From nick.kirk3 <@t> btopenworld.com Mon Mar 1 00:03:00 2004 From: nick.kirk3 <@t> btopenworld.com (=?iso-8859-1?q?NICK=20KIRK?=) Date: Fri Sep 16 15:22:37 2005 Subject: [Histonet] Prostate Needle Biopsies In-Reply-To: <1db.1b50c90c.2d7406c3@aol.com> Message-ID: <20040301060300.81766.qmail@web86311.mail.ukl.yahoo.com> I agree with the sentiments below, it's difficult enough searching for a translucent biopsy in a clear fluid, searching for a blue biopsy in a blue fluid (or whatever colour the ink is) would be even more difficult. We address the issue of prostate biopsies by giving the Urology department a bag of biopsy sponges. When they take the needle cores they then place them on a single damp biopsy sponge, when all the biopsies have been taken, they then place the sponge into a formalin pot. 99% of the biopsies stay on the sponges and we only get the occasional "stray", which is picked up easily, as we routinely check the pot and the inside of the lid for any missed biopsies. (It's surprising what will sometimes cling to a biopsy pot lid) As for the other issues, well we routinely receive two biopsy pots, one sampled from the left and the second from the right in annotated pots. Each request card details the number of cores in each pot. Then at the macroscopic description phase, the cores are counted and any discrepancies noted and detailed in the report itself. All the cores from each pot are placed in a single cassette and processed as per normal. The resultant blocks are then serially cut with as many sections as possible on each slide. We stain slides 1, 3, 5, 7 and 9 with H&E, sections 2 and 4 are unstained and kept and filed, sections 6 and 8 are placed onto coated slides and the majority of these are then immunostained for high molecular weight cytokeratin. However, we recently did an audit on this process due to the increasing numbers and time consuming nature of the work, to see if this was an effective method. We stained the slides as per normal and gave slides 1, 3 and 5 to the Pathologist to read and diagnose. They then looked at slides 7 and 9 to see if these slides made any difference to their diagnosis. The idea being that if slides 7 and 9 made no difference to their diagnosis we would change our working practice accordingley. The results showed overwhelmingly that slides 7 and 9 made no difference to the diagnosis in over 98% of cases and in the remaining 2% they didn't change the diagnosis from benign to malignant, just a different degree of benign disease. Due to this data, we are currently changing our working practice so that we don't have to cut so many sections, this will be more cost and time effective and should have little or no effect on the end diagnosis. After all, if we don't cut so many sections, there will still be the oppertunity to cut further sections should the pathologist be uncertain of the diagnosis. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England RSRICHMOND@aol.com wrote: Becky Garrison in Jacksonville FL observes: >>Just this week, our Urology Center, in reviewing its QA measures, wanted to know if it's OK for them to ink the specimen by color to reduce specimen identification mixups. My first reaction was no because of concerns that unknown inks might interfere with fixation and ultimately IHC and the possibility of crush artefact introduced by "non-pathology" hands.<< I agree - no. I want them to handle the specimen as little as possible - the ink needs to be added after fixation. They could use colored pens to mark the outsides of the containers with a set of laboratory-prescribed colors that the lab would use to select cassette and slide frosting colors. I know that there are services that pre-ink the specimens, but I've never actually seen it done. Bob Richmond Samurai Pathologist Gastonia NC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Inga.Hansson <@t> neuro.uu.se Mon Mar 1 03:38:38 2004 From: Inga.Hansson <@t> neuro.uu.se (Inga Hansson) Date: Fri Sep 16 15:22:37 2005 Subject: [Histonet] apoptosis Message-ID: Hi everyone! Is anyone using Apoptag kit on mouse cryos and have a problem with weak signal? Any suggestions would be appreciated! Thanks Inga -- Inga Hansson Dept. neuroscience, div. neurobiology PO Box 587, BMC Husargatan 3 S-751 23 Uppsala SWEDEN Phone: +46(18) 471 4384 Fax: +46(18)559017 From shaumik.adhya <@t> ucl.ac.uk Mon Mar 1 07:29:09 2004 From: shaumik.adhya <@t> ucl.ac.uk (Shaumik Adhya) Date: Fri Sep 16 15:22:37 2005 Subject: [Histonet] histological detection of protein carbonyl groups Message-ID: <1078147749.40433aa5a1b07@www.webmail.ucl.ac.uk> Hi Histonetters, Does anyone know of a stain for detecting carbonyl groups on proteins as a result of protein oxidation? All the methods I can find deal with carbonyl content of proteins in minced samples, not tissue sections. I'd be grateful for any ideas. Shaumik From yangpw <@t> umich.edu Mon Mar 1 09:31:03 2004 From: yangpw <@t> umich.edu (yangpw@umich.edu) Date: Fri Sep 16 15:22:37 2005 Subject: [Histonet] rat probe for insitu on mouse tissue? Message-ID: <1078155063.40435737a8c60@mail.umich.edu> Good morning, everyone, I have been doing double insitu on rat brain tissue (c-fos RNA rat probe radioactive and enk RNA rat probe non-radioactive). And the staining works fine. Recently, we are begining to try in situ on mouse brain tissue. Can I just use rat RNA probes on mouse tissue? I have blasted the probes sequence, they match rat c-fos/enk sequence very well, and match mouse c-fos/enk about 90%. Is this kind of match good enough to get good signal on mouse tissue? Does anyone have any suggestion? Do I need to reduce the strigency of wash to retain more signal in this case? If it is not good to use rat probe, can anyone recommend any good mouse probe for c-fos and enkephalin? Thanks very much! Pengwei Yang BioPsychology Program University of Michigan From lange <@t> kennedykrieger.org Mon Mar 1 09:51:07 2004 From: lange <@t> kennedykrieger.org (Mollie Lange) Date: Fri Sep 16 15:22:37 2005 Subject: [Histonet] MeCP2 antibody Message-ID: I am looking for a non-rabbit antibody to MeCP2 (methyl cpg binding protein 2) for immunohistochemistry on frozen sections. Does anybody know of one? Thanks, ML From georgecole <@t> ev1.net Mon Mar 1 10:29:48 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Going to Toronto!!! Message-ID: <000001c3ffaa$6ae08c90$034dbad0@hppav> Histonetters, T O R O N T O !!!! I have received an invitation to present a report on the new methods of muscle and nerve biopsies that have been sent all over the place----109 sent so far----at the National Histotechnology Convention in Toronto next September. I will look forward to meeting those of you who can come to that big get-together. I spoke at the 1978 convention in San Francisco on silver techniques. That was a speech-only presentation before the whole meeting. This time, there will he a lab set up for the muscle and nerve confab. I'm already putting a video together that will project on a large screen, and I mean to make most of it live so we can get down to practical biopsy performance. It will be a good chance to get acquainted, and to work out practical methods to improve the yield of information from muscle and nerve biopsy tissues. It will be a three hour lab. I'm already practicing my presentation on my wife. I've gone through it 32 times already for her. My, It'll be nice to present to folks that don't keep turning the TV up loud while I'm talking. I have been reading every message on the Histonet since last July, when I joined. And I have taken a cue from the messages you fellow techs send to each other. I have reduced my total 3 hour presentation down to the initial letters of every word----you know, like all of you immuno processing techs ---it should be just an elegant up-to-the-minute presentation. But truly folks, I will look forward to doing the best I can to get some of these methods into public domain for the good of the patients .I've been having the time of my life communicating with you folks all over the world. Toronto will be the peek capper to my histoteching. I already sleep hovering 14 inches over the bed.---tension?--- naaaahhh---but If I can just bring it off in Toronto, maybe I can look forward to coming in for a landing.------ See you there, I hope! georgecole@ev1.net From tpmorken <@t> labvision.com Mon Mar 1 10:43:29 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Going to Toronto!!! Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2177358@usca0082k08.labvision.apogent.com> George wrote:<> And only 29 weeks left 'till NSH..... Tim Morken -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Monday, March 01, 2004 8:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Going to Toronto!!! Histonetters, T O R O N T O !!!! I have received an invitation to present a report on the new methods of muscle and nerve biopsies that have been sent all over the place----109 sent so far----at the National Histotechnology Convention in Toronto next September. I will look forward to meeting those of you who can come to that big get-together. I spoke at the 1978 convention in San Francisco on silver techniques. That was a speech-only presentation before the whole meeting. This time, there will he a lab set up for the muscle and nerve confab. I'm already putting a video together that will project on a large screen, and I mean to make most of it live so we can get down to practical biopsy performance. It will be a good chance to get acquainted, and to work out practical methods to improve the yield of information from muscle and nerve biopsy tissues. It will be a three hour lab. I'm already practicing my presentation on my wife. I've gone through it 32 times already for her. My, It'll be nice to present to folks that don't keep turning the TV up loud while I'm talking. I have been reading every message on the Histonet since last July, when I joined. And I have taken a cue from the messages you fellow techs send to each other. I have reduced my total 3 hour presentation down to the initial letters of every word----you know, like all of you immuno processing techs ---it should be just an elegant up-to-the-minute presentation. But truly folks, I will look forward to doing the best I can to get some of these methods into public domain for the good of the patients .I've been having the time of my life communicating with you folks all over the world. Toronto will be the peek capper to my histoteching. I already sleep hovering 14 inches over the bed.---tension?--- naaaahhh---but If I can just bring it off in Toronto, maybe I can look forward to coming in for a landing.------ See you there, I hope! georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Mon Mar 1 10:51:36 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Going to Toronto!!! Message-ID: <566FB0B522443D43AF02D2ADBE35A6F063598B@UTHEVS3.mail.uthouston.edu> George Here in Houston we have rules that apply if you are going to have your wife as a critic of a scientific presentation that you are planning to give: 1. She should have a cattle prod to indicate times when you are talking too much. 2. The presentation should only be rehearsed a maximum of 3 times. 3. You owe her big!! (Now I think of it these are rules that my wife instituted). Good luck on your presentation. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of George Cole Sent: Monday, March 01, 2004 10:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Going to Toronto!!! Histonetters, T O R O N T O !!!! I have received an invitation to present a report on the new methods of muscle and nerve biopsies that have been sent all over the place----109 sent so far----at the National Histotechnology Convention in Toronto next September. I will look forward to meeting those of you who can come to that big get-together. I spoke at the 1978 convention in San Francisco on silver techniques. That was a speech-only presentation before the whole meeting. This time, there will he a lab set up for the muscle and nerve confab. I'm already putting a video together that will project on a large screen, and I mean to make most of it live so we can get down to practical biopsy performance. It will be a good chance to get acquainted, and to work out practical methods to improve the yield of information from muscle and nerve biopsy tissues. It will be a three hour lab. I'm already practicing my presentation on my wife. I've gone through it 32 times already for her. My, It'll be nice to present to folks that don't keep turning the TV up loud while I'm talking. I have been reading every message on the Histonet since last July, when I joined. And I have taken a cue from the messages you fellow techs send to each other. I have reduced my total 3 hour presentation down to the initial letters of every word----you know, like all of you immuno processing techs ---it should be just an elegant up-to-the-minute presentation. But truly folks, I will look forward to doing the best I can to get some of these methods into public domain for the good of the patients .I've been having the time of my life communicating with you folks all over the world. Toronto will be the peek capper to my histoteching. I already sleep hovering 14 inches over the bed.---tension?--- naaaahhh---but If I can just bring it off in Toronto, maybe I can look forward to coming in for a landing.------ See you there, I hope! georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From a-lisowski <@t> northwestern.edu Mon Mar 1 13:40:58 2004 From: a-lisowski <@t> northwestern.edu (a-lisowski@northwestern.edu) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Re: Histonet Digest, Vol 4, Issue 1 Message-ID: <200403011945.i21Jjitt009918@hecky.it.northwestern.edu> ==============Original message text=============== On Mon, 01 Mar 2004 12:00:04 pm CST 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/histonetor, 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. (no subject) (Paula Wilder) 2. Acetylcholinesterase -Kit? (Gudrun Lang) 3. Hi everyone! (Teresa Dominguez) 4. Prostate Needle Biopsies (RSRICHMOND@aol.com) 5. RE: Hi everyone! (Weems, Joyce) 6. Inga Hansson and ED1 antibody (Sharon Cooperman) 7. Re: Prostate Needle Biopsies (NICK KIRK) 8. apoptosis (Inga Hansson) 9. histological detection of protein carbonyl groups (Shaumik Adhya) 10. rat probe for insitu on mouse tissue? (yangpw@umich.edu) 11. MeCP2 antibody (Mollie Lange) 12. Going to Toronto!!! (George Cole) 13. RE: Going to Toronto!!! (Morken, Tim - Labvision) 14. RE: Going to Toronto!!! (Barry R Rittman) ---------------------------------------------------------------------- Message: 1 Date: Sun, 29 Feb 2004 18:21:12 +0000 From: "Paula Wilder" Subject: [Histonet] (no subject) To: ccdub@earthlink.net, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; format=flowed Hi Cindy! We also receive quite a few prostate biopsies A-M, but mostly A-F. Anyway, the policy at our institution is no more than 3 biopsy pieces per cassette, so if an A prostate biopsy contained 7 cores, then the PAs would make two cassettes, one with 3 and other with 4 pieces. As a tech, I find this a little difficult, since both A blocks would have to be embedded together to ensure that 7 pieces were indeed found. The PAs also "ink" the tips of each biopsy with a color dye. We use yellow, black, blue and green. We found that the red ink color bled out to other biopsies. We do this because we sometimes accession several prostate cases back to back, and although they are grossed at different work stations, this is another way to ensure that the accessioning is correct and that patients did not inadvertently get reversed. Paula Wilder St. Joseph Medical Center Towson, MD. 21204 _________________________________________________________________ Find and compare great deals on Broadband access at the MSN High-Speed Marketplace. http://click.atdmt.com/AVE/go/onm00200360ave/direct/01/ ------------------------------ Message: 2 Date: Sun, 29 Feb 2004 20:16:55 +0100 From: "Gudrun Lang" Subject: [Histonet] Acetylcholinesterase -Kit? To: "Histonetliste" Message-ID: <001601c3fef8$98178a90$eeeea8c0@SERVER> Content-Type: text/plain; charset="iso-8859-1" Can anyone recommend a Kit to perform Acetylcholinesterase on rectum biopsies? Can anyone tell me a substitute for iso-OMPA with this reaction? thanks in advance Gudrun Lang, Austria ------------------------------ Message: 3 Date: Sun, 29 Feb 2004 23:24:13 -0300 From: Teresa Dominguez Subject: [Histonet] Hi everyone! To: histonet@lists.utsouthwestern.edu Message-ID: <000a01c3ff34$4bafcaa0$277f46c8@FAMILIA> Content-Type: text/plain; charset=iso-8859-1 Hi All, I am a new suscriber in this list, I work in a Pathology Lab at the Hospital of the city where I live. I am glade to be here, and I would like to share with all of you my experiences, and ask to you whatever I need as an advice. See you in the list messages, bye for now, Maria P.S: sorry if I write something wrong, I am still studying English... H.T Maria T Dominguez Anatomy Pathology Service Hospital Regional Río Grande, Río Grande, Tierra del Fuego, Argentina 54- 02964-422086/88 Ext. 143 ------------------------------ Message: 4 Date: Sun, 29 Feb 2004 22:23:47 EST From: RSRICHMOND@aol.com Subject: [Histonet] Prostate Needle Biopsies To: histonet@lists.utsouthwestern.edu Message-ID: <1db.1b50c90c.2d7406c3@aol.com> Content-Type: text/plain; charset="US-ASCII" Becky Garrison in Jacksonville FL observes: >>Just this week, our Urology Center, in reviewing its QA measures, wanted to know if it's OK for them to ink the specimen by color to reduce specimen identification mixups. My first reaction was no because of concerns that unknown inks might interfere with fixation and ultimately IHC and the possibility of crush artefact introduced by "non-pathology" hands.<< I agree - no. I want them to handle the specimen as little as possible - the ink needs to be added after fixation. They could use colored pens to mark the outsides of the containers with a set of laboratory-prescribed colors that the lab would use to select cassette and slide frosting colors. I know that there are services that pre-ink the specimens, but I've never actually seen it done. Bob Richmond Samurai Pathologist Gastonia NC ------------------------------ Message: 5 Date: Sun, 29 Feb 2004 22:39:37 -0500 From: "Weems, Joyce" Subject: RE: [Histonet] Hi everyone! To: 'Teresa Dominguez ' , "'histonet-bounces@lists.utsouthwestern.edu '" , "'histonet@lists.utsouthwestern.edu '" Message-ID: <8C54DF3F71F6E34A80B1DE628C7EF4DD01DD63E1@exch2.sjha.org> Content-Type: text/plain; charset="iso-8859-1" Welcome, Maria! Don't worry about your English. We're still studying too! And we still get it wrong. Just hop in when you have a question. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu To: histonet@lists.utsouthwestern.edu Sent: 2/29/2004 9:24 PM Subject: [Histonet] Hi everyone! Hi All, I am a new suscriber in this list, I work in a Pathology Lab at the Hospital of the city where I live. I am glade to be here, and I would like to share with all of you my experiences, and ask to you whatever I need as an advice. See you in the list messages, bye for now, Maria P.S: sorry if I write something wrong, I am still studying English... H.T Maria T Dominguez Anatomy Pathology Service Hospital Regional Río Grande, Río Grande, Tierra del Fuego, Argentina 54- 02964-422086/88 Ext. 143_______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histon Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. ------------------------------ Message: 6 Date: Sun, 29 Feb 2004 22:41:34 -0500 From: Sharon Cooperman Subject: [Histonet] Inga Hansson and ED1 antibody To: Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi. I'm writing to say thank you to Inga Hansson who told me about using the ED1 antibody from Serotec for microglia on FFPE tissue with citrate antigen retrieval. It also works great on bone marrow macrophages and knowing about it saved me a huge amount of time. I can't find Inga's email address to email her directly, so if you're out there Inga, thanks. -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 ------------------------------ Message: 7 Date: Mon, 1 Mar 2004 06:03:00 +0000 (GMT) From: NICK KIRK Subject: Re: [Histonet] Prostate Needle Biopsies To: RSRICHMOND@aol.com, Histonet@lists.utsouthwestern.edu Message-ID: <20040301060300.81766.qmail@web86311.