in perpetuating world dominance by Christianity or any other organized religion. Cheers, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Donna Willis Sent: Friday, December 23, 2005 9:15 AM To: 'Dawson, Glen'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Glen, Hope you day gets better and you have time to enjoy the holidays. Merry Christmas and Happy New Year Everyone, Donna Willis, HT/HTL(ASCP) Milestone Medical North American Application Manager Gulf Coast Sales Rep -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dawson, Glen Sent: Friday, December 23, 2005 9:18 AM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry You're right. Sorry...it's already been a rough morning. Comment away. Glen D. -----Original Message----- From: Robyn Vazquez [mailto:vazquezr@ohsu.edu] Sent: Friday, December 23, 2005 8:44 AM To: GDawson@dynacaremilwaukee.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Happy or Merry Who said it is the main topic, it is simply holiday cheer amongst histonetters. Why ruin it for us with your comments? Robyn _______________________________________________ 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 t3zprestegard <@t> mmm.com Sun Dec 25 12:35:14 2005 From: t3zprestegard <@t> mmm.com (t3zprestegard@mmm.com) Date: Sun Dec 25 13:12:13 2005 Subject: [Histonet] Terri A. Prestegard/US-Corporate/3M/US is out of the office. Message-ID: I will be out of the office starting 12/25/2005 and will not return until 01/03/2006. I will respond to your message when I return. From LJApuzzio <@t> msn.com Mon Dec 26 06:21:00 2005 From: LJApuzzio <@t> msn.com (Louis Apuzzio) Date: Mon Dec 26 06:21:13 2005 Subject: [Histonet] Histology Employment Message-ID: Hello Histoneters; Currently seeking available opportunity in Histology. Small private practice and or Derm Lab, would be ideal. Please contact ljapuzzio@msn.com Thank you. From asmith <@t> mail.barry.edu Mon Dec 26 09:26:10 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Mon Dec 26 09:29:46 2005 Subject: [Histonet] Happy or Merry Christmas. Message-ID: <5D2189E74151CC42BEC02906BA8996322B9140@exchsrv01.barrynet.barry.edu> "Happy Holidays," is simply an attempt to be inclusive. To strangers, I say, "Happy Holidays." To those I know are Christian, or to strangers who wish me a Merry Christmas, I say, "Merry Christmas." To Jewish friends, I say, "Happy Hanukkah." I also ask my Muslim teaching assistant when Ramadan starts so that I know when to say, "Ramadan Mobarak!" to my Muslim students. For my Baha'i friends, I wait until the 21st of March to wish them a happy Nawruz. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Saturday, December 24, 2005 2:33 AM To: Histonet Subject: Re: [Histonet] Happy or Merry Christmas. Strange isn't it? The message was, as far as I recall, "Peace on earth, goodwill toward men" What a pity some can't show it! Snide remarks because someone expresses a wish for happiness within some other tradition than Christmas? That is nonsense, and completely unchristian. Good wishes should be accepted for what they are - a spiritual wish or blessing. As a committed atheist I accept them as such, so thank you all. Happy Holidays - Christmas, Yule, Saturnalia, Solstice, Hanukkah and a myriad of other midwinter celebrations ancient and modern. Bryan Llewellyn _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From histosci <@t> shentel.net Tue Dec 27 09:04:43 2005 From: histosci <@t> shentel.net (HSRL) Date: Tue Dec 27 09:05:03 2005 Subject: [Histonet] Milestone Microwave Message-ID: <000001c60af6$dec2d1a0$0200a8c0@HSRLMAIN> Dear Netters, I hope everyone had a great Christmas. We are thinking of purchasing a Milestone Microwave Tissue Processor. We are a contract lab who deals primarily with animal models. So far, we have received very little input on this processor from the animal/research community. Does anyone have any comments on the Milestone? Thanks, Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org From vazquezr <@t> ohsu.edu Tue Dec 27 10:13:13 2005 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Tue Dec 27 10:13:03 2005 Subject: [Histonet] Re: Happy or Merry Christmas. Message-ID: John, My sincere apologies, I stand corrected, it is Christmas cheer....thanks Robyn From vrn.nair <@t> gmail.com Tue Dec 27 11:50:00 2005 From: vrn.nair <@t> gmail.com (Vandana Nair) Date: Tue Dec 27 11:50:11 2005 Subject: [Histonet] regarding employment opportunities Message-ID: <34dab9490512270950l3c2b6270k518f02ec65cb3bce@mail.gmail.com> Dear Friends, I am a recent graduate. I have done my Masters in Biomedical Engineering with a thesis concentration in Biological sciences.I have research experience mainly in the field of immunohistochemistry, small animal surgery, tissue fixation, microtomy.I am looking for employment in hospitals or research centers, I would be much obliged if you all could help me in this regard. Please email me on vrn.nair@gmail.com . I can send my resume to u all. looking forward to ur replies. Thanks, Vandana From dsantana <@t> pmaonline.com Tue Dec 27 11:32:24 2005 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Tue Dec 27 11:50:53 2005 Subject: [Histonet] artifact Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB04D7E2B3@MAILPMA> a couple of weeks ago I sent out an e mail about an artifact I was getting on my GI bx's. I have done most of the suggestions. From diluting more in the earlier part of the process ( 70% /80%) to changing from 2 clear rite 22 mins ea, to 3 clear rites 22 mins ea. Also covering the lid on our Sakura processor with a towel. Nothing has changed, still its random. Mon, Tues, Fri no artifact. Wed, thus and today (toes) I do. If I have no problems, I don't make any attention changes, if I do I do another. Is there any more suggestions or any place I can send slides to that maybe able to help? My pathologist does not agree that it can be happening during the extraction of the bx's. It is driving me crazy! Diane Santana From linhines <@t> yahoo.com Tue Dec 27 12:03:41 2005 From: linhines <@t> yahoo.com (Linda Hines) Date: Tue Dec 27 12:03:49 2005 Subject: [Histonet] RE: Histo-Techs ON CALL Message-ID: <20051227180341.61548.qmail@web33005.mail.mud.yahoo.com> Happy New Year, Histonetters, Just a reminder : Histo-techs ON CALL is available. Providing Histologist to fill vacancies during staffing shortages. Short Term and Long Term availability. Our Technologists are experienced HT's and registered (ASCP). MOHS Routine Histology Special Stains Immunohistochemical Staining For questions or scheduling please call: Thank-you and have a fantastic year! Linda Hines HT (ASCP) Phone (480) 252-0050 Phoenix, AZ 85053 email: linhines@yahoo.com --------------------------------- Yahoo! for Good - Make a difference this year. From dsantana <@t> pmaonline.com Tue Dec 27 14:33:36 2005 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Tue Dec 27 14:28:42 2005 Subject: [Histonet] Xylene VS substitute Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB04D7E2C0@MAILPMA> Hi, I have another question, does most everyone use Xylene and has anyone successfully switched to a substitute. Diane From asmith <@t> mail.barry.edu Tue Dec 27 16:51:26 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Tue Dec 27 16:55:04 2005 Subject: [Histonet] Xylene VS substitute Message-ID: <5D2189E74151CC42BEC02906BA8996322B9145@exchsrv01.barrynet.barry.edu> I was taught with xylene, and I still use xylene. However, I now use it in a fume hood. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Santana, Diane Sent: Tuesday, December 27, 2005 3:34 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Xylene VS substitute Hi, I have another question, does most everyone use Xylene and has anyone successfully switched to a substitute. Diane _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From TJJ <@t> Stowers-Institute.org Tue Dec 27 17:28:20 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Dec 27 17:28:40 2005 Subject: [Histonet] RE: Histonet Digest, Vol 25, Issue 33 Message-ID: Diane, that's an annoying artifact, for sure. I think it might be useful to record the case numbers of the biopsies that show the artifact, and see if there is any correlation to staffing in the GI suite that is doing these. If they all come from one room, or one physician...it's drying artifact happening before fixation. Are they being grossed by one person/pathologist when you see the artifact, and by someone else when you don't? It will take some detective work to find out if it is a personnel issue. On the other hand, I have seen the same type of artifact on bone marrow samples that were most likely inadequately fixed prior to decalcification. Are these samples being grossed anywhere that they are subjected to decalcified samples (hence they may be coming in contact with a HCl-based decal solution before adequate fixation). It's a stretch, but something else to muddy the waters, for sure. We've also seen that artifact on placenta samples. Again, my instinct tells me it's drying artifact that causes it. I'd be willing to put a couple dollars on it. If you are using heat on your processor during the alcohol dehydration and Clear-Rite steps, maybe lower the temp to 25 degrees and see if that helps? Good luck, and I applaud your determination. Teri Johnson Stowers Institute for Medical Research Kansas City, MO 64110 From TJJ <@t> Stowers-Institute.org Tue Dec 27 17:28:41 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Tue Dec 27 17:30:21 2005 Subject: [Histonet] Re: processing artifact on GI biopsies Message-ID: Ugh, I forgot to change the subject line on my previous response...sorry! (Where IS that unsend button when you need it...) ~tj -----Original Message----- From: Johnson, Teri Sent: Tue 12/27/2005 5:28 PM To: histonet@lists.utsouthwestern.edu; dsantana@pmaonline.com Subject: RE: Histonet Digest, Vol 25, Issue 33 Diane, that's an annoying artifact, for sure. I think it might be useful to record the case numbers of the biopsies that show the artifact, and see if there is any correlation to staffing in the GI suite that is doing these. If they all come from one room, or one physician...it's drying artifact happening before fixation. Are they being grossed by one person/pathologist when you see the artifact, and by someone else when you don't? It will take some detective work to find out if it is a personnel issue. On the other hand, I have seen the same type of artifact on bone marrow samples that were most likely inadequately fixed prior to decalcification. Are these samples being grossed anywhere that they are subjected to decalcified samples (hence they may be coming in contact with a HCl-based decal solution before adequate fixation). It's a stretch, but something else to muddy the waters, for sure. We've also seen that artifact on placenta samples. Again, my instinct tells me it's drying artifact that causes it. I'd be willing to put a couple dollars on it. If you are using heat on your processor during the alcohol dehydration and Clear-Rite steps, maybe lower the temp to 25 degrees and see if that helps? Good luck, and I applaud your determination. Teri Johnson Stowers Institute for Medical Research Kansas City, MO 64110 From Dndsomi <@t> aol.com Tue Dec 27 17:30:43 2005 From: Dndsomi <@t> aol.com (Dndsomi@aol.com) Date: Tue Dec 27 17:30:57 2005 Subject: [Histonet] Xylene VS substitute Message-ID: <26a.32403e0.30e328a3@aol.com> After 14 years, I switched to Formula 83 Jan. 1, 2005 and didn't tell our pathologists until May !!!!!!! They had no idea and are very pleased with our slides. Diane From tahseen <@t> brain.net.pk Tue Dec 27 10:04:37 2005 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Tue Dec 27 18:05:33 2005 Subject: [Histonet] Xylene VS substitute References: <4C96AA62BADFD81180CB0002A5AD67FB04D7E2C0@MAILPMA> Message-ID: <002d01c60aff$3df5e060$972bfea9@m7c0y4> I still use xylene. Muhammad Tahseen, MLT(Punjab Medical Faculty) MT(JIMTEF)Japan Histology Supervisor SKMCH & RC Lahore Pakistan. ----- Original Message ----- From: Santana, Diane To: Sent: Wednesday, December 28, 2005 9:33 AM Subject: [Histonet] Xylene VS substitute > Hi, I have another question, does most everyone use Xylene and has anyone > successfully switched to a substitute. > Diane > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Tue Dec 27 18:34:40 2005 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Tue Dec 27 18:34:55 2005 Subject: [Histonet] Anti-betaGal antibody Message-ID: Any suggestions for an anti-bGal antibody for mouse paraffin or frozen section immunohistochemistry which will WORK? I want to make sure my records are accurate before I purchase another one :) Thanks! -- From lanbergld <@t> vcu.edu Tue Dec 27 19:17:29 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Tue Dec 27 19:17:41 2005 Subject: [Histonet] Wrinkling causes in the staining process? Message-ID: I am hoping that a microtomist far more experienced than I might know a likely cause for wrinkling in semi-thin (990nm)plastic sectioning, after the cutting process. The wrinkles will be profuse on one section, but its neighbor on the slide may be completely perfect. All sections will come from the very same ribbon. The wrinkles appear as extremely thin, spidery lines in an otherwise beautful section. Through much investigation into the matter, we are reaching the conclusion that the cause must be in the staining or mounting process and not the cutting, floating or drying process. 1. I dry the slides overnight on a low-to-very low heat. *The sections appear very nice after this step, under microscopic examination. 2. Next day I cover these with Toluidine Blue and set the stain, into the sections, briefly on a moderate heat. 70 - 80 degrees Celsius. 