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 I agree with the sentiments below, it's difficult enough searching for a translucent biopsy in a clear fluid, searching for a blue biopsy in a blue fluid (or whatever colour the ink is) would be even more difficult. We address the issue of prostate biopsies by giving the Urology department a bag of biopsy sponges. When they take the needle cores they then place them on a single damp biopsy sponge, when all the biopsies have been taken, they then place the sponge into a formalin pot. 99% of the biopsies stay on the sponges and we only get the occasional "stray", which is picked up easily, as we routinely check the pot and the inside of the lid for any missed biopsies. (It's surprising what will sometimes cling to a biopsy pot lid) As for the other issues, well we routinely receive two biopsy pots, one sampled from the left and the second from the right in annotated pots. Each request card details the number of cores in each pot. Then at the macroscopic description phase, the cores are counted and any discrepancies noted and detailed in the report itself. All the cores from each pot are placed in a single cassette and processed as per normal. The resultant blocks are then serially cut with as many sections as possible on each slide. We stain slides 1, 3, 5, 7 and 9 with H&E, sections 2 and 4 are unstained and kept and filed, sections 6 and 8 are placed onto coated slides and the majority of these are then immunostained for high molecular weight cytokeratin. However, we recently did an audit on this process due to the increasing numbers and time consuming nature of the work, to see if this was an effective method. We stained the slides as per normal and gave slides 1, 3 and 5 to the Pathologist to read and diagnose. They then looked at slides 7 and 9 to see if these slides made any difference to their diagnosis. The idea being that if slides 7 and 9 made no difference to their diagnosis we would change our working practice accordingley. The results showed overwhelmingly that slides 7 and 9 made no difference to the diagnosis in over 98% of cases and in the remaining 2% they didn't change the diagnosis from benign to malignant, just a different degree of benign disease. Due to this data, we are currently changing our working practice so that we don't have to cut so many sections, this will be more cost and time effective and should have little or no effect on the end diagnosis. After all, if we don't cut so many sections, there will still be the oppertunity to cut further sections should the pathologist be uncertain of the diagnosis. Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England RSRICHMOND@aol.com wrote: Becky Garrison in Jacksonville FL observes: >>Just this week, our Urology Center, in reviewing its QA measures, wanted to know if it's OK for them to ink the specimen by color to reduce specimen identification mixups. My first reaction was no because of concerns that unknown inks might interfere with fixation and ultimately IHC and the possibility of crush artefact introduced by "non-pathology" hands.<< I agree - no. I want them to handle the specimen as little as possible - the ink needs to be added after fixation. They could use colored pens to mark the outsides of the containers with a set of laboratory-prescribed colors that the lab would use to select cassette and slide frosting colors. I know that there are services that pre-ink the specimens, but I've never actually seen it done. Bob Richmond Samurai Pathologist Gastonia NC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Mon, 1 Mar 2004 10:38:38 +0100 From: Inga Hansson Subject: [Histonet] apoptosis To: histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset="us-ascii" ; format="flowed" Hi everyone! Is anyone using Apoptag kit on mouse cryos and have a problem with weak signal? Any suggestions would be appreciated! Thanks Inga -- Inga Hansson Dept. neuroscience, div. neurobiology PO Box 587, BMC Husargatan 3 S-751 23 Uppsala SWEDEN Phone: +46(18) 471 4384 Fax: +46(18)559017 ------------------------------ Message: 9 Date: Mon, 1 Mar 2004 13:29:09 +0000 From: Shaumik Adhya Subject: [Histonet] histological detection of protein carbonyl groups To: histonet@lists.utsouthwestern.edu Message-ID: <1078147749.40433aa5a1b07@www.webmail.ucl.ac.uk> Content-Type: text/plain; charset=ISO-8859-1 Hi Histonetters, Does anyone know of a stain for detecting carbonyl groups on proteins as a result of protein oxidation? All the methods I can find deal with carbonyl content of proteins in minced samples, not tissue sections. I'd be grateful for any ideas. Shaumik ------------------------------ Message: 10 Date: Mon, 1 Mar 2004 10:31:03 -0500 From: yangpw@umich.edu Subject: [Histonet] rat probe for insitu on mouse tissue? To: histonet@lists.utsouthwestern.edu Message-ID: <1078155063.40435737a8c60@mail.umich.edu> Content-Type: text/plain; charset=ISO-8859-1 Good morning, everyone, I have been doing double insitu on rat brain tissue (c-fos RNA rat probe radioactive and enk RNA rat probe non-radioactive). And the staining works fine. Recently, we are begining to try in situ on mouse brain tissue. Can I just use rat RNA probes on mouse tissue? I have blasted the probes sequence, they match rat c-fos/enk sequence very well, and match mouse c-fos/enk about 90%. Is this kind of match good enough to get good signal on mouse tissue? Does anyone have any suggestion? Do I need to reduce the strigency of wash to retain more signal in this case? If it is not good to use rat probe, can anyone recommend any good mouse probe for c-fos and enkephalin? Thanks very much! Pengwei Yang BioPsychology Program University of Michigan Hi, generally blast with a score of 90% would be a "go". As far as stringency washes, extend time or/and temperature by few degress so loosly bound probe will fell off. I have used probes designed for human with primate tissue, rat probes with mouse tissue and have v. good results. Hope this helps Andrew Lisowski Director Molecular Technologies Northwestern University Chicago a-lisowski@northwestern.edu ------------------------------ Message: 11 Date: Mon, 01 Mar 2004 10:51:07 -0500 From: "Mollie Lange" Subject: [Histonet] MeCP2 antibody To: Message-ID: Content-Type: text/plain; charset=US-ASCII I am looking for a non-rabbit antibody to MeCP2 (methyl cpg binding protein 2) for immunohistochemistry on frozen sections. Does anybody know of one? Thanks, ML ------------------------------ Message: 12 Date: Mon, 1 Mar 2004 08:29:48 -0800 From: "George Cole" Subject: [Histonet] Going to Toronto!!! To: Message-ID: <000001c3ffaa$6ae08c90$034dbad0@hppav> Content-Type: text/plain; charset="us-ascii" Histonetters, T O R O N T O !!!! I have received an invitation to present a report on the new methods of muscle and nerve biopsies that have been sent all over the place----109 sent so far----at the National Histotechnology Convention in Toronto next September. I will look forward to meeting those of you who can come to that big get-together. I spoke at the 1978 convention in San Francisco on silver techniques. That was a speech-only presentation before the whole meeting. This time, there will he a lab set up for the muscle and nerve confab. I'm already putting a video together that will project on a large screen, and I mean to make most of it live so we can get down to practical biopsy performance. It will be a good chance to get acquainted, and to work out practical methods to improve the yield of information from muscle and nerve biopsy tissues. It will be a three hour lab. I'm already practicing my presentation on my wife. I've gone through it 32 times already for her. My, It'll be nice to present to folks that don't keep turning the TV up loud while I'm talking. I have been reading every message on the Histonet since last July, when I joined. And I have taken a cue from the messages you fellow techs send to each other. I have reduced my total 3 hour presentation down to the initial letters of every word----you know, like all of you immuno processing techs ---it should be just an elegant up-to-the-minute presentation. But truly folks, I will look forward to doing the best I can to get some of these methods into public domain for the good of the patients .I've been having the time of my life communicating with you folks all over the world. Toronto will be the peek capper to my histoteching. I already sleep hovering 14 inches over the bed.---tension?--- naaaahhh---but If I can just bring it off in Toronto, maybe I can look forward to coming in for a landing.------ See you there, I hope! georgecole@ev1.net ------------------------------ Message: 13 Date: Mon, 1 Mar 2004 08:43:29 -0800 From: "Morken, Tim - Labvision" Subject: RE: [Histonet] Going to Toronto!!! To: histonet@lists.utsouthwestern.edu Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA2177358@usca0082k08.labvision.apogent.com> Content-Type: text/plain George wrote:<> And only 29 weeks left 'till NSH..... Tim Morken -----Original Message----- From: George Cole [mailto:georgecole@ev1.net] Sent: Monday, March 01, 2004 8:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Going to Toronto!!! Histonetters, T O R O N T O !!!! I have received an invitation to present a report on the new methods of muscle and nerve biopsies that have been sent all over the place----109 sent so far----at the National Histotechnology Convention in Toronto next September. I will look forward to meeting those of you who can come to that big get-together. I spoke at the 1978 convention in San Francisco on silver techniques. That was a speech-only presentation before the whole meeting. This time, there will he a lab set up for the muscle and nerve confab. I'm already putting a video together that will project on a large screen, and I mean to make most of it live so we can get down to practical biopsy performance. It will be a good chance to get acquainted, and to work out practical methods to improve the yield of information from muscle and nerve biopsy tissues. It will be a three hour lab. I'm already practicing my presentation on my wife. I've gone through it 32 times already for her. My, It'll be nice to present to folks that don't keep turning the TV up loud while I'm talking. I have been reading every message on the Histonet since last July, when I joined. And I have taken a cue from the messages you fellow techs send to each other. I have reduced my total 3 hour presentation down to the initial letters of every word----you know, like all of you immuno processing techs ---it should be just an elegant up-to-the-minute presentation. But truly folks, I will look forward to doing the best I can to get some of these methods into public domain for the good of the patients .I've been having the time of my life communicating with you folks all over the world. Toronto will be the peek capper to my histoteching. I already sleep hovering 14 inches over the bed.---tension?--- naaaahhh---but If I can just bring it off in Toronto, maybe I can look forward to coming in for a landing.------ See you there, I hope! georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 14 Date: Mon, 1 Mar 2004 10:51:36 -0600 From: "Barry R Rittman" Subject: RE: [Histonet] Going to Toronto!!! To: Message-ID: <566FB0B522443D43AF02D2ADBE35A6F063598B@UTHEVS3.mail.uthouston.edu> Content-Type: text/plain; charset="us-ascii" George Here in Houston we have rules that apply if you are going to have your wife as a critic of a scientific presentation that you are planning to give: 1. She should have a cattle prod to indicate times when you are talking too much. 2. The presentation should only be rehearsed a maximum of 3 times. 3. You owe her big!! (Now I think of it these are rules that my wife instituted). Good luck on your presentation. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of George Cole Sent: Monday, March 01, 2004 10:30 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Going to Toronto!!! Histonetters, T O R O N T O !!!! I have received an invitation to present a report on the new methods of muscle and nerve biopsies that have been sent all over the place----109 sent so far----at the National Histotechnology Convention in Toronto next September. I will look forward to meeting those of you who can come to that big get-together. I spoke at the 1978 convention in San Francisco on silver techniques. That was a speech-only presentation before the whole meeting. This time, there will he a lab set up for the muscle and nerve confab. I'm already putting a video together that will project on a large screen, and I mean to make most of it live so we can get down to practical biopsy performance. It will be a good chance to get acquainted, and to work out practical methods to improve the yield of information from muscle and nerve biopsy tissues. It will be a three hour lab. I'm already practicing my presentation on my wife. I've gone through it 32 times already for her. My, It'll be nice to present to folks that don't keep turning the TV up loud while I'm talking. I have been reading every message on the Histonet since last July, when I joined. And I have taken a cue from the messages you fellow techs send to each other. I have reduced my total 3 hour presentation down to the initial letters of every word----you know, like all of you immuno processing techs ---it should be just an elegant up-to-the-minute presentation. But truly folks, I will look forward to doing the best I can to get some of these methods into public domain for the good of the patients .I've been having the time of my life communicating with you folks all over the world. Toronto will be the peek capper to my histoteching. I already sleep hovering 14 inches over the bed.---tension?--- naaaahhh---but If I can just bring it off in Toronto, maybe I can look forward to coming in for a landing.------ See you there, I hope! georgecole@ev1.net _______________________________________________ Histonet mailing list Histonet@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 4, Issue 1 ************************************** ===========End of original message text=========== From rocan <@t> mac.com Mon Mar 1 15:47:45 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] anti-collagen I and III Message-ID: <131B7D55-6BCA-11D8-AA47-000A9589219E@mac.com> Hi, Can anyone suggest good anti-collagen antibodies for mouse tissue IHC? We are interested in collagens I, III and IV. Polysciences has some affinity purified rabbit polyclonals that should react to most mammalian collagens. Has anyone used their anti-collagen antibodies? Any input is greatly appreciated, Thank you M. Rocio Sierra-Honigmann, MD, Ph.D. Cedars Sinai Research Institute, CSMC 8700 Beverly Blvd. D-1091 Los Angeles, 90048 Tel. 310-423-1882 Fax 310-423-2325 From hylanderl <@t> comcast.net Mon Mar 1 16:23:02 2004 From: hylanderl <@t> comcast.net (Linda Hylander) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] TrkA antibodies Message-ID: <000b01c3ffdb$c307e0e0$bf6a3c18@ne2.client2.attbi.com> Question, Can anyone recommend a commercially available anti-TrkA (tyrosine kinase) monoclonal antibody that works in frozen sections for IHC? Any positive cell lines and/or tissues is greatly appreciated. Linda Hylander ImmunoGen Inc. From joeamateur <@t> hotmail.com Mon Mar 1 16:05:39 2004 From: joeamateur <@t> hotmail.com (Jack England) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Advice. Message-ID: I'm with Jackie on the freeze/thaw thing. I worked in antibody manufacturing for several years, and while it was common to store antibodies frozen (generally -20 C), the critical thing we tried to avoid was multiple freeze/thaw cycles. We started to see fracturing of the antibodies after about 3-5 freeze/thaw cycles as well. So, I'd certainly recommend aliquotting, *especially* if you've got a small amount of customized Ab (monoclonal, custom conjugate, etc.) that you're using. --Aloha to all, Jack England _________________________________________________________________ Say “good-bye” to spam, viruses and pop-ups with MSN Premium -- free trial offer! http://click.atdmt.com/AVE/go/onm00200359ave/direct/01/ From JWEEMS <@t> sjha.org Mon Mar 1 17:23:02 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Loss of Heterozygosity Message-ID: <8C54DF3F71F6E34A80B1DE628C7EF4DD01DD63EB@exch2.sjha.org> I've been commissioned to find a reference lab performing LOH on 1p and 19q - brain tissue? Is anyone familiar with this testing? Thanks for your help! j 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 syedab <@t> totalise.co.uk Tue Mar 2 05:16:24 2004 From: syedab <@t> totalise.co.uk (Anila Syed) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Help to find ataxin 3 antibody References: <000b01c3ffdb$c307e0e0$bf6a3c18@ne2.client2.attbi.com> Message-ID: <00a701c40047$cc63ff40$80c401a3@clneuro.ox.ac.uk> Hi all, We need some help to find anti ataxin-3 or anti sca-3 I have the chemicon one. Does anyone know of another supplier? Thanks Anila Syed --- Outgoing mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.602 / Virus Database: 383 - Release Date: 01/03/04 From Myri37 <@t> aol.com Tue Mar 2 07:58:37 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] acid decalcification Message-ID: <7D83D6B2.095C32EB.0005167B@aol.com> Hello everyone I would like to decalcify teeh in sodium formiate and acid formique but do you think if i should add a fixative to my solution, like glutarald?hye or formaline or paraformald?hyde, if you did use it, which purcentage did you use please ? After decalcification do you rinse your samples (bones or teeth) in a special solution before dehydrating ? Thank you very much in advance Myriam Natural Implant From Barry.R.Rittman <@t> uth.tmc.edu Tue Mar 2 08:27:15 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] acid decalcification Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FCD8@UTHEVS3.mail.uthouston.edu> Myriam If you add either formalin or glutaraldehyde they will dramatically slow down the mineralization process. If you have very large specimens and are worried about maceration you can always remove the tissue from the decalcification solution and "refix" for a short time in your original fixative. After acid demineralization, you need to wash well in running tap water to remove all traces of the acid so that they will not contaminate the processing solutions (or even remain in the tissue when it is in the paraffin block). Traces of acid in the processed paraffin block will cause all sort of problems. Hope that this helps Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Myri37@aol.com Sent: Tuesday, March 02, 2004 7:59 AM To: histonet@pathology.swmed.edu Subject: [Histonet] acid decalcification Hello everyone I would like to decalcify teeh in sodium formiate and acid formique but do you think if i should add a fixative to my solution, like glutarald?hye or formaline or paraformald?hyde, if you did use it, which purcentage did you use please ? After decalcification do you rinse your samples (bones or teeth) in a special solution before dehydrating ? Thank you very much in advance Myriam Natural Implant _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From spoulos <@t> saa.ars.usda.gov Tue Mar 2 08:35:09 2004 From: spoulos <@t> saa.ars.usda.gov (Sylvia Poulos) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] bench top fume hoods Message-ID: Hi everyone, Just wondering if anyone has had any experience with benchtop fume hoods. I'm looking into purchasing one primarily because of xylene, formaldehyde, and paraformaldehyde and would appreciate any thoughts about them. Thanks again for sharing all your wisdom! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) From StarkusL <@t> ummhc.org Tue Mar 2 08:41:14 2004 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] bench top fume hoods Message-ID: Sylvia, Try the Hacker DownDraft fume hood. It's affordable, movable and works GREAT! -----Original Message----- From: Sylvia Poulos [mailto:spoulos@saa.ars.usda.gov] Sent: Tuesday, March 02, 2004 9:35 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bench top fume hoods Hi everyone, Just wondering if anyone has had any experience with benchtop fume hoods. I'm looking into purchasing one primarily because of xylene, formaldehyde, and paraformaldehyde and would appreciate any thoughts about them. Thanks again for sharing all your wisdom! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From yangpw <@t> umich.edu Tue Mar 2 08:43:34 2004 From: yangpw <@t> umich.edu (yangpw@umich.edu) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Fwd: rat probe for insitu on mouse tissue? Message-ID: <1078238614.40449d9646a1e@mail.umich.edu> Looking for help! Any suggestion? Any comment? THanks in advance. Pengwei ----- Forwarded message from yangpw@umich.edu ----- Date: Mon, 1 Mar 2004 10:31:03 -0500 From: yangpw@umich.edu Reply-To: yangpw@umich.edu Subject: rat probe for insitu on mouse tissue? To: histonet@lists.utsouthwestern.edu Good morning, everyone, I have been doing double insitu on rat brain tissue (c-fos RNA rat probe radioactive and enk RNA rat probe non-radioactive). And the staining works fine. Recently, we are begining to try in situ on mouse brain tissue. Can I just use rat RNA probes on mouse tissue? I have blasted the probes sequence, they match rat c-fos/enk sequence very well, and match mouse c-fos/enk about 90%. Is this kind of match good enough to get good signal on mouse tissue? Does anyone have any suggestion? Do I need to reduce the strigency of wash to retain more signal in this case? If it is not good to use rat probe, can anyone recommend any good mouse probe for c-fos and enkephalin? Thanks very much! Pengwei Yang BioPsychology Program University of Michigan ----- End forwarded message ----- From juan.gutierrez <@t> christushealth.org Tue Mar 2 08:53:44 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] bench top fume hoods Message-ID: I have used two kinds so far with good results from both. One is the Thermo(Shandon), I believe they're called Hyperclean something. They have different sizes. The one I have now is by Labconco, but I don't know where we got it from (it was here before I got here). It also works very good. I also looked in the Mopec catalog and found some very similar, if not the same, as the Thermo ones you might give them a try. You can always try them out before you buy them, and I recomend you do. Thermo- 1-800-547-7429 Mopec- 1-800-362-8491 Fisher- 1-800-640-0640 p.s. Labconco is distributed by Fisher Scientific. Juan -----Original Message----- From: Sylvia Poulos [mailto:spoulos@saa.ars.usda.gov] Sent: Tue 3/2/2004 8:35 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] bench top fume hoods Hi everyone, Just wondering if anyone has had any experience with benchtop fume hoods. I'm looking into purchasing one primarily because of xylene, formaldehyde, and paraformaldehyde and would appreciate any thoughts about them. Thanks again for sharing all your wisdom! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From skroeker <@t> cellsignal.com Tue Mar 2 10:11:55 2004 From: skroeker <@t> cellsignal.com (Kroeker, Sally) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Job Opportunities Message-ID: <53606486-6C64-11D8-9ABE-00039380009A@cellsignal.com> Cell Signaling Technology, Inc, in Beverly, MA has 2 exciting job opportunities in the Boston area. Please visit: www.cellsignal.com for information about CST. Research Associate - Immunohistochemistry A research associate is needed in the IHC group to participate in the validation of signal transduction antibodies for use in IHC. Responsibilities will include protocol development and optimization, screening antibodies for IHC suitability and developing model systems for such testing. Other responsibilities may emerge as the company continues to grow. This candidate should be highly motivated and organized and have an MS or BS with equivalent experience. Experience with immunohistochemistry is required. Other applicable techniques include but are not limited to Western blotting and tissue culture. Knowledge of signal transduction is preferred. Job Code: IHC01 Research Associate Cell Signaling Technology (CST) is looking for a Research Associate to perform basic and clinical research identifying predictive biomarkers for cancer drug candidates. The individual will work closely with scientific collaborators in analyzing model cell lines and xenograft samples. The techniques to be used include immunohistochemistry, Western blotting and cell culture. Must have broad laboratory experience with expertise in cell biology techniques and immunohistochemistry. Knowledge of pre-clinical research and drug development would be helpful. Applicant should be highly motivated, have an interest in cancer biology and drug development. Job Code: COSRA107 Reference the JOB CODE in the Subject Line and forward your cover letter and resume to: hire@cellsignal.com From gerry <@t> houston.rr.com Tue Mar 2 10:40:40 2004 From: gerry <@t> houston.rr.com (gerry@houston.rr.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Histology Position Message-ID: <323910-22004322164040420@M2W048.mail2web.com> Park Plaza Hospital, adjacent to the Houston Medical Center in the Museum District, has a full time position open for a histology technologist, full time, Monday - Friday, 5:30 am - 1:00 pm. Interested parties may email Sandy Nelson at sandy.nelson@tenethealth.com. -------------------------------------------------------------------- mail2web - Check your email from the web at http://mail2web.com/ . From lesley <@t> vancouverbc.net Tue Mar 2 10:59:24 2004 From: lesley <@t> vancouverbc.net (Lesley Weston) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] bench top fume hoods In-Reply-To: Message-ID: They're not as effective as the vented type, but a great deal better than nothing. If you can persuade your people to install a vented hood (they're much more expensive), then that's the best solution. Otherwise, the benchtop kind will provide some protection. Lesley Weston. on 02/03/2004 6:35 AM, Sylvia Poulos at spoulos@saa.ars.usda.gov wrote: > Hi everyone, > Just wondering if anyone has had any experience with benchtop fume > hoods. I'm looking into purchasing one primarily because of xylene, > formaldehyde, and paraformaldehyde and would appreciate any thoughts > about them. Thanks again for sharing all your wisdom! Sylvia > > Sylvia P. Poulos > USDA-ARS-Animal Physiology Research Unit > Athens, GA 30605 > 706-583-8279 > 706-542-0399 (fax) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Tue Mar 2 12:54:12 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] anti-collagen I and III In-Reply-To: <131B7D55-6BCA-11D8-AA47-000A9589219E@mac.com> Message-ID: I think the best source for collagen antibodies is the U of Iowa Hybridoma Bank for any species, they are monoclonals though so you would have to do mouse on mouse methods. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of rocan@mac.com Sent: Monday, March 01, 2004 2:48 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] anti-collagen I and III Hi, Can anyone suggest good anti-collagen antibodies for mouse tissue IHC? We are interested in collagens I, III and IV. Polysciences has some affinity purified rabbit polyclonals that should react to most mammalian collagens. Has anyone used their anti-collagen antibodies? Any input is greatly appreciated, Thank you M. Rocio Sierra-Honigmann, MD, Ph.D. Cedars Sinai Research Institute, CSMC 8700 Beverly Blvd. D-1091 Los Angeles, 90048 Tel. 310-423-1882 Fax 310-423-2325 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GArriaga <@t> chw.edu Tue Mar 2 13:25:32 2004 From: GArriaga <@t> chw.edu (Arriaga, Gustavo) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] quantum dot secondary antibody conjugates Message-ID: <4A778810F700DA4A984A7753319D7EB4016B2D41@srdh-msg-01.chw.edu> Does anybody have experience labeling with the new quantum dot secondary conjugates from the Quantum Dot Corporation? The brightness, resolution, excitation range, narrow near IR emission and photostability make them really attractive for many purposes. In fact, they sound almost too good to be true. So, before trying them out on our own slides I was wondering if anybody could confirm the hype or relate bad experiences with them. Gustavo Arriaga Research Technician Barrow Neurological Institute From caroline.stott <@t> anatomy.otago.ac.nz Tue Mar 2 13:54:58 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] acid decalcification Message-ID: <5.2.1.1.0.20040303084920.02574b40@anatomy.otago.ac.nz> Hi Myriam We generally use Formic acid for decalcifying. I agree that you should not use any fixative as it will slow the process down. You will need to check the acid solution every few days for the amount of mineral left in the tissue. Regular changes of acid solution are needed as the solution becomes saturated. Once there is no mineral left in the tissue we wash in running water for several hours to remove the acid. Some tissues however may require a neutralising step before processing. Hope that helps. Caroline Hello everyone I would like to decalcify teeh in sodium formiate and acid formique but do you think if i should add a fixative to my solution, like glutarald?hye or formaline or paraformald?hyde, if you did use it, which purcentage did you use please ? After decalcification do you rinse your samples (bones or teeth) in a special solution before dehydrating ? Thank you very much in advance Myriam Natural Implant et Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From rocan <@t> mac.com Tue Mar 2 14:51:39 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] UI-hybridoma bank-anti-collagen III Message-ID: <6765DC26-6C8B-11D8-A603-000A9589219E@mac.com> Patsy, I have the antibodies from the bank but when I was looking at the spec sheet I noticed that the Col III antibody (3B2) is listed as reactive to chicken collagen only in the Species Specificity. Have you used this antibody on non-avian tissues? From GArriaga <@t> chw.edu Tue Mar 2 15:39:24 2004 From: GArriaga <@t> chw.edu (Arriaga, Gustavo) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] (no subject) Message-ID: <4A778810F700DA4A984A7753319D7EB4016B2D42@srdh-msg-01.chw.edu> I just registered for the listserve and have had trouble posting. sorry if you receive duplicates of this message. Does anybody have experience labeling with the new quantum dot secondary conjugates from the Quantum Dot Corporation? The brightness, resolution, excitation range, narrow near IR emission and photostability make them really attractive for many purposes. In fact, they sound almost too good to be true. So, before trying them out on our own slides I was wondering if anybody could confirm the hype or relate bad experiences with them. Gustavo Arriaga Research Technician Barrow Neurological Institute From GArriaga <@t> chw.edu Tue Mar 2 15:41:02 2004 From: GArriaga <@t> chw.edu (Arriaga, Gustavo) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] quantum dot secondary antibody conjugates Message-ID: <4A778810F700DA4A984A7753319D7EB4016B2D43@srdh-msg-01.chw.edu> I just registered for the listserve and have had trouble posting. sorry if you receive duplicates of this message. Does anybody have experience labeling with the new quantum dot secondary conjugates from the Quantum Dot Corporation? The brightness, resolution, excitation range, narrow near IR emission and photostability make them really attractive for many purposes. In fact, they sound almost too good to be true. So, before trying them out on our own slides I was wondering if anybody could confirm the hype or relate bad experiences with them. Gustavo Arriaga Research Technician Barrow Neurological Institute From pruegg <@t> colobio.com Tue Mar 2 18:12:40 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Cartilage Marker for IHC? In-Reply-To: Message-ID: Jennifer, Collagen IV is a good marker for cartilage. I get it from the U of Iowa Hybridoma Bank. Also link protein is another good marker for cartilage from Iowa. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jennifer Hoover Sent: Tuesday, February 17, 2004 7:49 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cartilage Marker for IHC? Histonet, I am looking for suggestions for an IHC marker that would be constitutively expressed in the cartilage of rat knee joints. Can anyone recommend a clone and the Company to purchase the antibody from. Thank you very much! Jennifer Hoover Bone and Inflammation Research Eli Lilly and Company _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From doscwk <@t> nus.edu.sg Tue Mar 2 19:26:25 2004 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Bench top fume hoods Message-ID: Hi Sylvia, Our lab has 2 bench top fume hoods and we're having a problem with the filters getting saturated very quickly as our usage of solvents is quite heavy. We've wrote to the building's management for permission to install a ducted fume hood but was rejected as our basement lab isn't meant to be used for such work and didn't come with an installed hood. I think it would depend on your type of usage. For heavy chemical usage, a ducted hood is more suitable. If not, like us, you'll have a problem with frequent changes of filters and they don't come cheap either. Regards Julee Chan Musculoskeletal Tissue Lab Orthopaedic Surgery National University of Singapore Sylvia wrote: Hi everyone, Just wondering if anyone has had any experience with benchtop fume hoods. I'm looking into purchasing one primarily because of xylene, formaldehyde, and paraformaldehyde and would appreciate any thoughts about them. Thanks again for sharing all your wisdom! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) From GArriaga <@t> chw.edu Tue Mar 2 12:22:20 2004 From: GArriaga <@t> chw.edu (Arriaga, Gustavo) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] quantum dot secondary antibody conjugates Message-ID: <4A778810F700DA4A984A7753319D7EB4016B2D40@srdh-msg-01.chw.edu> Does anybody have experience labeling with the new quantum dot secondary conjugates from the Quantum Dot Corporation? The brightness, resolution, excitation range, narrow near IR emission and photostability make them really attractive for many purposes. In fact, they sound almost too good to be true. So, before trying them out on our own slides I was wondering if anybody could confirm the hype or relate bad experiences with them. Gustavo Arriaga Research Technician Barrow Neurological Institute From arename <@t> hotmail.com Wed Mar 3 05:44:18 2004 From: arename <@t> hotmail.com (hanim shueb) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] TUNEL (PROMEGA) Message-ID: I'm doing TUNEL staining on mouse brains (formaldehyde fixed, paraffin section) right now. I have problem with non-specific binding of the cells; ie almost all cells stained positive. I used the protocol given by Promega with some modifications; I performed microwave antigen retrieval and block the tissues with 20% NGS/1%BSA in PBS before adding TdT reaction mix. Can anyone help me to solve the non-specific bindings I keep getting? many thanks, Hanim Shueb Western Australia _________________________________________________________________ Download the latest MSN Messenger http://messenger.msn.com.my From PKrueger <@t> Lifespan.org Wed Mar 3 06:42:03 2004 From: PKrueger <@t> Lifespan.org (Krueger, Paula M) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] anti-collagen I and III for sheep Message-ID: <85151E3A0A49DF498F39F19EE56FEA8D01D034BA@lsexch2.lsmaster.lifespan.org> To all, I, too, am looking for collagen I and III antibodies that will work on sheep tissues. I contacted the UI Hybridoma Bank but their collagen III is avian and they only have PRO-collagen I for sheep. Does anyone know of any additional sources? Thanks for your help, Paula Krueger Sr. Research Asst./Lab Manager Collis Cardiac Surgical Research Laboratory Aldrich, 723 Rhode Island Hospital Providence, Rhode Island 02903 Ph: 401-444-1668 FAX: 401-444-5153 From Jeannette.Mitchell <@t> vtmednet.org Wed Mar 3 07:20:47 2004 From: Jeannette.Mitchell <@t> vtmednet.org (Mitchell, Jeannette M.) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] (no subject) Message-ID: Does anyone know of someone who supplies Her2 Neu Probes. We really don't want to be tied into kits. We prefer just the probe. Jeannette Mitchell FAHC Burlington,Vt Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. From David.Edmondson <@t> christie-tr.nwest.nhs.uk Wed Mar 3 08:38:41 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] bench top fume hoods Message-ID: We too have the Thermo Shandon hood,on our staining machine, which has a charcoal pack, which is saturated quickley, so we may end up encouraging solvent vapours rather than reducing them if we do not change them frequently enough Dave -----Original Message----- From: Sylvia Poulos [mailto:spoulos@saa.ars.usda.gov] Sent: 02 March 2004 14:35 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bench top fume hoods Hi everyone, Just wondering if anyone has had any experience with benchtop fume hoods. I'm looking into purchasing one primarily because of xylene, formaldehyde, and paraformaldehyde and would appreciate any thoughts about them. Thanks again for sharing all your wisdom! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Ronnie_Houston <@t> bshsi.com Wed Mar 3 09:10:19 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] (no subject) Message-ID: <530361BF03351B4CAE5270A05D3037B503673105@bsrexms01.BSHSIR.COM> Zymed Laboratories 800 874 4494 Her2 Probe Cat# 84-0100 Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 (804) 287 7906 - fax ronnie_houston@bshsi.com -----Original Message----- From: Mitchell, Jeannette M. [mailto:Jeannette.Mitchell@vtmednet.org] Sent: Wednesday, March 03, 2004 8:21 AM To: histonet@pathology.swmed.edu Subject: [Histonet] (no subject) Does anyone know of someone who supplies Her2 Neu Probes. We really don't want to be tied into kits. We prefer just the probe. Jeannette Mitchell FAHC Burlington,Vt Confidentiality Notice: This message, and any attachments, may contain information that is confidential, privileged, and/or protected from disclosure under state and federal laws that deal with the privacy and security of medical information. If you received this message in error or through inappropriate means, please reply to this message to notify the Sender that the message was received by you in error, and then permanently delete this message from all storage media, without forwarding or retaining a copy. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From shaumik.adhya <@t> ucl.ac.uk Wed Mar 3 10:26:09 2004 From: shaumik.adhya <@t> ucl.ac.uk (Shaumik Adhya) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] plea for use of subject headers Message-ID: <1078331169.404607212af8e@www.webmail.ucl.ac.uk> Hi histonetters, as a fairly new person to the list, and indeed to histotechnology, I can't really add much to a lot of the posts flying around. It'd be great if people could use subjects so that the list can work out whether or not they want to read the mail. thanks, especially if you actually read this, Shaumik From lleach <@t> uic.edu Wed Mar 3 11:57:06 2004 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] chondroitinase usage Message-ID: <6.0.1.1.2.20040303115222.0233afc0@tigger.uic.edu> Dear Histonetters, I'm using an Versican antibody that requires Chondroitinase treatment prior to staining, does anyone have any suggestions? There are many to choose from. Thank you, Lu Leach Director Research Pathology Core University of Illinois at Chicago 312-996-3869 lleach@uic.edu From sladd <@t> hsc.usf.edu Wed Mar 3 12:33:48 2004 From: sladd <@t> hsc.usf.edu (Sharron Ladd) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] chondroitinase usage In-Reply-To: <6.0.1.1.2.20040303115222.0233afc0@tigger.uic.edu> References: <6.0.1.1.2.20040303115222.0233afc0@tigger.uic.edu> Message-ID: <4046250C.6010107@hsc.usf.edu> Chondroitinase ABC from Associates of Cape Cod (Seikagaku) 1-800-237-4512 Lu Leach wrote: > Dear Histonetters, > > I'm using an Versican antibody that requires Chondroitinase treatment > prior to staining, does anyone have any suggestions? There are many to > choose from. > > Thank you, > > Lu Leach > Director Research Pathology Core > University of Illinois at Chicago > 312-996-3869 > lleach@uic.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From jlf <@t> jax.org Wed Mar 3 12:42:01 2004 From: jlf <@t> jax.org (Judi Ford) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] suscribe digest Message-ID: <5.1.0.14.0.20040303134047.00a6c608@aretha.jax.org> From ramona.tolliver <@t> yale.edu Wed Mar 3 13:16:27 2004 From: ramona.tolliver <@t> yale.edu (Ramona Tolliver) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Yale University - Opening Message-ID: <6.0.1.1.2.20040303141428.01d574d8@email.med.yale.edu> Good afternoon, Yale University has a 2nd Shift Histology Supervisor Position available from 3:00 -11:00 p.m., Monday-Friday. If you know of anyone who may be interested in a great opportunity with advancement potential, please forward this information on to them. The posting is located at : www.yale.edu/jobs (click on external candidates and do a job search for requisition MSGM11927). Salary starts at 65K and is commensurate with experience. Relocation assistance is available. Thank you for your time! Ramona Tolliver Ramona E. Tolliver Human Resource Manager Yale University School of Medicine Department of Pathology 310 Cedar Street New Haven, CT 06520-8023 Telephone: (203) 785-6689 Fax: (203) 785-7303 From rjr6 <@t> psu.edu Wed Mar 3 14:04:31 2004 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Potomac Horse Fever Message-ID: <003301c4015a$bdaecb10$8861ba92@padlspsu.psu.edu> Does anyone do or have a stain for Potomac Horse Fever? Thank you, Roberta Horner Penn State University Animal Diagnostic Lab From jeannie_heck <@t> yahoo.com Wed Mar 3 14:41:53 2004 From: jeannie_heck <@t> yahoo.com (Jeannie Heck) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Formaldehyde Exposure Plan Message-ID: <20040303204153.46759.qmail@web41605.mail.yahoo.com> I have been asked to update the policy for storage and handling of formaldehyde. I have a copy of 29 CFR 1910.1048 and would appreciate all comments and suggestions for help with this policy. Thanks in advance. Jeannie Heck HT (ASCP) Hutchinson Hospital Hutchinson Kansas heckj@hhosp.com --------------------------------- Do you Yahoo!? Yahoo! Search - Find what you’re looking for faster. From HACKERLAB <@t> aol.com Wed Mar 3 15:01:00 2004 From: HACKERLAB <@t> aol.com (HACKERLAB@aol.