3. I rinse lightly with deionized water and then a gentle wash of 70% EtOH. 4. Complete destaining by submerging in Coplins of 100% EtOH. 5. Air dry. 6. A drop or two of xylene on the stained sections, then place face down onto a cover slip that has a BB-sized drop of Permount. Is anything about this process likely to cause a wrinkling? If so, is there an alternative step that's worth trying? Any input will be greatly appreciated. Thanks so very much. From jkiernan <@t> uwo.ca Tue Dec 27 23:45:48 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Dec 27 23:45:29 2005 Subject: [Histonet] Wrinkling causes in the staining process? References: Message-ID: <43B2268C.8A8E88D@uwo.ca> Dear Lawrence D Lanberg, You don't specify the type of tissue or plastic, or the sizes of the sections. Are these tiny little squares cut from blocks also used for ultrathin sections for EM? Or are they bigger sections, intended only for light microscopy? Has the tissue passed through glutaraldehyde, osmium tetroxide etc (for EM) or some simpler fixation for LM? You mention ribbons, a term usually applied to adhering paraffin sections coming off a steel knife edge. In your case, does ribbons mean lines of sections floating on the trough behind a glass or diamond knife? If these are semi-thin sections of specimens prepared for EM (embedded in a cross-linked epoxy resin), the lines that you describe might be cracks rather than wrinkles. Air drying is usually OK for semi-thin epoxy sections, but cracking happens when sections of any thickness air-dry too quickly, whatever the embedding medium (if any). In your procedure, air drying after 100% alcohol (Step 5) could well be the culprit, and it's not needed. Move the slide from the last 100% alcohol through 2 Coplin jars of xylene, and apply the coverslip, using Permount or any other resinous mountant. If this doesn't solve the problem, look at the sections with a microscope (wet and dry) at every step! Find out when the wrinkles or cracks develop. Optical trickery can reveal physical defects that are invisible with the ideal (Kohler) illumination that we use for transparent, stained slides. To trace a physical artifact, change first the illumination. Lower the condenser a bit. Close down the substage diaphragm. Intrude a 5-10mm object (a pea on a pin) between the field iris and the condenser, a bit off-centre. That trick can provide an almost Nomarski effect. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Lawrence D Lanberg/O/VCU wrote: > > I am hoping that a microtomist far more experienced than I might know a > likely cause for wrinkling in semi-thin (990nm)plastic sectioning, after > the cutting process. The wrinkles will be profuse on one section, but its > neighbor on the slide may be completely perfect. All sections will come > from the very same ribbon. The wrinkles appear as extremely thin, spidery > lines in an otherwise beautful section. > > Through much investigation into the matter, we are reaching the conclusion > that the cause must be in the staining or mounting process and not the > cutting, floating or drying process. > > 1. I dry the slides overnight on a low-to-very low heat. *The sections > appear very nice after this step, under microscopic examination. > 2. Next day I cover these with Toluidine Blue and set the stain, into the > sections, briefly on a moderate heat. 70 - 80 degrees Celsius. > 3. I rinse lightly with deionized water and then a gentle wash of 70% EtOH. > 4. Complete destaining by submerging in Coplins of 100% EtOH. > 5. Air dry. > 6. A drop or two of xylene on the stained sections, then place face down > onto a cover slip that has a BB-sized drop of Permount. > > Is anything about this process likely to cause a wrinkling? If so, is there > an alternative step that's worth trying? Any input will be greatly > appreciated. Thanks so very much. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Dec 27 23:46:13 2005 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Dec 27 23:45:53 2005 Subject: [Histonet] Wrinkling causes in the staining process? References: Message-ID: <43B226A5.28A368F5@uwo.ca> Dear Lawrence D Lanberg, You don't specify the type of tissue or plastic, or the sizes of the sections. Are these tiny little squares cut from blocks also used for ultrathin sections for EM? Or are they bigger sections, intended only for light microscopy? Has the tissue passed through glutaraldehyde, osmium tetroxide etc (for EM) or some simpler fixation for LM? You mention ribbons, a term usually applied to adhering paraffin sections coming off a steel knife edge. In your case, does ribbons mean lines of sections floating on the trough behind a glass or diamond knife? If these are semi-thin sections of specimens prepared for EM (embedded in a cross-linked epoxy resin), the lines that you describe might be cracks rather than wrinkles. Air drying is usually OK for semi-thin epoxy sections, but cracking happens when sections of any thickness air-dry too quickly, whatever the embedding medium (if any). In your procedure, air drying after 100% alcohol (Step 5) could well be the culprit, and it's not needed. Move the slide from the last 100% alcohol through 2 Coplin jars of xylene, and apply the coverslip, using Permount or any other resinous mountant. If this doesn't solve the problem, look at the sections with a microscope (wet and dry) at every step! Find out when the wrinkles or cracks develop. Optical trickery can reveal physical defects that are invisible with the ideal (Kohler) illumination that we use for transparent, stained slides. To trace a physical artifact, change first the illumination. Lower the condenser a bit. Close down the substage diaphragm. Intrude a 5-10mm object (a pea on a pin) between the field iris and the condenser, a bit off-centre. That trick can provide an almost Nomarski effect. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Lawrence D Lanberg/O/VCU wrote: > > I am hoping that a microtomist far more experienced than I might know a > likely cause for wrinkling in semi-thin (990nm)plastic sectioning, after > the cutting process. The wrinkles will be profuse on one section, but its > neighbor on the slide may be completely perfect. All sections will come > from the very same ribbon. The wrinkles appear as extremely thin, spidery > lines in an otherwise beautful section. > > Through much investigation into the matter, we are reaching the conclusion > that the cause must be in the staining or mounting process and not the > cutting, floating or drying process. > > 1. I dry the slides overnight on a low-to-very low heat. *The sections > appear very nice after this step, under microscopic examination. > 2. Next day I cover these with Toluidine Blue and set the stain, into the > sections, briefly on a moderate heat. 70 - 80 degrees Celsius. > 3. I rinse lightly with deionized water and then a gentle wash of 70% EtOH. > 4. Complete destaining by submerging in Coplins of 100% EtOH. > 5. Air dry. > 6. A drop or two of xylene on the stained sections, then place face down > onto a cover slip that has a BB-sized drop of Permount. > > Is anything about this process likely to cause a wrinkling? If so, is there > an alternative step that's worth trying? Any input will be greatly > appreciated. Thanks so very much. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lanbergld <@t> vcu.edu Wed Dec 28 07:47:11 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Wed Dec 28 07:47:22 2005 Subject: [Histonet] Wrinkling causes in the staining process? Message-ID: I've read your reply - and I thank you. When I begin, shortly, I am going to try not drying between the EtOH and xylene. I did mention the sections are 990 nanometers thick, which are used for light microscopy. I use Spurr's Formula to make the 'plastic'. These are mice retina sectioning. Thanks again. I'll see what comes about today. From lanbergld <@t> vcu.edu Wed Dec 28 07:50:26 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Wed Dec 28 07:50:35 2005 Subject: [Histonet] Wrinkling causes in the staining process? Message-ID: I've read your relpy and I thank you. Every little tip I get I utilize, or, at least try one time. Might you be suggesting that the Permount be used without xylene? From mcauliff <@t> umdnj.edu Wed Dec 28 08:30:54 2005 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Wed Dec 28 08:32:32 2005 Subject: [Histonet] Wrinkling causes in the staining process? In-Reply-To: References: Message-ID: <43B2A19E.3080707@umdnj.edu> Hi Lawrence: I agree with John Kiernan's comments and I have several points to add. If you are cutting with a glass knife you may have small cracks (as John mentioned) because your knife is not as sharp as necessary for optimum sections. A second possibility is that the xylene-based mounting medium is reacting with the epoxy section and causing wrinkling. Many people use a drop of the same epoxy resin the sections are embedded in as a mounting medium. Geoff Lawrence D Lanberg/O/VCU wrote: >I am hoping that a microtomist far more experienced than I might know a >likely cause for wrinkling in semi-thin (990nm)plastic sectioning, after >the cutting process. The wrinkles will be profuse on one section, but its >neighbor on the slide may be completely perfect. All sections will come >from the very same ribbon. The wrinkles appear as extremely thin, spidery >lines in an otherwise beautful section. > >Through much investigation into the matter, we are reaching the conclusion >that the cause must be in the staining or mounting process and not the >cutting, floating or drying process. > >1. I dry the slides overnight on a low-to-very low heat. *The sections >appear very nice after this step, under microscopic examination. >2. Next day I cover these with Toluidine Blue and set the stain, into the >sections, briefly on a moderate heat. 70 - 80 degrees Celsius. >3. I rinse lightly with deionized water and then a gentle wash of 70% EtOH. >4. Complete destaining by submerging in Coplins of 100% EtOH. >5. Air dry. >6. A drop or two of xylene on the stained sections, then place face down >onto a cover slip that has a BB-sized drop of Permount. > >Is anything about this process likely to cause a wrinkling? If so, is there >an alternative step that's worth trying? Any input will be greatly >appreciated. Thanks so very much. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff@umdnj.edu ********************************************** From ewrona <@t> yahoo.com Wed Dec 28 10:16:40 2005 From: ewrona <@t> yahoo.com (Erin Wrona) Date: Wed Dec 28 10:16:52 2005 Subject: [Histonet] High melting point temperature Message-ID: <20051228161640.51475.qmail@web52511.mail.yahoo.com> Does anyone know of a really high melting point paraffin for embedding, and who sells it? We are working on a project and need the highest we can find. Thanks, Erin Wrona San Francisco From ROrr <@t> enh.org Wed Dec 28 10:29:10 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Dec 28 10:29:25 2005 Subject: [Histonet] QIHC exam Message-ID: Hello everyone, For all you who have taken the new version QIHC WRITTEN exam, what was the most valuable study guide you used? I am not asking for anyone to tell me what is ON the test...I would appreciate knowing what the number one guide used that was the most effective. Due to a death in my family I was unable to attend this year's NSH and the QIHC seminar given by Ethel and Patsy. So I'm taking it "cold"...I have a good idea in MY mind what I know and what I don't know... it's the stuff I don't know that I don't know that I'm a bit nervous about...you know? Thank muchly, Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From ewrona <@t> yahoo.com Wed Dec 28 10:58:14 2005 From: ewrona <@t> yahoo.com (Erin Wrona) Date: Wed Dec 28 10:58:22 2005 Subject: [Histonet] deep embedding molds Message-ID: <20051228165814.23668.qmail@web52515.mail.yahoo.com> Hi all, I am looking for deep embedding molds, as deep as one would get with the old "L" type forms. Does anyone know of a source? Thanks, Erin Wrona San Francisco From jcline <@t> wchsys.org Wed Dec 28 11:20:03 2005 From: jcline <@t> wchsys.org (Joyce Cline) Date: Wed Dec 28 11:20:16 2005 Subject: [Histonet] Xylene vs Substitute/Santana, Diane Message-ID: <000701c60bd2$f1a296c0$1d2a14ac@wchsys.org> I have used a substitute for over 10 years. I have just recently switched to Formula 83 by CBG. I use it on my processor and for my staining. The results are great. Formula 83 dries faster than other subs I have tried. ***** 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 TJJ <@t> Stowers-Institute.org Wed Dec 28 12:06:19 2005 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Dec 28 12:06:44 2005 Subject: [Histonet] Re: QIHC exam Message-ID: Becky, I just recently took and passed it. I'd say the Dako handbook is probably the one most helpful, comprehensive study aid for the exam. Good luck! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From denise.woodward <@t> uconn.edu Wed Dec 28 13:01:24 2005 From: denise.woodward <@t> uconn.edu (Woodward, Denise) Date: Wed Dec 28 13:01:37 2005 Subject: [Histonet] Pasturella and Vibrio antibodies Message-ID: Hello all, I'm looking for two antibodies for IHC on Bovine lung tissues: Pasturella and Vibrio. Anyone out there have a clue where I can get them?? Much thanks and Happy New Year to all, Denise Long Woodward Dept. of Pathobiology and Veterinary Sciences University of Connecticut 31 N. Eagleville Road Storrs, Connecticut 06269-3089 (860) 486-0851 From mclarke <@t> allsaintshealthcare.org Wed Dec 28 13:01:39 2005 From: mclarke <@t> allsaintshealthcare.org (Clarke, Mary) Date: Wed Dec 28 13:01:46 2005 Subject: [Histonet] Xylene vs substitute Message-ID: <6FE2A453DA437F4D96FAAF363F20ABB201578530@WFEXBE04.wfsi.priv> We are currently using Richard-Allan Citrus Clearing agent and are very happy with it. We use for processing and staining with good results. Terri Clarke Histology Supervisor All Saints Laboratory 3801 Spring Street Racine, Wi 53405 mclarke@allsaintshealthcare.org 262-687-6609 - Privileged/Confidential information may be contained in this message. The information contained in this message is intended only for the use of the recipient(s) named above and their co-workers who are working on the same matter. The recipient of this information is prohibited from disclosing the information to any other party unless this disclosure has been authorized in advance. If you are not intended recipient of this message or any agent responsible for delivery of the message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents of this message is strictly prohibited. You should immediately destroy this message and kindly notify the sender by reply E-Mail. Please advise immediately if you or your employer does not consent to Internet E-Mail for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the firm shall be understood as neither given nor endorsed by it. From pruegg <@t> ihctech.net Wed Dec 28 13:56:28 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Wed Dec 28 13:56:28 2005 Subject: [Histonet] Re: QIHC exam In-Reply-To: Message-ID: <200512281956.jBSJu6JJ011405@pro12.abac.com> Members of the NSH IHC Resource Group can access the power point presentation on taking the QIHC Exam given by Patsy Ruegg and Ethel Macrea on our website at www.ihcrg.org join the group at the same website online, membership with NSH is required. We do list several study guides for this exam which include the DAKO IHC Handbook. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Johnson, Teri Sent: Wednesday, December 28, 2005 11:06 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: QIHC exam Becky, I just recently took and passed it. I'd say the Dako handbook is probably the one most helpful, comprehensive study aid for the exam. Good luck! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PatPatterson <@t> mhd.com Wed Dec 28 14:54:18 2005 From: PatPatterson <@t> mhd.com (Patterson, Pat) Date: Wed Dec 28 14:54:29 2005 Subject: [Histonet] Tissue Processors Message-ID: <293C7C19EFF7D611AE1A0002A53F8114164ABFDC@omega.mhd.com> Hi to All! My memory has faded since NSH S/C - I believe there were at least 2 tissue processors displayed that had 2 retorts so you could run two different programs on the same machine at the same time. VisionBiosystems' Peloris was one. Who was the other company? Pat Patterson Methodist Dallas Medical Center patpatterson@mhd.com *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. From leahcox27 <@t> yahoo.com Thu Dec 29 07:46:02 2005 From: leahcox27 <@t> yahoo.com (Leah Cox) Date: Thu Dec 29 07:46:11 2005 Subject: [Histonet] Histology Position in Seattle Message-ID: <20051229134602.35830.qmail@web50308.mail.yahoo.com> Hello Histonetters, A small friendly lab is hiring a Histotech in Seattle. 6am-2pm. If interested or have questions, please contact or send resume to leahcox27@yahoo.com. Thank you, Leah Cox --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From histosci <@t> shentel.net Thu Dec 29 09:40:39 2005 From: histosci <@t> shentel.net (HSRL) Date: Thu Dec 29 09:40:56 2005 Subject: [Histonet] Used Cassette Printers Message-ID: <000401c60c8e$38cb3a50$0200a8c0@HSRLMAIN> Hey Vendors, Do any of you happen to have a used cassette printer for sale? Preferably NOT a Thermo. If so, please give me an e-mail at tomgalati@hsrl.org. Thanks. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org From funderwood <@t> mcohio.org Thu Dec 29 09:53:28 2005 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Thu Dec 29 09:56:28 2005 Subject: [Histonet] deep embedding molds Message-ID: Sakura makes them. They are more than twice as deep as the usual molds. I bought them through VWR. Cat# 25608-834. Fred >>> Erin Wrona 12/28/05 11:58AM >>> Hi all, I am looking for deep embedding molds, as deep as one would get with the old "L" type forms. Does anyone know of a source? Thanks, Erin Wrona San Francisco _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cwscouten <@t> myneurolab.com Thu Dec 29 09:56:36 2005 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Thu Dec 29 09:57:18 2005 Subject: [Histonet] Tissue Processors Message-ID: <5784D843593D874C93E9BADCB87342AB916A4F@tpiserver03.Coretech-holdings.com> The Medite System from Vibratome Company. See the link below. I think you talked to me and to Barbara Oberheide from Medite. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=5 11101&catdesc=Histology+Equipment&CatThreeID=595&CatOneID=4&subcatdesc=T issue+Processing+Systems&idsubcategory=180 You liked our system. Cordially, Charles W. Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300 x 342 FAX 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patterson, Pat Sent: Wednesday, December 28, 2005 2:54 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] Tissue Processors Hi to All! My memory has faded since NSH S/C - I believe there were at least 2 tissue processors displayed that had 2 retorts so you could run two different programs on the same machine at the same time. VisionBiosystems' Peloris was one. Who was the other company? Pat Patterson Methodist Dallas Medical Center patpatterson@mhd.com *********************************************************************** This electronic transmission contains information from Methodist Health System and should be considered confidential and privileged. The information contained in the above messages is intended only for the use of the individual(s) and entity(ies) named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution, or use of this information is prohibited. If you receive this transmission in error, please notify the sender immediately by return e-mail. Methodist Health System, its subsidiaries and affiliates hereby claim all applicable privileges related to the transmission of this communication. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From eileen_dusek <@t> yahoo.com Thu Dec 29 12:55:50 2005 From: eileen_dusek <@t> yahoo.com (eileen dusek) Date: Thu Dec 29 12:55:58 2005 Subject: [Histonet] TBS processor Message-ID: <20051229185550.2937.qmail@web30515.mail.mud.yahoo.com> Happy Holidays Is anyone using a TBS tissue processor? We are trying one and would love input pros and cons. Eileen Dusek Edward Hospital 630-527-3545 --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From asmith <@t> mail.barry.edu Thu Dec 29 15:06:56 2005 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Thu Dec 29 15:10:40 2005 Subject: [Histonet] online journals Message-ID: <5D2189E74151CC42BEC02906BA8996322B9148@exchsrv01.barrynet.barry.edu> A page charge to authors means that a paper is published or not according to whether or not the author can afford to publish. That's a rather poor way to referee science. A charge for subscription or download means that a paper is read only if enough people feel it is worth the cost of butying it. That's very mercenary, but it does give some indication of the value of the paper. Currently, publishers are very foolish about download fees. $25 to $30 a paper is unaffordable. At that price, I download about 1 paper a year. Thus, if my library doesn't have the journal, I won't cite the paper. If download fees were $5 paper, I'd probably download around 10 papers a year and the publishers' revenue would double. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malcolm McCallum Sent: Tuesday, December 20, 2005 1:11 PM To: parc@listserv.uga.edu; ecolog-l@listserv.umd.edu; tws-l@list.wildlife.org; histonet@lists.utsouthwestern.edu Subject: [Histonet] online journals Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) From Adesupod <@t> aol.com Thu Dec 29 16:09:25 2005 From: Adesupod <@t> aol.com (Adesupod@aol.com) Date: Thu Dec 29 16:09:35 2005 Subject: [Histonet] Floater's Threshold Message-ID: Hi, Please I need an infomations on the floater's threshold in the Pathology laboratory. I will appreciate any infomations on this topic of floater's threshold. You guys are doing a wonderful job. You are the best. Histologically Yours, Adesupo Adesuyi. From dellav <@t> musc.edu Thu Dec 29 16:19:53 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Thu Dec 29 16:20:45 2005 Subject: [Histonet] online journals Message-ID: I would like to inform that Histonet community that the NSH is working to put the Journal of Histotechnology online which is scheduled to occur sometime in 2006. Complete articles of the JOH will be available free to NSH members. when considered along with other member benefits, this would seem to offer the greatest value for your dollar. Non-members will be able to obtain articles electronically for $15 Non-members who have a medical library available to them may want to ask their librarian to consider an electronic subscription to JOH. Even those who do not have access to a medical library can have their facility subscribe so that the pathologists and staff can access the articles Authors of articles appearing in JOH will not be charged anything to make their articles available electronically I understand that download fees can be daunting and counterproductive to disseminating important information. However I would question the fairness of having the society's members underwrite the expense of allowing non-members to have free access so I believe that some fee structure is appropriate. The goal of the NSH leadership is to make articles contained in the Journal of Histotechnology widely available. Vinnie Della Speranza President, National Society for Histotechnology >>> "Smith, Allen" 12/29/05 04:06PM >>> A page charge to authors means that a paper is published or not according to whether or not the author can afford to publish. That's a rather poor way to referee science. A charge for subscription or download means that a paper is read only if enough people feel it is worth the cost of butying it. That's very mercenary, but it does give some indication of the value of the paper. Currently, publishers are very foolish about download fees. $25 to $30 a paper is unaffordable. At that price, I download about 1 paper a year. Thus, if my library doesn't have the journal, I won't cite the paper. If download fees were $5 paper, I'd probably download around 10 papers a year and the publishers' revenue would double. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malcolm McCallum Sent: Tuesday, December 20, 2005 1:11 PM To: parc@listserv.uga.edu; ecolog-l@listserv.umd.edu; tws-l@list.wildlife.org; histonet@lists.utsouthwestern.edu Subject: [Histonet] online journals Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Thu Dec 29 16:40:48 2005 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Dec 29 16:40:58 2005 Subject: [Histonet] Floater's Threshold In-Reply-To: Message-ID: <20051229224048.9007.qmail@web61224.mail.yahoo.com> Hi Adesupo: I have to confess that I am puzzled by your question. Do you mean "how many floaters are acceptable". If that is the meaning of your question then my answer is none! This is an unacceptable mistake always due to carelessness. It is always identified by the pathologists and will require to compare the section to the block to determine if the floater was originated while embedding (the floater will be in the block) or if it was due to a mistake in the water bath. In either circumstance counseling and retraining are of the essence. The whole protocol has to be reviewed. Now if you refer to "threshold" as to how many times can it occur to determine counseling I had answered that (counseling is mandatory). How many counseling had to be done by the same histotech to cause different types of sanction, including termination, that will depend on your lab specific policies. I hope this will help you! Ren? J. Adesupod@aol.com wrote: Hi, Please I need an infomations on the floater's threshold in the Pathology laboratory. I will appreciate any infomations on this topic of floater's threshold. You guys are doing a wonderful job. You are the best. Histologically Yours, Adesupo Adesuyi. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Shopping Find Great Deals on Holiday Gifts at Yahoo! Shopping From rkrug <@t> sial.com Thu Dec 29 17:07:05 2005 From: rkrug <@t> sial.com (Robert Krug) Date: Thu Dec 29 17:07:18 2005 Subject: [Histonet] online journals In-Reply-To: Message-ID: Vinnie: Would past years be accessible and would there be a search function? Placing the years on line would be a great idea. Best Regards, Robert Krug St Louis, Missouri "Vinnie Della Speranza" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/29/2005 04:19 PM To , cc histonet@lists.utsouthwestern.edu Subject RE: [Histonet] online journals I would like to inform that Histonet community that the NSH is working to put the Journal of Histotechnology online which is scheduled to occur sometime in 2006. Complete articles of the JOH will be available free to NSH members. when considered along with other member benefits, this would seem to offer the greatest value for your dollar. Non-members will be able to obtain articles electronically for $15 Non-members who have a medical library available to them may want to ask their librarian to consider an electronic subscription to JOH. Even those who do not have access to a medical library can have their facility subscribe so that the pathologists and staff can access the articles Authors of articles appearing in JOH will not be charged anything to make their articles available electronically I understand that download fees can be daunting and counterproductive to disseminating important information. However I would question the fairness of having the society's members underwrite the expense of allowing non-members to have free access so I believe that some fee structure is appropriate. The goal of the NSH leadership is to make articles contained in the Journal of Histotechnology widely available. Vinnie Della Speranza President, National Society for Histotechnology >>> "Smith, Allen" 12/29/05 04:06PM >>> A page charge to authors means that a paper is published or not according to whether or not the author can afford to publish. That's a rather poor way to referee science. A charge for subscription or download means that a paper is read only if enough people feel it is worth the cost of butying it. That's very mercenary, but it does give some indication of the value of the paper. Currently, publishers are very foolish about download fees. $25 to $30 a paper is unaffordable. At that price, I download about 1 paper a year. Thus, if my library doesn't have the journal, I won't cite the paper. If download fees were $5 paper, I'd probably download around 10 papers a year and the publishers' revenue would double. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malcolm McCallum Sent: Tuesday, December 20, 2005 1:11 PM To: parc@listserv.uga.edu; ecolog-l@listserv.umd.edu; tws-l@list.wildlife.org; histonet@lists.utsouthwestern.edu Subject: [Histonet] online journals Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lanbergld <@t> vcu.edu Thu Dec 29 18:23:24 2005 From: lanbergld <@t> vcu.edu (Lawrence D Lanberg/O/VCU) Date: Thu Dec 29 18:23:36 2005 Subject: [Histonet] Using Embedding Medium in Place of Permount Message-ID: I used the suggestions given to me by several of you: Mounting my slides with excess Spurr's Formula I had (tucked away in the freezer)instead of the usual Permount. For each specimen cut and stained, I mounted one slide with Permount and another slide with my excess emebedding formula. There was actually quite a striking difference in amount of wrinkles, between the two types of mounts. So far I am extremely pleased with the difference, but for the life of me I cannot understand why there'd be such a difference. I thank you all for your suggestion. Hope my luck holds out! From ali.moussavi <@t> molbio.gu.se Fri Dec 30 04:15:32 2005 From: ali.moussavi <@t> molbio.gu.se (Ali Moussavi Nik) Date: Fri Dec 30 04:18:04 2005 Subject: [Histonet] problem with autoflurescence Message-ID: <000e01c60d29$f8120ab0$274ef182@alis> Dear All Happy new year I am doing Immunohistochemistry on 4% PFA fixed, vibrotome sectioned embryonic brain. My antibody is FITC conjugated I have a problem with the background green Autoflorescence of the PFA . Can anyone propose me other method of fixation for embryonic brain. Thanks in advance. Best wishes for the new year. Ali Seyed Ali Moussavi Nik D.V.M , PhD Student Department of Cell and Molecular Biology, Gothenburg University Box 462 S-405 30 G?teborg Sweden Phone: +46(031) 7733895 Fax: +46(031) 7733801 www.molbio.gu.se From jotahal <@t> ftvs.cuni.cz Fri Dec 30 06:28:07 2005 From: jotahal <@t> ftvs.cuni.cz (=?iso-8859-2?Q?Jakub_Ot=E1hal?=) Date: Fri Dec 30 06:28:17 2005 Subject: [Histonet] Quantum dot immunohistochemistry Message-ID: <002b01c60d3c$7d6110f0$8428e793@kubanotes> Dear Histonet users, I would like to ask you for advice. I am starting with quantum dot conjugated secondary antibody immunohistochemistry. I have tried one trial with the same protocol I use with Alexa Fluor. Secondary antibody concentration 1:100 (1:50 is recommended by chemicon), but I have terrible background. Does anybody have idea what is "best" working dilution? Thank you very much Happy New Year Jakub Otahal -------------------------------------------------------------------------- Jakub Otahal MD,PhD Department of Developemental Epileptology Institute of Physiology Czech Academy of Sciences Videnska 1083 142 20 Prague 4 Czech Republic tel: +420 241062495 jotahal@ftvs.cuni.cz ------------------------------------------------------------------------- From dellav <@t> musc.edu Fri Dec 30 08:54:11 2005 From: dellav <@t> musc.edu (Vinnie Della Speranza) Date: Fri Dec 30 08:54:53 2005 Subject: [Histonet] online journals Message-ID: Robert, getting past articles scanned and on line will take some doing but this is our plan. and yes, we recognize that a search function would be advantageous so this is our goal. the question is how quickly we can implement this. I'm unable to give you a timeframe at this time but we are actively working toward this goal. Vinnie >>> Robert Krug 12/29/05 06:07PM >>> Vinnie: Would past years be accessible and would there be a search function? Placing the years on line would be a great idea. Best Regards, Robert Krug St Louis, Missouri "Vinnie Della Speranza" Sent by: histonet-bounces@lists.utsouthwestern.edu 12/29/2005 04:19 PM To , cc histonet@lists.utsouthwestern.edu Subject RE: [Histonet] online journals I would like to inform that Histonet community that the NSH is working to put the Journal of Histotechnology online which is scheduled to occur sometime in 2006. Complete articles of the JOH will be available free to NSH members. when considered along with other member benefits, this would seem to offer the greatest value for your dollar. Non-members will be able to obtain articles electronically for $15 Non-members who have a medical library available to them may want to ask their librarian to consider an electronic subscription to JOH. Even those who do not have access to a medical library can have their facility subscribe so that the pathologists and staff can access the articles Authors of articles appearing in JOH will not be charged anything to make their articles available electronically I understand that download fees can be daunting and counterproductive to disseminating important information. However I would question the fairness of having the society's members underwrite the expense of allowing non-members to have free access so I believe that some fee structure is appropriate. The goal of the NSH leadership is to make articles contained in the Journal of Histotechnology widely available. Vinnie Della Speranza President, National Society for Histotechnology >>> "Smith, Allen" 12/29/05 04:06PM >>> A page charge to authors means that a paper is published or not according to whether or not the author can afford to publish. That's a rather poor way to referee science. A charge for subscription or download means that a paper is read only if enough people feel it is worth the cost of butying it. That's very mercenary, but it does give some indication of the value of the paper. Currently, publishers are very foolish about download fees. $25 to $30 a paper is unaffordable. At that price, I download about 1 paper a year. Thus, if my library doesn't have the journal, I won't cite the paper. If download fees were $5 paper, I'd probably download around 10 papers a year and the publishers' revenue would double. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Malcolm McCallum Sent: Tuesday, December 20, 2005 1:11 PM To: parc@listserv.uga.edu; ecolog-l@listserv.umd.edu; tws-l@list.wildlife.org; histonet@lists.utsouthwestern.edu Subject: [Histonet] online journals Hi Which of the below three options do you think is best regarding a journal with both print and online versions???? 1) Journal is open access, the publisher will charge page charges to authors. 2) No page charges (except color plates), but the publisher charges download fees and only the abstracts are open access. 3) Page charges are optional for the author, if the author pays the entire article is open access, if not then only the abstract will be accessible without paying a download fee. In case someone is unaware, If an article is entirely open access then anyone can read it or download it online. IF only the abstract is available then you can read the abstract online, but must pay a fee to download the entire article. Thanks for the feedback, its actually very important! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information transmitted is intended only for the person or entity to which it is addressed and may contain confidential, and/or privileged material. No confidentiality or privilege is waived or lost by any errant transmission. If you receive this message in error, please immediately delete it and all copies of it from your system and notify the sender. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. Barry University - Miami Shores, FL (http://www.barry.edu) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ 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 ROrr <@t> enh.org Fri Dec 30 10:08:47 2005 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Dec 30 10:08:56 2005 Subject: [Histonet] seeking Deb Croegaert Message-ID: Hi everyone... Deb Croegaert wrote an article in the Spring '05 issue of Histo Logic and I had a few questions for her. If any of you at U of Iowa know her, could you please ask her to email me? I tried using her email address from the article and it keeps coming back to me undeliverable. Many thanks to my friends in Iowa City! Becky Becky Orr CLA,HT(ASCP) IHC Lead Evanston Northwestern Healthcare 847-570-2771 From godsgirlnow <@t> msn.com Fri Dec 30 11:22:52 2005 From: godsgirlnow <@t> msn.com (Roxanne Soto) Date: Fri Dec 30 11:23:04 2005 Subject: [Histonet] Floater's Threshold In-Reply-To: Message-ID: In my lab, there is no threshold........ Roxanne Soto ______________________________________________________________ From: Adesupod@aol.com To: histonet@lists.utsouthwestern.edu CC: Adesupod@aol.com Subject: [Histonet] Floater's Threshold Date: Thu, 29 Dec 2005 17:09:25 EST > > Hi, > Please I need an infomations on the floater's threshold in the >Pathology laboratory. I will appreciate any infomations on this topic of floater's >threshold. > You guys are doing a wonderful job. You are the best. > > Histologically Yours, > Adesupo Adesuyi. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From linhines <@t> yahoo.com Fri Dec 30 11:54:24 2005 From: linhines <@t> yahoo.com (Linda Hines) Date: Fri Dec 30 11:54:34 2005 Subject: [Histonet] Re: Histo-Techs ON CALL Message-ID: <20051230175424.4449.qmail@web33007.mail.mud.yahoo.com> Hello everyone, We are currently seeking histologists for the state of Florida. To do temporary assignments. This does require a Florida state license and HT from a national registry (ASCP). Anyone that is interested please contact me with your availability and your salary expectations. Thank-you and Have a Happy New Year! Linda Hines HT (ASCP) Histo-Techs ON CALL Phone: 480-252-0050 email: linhines@yahoo.com --------------------------------- Yahoo! DSL Something to write home about. Just $16.99/mo. or less From pruegg <@t> ihctech.net Fri Dec 30 13:02:51 2005 From: pruegg <@t> ihctech.net (patsy ruegg) Date: Fri Dec 30 13:02:53 2005 Subject: [Histonet] expired antibodies Message-ID: <200512301902.jBUJ2TfH075326@pro12.abac.com> Just a reminder! If you have to get rid of expired antibodies because of CAP, I would be happy to take them off your hands. I can provide you with a fedx acct. # to cover shipping cost. Thank you and Happy New Year, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech 12635 Montview Blvd. #216 Aurora, CO 80010 720-859-4060 fax 720-859-4110 pruegg@ihctech.net www.ihctech.net From Melissa.Gonzalez <@t> cellgenesys.com Fri Dec 30 13:55:47 2005 From: Melissa.Gonzalez <@t> cellgenesys.com (Melissa Gonzalez) Date: Fri Dec 30 13:56:00 2005 Subject: [Histonet] RE:Quantum dot IHC Message-ID: Jakub, I would suggest diluting further in ranges until you still see the unspecific background disappear but retain the "real" staining. Also, ofcourse, make sure the secondary is compatible with the primary (ie-cross-reactivity between species). I am actually curious, you are the first person I have communicated with that has given Quantum dots a try. What source are you using to visualize the staining? How do you differentiate the different wavelengths using these products? I am so happy with Alexa Fluors that I haven't yet given the Quantum dots a go. Good luck, Happy New Year to everyone, Melissa Message: 9 Date: Fri, 30 Dec 2005 13:28:07 +0100 From: Jakub Ot?hal Subject: [Histonet] Quantum dot immunohistochemistry To: Message-ID: <002b01c60d3c$7d6110f0$8428e793@kubanotes> Content-Type: text/plain; charset="iso-8859-2" Dear Histonet users, I would like to ask you for advice. I am starting with quantum dot conjugated secondary antibody immunohistochemistry. I have tried one trial with the same protocol I use with Alexa Fluor. Secondary antibody concentration 1:100 (1:50 is recommended by chemicon), but I have terrible background. Does anybody have idea what is "best" working dilution? Thank you very much Happy New Year Jakub Otahal ------------------------------------------------------------------------ -- Jakub Otahal MD,PhD Department of Developemental Epileptology Institute of Physiology Czech Academy of Sciences Videnska 1083 142 20 Prague 4 Czech Republic tel: +420 241062495 jotahal@ftvs.cuni.cz ------------------------------------------------------------------------ - From jseaton <@t> wlgore.com Fri Dec 30 17:03:10 2005 From: jseaton <@t> wlgore.com (Janella Seaton) Date: Fri Dec 30 17:03:25 2005 Subject: [Histonet] Janella Seaton/WLGORE is out of the office. Message-ID: I will be out of the office starting 12/30/2005 and will not return until 01/03/2006. I will respond to your message when I return. From foley1 <@t> niehs.nih.gov Thu Dec 29 11:40:59 2005 From: foley1 <@t> niehs.nih.gov (foley1) Date: Fri Jan 6 10:06:19 2006 Subject: [Histonet] Processing/embedding question - your opinion Message-ID: Does anyone routinely allow for the hot wax to drain off multiple cassettes of processed tissue and be held at room temperature for multiple days (6 days) before embedding? What would the consequences be to morphology, possible immunohistochemistry and molecular (DNA) studies?