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Hacker has a new home Message-ID: <1a1.212bc4f9.2d77a18c@aol.com> Dear HistoNetter; HACKER Instruments and Industries Inc has a new home! Beginning March 8th, 2004 our corporate headquarters will be located in Winnsboro, South Carolina. Please note our new address & telephone numbers: HACKER Instruments & Industries Inc 1132 Kincaid Bridge Road PO Box 1176 Winnsboro, SC Tel: (803) 712-6100 Fax:(803) 712-6110 1-800-4-HACKER hackerlab@aol.com www.hackerinstruments.com Thanks for your attention. Please call if you have any questions. We will be at our Fairfield, NJ location thru Friday, March 5th (973) 226-8450. Best regards, Elfi Hacker HACKER Instruments & Industries Inc From HACKERLAB <@t> aol.com Wed Mar 3 15:01:00 2004 From: HACKERLAB <@t> aol.com (HACKERLAB@aol.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Hacker has a new home Message-ID: <1a1.212bc4f9.2d77a18c@aol.com> Dear HistoNetter; HACKER Instruments and Industries Inc has a new home! Beginning March 8th, 2004 our corporate headquarters will be located in Winnsboro, South Carolina. Please note our new address & telephone numbers: HACKER Instruments & Industries Inc 1132 Kincaid Bridge Road PO Box 1176 Winnsboro, SC Tel: (803) 712-6100 Fax:(803) 712-6110 1-800-4-HACKER hackerlab@aol.com www.hackerinstruments.com Thanks for your attention. Please call if you have any questions. We will be at our Fairfield, NJ location thru Friday, March 5th (973) 226-8450. Best regards, Elfi Hacker HACKER Instruments & Industries Inc From ESmith <@t> CuraGen.com Wed Mar 3 16:18:44 2004 From: ESmith <@t> CuraGen.com (Smith, Emily) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Confirming IHC staining as "real" Message-ID: Hi All, What techniques are generally used to confirm immunohistochemistry staining as real? If a tissue stains with a labeled primary antibody what follow up assays and experiments are done to confirm the staining is specific? For example, we have tried titering the primary antibody concentration to see the staining intensity reduced. We have tried to "compete" the staining away with unlabeled antibody or with the peptide. Has anyone tried these or similar approaches? How were the results? Thanks for your help. ~Emily CuraGen Corporation From rocan <@t> mac.com Wed Mar 3 23:46:38 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Confirming IHC staining as "real" In-Reply-To: References: Message-ID: <4DFA846E-6D9F-11D8-9FA4-000A9589219E@mac.com> Emily, I am always surprised how much IHC goes on without these types of controls. I think a lot of IHC is meaningless unless you do these types of controls whenever such controls are possible. In the case of an antigen of which we have recombinant form of the protein, we compete with the antigen and get clean results. I think if your primary was made against a peptide, competing with soluble peptide is always a very convincing control. These "soluble antigen" approaches will show specificity. Another way to have some piece of mind when you don't have antigen for competing is and you are using an antibody against specific structures or cell types in a tissue is of course to make sure other cell types or structures are not staining along. If your primary is labeled, the competition with an excess of at least 10 fold or more unlabeled Ab is a good control as well. However, competing with unlabeled antibody is not going to give you specificity. If the labeled antibody is binding to non-specific structures, you unlabeled antibody is probably also going to bind to them. Rocio On Mar 3, 2004, at 2:18 PM, Smith, Emily wrote: > Hi All, > What techniques are generally used to confirm immunohistochemistry > staining as real? If a tissue stains with a labeled primary antibody > what follow up assays and experiments are done to confirm the staining > is specific? > For example, we have tried titering the primary antibody concentration > to see the staining intensity reduced. We have tried to "compete" the > staining away with unlabeled antibody or with the peptide. > Has anyone tried these or similar approaches? How were the results? > Thanks for your help. > ~Emily > CuraGen Corporation > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ESerrano <@t> imim.es Thu Mar 4 01:29:02 2004 From: ESerrano <@t> imim.es (Serrano del Pozo, Erika) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: flow citometry protocol Message-ID: <10708933FD0CD511BCFE000102BE5F72025FACF2@hpserv.imim.es> > Hi histotechs, > > I'm working with injured mice muscle, so it?s full of inflammatory cells > at 2-5 days after injury. I?d like to know if anyone of you knows how to > separate these inflammatory cells from a whole muscle to perform a flow > citometry assay or where can I find a protocol to do it. Since now I've > been doing IHC in frozen tissue because I only needed to check there were > macrophages and T-lymphocytes, but now I need to count them and I don't > know how to obtain "clean" cells to perform the experiment, I couldn?t > find anything on the internet. > > Thanks in advance, > > > Erika Serrano > > Centre de Regulaci? Gen?mica > Differenciation and Cancer Programme > Barcelona > > From STapper <@t> slhduluth.com Thu Mar 4 06:12:28 2004 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] reticulin procedure Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D6716C7@slhw2smail01.slhdomain.com> Does anyone perform a reticulin stain using an acidulated Potassium permanganate? I would be interested in your procedure! Thanks! Sheila Tapper St. Luke's Hospital Duluth, MN CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. From David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Mar 4 08:25:45 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] reticulin procedure Message-ID: 250ml 0.25% Pot permang and 10ml of 3% Sulphuric acid, 5 mins oxidation then Oxalic acid prior to sensitisation and silver staining. Dave Manchester UK -----Original Message----- From: Tapper, Sheila [mailto:STapper@slhduluth.com] Sent: 04 March 2004 12:12 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] reticulin procedure Does anyone perform a reticulin stain using an acidulated Potassium permanganate? I would be interested in your procedure! Thanks! Sheila Tapper St. Luke's Hospital Duluth, MN CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Rushworth <@t> mail.bhrv.nwest.nhs.uk Thu Mar 4 09:00:18 2004 From: David.Rushworth <@t> mail.bhrv.nwest.nhs.uk (David Rushworth) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] sirius red connective tissue stain[Scanned] Message-ID: <1030B679AD69D6119C3F00080210DD9D0103F87C@bhrv-nt-11.bhrv.nwest.nhs.uk> Can someone please supply method for Sirius red-fast green stain for connective tissue. Thanks in anticipation. Dave. From vbaker60 <@t> yahoo.com Thu Mar 4 09:01:43 2004 From: vbaker60 <@t> yahoo.com (Victoria Baker) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Research labs - charges and a question about outside contracting Message-ID: <20040304150143.63346.qmail@web12103.mail.yahoo.com> Good Morning Everyone, I have been asked to look around at what research labs are currently charging for Histology work. Our lab is NCI funded (partially) the balance of our funding comes from donations, grants and some private contract work. Currently we do the following procedures in our facility and I have attached what the fees we charge are. Basic H&E with pathology review - $9.00/slide Basic H&E without pathology review - $4.50/slide IHC (ES automated)with pathology review - $25.00/slide* Manual IHC with pathology review - $45.00 (w/o $40.00) IHC without pathology review - $20.00/slide ISH with pathology review - $45.00/slide FISH with pathology review/report - $45.00/slide Immunofluour. - cells & tissue $30.00/antibody Laser scanning cytometry with analysis - $15.00/per hour Cell staining for flow cytometry including PI - $45.00/sample Flow cytometry/cell cycle analysis and report - $15.00 per hour. We have several more procedures that we do, but they are not that common. *These are antibodies like PCNA, Vimentin etc done on mostly rat and human samples. Our mouse and other animal models have to be done manually because of cross reactivity issues. We have a Ventana ES and a Ventana Discovery stainer in our lab and I am the only Histologist in house. I have 1.5 people that can help with certain aspects of the staining but are not Histology trained. Our our prices in line with labs similar to ours? Is anyone currently using outside contractors to do some of their Histology work? If so which work? Any input would be greatly appreciated. Thank you. Vikki Baker Institute for Cancer Prevention Valhalla, New York 10595 __________________________________ Do you Yahoo!? Yahoo! Search - Find what you’re looking for faster http://search.yahoo.com From funderwood <@t> mcohio.org Thu Mar 4 09:38:09 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] frozen sections for fat stain Message-ID: Hi Histonetters. What is the best way to store frozen sections (on formalin fixed tissue). I'll be doing an oil red O but need to store them over the weekend. Thanks, Fred Underwood Mont. County Coroner Dayton, OH From tpmorken <@t> labvision.com Thu Mar 4 10:36:04 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Confirming IHC staining as "real" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA217738E@usca0082k08.labvision.apogent.com> Probably the best is to complex it with the known antigen before putting it on the slide and then it should be negative. If you don't have the antigen, then you need a known control and some info on what the ab is supposed to stain. Negative controls should include 1) no primary or serum (buffer only), 2) normal serum from host animal of your primary 3) nonsense antibodies (aren't supposed to stain the target tissue - use ab's from the same host and preferably same Ig type) and 4) non-target tissues incubated with your primary(should be negative). Tim Morken Lab Vision / Neomarkers www.labvision.com -----Original Message----- From: Smith, Emily [mailto:ESmith@CuraGen.com] Sent: Wednesday, March 03, 2004 2:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Confirming IHC staining as "real" Hi All, What techniques are generally used to confirm immunohistochemistry staining as real? If a tissue stains with a labeled primary antibody what follow up assays and experiments are done to confirm the staining is specific? For example, we have tried titering the primary antibody concentration to see the staining intensity reduced. We have tried to "compete" the staining away with unlabeled antibody or with the peptide. Has anyone tried these or similar approaches? How were the results? Thanks for your help. ~Emily CuraGen Corporation _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Mar 4 10:45:18 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] frozen sections for fat stain Message-ID: I remember wrapping slides in tinfoil and putting them in the freezer,in a slide transport box and using them as control slides for Oil Red O for many weeks Dave Manchester UK -----Original Message----- From: Fred Underwood [mailto:funderwood@mcohio.org] Sent: 04 March 2004 15:38 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] frozen sections for fat stain Hi Histonetters. What is the best way to store frozen sections (on formalin fixed tissue). I'll be doing an oil red O but need to store them over the weekend. Thanks, Fred Underwood Mont. County Coroner Dayton, OH _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Thu Mar 4 13:01:22 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Article request Message-ID: Hi All Does anyone have a copy of Journal of Histotechnology 1988, V11, pg 61-63. Article by C. Sanderson and J. Emmanual. If so could you fax to (212) 263-2041? Please let me know...... Thanks Luis From Pathologyarts <@t> aol.com Thu Mar 4 13:08:59 2004 From: Pathologyarts <@t> aol.com (Pathologyarts@aol.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Looking for a 1 Oz. specimen bottle Message-ID: Looking for a 1 Oz. specimen bottle, Not the common size though. This NBF prefilled specimen container/bottle is 7 cm tall and 2.7 cm wide at the base; the white plastic lid is 3.4 cm wide. I need this specifically for our label. If anyone can help with a vendor or two, I would be very grateful. Thank you From djohnson14 <@t> hotmail.com Thu Mar 4 13:01:14 2004 From: djohnson14 <@t> hotmail.com (Dave Johnson) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Article request Message-ID: Call Cunningham Associates, they are the publisher of the Journal of Histo 201-767-4170 Dave Johnson Pacific SW Lab Equipment >From: Luis Chiriboga >To: Histonet >Subject: [Histonet] Article request >Date: Thu, 04 Mar 2004 14:01:22 -0500 > >Hi All >Does anyone have a copy of Journal of Histotechnology 1988, V11, pg 61-63. >Article by C. Sanderson and J. Emmanual. If so could you fax to (212) >263-2041? Please let me know...... >Thanks > Luis >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Find things fast with the new MSN Toolbar – includes FREE pop-up blocking! http://clk.atdmt.com/AVE/go/onm00200414ave/direct/01/ From AnthonyH <@t> chw.edu.au Thu Mar 4 15:47:29 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] reticulin procedure Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E14C@simba.kids> Sheila, Our procedure is as follows: Silver Reticulin Stain Principle: The mechanism of staining is: a) Sensitisation of the tissue by an oxidising agent, b) Deposition of the positively charged diamine silver ions, c) Reduction of the silver ions, d) Toning with gold chloride to increase contrast and clarity, The following technique uses a volumetric rather than a titrated silver diamine solution. Oxidation (periodic acid and chromium trioxide can be used as well as potassium permanganate) produces reducing groups in reticulin (aldehydes), which are the groups subsequently, demonstrated by the sensitisation and reduction steps in the technique. If non-acidified pot permanganate is used, alkene groups in lipofuscins can also give rise to aldehyde groups and stain black. These are blocked if the pH of potassium permanganate is under 1.5. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Liver Reagents: 1. Acidified permanganate solution: 0.25% Potassium Permanganate 20ml 0.25% Sulphuric acid 20ml 2. 2% Oxalic Acid 3. 4% aqueous iron alum (ferric ammonium sulphate). 4. 8% Ammonium Nitrate (16g in 200ml) - Warning: Strong Oxidizer - see MSDS 5. 4% sodium hydroxide (8g in 200ml) - Warning: Corrosive - see MSDS 6. 10% silver nitrate - Warning: Corrosive - see MSDS 7. Silver Diamine solution: prepare in the following order just prior to use. 8% ammonium nitrate 7ml Distilled water 35ml 4% sodium hydroxide 8ml 10% silver nitrate 3.8ml 8. Reducing Solution (10% formalin) Warning: Toxic - see MSDS Distilled water 90ml 40% Formaldehyde 10ml 9. 0.1% Gold Chloride 10. 2% Sodium Thiosulphate 11. Kernechtrot Counterstain Procedure: 1. Bring sections to distilled water. 2. Acidified Potassium permanganate 5 min. 3. Running Tap water Wash 4. 2% oxalic acid 1 min 5. Running Tap water Wash 6. 4% Iron alum 5 min 7. Distilled water Rinse rapidly 8. Silver Diamine solution 6 min 9. Rinse in distilled water Quick dip 10. Reducing solution 2 min 11. Distilled water Wash 12. 0.2% Gold chloride 30-60 sec(Tone) 13. Distilled water Rinse 14. 2% Sodium Thiosulphate 1-2 min 15. Rinse in water 2 min 16. Counterstain in Kernechtrot 3 min 17. Rinse in water 2 min 18. Absolute alcohol x 2 Wash 19. Clear and mount. Results: Reticulin fibres Black Collagen Brown Notes: Use chemically clean glassware. Step 10. Reducing solution - slides must be agitated constantly until entire section goes brown. Reference: 1. Naoumenko,J.,Feigin,I., (1974) Stain Tech 49(3):153-155. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html -----Original Message----- From: Tapper, Sheila [mailto:STapper@slhduluth.com] Sent: Thursday, 4 March 2004 11:12 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] reticulin procedure Does anyone perform a reticulin stain using an acidulated Potassium permanganate? I would be interested in your procedure! Thanks! Sheila Tapper St. Luke's Hospital Duluth, MN CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From JSCHUMA1 <@t> FAIRVIEW.ORG Thu Mar 4 16:19:51 2004 From: JSCHUMA1 <@t> FAIRVIEW.ORG (JENNIFER SCHUMACHER) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Certification Message-ID: I heard through the grapevine that there was legislation recently passed that histology technicians will be required to be boarded. I know about the changes in routes to become boarded (no longer OJT after this year), and I know some institutions require boards . . . does anyone have any information about boarding becoming mandatory? Did someone just have their wires crossed? Thanks. Jen The information transmitted in this e-mail is intended only for the person or entity to which it is addressed and may contain confidential and/or privileged material, including “protected health information.” If you are not the intended recipient, you are hereby notified that any review, retransmission, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. <<<>>> From AJohnson <@t> system1.net Thu Mar 4 16:33:03 2004 From: AJohnson <@t> system1.net (Amy Johnson) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Supervisor Position in MidWest Message-ID: Seeking an experienced Histology Supervisor for the Mid West. Eligible candidates must have either HT or HTL with a minimum of 3-6 years experience in the lab at least 1-2 of those in a leadership role. 50-50 administrative and benchwork. (some grossing) This is a high volume, high energy lab with growth potential. For further information or to be considered, please contact Amy Johnson at Toll Free: 866-797-8361 via Email: ajohnson@system1.net Have a Great Day! Amy Johnson System 1 Search (864) 627-0012 (864) 627-0013 Fax From billions <@t> public1.sz.js.cn Thu Mar 4 20:09:34 2004 From: billions <@t> public1.sz.js.cn (Sinoera Tech) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Re: Try Message-ID: <013b01c40256$e9149620$a001a8c0@ming> Dear Histonetter, From c.m.vanderloos <@t> amc.uva.nl Fri Mar 5 03:34:52 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] RE: Confirming IHC staining as "real" Message-ID: <5cdc545cafd2.5cafd25cdc54@amc.uva.nl> Hi All, Interesting problem we have here. Feels a bit like the chicken and the egg problem! I will try to shed some light in it. The procedure to complex an antibody with its original antigen or peptide is to my opinion a technique from an old dusty box. A technique from the time we were still dealing with POLYCLONAL antibodies that were not so monospecific and sometimes needed to be absorbed to reduce background staining. Doing such a blocking experiment with a MONOCLONAL antibody is however quite useless. Once the peptide blocks your staining for 100%, you end up with a feeling of a "good antibody" whereas one that isn't blocking that well is considered to be a "bad one". In fact, neither results of the blocking experiment give any information about the "real" specificity of the antibody. Three good reasons: 1. To produce monoclonal antibodies applicable on FFPE sections, antigens are processed through formalin, alcohol, xylene before immunization of mice. This strategy may introduce a conformational change of an antigen/epitope. For that reason those antibodies will not, or just partly, recognize a purified antigen in its original state. 2. Peptides used for blocking experiments may be recombinant proteins. These aren't containing the characteristic sugar chains being the target of many antibodies. 3. A monoclonal antibody usually reacts with a very small sequence composed of only a few amino acids or a few sugar moieties. When performing IHC we all hope such a sequence is characteristic for a certain cell type or protein we wish to visualize. However, because the sequence is so small there is quite a good chance that other cell types or proteins contain something a similar sequence. If such a sequence is also present at other cells or proteins, we observe positive staining we don't want to see; let's call it "unwanted specific staining". This is not "background staining" because the reaction between antibody-antigen is after all still "specific". For further reading on these issues read: Willingham, JHC 47:1233-1235, 1999: "Conditional epitopes: Is your antibody always specific?" For the three reasons above I believe that blocking tests doesn't give any insight to the specificity of an antibody. And, that was the initial question to this list. Apart from the controls as suggested by Tim, you better select a number of good positive and negative control tissues based on literature data. Not necessarily based on IHC, but perhaps with other detection methods like ELISA, FACS, Western blotting. Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From "Morken, Tim - Labvision" Date Thu, 04 Mar 2004 08:36:04 -0800 To Histonet@pathology.swmed.edu Subject RE: [Histonet] Confirming IHC staining as "real" Probably the best is to complex it with the known antigen before putting it on the slide and then it should be negative. If you don't have the antigen, then you need a known control and some info on what the ab is supposed to stain. Negative controls should include 1) no primary or serum (buffer only), 2) normal serum from host animal of your primary 3) nonsense antibodies (aren't supposed to stain the target tissue - use ab's from the same host and preferably same Ig type) and 4) non-target tissues incubated with your primary(should be negative). Tim Morken Lab Vision / Neomarkers www.labvision.com From la.sebree <@t> hosp.wisc.edu Fri Mar 5 08:13:36 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!!! Message-ID: I thought these communiqu?s should be shared with the Histonet list server. 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: lualhati harkins [mailto:leharkins@yahoo.com] Sent: Thursday, March 04, 2004 6:23 PM To: Patsy Ruegg; Rcartun@harthosp.org; deb.vaneyck@phci.org; ihcrg@yahoogroups.com Cc: brian.kueck@phci.org; david.ferguson@phci.org; katherine.bayliss@phci.org Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! HI Patsy, In addition to EGFr issue, i use Egfr antibody from Zymed and our pathologist like it. I also tried polyclonal from Bio Care and it is very good too. Currently, we do not have plans to evaluate and EGFR kit, we are confident with our developed protocol for the EGFr antibody we are currently using. We did pretty good with The CAP Survey samples using EGFR. We placed confidently in good percent of agreement with results from other labs. Thank you, Loui Patsy Ruegg wrote: Yes the rab. monoclonal egfr I am using is a sample from Lab Vision. Have you used it? Any pointers? 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 HP-303-644-4538 hm email pruegg@msn.com wk email pruegg@colobio.com 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 Follows---- From: "Richard Cartun" To: ,, CC: ,, Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! Date: Thu, 04 Mar 2004 14:55:46 -0500 Yes, and it's very expensive! We evaluated their EGFr kit a while ago along with two other mAbs (BioGenex and Zymed). We actually prefer the Zymed antibody. Patsy, is the rabbit mAb that you are using from Lab Vision? Rich Cartun >>> "Patsy Ruegg" 03/04/04 02:11PM >>> Deb, This is a very important issue and I hope that we all get fed back on this. I am using the rabbit monoclonal egfr antibody on some research samples, but not the kit. I have very little data yet to report. Is DAKO selling the EGFR kit? 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 HP-303-644-4538 hm email pruegg@msn.com wk email pruegg@colobio.com 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 Follows---- From: "Van Eyck, Deb" To: "'ihcrg'" CC: "Kueck, Brian D." , "Bayliss, Katherine" , "Ferguson, David J." Subject: [IHCRG] Very important question!!!!!!!!!!!!!!!! Date: Thu, 4 Mar 2004 10:26:38 -0600 Hi and I would like feedback from clinical folks out there--------have you started running your EGFR kits on patients yet???? How is the staining???? I ran a number of random cases and seem to be getting darker staining and more cytoplasmic reaction than suggested. My negs are great and clean-kit exactly followed. My cervix is much darker than in picture in old guide and the diffuse cytoplasmic staining predominated most cases making the membrane staining hard to evaluate. What is your experience?? Most of my cases are 6 months or less tissue cases. This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. [Non-text portions of this message have been removed] Yahoo! Groups Links _________________________________________________________________ Create a Job Alert on MSN Careers and enter for a chance to win $1000! http://msn.careerbuilder.com/promo/kaday.htm?siteid=CBMSN_1K&sc_extcmp=JS_JASweep_MSNHotm2 Yahoo! Groups Links _________________________________________________________________ Learn how to help protect your privacy and prevent fraud online at Tech Hacks & Scams. http://special.msn.com/msnbc/techsafety.armx Yahoo! Groups SponsorADVERTISEMENT Click Here --------------------------------- Yahoo! Groups Links To visit your group on the web, go to: http://groups.yahoo.com/group/IHCRG/ To unsubscribe from this group, send an email to: IHCRG-unsubscribe@yahoogroups.com Your use of Yahoo! Groups is subject to the Yahoo! Terms of Service. **************************************************************** Please note: Protocol and technical information provided by the undersigned are developed on site with conditions limited to fixation of tissues,reagents,control tissues and execution of protocol which is characteristic only of our laboratory and therefore we do not bear neither responsibility nor liability for results outside of our laboratory. It is our courteous gesture to share information that we deemed might be useful to laboratorians and technologists. ****************************************************************** Loui Harkins , M.S. QIHC Biologist, Immunohistochemistry/Path.& lab. Medicine DEpt. Of Veterans Affairs,700 South, 19th Street Birmingham, Alabama 35233 (Research Consultant) Dept .of Surgery, Neurosurgery Division University of Alabama in Birmingham 205-933-8101 ext.6719; fax 205-558-4817 or 558-7034 --------------------------------- Do you Yahoo!? Yahoo! Search - Find what you're looking for faster. [Non-text portions of this message have been removed] Yahoo! Groups Links <*> To visit your group on the web, go to: http://groups.yahoo.com/group/IHCRG/ <*> To unsubscribe from this group, send an email to: IHCRG-unsubscribe@yahoogroups.com <*> Your use of Yahoo! Groups is subject to: http://docs.yahoo.com/info/terms/ From lleach <@t> uic.edu Fri Mar 5 09:21:40 2004 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Collagen I antibody Message-ID: <6.0.1.1.2.20040305091540.02354ad0@tigger.uic.edu> Hi all, I need to locate a Collagen I antibody that will work in FFPE tissue. I am currently using one from Cedarlane with inconsistent results. Can anyone suggest a replacement? Thank you, Lu Leach Director Research Pathology Core University of Illinois at Chicago 1819 West Polk Street Chicago, Illinois 60612 312-996-3869 lleach@uic.edu From la.sebree <@t> hosp.wisc.edu Fri Mar 5 09:21:51 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!!! Message-ID: FYI 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: Mary Cheles [mailto:mynerva7@msn.com] Sent: Friday, March 05, 2004 8:54 AM To: 'Van Eyck Deb'; Rena Fail Cc: Ihcrg Subject: Re: [IHCRG] Very important question!!!!!!!!!!!!!!!! Hello All, I will preface my remarks by stating that I am employed by DakoCytomation as the US Manager for pharmacoDiagnostics which includes the EGFR pharmDx kit. I am more than happy to answer any questions that you may have either by email or phone. I would like to take this opportunity to respond to some of the questions that have already been posed. The EGFR pharmDx kit received FDA approval in conjunction with Erbitux on Feb 12, 2004. Erbitux is a new monoclonal antibody therapy that binds to the extracellular domain of EGFR and is used for metastatic colorectal cancer patients who are refractory to irinotecan therapy. The EGFR pharmDx kit's FDA approval is to aid in identifying colorectal cancer patients eligible for Erbitux therapy. The EGFR pharmDx assay was used to determine eligibility of all patients entered into all the Erbitux clinical trials (entrance into the trial required positive EGFR expression). The clone in the EGFR pharmDx kit (2-18C9) is a monoclonal and due to its use in the clinical trials is directly linked to patient outcome. Therefore, matching an alternative antibody to its sensitivity and specificity with total precision would not easily be accomplished. The staining pattern for EGFR is primarily membranous (complete and incomplete) although some staining may be observed in the cytoplasm and extracellular spaces. Also contributing to a 'different' look than we are accustomed to with HER2 staining is that they are many normal cells that will mark with EGFR (epithelial & non-epithelial). Colorectal tumors tend to stain heterogeneously - this is part of the biology and again presents a different look than what we are used to seeing. Bristol Myers Squibb (a large pharmaceutical company & the US distributor of Erbitux) is working closely with your oncologists. Their FDA approved package insert does specify that EGFR testing with the DakoCytomation EGFR pharmDx assay is required to determine patient eligibility for this therapy. Therefore, most labs are receiving very specific requests for this test. Certainly for this specific purpose, the kit should be used to comply with the FDA approval. The EGFR pharmDx kit can obviously used to stain for EGFR for other reasons as well although labs may opt to use a different EGFR antibody for that use. It would entail having two EGFR antibodies/protocols on-line in the lab and clearly differentiating when to use which. Regarding fixation and processing... The Package Insert only states that "Biopsy specimens must be handled to preserve the tissue for IHC staining. Standard methods of tissue processing should be used for all specimens." There is a list of 6 fixatives which have been validated for use. I would be happy to email an electronic copy of the Package Insert to anyone interested in reviewing it. Specifically to Deb, the heavy cytomplasmic staining that you have seen is uncharacteristic. I know that our Technical Service Group is working with you to identify what is causing this in your lab. Please feel free to contact me for additional information. Mary Cheles, MPH, HTL, DLM (ASCP) 805-566-5492 mary.cheles@dakocytomation.com ----- Original Message ----- From: Rena Fail To: 'Van Eyck Deb' Cc: Ihcrg Sent: Friday, March 05, 2004 3:15 AM Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! We are also using Zymeds EGFR with very good results. The kit from DAKO is expensive, but we do expect to have request for this test from the oncologist. We have not yet tested the kit. Rena Fail HT(QIHC)ASCP -----Original Message----- From: Van Eyck Deb [mailto:deb.vaneyck@phci.org] Sent: Thursday, March 04, 2004 11:27 AM To: 'ihcrg' Cc: Kueck Brian D; Bayliss Katherine; Ferguson David J Subject: [IHCRG] Very important question!!!!!!!!!!!!!!!! Hi and I would like feedback from clinical folks out there--------have you started running your EGFR kits on patients yet???? How is the staining???? I ran a number of random cases and seem to be getting darker staining and more cytoplasmic reaction than suggested. My negs are great and clean-kit exactly followed. My cervix is much darker than in picture in old guide and the diffuse cytoplasmic staining predominated most cases making the membrane staining hard to evaluate. What is your experience?? Most of my cases are 6 months or less tissue cases. This information is confidential and intended solely for the use of the individual or entity to whom it is addressed. If you have received this email in error please notify the sender or our Customer Support Center at (262) 928-2777. We have scanned this email and its attachments for malicious content. However, the recipient should check this email and any attachments for the presence of viruses. ProHealth Care accepts no liability for any damage caused by any virus transmitted by this email. [Non-text portions of this message have been removed] Yahoo! Groups Links Yahoo! Groups Links [Non-text portions of this message have been removed] Yahoo! Groups Links <*> To visit your group on the web, go to: http://groups.yahoo.com/group/IHCRG/ <*> To unsubscribe from this group, send an email to: IHCRG-unsubscribe@yahoogroups.com <*> Your use of Yahoo! Groups is subject to: http://docs.yahoo.com/info/terms/ From ploykasek <@t> phenopath.com Fri Mar 5 09:55:56 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!!! In-Reply-To: Message-ID: Also, in regards to the use of the Dako EGFR pharmDx kit, the patients may not be reimbursed for their therapy if the EGFR testing is done with something other than the kit. This therapy is approximately $5-6,000 for the 1st month, and maintenance is $3-4,000 per month.>As you can see, it becomes an important patient care issue. The patients are the people we are really working for, after all. Patti Loykasek PhenoPath Laboratories Seattle, WA FYI > > 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: Mary Cheles [mailto:mynerva7@msn.com] > Sent: Friday, March 05, 2004 8:54 AM > To: 'Van Eyck Deb'; Rena Fail > Cc: Ihcrg > Subject: Re: [IHCRG] Very important question!!!!!!!!!!!!!!!! > > > Hello All, > > I will preface my remarks by stating that I am employed by DakoCytomation as > the US Manager for pharmacoDiagnostics which includes the EGFR pharmDx kit. I > am more than happy to answer any questions that you may have either by email > or phone. I would like to take this opportunity to respond to some of the > questions that have already been posed. The EGFR pharmDx kit received FDA > approval in conjunction with Erbitux on Feb 12, 2004. Erbitux is a new > monoclonal antibody therapy that binds to the extracellular domain of EGFR and > is used for metastatic colorectal cancer patients who are refractory to > irinotecan therapy. > > The EGFR pharmDx kit's FDA approval is to aid in identifying colorectal cancer > patients eligible for Erbitux therapy. The EGFR pharmDx assay was used to > determine eligibility of all patients entered into all the Erbitux clinical > trials (entrance into the trial required positive EGFR expression). The clone > in the EGFR pharmDx kit (2-18C9) is a monoclonal and due to its use in the > clinical trials is directly linked to patient outcome. Therefore, matching an > alternative antibody to its sensitivity and specificity with total precision > would not easily be accomplished. > > The staining pattern for EGFR is primarily membranous (complete and > incomplete) although some staining may be observed in the cytoplasm and > extracellular spaces. Also contributing to a 'different' look than we are > accustomed to with HER2 staining is that they are many normal cells that will > mark with EGFR (epithelial & non-epithelial). Colorectal tumors tend to stain > heterogeneously - this is part of the biology and again presents a different > look than what we are used to seeing. > > Bristol Myers Squibb (a large pharmaceutical company & the US distributor of > Erbitux) is working closely with your oncologists. Their FDA approved package > insert does specify that EGFR testing with the DakoCytomation EGFR pharmDx > assay is required to determine patient eligibility for this therapy. > Therefore, most labs are receiving very specific requests for this test. > Certainly for this specific purpose, the kit should be used to comply with the > FDA approval. The EGFR pharmDx kit can obviously used to stain for EGFR for > other reasons as well although labs may opt to use a different EGFR antibody > for that use. It would entail having two EGFR antibodies/protocols on-line in > the lab and clearly differentiating when to use which. > > Regarding fixation and processing... The Package Insert only states that > "Biopsy specimens must be handled to preserve the tissue for IHC staining. > Standard methods of tissue processing should be used for all specimens." > There is a list of 6 fixatives which have been validated for use. I would be > happy to email an electronic copy of the Package Insert to anyone interested > in reviewing it. > > Specifically to Deb, the heavy cytomplasmic staining that you have seen is > uncharacteristic. I know that our Technical Service Group is working with you > to identify what is causing this in your lab. > > Please feel free to contact me for additional information. > Mary Cheles, MPH, HTL, DLM (ASCP) > 805-566-5492 > mary.cheles@dakocytomation.com > > ----- Original Message ----- > From: Rena Fail > To: 'Van Eyck Deb' > Cc: Ihcrg > Sent: Friday, March 05, 2004 3:15 AM > Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! > > > > We are also using Zymeds EGFR with very good results. The kit from DAKO > is expensive, but we do expect to have request for this test from the > oncologist. We have not yet tested the kit. > > Rena Fail HT(QIHC)ASCP > > -----Original Message----- > From: Van Eyck Deb [mailto:deb.vaneyck@phci.org] > Sent: Thursday, March 04, 2004 11:27 AM > To: 'ihcrg' > Cc: Kueck Brian D; Bayliss Katherine; Ferguson David J > Subject: [IHCRG] Very important question!!!!!!!!!!!!!!!! > > > Hi and I would like feedback from clinical folks out there--------have > you started running your EGFR kits on patients yet???? How is the > staining???? I ran a number of random cases and seem to be getting > darker staining and more cytoplasmic reaction than suggested. My negs > are great and clean-kit exactly followed. My cervix is much darker than > in picture in old guide and the diffuse cytoplasmic staining > predominated most cases making the membrane staining hard to evaluate. > What is your experience?? Most of my cases are 6 months or less tissue > cases. > > > This information is confidential and intended solely for the use of the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for > the presence of viruses. ProHealth Care accepts no liability for any > damage caused by any virus transmitted by this email. > > > > [Non-text portions of this message have been removed] > > > > > > Yahoo! Groups Links > > > > > > > > > Yahoo! Groups Links > > > > > > > > [Non-text portions of this message have been removed] > > > > > Yahoo! Groups Links > > <*> To visit your group on the web, go to: > http://groups.yahoo.com/group/IHCRG/ > > <*> To unsubscribe from this group, send an email to: > IHCRG-unsubscribe@yahoogroups.com > > <*> Your use of Yahoo! Groups is subject to: > http://docs.yahoo.com/info/terms/ > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Jackie.O'Connor <@t> abbott.com Fri Mar 5 10:50:52 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Another plea for human vs mouse Message-ID: Hi Y'all - I've thrown this question out before, but still don't have an answer. Please oh please, if you have an idea, please share! I'm trying to find an antibody that will specifically identify any and ALL human tissue but will not cross react with mouse. My boss is thinking along the lines of a ribosomal protein - I just look at him like a deer caught in the headlights. We're working with human xenografts in mice. Since we work with so many different tumor lines, from melanoma to prostate, we would like a type of broad spectrum antibody that WILL stain human cells but will NOT stain mouse stroma, etc. Any clues?? Jackie O' Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics Jackie.O'Connor@abbott.com From dgnajarian <@t> comcast.net Fri Mar 5 11:09:01 2004 From: dgnajarian <@t> comcast.net (Dennis Najarian) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Collagen I antibody References: <6.0.1.1.2.20040305091540.02354ad0@tigger.uic.edu> Message-ID: <000801c402d4$8eb3b200$6401a8c0@natm.corp.chem> Chemicon offers a few for use in IHC. I am not sure if it works on FFPE tissue. Theit tech support dept offers a 50% rebate for any antibody that might not have been tested on certain tissues/applications. ----- Original Message ----- From: "Lu Leach" To: Sent: Friday, March 05, 2004 7:21 AM Subject: [Histonet] Collagen I antibody > Hi all, > > I need to locate a Collagen I antibody that will work in FFPE tissue. I am > currently using one from Cedarlane with inconsistent results. Can anyone > suggest a replacement? > > Thank you, > > Lu Leach > Director Research Pathology Core > University of Illinois at Chicago > 1819 West Polk Street > Chicago, Illinois 60612 > 312-996-3869 > lleach@uic.edu > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From whitmang <@t> trinity-health.org Fri Mar 5 10:36:42 2004 From: whitmang <@t> trinity-health.org (Georgine Whitman) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] keeping fingernail and toenail on the slide Message-ID: we are having a problem keeping the nail tissue on our slides. we have used plus slides, plus slides with albumin on it, albumin on it and in the oven for about 5 minutes. most of the time the tissue stays on the h&e slide. but with the pas notttttttttttttttt. We use koh to soften the nail prior to processing and before we cut the block. We also have soaked it in decal before cutting the block. Thanks Georgine Livonia MI From japoteete <@t> saintfrancis.com Fri Mar 5 11:05:31 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: We had a very enlightening call from the Erbitux drug rep last week, and from her statement, the drug will not even be offered to any patient without documentation that the DAKO kit was used. I have had to supply that information to our research people who enter patients on the protocols Possibly it might be due to the "newness" of this kit, but probably, from Patti's numbers, reimbursement will be a big issue. We were also advised to use charge code 88342-22 to increase our reimbursement. Jacquie Poteete MT(ASCP)QIHC, Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Sebree Linda A. [SMTP:la.sebree@hosp.wisc.edu] > Sent: Friday, March 05, 2004 8:14 AM > To: Histonet (E-mail) > Subject: [Histonet] FW: [IHCRG] Very important > question!!!!!!!!!!!!!!!! > > I thought these communiqu?s should be shared with the Histonet list > server. > > 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: lualhati harkins [mailto:leharkins@yahoo.com] > Sent: Thursday, March 04, 2004 6:23 PM > To: Patsy Ruegg; Rcartun@harthosp.org; deb.vaneyck@phci.org; > ihcrg@yahoogroups.com > Cc: brian.kueck@phci.org; david.ferguson@phci.org; > katherine.bayliss@phci.org > Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! > > > HI Patsy, > In addition to EGFr issue, i use Egfr antibody from Zymed and our > pathologist like it. I also tried polyclonal from Bio Care and it is very > good too. Currently, we do not have plans to evaluate and EGFR kit, we are > confident with our developed protocol for the EGFr antibody we are > currently using. We did pretty good with The CAP Survey samples using > EGFR. We placed confidently in good percent of agreement with results from > other labs. > Thank you, > Loui > > Patsy Ruegg wrote: > Yes the rab. monoclonal egfr I am using is a sample from Lab Vision. Have > > you used it? Any pointers? > 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 > HP-303-644-4538 > > hm email pruegg@msn.com > wk email pruegg@colobio.com > > 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 Follows---- > From: "Richard Cartun" > To: ,, > CC: > ,, rg> > Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! > Date: Thu, 04 Mar 2004 14:55:46 -0500 > > Yes, and it's very expensive! We evaluated their EGFr kit a while ago > along with two other mAbs (BioGenex and Zymed). We actually prefer the > Zymed antibody. Patsy, is the rabbit mAb that you are using from Lab > Vision? > > Rich Cartun > > >>> "Patsy Ruegg" 03/04/04 02:11PM >>> > Deb, > This is a very important issue and I hope that we all get fed back on > this. > I am using the rabbit monoclonal egfr antibody on some research > samples, > but not the kit. I have very little data yet to report. Is DAKO > selling > the EGFR kit? > 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 > HP-303-644-4538 > > hm email pruegg@msn.com > wk email pruegg@colobio.com > > 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 Follows---- > From: "Van Eyck, Deb" > To: "'ihcrg'" > CC: "Kueck, Brian D." , "Bayliss, Katherine" > , "Ferguson, David J." > > Subject: [IHCRG] Very important question!!!!!!!!!!!!!!!! > Date: Thu, 4 Mar 2004 10:26:38 -0600 > > Hi and I would like feedback from clinical folks out there--------have > you > started running your EGFR kits on patients yet???? How is the > staining???? > I ran a number of random cases and seem to be getting darker staining > and > more cytoplasmic reaction than suggested. My negs are great and > clean-kit > exactly followed. My cervix is much darker than in picture in old > guide and > the diffuse cytoplasmic staining predominated most cases making the > membrane > staining hard to evaluate. What is your experience?? Most of my cases > are 6 > months or less tissue cases. > > > This information is confidential and intended solely for the use of > the > individual or entity to whom it is addressed. > If you have received this email in error please notify the sender or > our > Customer Support Center at (262) 928-2777. > > We have scanned this email and its attachments for malicious content. > However, the recipient should check this email and any attachments for > the > presence of viruses. > ProHealth Care accepts no liability for any damage caused by any virus > transmitted by this email. > > > > [Non-text portions of this message have been removed] > > > > > > Yahoo! Groups Links > > > > > _________________________________________________________________ > Create a Job Alert on MSN Careers and enter for a chance to win $1000! > > http://msn.careerbuilder.com/promo/kaday.htm?siteid=CBMSN_1K&sc_extcmp=JS_ > JASweep_MSNHotm2 > > > > > > Yahoo! Groups Links > > > > > _________________________________________________________________ > Learn how to help protect your privacy and prevent fraud online at Tech > Hacks & Scams. http://special.msn.com/msnbc/techsafety.armx > > > Yahoo! Groups SponsorADVERTISEMENT > Click Here > > --------------------------------- > Yahoo! Groups Links > > To visit your group on the web, go to: > http://groups.yahoo.com/group/IHCRG/ > > To unsubscribe from this group, send an email to: > IHCRG-unsubscribe@yahoogroups.com > > Your use of Yahoo! Groups is subject to the Yahoo! Terms of Service. > > > > **************************************************************** > Please note: Protocol and technical information provided by the > undersigned are developed on site with conditions limited to fixation of > tissues,reagents,control tissues and execution of protocol which is > characteristic only of our laboratory and therefore we do not bear neither > responsibility nor liability for results outside of our laboratory. It is > our courteous gesture to share > information that we deemed might be useful to laboratorians and > technologists. > ****************************************************************** > Loui Harkins , M.S. QIHC > Biologist, Immunohistochemistry/Path.& lab. Medicine > DEpt. Of Veterans Affairs,700 South, 19th Street > Birmingham, Alabama 35233 > (Research Consultant) Dept .of Surgery, Neurosurgery Division > University of Alabama in Birmingham > 205-933-8101 ext.6719; fax 205-558-4817 or 558-7034 > > --------------------------------- > Do you Yahoo!? > Yahoo! Search - Find what you're looking for faster. > > [Non-text portions of this message have been removed] > > > > > Yahoo! Groups Links > > <*> To visit your group on the web, go to: > http://groups.yahoo.com/group/IHCRG/ > > <*> To unsubscribe from this group, send an email to: > IHCRG-unsubscribe@yahoogroups.com > > <*> Your use of Yahoo! Groups is subject to: > http://docs.yahoo.com/info/terms/ > > > > _______________________________________________ > 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 DDDeltour <@t> sig.med.navy.mil Fri Mar 5 11:20:42 2004 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] keeping fingernail and toenail on the slide Message-ID: Don't you just love it when they order those??? I have to leave them in the oven for hours before it will stay on the slide. Good luck -----Original Message----- From: Georgine Whitman To: histonet@pathology.swmed.edu Sent: 3/5/04 5:36 PM Subject: [Histonet] keeping fingernail and toenail on the slide we are having a problem keeping the nail tissue on our slides. we have used plus slides, plus slides with albumin on it, albumin on it and in the oven for about 5 minutes. most of the time the tissue stays on the h&e slide. but with the pas notttttttttttttttt. We use koh to soften the nail prior to processing and before we cut the block. We also have soaked it in decal before cutting the block. Thanks Georgine Livonia MI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. From MGomez <@t> ameripath.com Fri Mar 5 11:47:43 2004 From: MGomez <@t> ameripath.com (MGomez@ameripath.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] using microwave to dry paraffin slides Message-ID: Good morning Histonetters: I'm currently using the microwave for 3' instead of the conventional oven to dry paraffin sections. Occasionally the Hematoxylin staining is uneven and the cells look pale blue. Has anyone experienced this and if so do you have any recommendations? Thank you very much, Milton From ploykasek <@t> phenopath.com Fri Mar 5 12:02:08 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! In-Reply-To: Message-ID: I will add that I know our insurance carriers will not reimburse for the modifier 22 with IHC. I am sure that this is variable across the country, and can only relate our experience. Patti Loykasek PhenoPath Laboratories Seattle, WA> We had a very enlightening call from the Erbitux drug rep last week, and > from her statement, the drug will not even be offered to any patient without > documentation that the DAKO kit was used. I have had to supply that > information to our research people who enter patients on the protocols > Possibly it might be due to the "newness" of this kit, but probably, from > Patti's numbers, reimbursement will be a big issue. We were also advised to > use charge code 88342-22 to increase our reimbursement. > > Jacquie Poteete MT(ASCP)QIHC, Lead Technologist, IHC Laboratory > Saint Francis Hospital, Tulsa, OK > japoteete@saintfrancis.com > >> -----Original Message----- >> From: Sebree Linda A. [SMTP:la.sebree@hosp.wisc.edu] >> Sent: Friday, March 05, 2004 8:14 AM >> To: Histonet (E-mail) >> Subject: [Histonet] FW: [IHCRG] Very important >> question!!!!!!!!!!!!!!!! >> >> I thought these communiqu?s should be shared with the Histonet list >> server. >> >> 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: lualhati harkins [mailto:leharkins@yahoo.com] >> Sent: Thursday, March 04, 2004 6:23 PM >> To: Patsy Ruegg; Rcartun@harthosp.org; deb.vaneyck@phci.org; >> ihcrg@yahoogroups.com >> Cc: brian.kueck@phci.org; david.ferguson@phci.org; >> katherine.bayliss@phci.org >> Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! >> >> >> HI Patsy, >> In addition to EGFr issue, i use Egfr antibody from Zymed and our >> pathologist like it. I also tried polyclonal from Bio Care and it is very >> good too. Currently, we do not have plans to evaluate and EGFR kit, we are >> confident with our developed protocol for the EGFr antibody we are >> currently using. We did pretty good with The CAP Survey samples using >> EGFR. We placed confidently in good percent of agreement with results from >> other labs. >> Thank you, >> Loui >> >> Patsy Ruegg wrote: >> Yes the rab. monoclonal egfr I am using is a sample from Lab Vision. Have >> >> you used it? Any pointers? >> 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 >> HP-303-644-4538 >> >> hm email pruegg@msn.com >> wk email pruegg@colobio.com >> >> 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 Follows---- >> From: "Richard Cartun" >> To: ,, >> CC: >> ,,> rg> >> Subject: RE: [IHCRG] Very important question!!!!!!!!!!!!!!!! >> Date: Thu, 04 Mar 2004 14:55:46 -0500 >> >> Yes, and it's very expensive! We evaluated their EGFr kit a while ago >> along with two other mAbs (BioGenex and Zymed). We actually prefer the >> Zymed antibody. Patsy, is the rabbit mAb that you are using from Lab >> Vision? >> >> Rich Cartun >> >>>>> "Patsy Ruegg" 03/04/04 02:11PM >>> >> Deb, >> This is a very important issue and I hope that we all get fed back on >> this. >> I am using the rabbit monoclonal egfr antibody on some research >> samples, >> but not the kit. I have very little data yet to report. Is DAKO >> selling >> the EGFR kit? >> 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 >> HP-303-644-4538 >> >> hm email pruegg@msn.com >> wk email pruegg@colobio.com >> >> 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 Follows---- >> From: "Van Eyck, Deb" >> To: "'ihcrg'" >> CC: "Kueck, Brian D." , "Bayliss, Katherine" >> , "Ferguson, David J." >> >> Subject: [IHCRG] Very important question!!!!!!!!!!!!!!!! >> Date: Thu, 4 Mar 2004 10:26:38 -0600 >> >> Hi and I would like feedback from clinical folks out there--------have >> you >> started running your EGFR kits on patients yet???? How is the >> staining???? >> I ran a number of random cases and seem to be getting darker staining >> and >> more cytoplasmic reaction than suggested. My negs are great and >> clean-kit >> exactly followed. My cervix is much darker than in picture in old >> guide and >> the diffuse cytoplasmic staining predominated most cases making the >> membrane >> staining hard to evaluate. What is your experience?? Most of my cases >> are 6 >> months or less tissue cases. >> >> >> This information is confidential and intended solely for the use of >> the >> individual or entity to whom it is addressed. >> If you have received this email in error please notify the sender or >> our >> Customer Support Center at (262) 928-2777. >> >> We have scanned this email and its attachments for malicious content. >> However, the recipient should check this email and any attachments for >> the >> presence of viruses. >> ProHealth Care accepts no liability for any damage caused by any virus >> transmitted by this email. >> >> >> >> [Non-text portions of this message have been removed] >> >> >> >> >> >> Yahoo! Groups Links >> >> >> >> >> _________________________________________________________________ >> Create a Job Alert on MSN Careers and enter for a chance to win $1000! >> >> http://msn.careerbuilder.com/promo/kaday.htm?siteid=CBMSN_1K&sc_extcmp=JS_ >> JASweep_MSNHotm2 >> >> >> >> >> >> Yahoo! Groups Links >> >> >> >> >> _________________________________________________________________ >> Learn how to help protect your privacy and prevent fraud online at Tech >> Hacks & Scams. http://special.msn.com/msnbc/techsafety.armx >> >> >> Yahoo! Groups SponsorADVERTISEMENT >> Click Here >> >> --------------------------------- >> Yahoo! Groups Links >> >> To visit your group on the web, go to: >> http://groups.yahoo.com/group/IHCRG/ >> >> To unsubscribe from this group, send an email to: >> IHCRG-unsubscribe@yahoogroups.com >> >> Your use of Yahoo! Groups is subject to the Yahoo! Terms of Service. >> >> >> >> **************************************************************** >> Please note: Protocol and technical information provided by the >> undersigned are developed on site with conditions limited to fixation of >> tissues,reagents,control tissues and execution of protocol which is >> characteristic only of our laboratory and therefore we do not bear neither >> responsibility nor liability for results outside of our laboratory. It is >> our courteous gesture to share >> information that we deemed might be useful to laboratorians and >> technologists. >> ****************************************************************** >> Loui Harkins , M.S. QIHC >> Biologist, Immunohistochemistry/Path.& lab. Medicine >> DEpt. Of Veterans Affairs,700 South, 19th Street >> Birmingham, Alabama 35233 >> (Research Consultant) Dept .of Surgery, Neurosurgery Division >> University of Alabama in Birmingham >> 205-933-8101 ext.6719; fax 205-558-4817 or 558-7034 >> >> --------------------------------- >> Do you Yahoo!? >> Yahoo! Search - Find what you're looking for faster. >> >> [Non-text portions of this message have been removed] >> >> >> >> >> Yahoo! Groups Links >> >> <*> To visit your group on the web, go to: >> http://groups.yahoo.com/group/IHCRG/ >> >> <*> To unsubscribe from this group, send an email to: >> IHCRG-unsubscribe@yahoogroups.com >> >> <*> Your use of Yahoo! Groups is subject to: >> http://docs.yahoo.com/info/terms/ >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> >> > ********* Email Confidentiality Statement ********* > Visit http://www.saintfrancis.com/emailconf.asp > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Bauer.Karen <@t> mayo.edu Fri Mar 5 12:13:50 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] keeping fingernail and toenail on the slide Message-ID: <8C6E05FA69571948B461F1327CBB893E16C39F@lmmail2.ad.lmmhs.org> On all finger/toe nails, we take plus slides and smear a little Elmers Glue on them. Cut, apply sections and heat normally with the rest of the slides. (We always cut a few extras, just in case they order a GMS and PAS.) They actually stay on the slides quite well, although we get an occasional stubborn one once in a while. We do our GMS stain in the microwave, and they stay on during the heat an all. Give it a try... Good Luck!! Karen Bauer HT(ASCP) Histology Supervisor Department of Pathology Luther Hospital Eau Claire, WI -----Original Message----- From: Georgine Whitman [mailto:whitmang@trinity-health.org] Sent: Friday, March 05, 2004 10:37 AM To: histonet@pathology.swmed.edu Subject: [Histonet] keeping fingernail and toenail on the slide we are having a problem keeping the nail tissue on our slides. we have used plus slides, plus slides with albumin on it, albumin on it and in the oven for about 5 minutes. most of the time the tissue stays on the h&e slide. but with the pas notttttttttttttttt. We use koh to soften the nail prior to processing and before we cut the block. We also have soaked it in decal before cutting the block. Thanks Georgine Livonia MI _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From a.hannamitchell <@t> ucc.ie Fri Mar 5 12:18:46 2004 From: a.hannamitchell <@t> ucc.ie (Hanna-Mitchell, Ann) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Re removal of unfixed 'dorsal root(+DRG) -spinal cord' section from adult rat Message-ID: <9FBB394A25826C46B2C6F0EBDAD42755059C255F@xch2.ucc.ie> Good Evening (from Ireland), I am a first time user , so please bear with me. I was wondering whether anyone could advice me on the procedure for partial laminectomy ( I hope that this is the correct term) , in order to expose the dorsal root ganglion . I am able to remove the spinal cord from the adult rat, but I should like to get a section with the dorsal root plus the ganglion attached. I perfuse to remove blood but do not fix the tissue. Thank You, Ann T. Hanna-Mitchell PhD Dr. Ann T. Hanna-Mitchell Department of Anatomy BioSciences Institute University College Cork Ireland. Phone +353 21 4902246/+353 21 4901309 E mail a.hannamitchell@ucc.ie/athm@ucc.ie From shive003 <@t> umn.edu Fri Mar 5 13:01:36 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Canine Herpesvirus antibody Message-ID: <03d101c402e4$48734750$78065486@vdl220FAC> Does anyone know of a commercial source for an antibody to Canine Herpesvirus that can be used for IHC purposes (on FFPE tissue)? Thank you very much in advance. Jan Shivers Univ. of Minnesota Vet. Diag. Lab shive003@tc.umn.edu From japoteete <@t> saintfrancis.com Fri Mar 5 14:09:52 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] AFB Control Message-ID: Hello all, Our Histology department has exhausted their supply of AFB control blocks. I'm sure I saw a method of inoculating tissue to prepare this control, but I can't find it. They would appreciate any help anyone can give them. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com ********* Email Confidentiality Statement ********* Visit http://www.saintfrancis.com/emailconf.asp From DDittus787 <@t> aol.com Fri Mar 5 14:05:32 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: <536EBFBC.3D81F4F6.0A1F969F@aol.com> I have heard this stuff before ie: herceptest,however i am very aware of the hcfa form used in billing (it is universal-all over the country)and nowhere does it ask what test method!!!! now i think it might be wise to run some correlation studies with the dako kit, but the antibody egfr has been around longer than erbitux and from many manufacturers, and all that said one has to do whats best for ones lab and with the pathologists approval. i run a different clone for her2 and i run cd117 for another monoclonal therapy(gleevic) and all patients are receiving according to their doctors whatever they need and were getting paid, and yes there is full disclosure on our reports. just my 2 cents dana From jcline <@t> wchsys.org Fri Mar 5 14:44:10 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] nail problem Message-ID: <002c01c402f2$9d27f800$1d2a14ac@wchsys.org> To Georgine Whitman I use Stay-On by Surgipath and I just put some on a Kimwipe and wipe the slide with Stayon before I pick up my section. The waterbath also has Stayon in it. From TJJ <@t> Stowers-Institute.org Fri Mar 5 14:36:41 2004 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] TUNEL staining in whole mount chick embryos Message-ID: Has anybody done any TUNEL staining in whole mount chick or mouse embryos? If so, can you give me your technique or tips for doing such? Thanks! Teri Johnson Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, Missouri 64110 tjj@stowers-institute.org From Ronnie_Houston <@t> bshsi.com Fri Mar 5 14:40:09 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: <530361BF03351B4CAE5270A05D3037B503673124@bsrexms01.BSHSIR.COM> The clinical trials that gained FDA approval for Erbitux were based on positivity with the DakoCytomation EGFR pharmDx kit. It gives some creedence to the story that insurance companies will not pay for the drug unless this specific kit is used; certainly in the embryonic status of the drug's usage. It should be added though, that during the clinical trials no-one with a negative result by IHC was included in the study. Oncologists are asking whether there is a need to order the test. Perhaps those "negative" patients will also beneficially respond to Erbitux. There is no evidence to suggest they won't. Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 (804) 287 7906 - fax ronnie_houston@bshsi.com -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Friday, March 05, 2004 3:06 PM To: ploykasek@phenopath.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! I have heard this stuff before ie: herceptest,however i am very aware of the hcfa form used in billing (it is universal-all over the country)and nowhere does it ask what test method!!!! now i think it might be wise to run some correlation studies with the dako kit, but the antibody egfr has been around longer than erbitux and from many manufacturers, and all that said one has to do whats best for ones lab and with the pathologists approval. i run a different clone for her2 and i run cd117 for another monoclonal therapy(gleevic) and all patients are receiving according to their doctors whatever they need and were getting paid, and yes there is full disclosure on our reports. just my 2 cents dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From PKubier <@t> propathlab.com Fri Mar 5 15:00:20 2004 From: PKubier <@t> propathlab.com (PKubier@propathlab.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Lack of IHC staining peripheral edge Message-ID: <795594536E92D4118C800050046449F301079944@mailhost.propathlab.com> I apologize if this is a duplicate message. I've had difficulty posting on the histonet. In our laboratory we have noticed a lack of immunohistochemistry staining on the peripheral edges of the tissue sections on occasion. It is not limited to a specific case, as it seems to occur on all the cases to some degree on the same IHC run. The repeat stains have been fine, no artifact. I thought that it was a drying artifact and therefore, intentionally allowed a slide to dry out completely before application of the primary antibody. The immuno stain result was fine. Can anyone share with me what causes the lack of staining along the peripheral edges during IHC staining? Thanks in advance, Patty Kubier 800-258-1253 ext. 2027 From japoteete <@t> saintfrancis.com Fri Mar 5 15:07:35 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: In our study of the kit, we included several patients who had positive results on the "old" EGFR. Of the three, 2 were negative with EGFRdx and 1 was positive. I'm certainly not brave enough to push my luck and accept the liability for allowing my results to be the basis for entering patients on a clinical trial or other treatment, based on results from the use of another clone. Our pathologists and oncologists refuse to accept anything other than the FDA approved kits, but I also realize that this can vary from institution to institution. Some accept other clones or procedures, some don't. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: Houston, Ronnie [SMTP:Ronnie_Houston@bshsi.com] > Sent: Friday, March 05, 2004 2:40 PM > To: 'DDittus787@aol.com'; ploykasek@phenopath.com; > histonet@pathology.swmed.edu > Subject: RE: [Histonet] FW: [IHCRG] Very important > question!!!!!!!!!!!!!!! ! > > > The clinical trials that gained FDA approval for Erbitux were based on > positivity with the DakoCytomation EGFR pharmDx kit. It gives some > creedence > to the story that insurance companies will not pay for the drug unless > this > specific kit is used; certainly in the embryonic status of the drug's > usage. > > It should be added though, that during the clinical trials no-one with a > negative result by IHC was included in the study. Oncologists are asking > whether there is a need to order the test. Perhaps those "negative" > patients > will also beneficially respond to Erbitux. There is no evidence to suggest > they won't. > > Ronnie > > Ronnie Houston > Director of Anatomic Pathology > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > (804) 287 7906 - fax > ronnie_houston@bshsi.com > > -----Original Message----- > From: DDittus787@aol.com [mailto:DDittus787@aol.com] > Sent: Friday, March 05, 2004 3:06 PM > To: ploykasek@phenopath.com; histonet@pathology.swmed.edu > Subject: Re: [Histonet] FW: [IHCRG] Very important > question!!!!!!!!!!!!!!! ! > > > I have heard this stuff before ie: herceptest,however i am very aware of > the > hcfa form used in billing (it is universal-all over the country)and > nowhere > does it ask what test method!!!! now i think it might be wise to run some > correlation studies with the dako kit, but the antibody egfr has been > around > longer than erbitux and from many manufacturers, and all that said one has > to do whats best for ones lab and with the pathologists approval. i run a > different clone for her2 and i run cd117 for another monoclonal > therapy(gleevic) and all patients are receiving according to their doctors > whatever they need and were getting paid, and yes there is full disclosure > on our reports. just my 2 cents > dana > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > __________________________________________________________________________ > ______________________________________________________ > __________________________________________________________________________ > ______________________________________________________ > > The information in this communication is intended to be confidential to > the Individual(s) and/or Entity to whom it is addressed. > It may contain information of a Privileged and/or Confidential nature, > which is subject to Federal and/or State privacy regulations. > In the event that you are not the intended recipient or the agent of the > intended recipient, do not copy or use the information > contained within this communication, or allow it to be read, copied or > utilized in any manner, by any other person(s). Should > this communication be received in error, please notify the sender > immediately either by response e-mail or by phone at 410-442-3250, > and permanently delete the original e-mail, attachment(s), and any copies. > > > > _______________________________________________ > 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 ploykasek <@t> phenopath.com Fri Mar 5 15:24:35 2004 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! In-Reply-To: <530361BF03351B4CAE5270A05D3037B503673124@bsrexms01.BSHSIR.COM> Message-ID: Mr. Houston has a good point. We have actually included this in a comment that will be on all of our EGFR reports. Patti Loykasek > > The clinical trials that gained FDA approval for Erbitux were based on > positivity with the DakoCytomation EGFR pharmDx kit. It gives some creedence > to the story that insurance companies will not pay for the drug unless this > specific kit is used; certainly in the embryonic status of the drug's usage. > > It should be added though, that during the clinical trials no-one with a > negative result by IHC was included in the study. Oncologists are asking > whether there is a need to order the test. Perhaps those "negative" patients > will also beneficially respond to Erbitux. There is no evidence to suggest > they won't. > > Ronnie > > Ronnie Houston > Director of Anatomic Pathology > Bon Secours HealthPartners Laboratories > 5801 Bremo Road > Richmond, VA 23226 > (804) 287 7972 > (804) 287 7906 - fax > ronnie_houston@bshsi.com > > -----Original Message----- > From: DDittus787@aol.com [mailto:DDittus787@aol.com] > Sent: Friday, March 05, 2004 3:06 PM > To: ploykasek@phenopath.com; histonet@pathology.swmed.edu > Subject: Re: [Histonet] FW: [IHCRG] Very important > question!!!!!!!!!!!!!!! ! > > > I have heard this stuff before ie: herceptest,however i am very aware of the > hcfa form used in billing (it is universal-all over the country)and nowhere > does it ask what test method!!!! now i think it might be wise to run some > correlation studies with the dako kit, but the antibody egfr has been around > longer than erbitux and from many manufacturers, and all that said one has > to do whats best for ones lab and with the pathologists approval. i run a > different clone for her2 and i run cd117 for another monoclonal > therapy(gleevic) and all patients are receiving according to their doctors > whatever they need and were getting paid, and yes there is full disclosure > on our reports. just my 2 cents > dana > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ______________________________________________________________________________ > __________________________________________________ > ______________________________________________________________________________ > __________________________________________________ > > The information in this communication is intended to be confidential to the > Individual(s) and/or Entity to whom it is addressed. > It may contain information of a Privileged and/or Confidential nature, which > is subject to Federal and/or State privacy regulations. > In the event that you are not the intended recipient or the agent of the > intended recipient, do not copy or use the information > contained within this communication, or allow it to be read, copied or > utilized in any manner, by any other person(s). Should > this communication be received in error, please notify the sender immediately > either by response e-mail or by phone at 410-442-3250, > and permanently delete the original e-mail, attachment(s), and any copies. > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From DDittus787 <@t> aol.com Fri Mar 5 15:46:34 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: <3A4AF447.5A9D3625.0A1F969F@aol.com> not to second guess you,but i have been told (2days ago) that labs are not reading results (kit or not) correctly and that this is not graded like her2 (even though it is called her1)so weak positivity is really positive not neg, so studies should be done carefully. dana From arunams <@t> interchange.ubc.ca Fri Mar 5 15:38:14 2004 From: arunams <@t> interchange.ubc.ca (arunams) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] looking for used equipment Message-ID: <30341848.1078522694672.JavaMail.myubc2@portal1.itservices.ubc.ca> Hi Everybody, Does any one have used histology equipment that they want to get rid of or can live without? I am specifically looking for a cryostat but would be interested in any other equipment too. Thanks for your help. Aruna Somasiri Vancouver BC. Canada From SCheasty <@t> ahs.llumc.edu Fri Mar 5 15:58:19 2004 From: SCheasty <@t> ahs.llumc.edu (Cheasty, Sandra) Date: Fri Sep 16 15:22:38 2005 Subject: [Histonet] Laboratory Furniture Resources Needed Message-ID: <2E50F33F91EEDA46A77BC3B2575BB09105877B@mars.llumc.edu> Anyone know of a company that is a source for pathologist's desks? Our pathologist's offices are moving and we need to buy desks that accommodate a double-headed scope seating arrangement. Their current offices have built-in desks with the double-headed scopes on an extra "peninsula" counter top that extends from one end of the desk and that is lower than the desk itself to accommodate the scope. Thanks, Sandy Confidentiality Note: The preceding e-mail message (including any attachments) contains information that may be confidential, protected by applicable legal privileges, or constitute non-public information. It is intended to be conveyed only to the designated recipient(s). If you are not an intended recipient of this message, please notify the sender by replying to this message and then delete it from your system. Use, dissemination, distribution or reproduction of this message by unintended recipients is not authorized and may be unlawful. From Ronnie_Houston <@t> bshsi.com Fri Mar 5 15:55:48 2004 From: Ronnie_Houston <@t> bshsi.com (Houston, Ronnie) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: <530361BF03351B4CAE5270A05D3037B503673126@bsrexms01.BSHSIR.COM> Where did this come from? The inclusion for Erbitux treatment depends on a POSITIVE reaction to the DakoCytomation EGFR pharmDx, no more no less. There has never been any discussion about grading the reaction either semi-quantitatively or otherwise. I suggest that those grading their test results are not using this kit. Dako make it very clear to their clients on how to interpret the test results. Ronnie Ronnie Houston Director of Anatomic Pathology Bon Secours HealthPartners Laboratories 5801 Bremo Road Richmond, VA 23226 (804) 287 7972 (804) 287 7906 - fax ronnie_houston@bshsi.com -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Friday, March 05, 2004 4:47 PM To: japoteete@saintfrancis.com; Ronnie_Houston@bshsi.com; ploykasek@phenopath.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! not to second guess you,but i have been told (2days ago) that labs are not reading results (kit or not) correctly and that this is not graded like her2 (even though it is called her1)so weak positivity is really positive not neg, so studies should be done carefully. dana ________________________________________________________________________________________________________________________________ ________________________________________________________________________________________________________________________________ The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at 410-442-3250, and permanently delete the original e-mail, attachment(s), and any copies. From PKubier <@t> propathlab.com Fri Mar 5 16:52:17 2004 From: PKubier <@t> propathlab.com (PKubier@propathlab.com) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Dako EGFR Kit Message-ID: <795594536E92D4118C800050046449F301079948@mailhost.propathlab.com> The use of the DakoCytomation EGFR kit seems to be a hot topic currently, and I'd like to have some feedback as to how to interpret the following excerpt from the Erbitux package insert, obtained from the following link through the FDA (see page 8 of this document) http://www.fda.gov/cder/foi/label/2004/125084lbl.pdf The excerpt is as follows "Patients enrolled in the clinical studies were required to have immunohistochemical evidence of positive EGFR expression using the DakoCytomation EGFR PharmDx test kit. Assessment for EGFR expression should be performed by laboratories with demonstrated proficiency in the specific technology being utilized. Improper assay performance, including use of suboptimally fixed tissue, failure to utilize specified reagents, deviation from specific assay instructions, and failure to include appropriate controls for assay validation, can lead to unreliable results. Refer to the DakoCytomation test kit package insert for full instructions on assay performance". Is this to be interpreted that only the Dako EGFR PharmDx kit is to be used for EGFR testing in patients who are to receive Erbitux, or can another EGFR clone be used as long as it has been validated for optimal use within the laboratory that is performing the testing? Or, is it acceptable to use another assay if it has been validated against the Dako EGFR Pharm Dx kit? Thanks, Patty Kubier ProPath Immunohistochemistry From Luis.Chiriboga <@t> med.nyu.edu Fri Mar 5 16:22:52 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Thanks & another question Message-ID: Hi everyone.... Thanks to all of you who responded to my article request.......& here's another question Does anyone know of a staining method that would demonstrate the limit dextrin, after amylase digestion in FFPE tissue..... Thanks WAY in advance.... Good Weekend to all.... Luis From bamur <@t> alaska.net Fri Mar 5 06:03:48 2004 From: bamur <@t> alaska.net (Barbara Murray) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Cryostats Message-ID: <000801c402aa$1fcf6d40$b68d70d1@d3b1l9> Greetings from Alaska! We are in the process of purchasing a new cryostat. I have information on three and they are: Vibratome 700 - open top cryostat, Leica 1850 and Sakura Cryo 3. I would like some feedback please if any of you have one of these cryostats and add to the list if there are more to choose from! This may have been posted before but I did not see it @ the time. Thanks so much for your input and have a great weekend! Barbara A. Murray,HT.(ASCP) The Alaska Native Medical Center Pathology/Histology Dept. Anchorage, Alaska (907-729-1804 From pathoprasad <@t> rediffmail.com Fri Mar 5 21:26:56 2004 From: pathoprasad <@t> rediffmail.com (prasad sbr prasad) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] AgNOR Message-ID: <20040306032656.24203.qmail@webmail18.rediffmail.com> Dear List, I shall be thankful to you if you send me the detaails of AgNOR staining and scoring methods. with regards, Prasad. Dr.CSBR.Prasad,MD., Al Hakeem Polyclinic, PO.BOX: 34985, Riyadh-11478, Saudi Arabia. ************************************************ ************************************************ From lpwenk <@t> covad.net Sat Mar 6 14:10:00 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] AFB Control References: Message-ID: <00ce01c403b7$01a67080$89584f44@domainnotset.invalid> Someone asked a similar question about 2 weeks ago, but about gram controls. I included in my response that the technique also works for AFB, fungus and spirochetes. I'm therefore reprinting my response from 2 weeks ago: = = = = = = = = = = Start with fresh, unfixed placenta or lung. - If lung, slightly edematous works best. - If placenta, press between towels several times, to try to get the excess RBCs out. Gross tissue into 2-3 mm cubes. Contact microbiology to let them know you are on your way (after previously talking with the supervisor days earlier about your needs). Have the microbiology prepare tubes of liquid incubating media (appropriate for the type(s) of micro-organism(s) you need). - Gram negative (E. coli works well) - Gram positive - AFB (non-pathogenic) - Fungus - Spirochetes (rarely is it ever syphilis. Usually some type of large spirochete, unfortunately. When large spirochetes are positive with the silver stains, the small syphilis are not being demonstrated yet.) (Helicobacter, as far as I know, cannot be grown in incubating media in routine microbiology labs.) Incubate tissue in culture media in 37 degree C. incubator overnight. Pour 10% NBF in tubes in morning, and allow to fix for 30-60 minutes. Pour out formalin/incubating media mixture, and pour in fresh NBF. Allow to fix all day. Place tissues in cassettes, label, process as usual. Embed. Write up cost containment report, on how you make X number of blocks of control tissues, which would equal Y number of slides. Which, if you had to buy them from an outside source would have cost you $Z amount of money. And the cost for you to do this procedures was a little bit of tech time, a few cassettes and some paraffin. Therefore, you just saved your facility lots of money! Document the collaboration between Histology and Microbiology. Everyone looks like a winner. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Poteete, Jacquie A." To: Sent: Friday, March 05, 2004 3:09 PM Subject: [Histonet] AFB Control > Hello all, > > Our Histology department has exhausted their supply of AFB control blocks. > I'm sure I saw a method of inoculating tissue to prepare this control, but I > can't find it. They would appreciate any help anyone can give them. > > Jacquie Poteete MT(ASCP)QIHC > Lead Technologist, IHC Laboratory > Saint Francis Hospital, Tulsa, OK > japoteete@saintfrancis.com > > > > > ********* Email Confidentiality Statement ********* > Visit http://www.saintfrancis.com/emailconf.asp > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From DDittus787 <@t> aol.com Sun Mar 7 15:09:32 2004 From: DDittus787 <@t> aol.com (DDittus787@aol.com) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! Message-ID: well,ronnie 2 reps were in my office telling me about reference labs using their kit and pathologists grading reactions, so either someone never told them, or they assumed,however they are using the dako kit. dana From paivi.parkkisenniemi <@t> lshp.fi Mon Mar 8 03:50:24 2004 From: paivi.parkkisenniemi <@t> lshp.fi (=?iso-8859-1?Q?Parkkisenniemi_P=E4ivi?=) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Softening of a fingernail Message-ID: <000001c404f2$c7fc8b70$0d14a8c0@patolvideolait> Hello, We have got a fingernail that we should soften before cutting. What would you recommen for softening method ? Few years ago we had a nail and I think that we used KOH, but I?m not sure because we dot?t have the recipe anymore. thank you in advance, P?ivi Parkkisenniemi Medical lab technologist Patologian osasto Lapin keskussairaala Rovaniemi FINLAND From DDDeltour <@t> sig.med.navy.mil Mon Mar 8 04:06:09 2004 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Softening of a fingernail Message-ID: I use warm soapy water. Sounds funny but it works. -----Original Message----- From: Parkkisenniemi P?ivi [mailto:paivi.parkkisenniemi@lshp.fi] Sent: Monday, March 08, 2004 10:50 AM To: 'histonetters' Subject: [Histonet] Softening of a fingernail Hello, We have got a fingernail that we should soften before cutting. What would you recommen for softening method ? Few years ago we had a nail and I think that we used KOH, but I?m not sure because we dot?t have the recipe anymore. thank you in advance, P?ivi Parkkisenniemi Medical lab technologist Patologian osasto Lapin keskussairaala Rovaniemi FINLAND _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon Mar 8 04:39:48 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Softening of a fingernail Message-ID: I think that a couple of percent phenol in aqueous solution softened nails, soak overnight. Dave Manchester UK -----Original Message----- From: Parkkisenniemi P?ivi [mailto:paivi.parkkisenniemi@lshp.fi] Sent: 08 March 2004 09:50 To: 'histonetters' Subject: [Histonet] Softening of a fingernail Hello, We have got a fingernail that we should soften before cutting. What would you recommen for softening method ? Few years ago we had a nail and I think that we used KOH, but I?m not sure because we dot?t have the recipe anymore. thank you in advance, P?ivi Parkkisenniemi Medical lab technologist Patologian osasto Lapin keskussairaala Rovaniemi FINLAND _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From benardsolomona <@t> yahoo.com Mon Mar 8 06:24:43 2004 From: benardsolomona <@t> yahoo.com (Solomon Adebayo) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Re(softening o Finger nails Message-ID: <20040308122443.75043.qmail@web20808.mail.yahoo.com> I want to suggest the use of 5% solution of phenol soaked overnight. The result will be considerable satisfactory. Benard Solomon University Of Ilorin Teaching Hosp. Kwara State Nigeria benardsolomona@yahoo.com www.histosearch.org/homepages/BenardSolomon __________________________________ Do you Yahoo!? Yahoo! Search - Find what you’re looking for faster http://search.yahoo.com From cdemarinis <@t> SARATOGACARE.ORG Mon Mar 8 06:12:24 2004 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis, Carolyn) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] softening toenails Message-ID: Putting the toenail in "Nair" hair remover solution works wonders. From StarkusL <@t> ummhc.org Mon Mar 8 06:53:44 2004 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Looking for a Mohs tech on Cape Cod in Massachusetts Message-ID: I am currently doing contract work for a Mohs surgeon on the Cape. He is looking for a full time Mohs tech. He's willing to train someone with histology experience but no Mohs experience. Please feel free to email me here for more information. From Nancy.Walker <@t> sanofi-synthelabo.com Mon Mar 8 08:48:54 2004 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] AP substrates Message-ID: Hello, Looking for the best (most sensitive) substrate for Alkaline Phoshatase. Heard great things about BCIP/TNBT but tried Chemicon's ready to use liquid, and saw nothing compared to good old BCIP/NBT. I see Abcam has also a ready to use BCIP/TNBT, and so does Moss Inc. Does anyone have any comments, or suggestions. thanks, Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From japoteete <@t> saintfrancis.com Mon Mar 8 08:40:58 2004 From: japoteete <@t> saintfrancis.com (Poteete, Jacquie A.) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Dako EGFR Kit Message-ID: Patty, our research people and Medical Oncologists will not accept our "other EGFR". When we did our verification, we had several specimens that tested positive with our other DAKO clone and did not test positive with the test kit clone. Most of the protocols we work with are extremely specific in regards to both the EGFRdx and Her-2 test KITS. Jacquie Poteete MT(ASCP)QIHC Lead Technologist, IHC Laboratory Saint Francis Hospital, Tulsa, OK japoteete@saintfrancis.com > -----Original Message----- > From: PKubier@propathlab.com [SMTP:PKubier@propathlab.com] > Sent: Friday, March 05, 2004 4:52 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Dako EGFR Kit > > The use of the DakoCytomation EGFR kit seems to be a hot topic currently, > and I'd like to have some feedback as to how to interpret the following > excerpt from the Erbitux package insert, obtained from the following link > through the FDA (see page 8 of this document) > > http://www.fda.gov/cder/foi/label/2004/125084lbl.pdf > > The excerpt is as follows "Patients enrolled in the clinical studies were > required to have immunohistochemical evidence of positive EGFR expression > using the DakoCytomation EGFR PharmDx test kit. Assessment for EGFR > expression should be performed by laboratories with demonstrated > proficiency > in the specific technology being utilized. Improper assay performance, > including use of suboptimally fixed tissue, failure to utilize specified > reagents, deviation from specific assay instructions, and failure to > include > appropriate controls for assay validation, can lead to unreliable results. > Refer to the DakoCytomation test kit package insert for full instructions > on > assay performance". > > Is this to be interpreted that only the Dako EGFR PharmDx kit is to be > used > for EGFR testing in patients who are to receive Erbitux, or can another > EGFR > clone be used as long as it has been validated for optimal use within the > laboratory that is performing the testing? Or, is it acceptable to use > another assay if it has been validated against the Dako EGFR Pharm Dx kit? > > Thanks, > > Patty Kubier > ProPath Immunohistochemistry > > > _______________________________________________ > 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 funderwood <@t> mcohio.org Mon Mar 8 09:37:34 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] softening toenails Message-ID: Fabric softner may work, also. >>> "Demarinis, Carolyn" 03/08/04 07:12AM >>> Putting the toenail in "Nair" hair remover solution works wonders. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kspencer <@t> utmem.edu Mon Mar 8 09:47:16 2004 From: kspencer <@t> utmem.edu (Kathleen A Spencer) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] croystat sectioning Message-ID: <101ee4c1022ebe.1022ebe101ee4c@utnet2.utmem.edu> I have a problem with cryostat sectioning that is driving me crazy. I hope someone out there can help because we have exhausted our brains coming up with solutions that have not helped. I am sectioning perfused rat brain on a cryostat, 20u thick, for IHC on floating sections. I have been doing this successfully for years. Now the problem is that they are curling up into tight little rolls in the PBS, which make them impossible to stain. They even curl as soon as they come off the knife edge, so I can't even put them on slides. We have: tried several different formulas for the fix, 3 different people doing the perfusion, 3 different kinds of PBS, warm PBS, cold PBS, all possible knife angles, different knives, different side of the knife, made the block colder, or warmer, thicker sections, thinner sections, cut fast, cut slow, NOTHING HELPS! When we cut fresh frozen brain to put on slides we do not have this problem. So, naturally we think it is the fix, or the method, or the person doing the perfusion. So, I had the expert do a perfusion and it still happened. At this point I just wanted to go out and get drunk and I am not a drinking person. Can anyone help? All advice is appreciated. Kathleen Spencer Lab Manager UTHSC From adelaur <@t> nimr.mrc.ac.uk Sun Mar 7 22:17:20 2004 From: adelaur <@t> nimr.mrc.ac.uk (April DeLaurier) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Anti-GFP antibodies for mouse wholemount Message-ID: Does anyone have any experience using anti-GFP antibodies on embryonic mouse wholemount specimens (up to E14.5)? Can anyone recommend any commercially available antibodies? Thanks, April -- April DeLaurier, PhD Division of Developmental Biology National Institute for Medical Research The Ridgeway, Mill Hill London NW7 1AA United Kingdom Tel: +44 208 959 3666 ext. 2095 Fax: +44 208 816 4477 E-mail: adelaur@nimr.mrc.ac.uk From kim_p_kowsz <@t> groton.pfizer.com Mon Mar 8 10:06:41 2004 From: kim_p_kowsz <@t> groton.pfizer.com (Kowsz, Kim P) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] muscle stain for image analysis Message-ID: Can someone please suggest a stain for skeletal muscle that will out-line each cross-sectional muscle fiber cell with a contrasting color. The goal is to allow a computer imaging system to easily identify each individual cell. Kim kowszkp@groton.pfizer.com LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. From MAUGER <@t> email.chop.edu Mon Mar 8 10:05:09 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] AP substrates Message-ID: Nancy, Dako has a BCIP/NBT/INT chromagen that looks like DAB. It is a dark brown color. I have had great results with it. Jo M >>> 03/08/04 09:48AM >>> Hello, Looking for the best (most sensitive) substrate for Alkaline Phoshatase. Heard great things about BCIP/TNBT but tried Chemicon's ready to use liquid, and saw nothing compared to good old BCIP/NBT. I see Abcam has also a ready to use BCIP/TNBT, and so does Moss Inc. Does anyone have any comments, or suggestions. thanks, Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179 fax :(33)561004001 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abright <@t> brightinstruments.com Mon Mar 8 10:33:44 2004 From: abright <@t> brightinstruments.com (Alan Bright) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] croystat sectioning Message-ID: Dear Kathleen, It seem like you have a static problem, do you have an anti-static brush or some other way of eliminating the static.? Best Regards Alan Bright Bright Instrument Co.Ltd. St Margaret's Way Huntingdon Cambridgeshire PE29 6EU England Tel No:+44 (0)1480 454528 Fax No:+44 (0)1480 456031 Email: abright@brightinstruments.com Web Site: www.brightinstruments.com -----Original Message----- From: Kathleen A Spencer [mailto:kspencer@utmem.edu] Sent: 08 March 2004 15:47 To: histonet@pathology.swmed.edu Subject: [Histonet] croystat sectioning I have a problem with cryostat sectioning that is driving me crazy. I hope someone out there can help because we have exhausted our brains coming up with solutions that have not helped. I am sectioning perfused rat brain on a cryostat, 20u thick, for IHC on floating sections. I have been doing this successfully for years. Now the problem is that they are curling up into tight little rolls in the PBS, which make them impossible to stain. They even curl as soon as they come off the knife edge, so I can't even put them on slides. We have: tried several different formulas for the fix, 3 different people doing the perfusion, 3 different kinds of PBS, warm PBS, cold PBS, all possible knife angles, different knives, different side of the knife, made the block colder, or warmer, thicker sections, thinner sections, cut fast, cut slow, NOTHING HELPS! When we cut fresh frozen brain to put on slides we do not have this problem. So, naturally we think it is the fix, or the method, or the person doing the perfusion. So, I had the expert do a perfusion and it still happened. At this point I just wanted to go out and get drunk and I am not a drinking person. Can anyone help? All advice is appreciated. Kathleen Spencer Lab Manager UTHSC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From David.Edmondson <@t> christie-tr.nwest.nhs.uk Mon Mar 8 10:45:12 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] croystat sectioning Message-ID: Kathleen Like the Bright man says, that works for stopping the section rolling up as it's cut, catch the leading edge as it comes off the blade and hold until the section is there in all its glory. You have been doing this for years. But do you mean that it is rolling up when places on the PBS? If so then all I have as an idea is to put them face down on the PBS. Rather than face-up Dave Manchester. UK -----Original Message----- From: Kathleen A Spencer [mailto:kspencer@utmem.edu] Sent: 08 March 2004 15:47 To: histonet@pathology.swmed.edu Subject: [Histonet] croystat sectioning I have a problem with cryostat sectioning that is driving me crazy. I hope someone out there can help because we have exhausted our brains coming up with solutions that have not helped. I am sectioning perfused rat brain on a cryostat, 20u thick, for IHC on floating sections. I have been doing this successfully for years. Now the problem is that they are curling up into tight little rolls in the PBS, which make them impossible to stain. They even curl as soon as they come off the knife edge, so I can't even put them on slides. We have: tried several different formulas for the fix, 3 different people doing the perfusion, 3 different kinds of PBS, warm PBS, cold PBS, all possible knife angles, different knives, different side of the knife, made the block colder, or warmer, thicker sections, thinner sections, cut fast, cut slow, NOTHING HELPS! When we cut fresh frozen brain to put on slides we do not have this problem. So, naturally we think it is the fix, or the method, or the person doing the perfusion. So, I had the expert do a perfusion and it still happened. At this point I just wanted to go out and get drunk and I am not a drinking person. Can anyone help? All advice is appreciated. Kathleen Spencer Lab Manager UTHSC _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Meera.Bansal <@t> chsli.org Mon Mar 8 10:59:31 2004 From: Meera.Bansal <@t> chsli.org (Bansal, Meera) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Shur/Mark cassette engraver Message-ID: <6D3CDB9369540244BFFB32DFEEB9360E745AEC@MMDCNT0BXVS002.chsli.org> We have a Shur/Mark 32 cassette engraver linked to our Copath v 2.2 anatomic pathology software. We have been using the Simport Histosette II casettes. Sometimes we find that the subdesignations for the cassettes do not fit correctly on the face of the cassette and part of it goes over the bottom edge. As a result, D1, O1 and Q1 all end up looking the same! Anyone else out there using the Shur/Mark? Which cassettes do you use? Thanks for your help. Meera Bansal,MD Mercy Medical Centre Rockville Centre, NY. Tel 516-705-2150 Fax 516-705-2691 From mcauliff <@t> umdnj.edu Mon Mar 8 13:49:47 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] muscle stain for image analysis In-Reply-To: References: Message-ID: <404CCE5B.4070400@umdnj.edu> PAS does a nice job of outlining smooth muscle cells in cross section, might be worth a try. Be advised that it stains the "cell coat" of the smooth muscle cell, not the membrane itself. Otherwise, immuno directed against a molecule unique to the cell membrane of skeletal muscle cells, assuming there is such a marker. Geoff Kowsz, Kim P wrote: >Can someone please suggest a stain for skeletal muscle that will out-line >each cross-sectional muscle fiber cell with a contrasting color. The goal >is to allow a computer imaging system to easily identify each individual >cell. > >Kim >kowszkp@groton.pfizer.com > > >LEGAL NOTICE >Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately. > >_______________________________________________ >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 Dave.Pizi <@t> carolinashealthcare.org Mon Mar 8 11:15:18 2004 From: Dave.Pizi <@t> carolinashealthcare.org (Pizi, Dave) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Shur/Mark cassette engraver Message-ID: <7E22BEDC45D51449B847E15983E99BAD721DDE@dcr-xchg-04.carolinas.org> Dr. Bansal We have 2 Shurmark cassette engravers and we have pretty good luck with Richard Allan cassettes. It sounds like the stop that the cassette rests against when they are being labeld needs adjusting. Also a new sytlus makes the numbers and letters crisp and easier to read. Dave Pizi Carolinas HealthCare System Charlotte, NC > -----Original Message----- > From: Bansal, Meera [SMTP:Meera.Bansal@chsli.org] > Sent: Monday, March 08, 2004 12:00 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Shur/Mark cassette engraver > > We have a Shur/Mark 32 cassette engraver linked to our Copath v 2.2 > anatomic pathology software. We have been using the Simport Histosette II > casettes. Sometimes we find that the subdesignations for the cassettes do > not fit correctly on the face of the cassette and part of it goes over the > bottom edge. As a result, D1, O1 and Q1 all end up looking the same! > Anyone else out there using the Shur/Mark? Which cassettes do you use? > Thanks for your help. > Meera Bansal,MD > Mercy Medical Centre > Rockville Centre, NY. > Tel 516-705-2150 > Fax 516-705-2691 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- This electronic message may contain information that is confidential and/or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you. From lange <@t> kennedykrieger.org Mon Mar 8 12:36:00 2004 From: lange <@t> kennedykrieger.org (Mollie Lange) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] re: cryostat sectioning Message-ID: Hi Kathleen, I feel your pain. I've been cutting cryostat sections for almost 30 years, and sometimes it can be completely maddening for no discernible reason. It could possibly be static as one Histonetter suggested. A Japanese fellow in our lab showed us that spraying Cytocool on the knife and roll plate really works. It's the ONLY thing that consistently works. Are your tissue blocks cryoprotected in sucrose? Have you tried a freshly made solution of your cryoprotectant in case the original was not made proplerly? We are a research lab, and we always cut our fixed frozen sections for immuno on a sliding microtome. We find this method much easier than the cryostat for fixed frozen. Do you have a sliding microtome available and have you tried it? Good luck, Mollie Lange From jcline <@t> wchsys.org Mon Mar 8 13:52:57 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] to barbara murray Message-ID: <005601c40546$f467a7e0$1d2a14ac@wchsys.org> I have two Leica 1850's and we are very satisfied with the model. From jcline <@t> wchsys.org Mon Mar 8 14:12:41 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Hello, Message-ID: <006101c40549$b5d05f60$1d2a14ac@wchsys.org> I have a Hacker H/I76 knife sharpener and redonditioner I would like to sell. It is in excellant condition. All accessories are there. Anyone interested or does it go the way of all the old fashioned histo ways. From kerney <@t> fas.harvard.edu Mon Mar 8 14:25:37 2004 From: kerney <@t> fas.harvard.edu (Ryan Kerney) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Leitz microtome servicing in Boston In-Reply-To: References: Message-ID: Hello everybody. Does anyone know of a good 3rd party shop that will service our Leitz rotary microtome in the Boston area (preferably but not required). It's a 20 year old Leitz 1512 that Leica won't touch. We've been having some problems with consistency in our cuts. The guide tracks may be sticking a little, although we've been diligent with oiling. Purchasing a new one is not an option. Thanks, Ryan From Sue.Kapoor <@t> uhsi.org Mon Mar 8 14:26:18 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] EGFR/Her1 Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F555B@khmcexch.uhsi.org> stupid me...I just found info explaining Her 1. ...funny how I've managed to go all these years without knowing this :~/ Sue Kapoor From Sue.Kapoor <@t> uhsi.org Mon Mar 8 14:18:17 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] EGFR/Her1 Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F555A@khmcexch.uhsi.org> About a month ago, I had a patient request a "Her 1", I couldn't find any reference to it...how did EGFR come about to being called "Her 1"?? Sue Kapoor, HT (ASCP) Histology Supervisor Kenosha Medical Center Kenosha, WI -----Original Message----- From: DDittus787@aol.com [mailto:DDittus787@aol.com] Sent: Friday, March 05, 2004 3:47 PM To: japoteete@saintfrancis.com; Ronnie_Houston@bshsi.com; ploykasek@phenopath.com; histonet@pathology.swmed.edu Subject: Re: [Histonet] FW: [IHCRG] Very important question!!!!!!!!!!!!!!! ! not to second guess you,but i have been told (2days ago) that labs are not reading results (kit or not) correctly and that this is not graded like her2 (even though it is called her1)so weak positivity is really positive not neg, so studies should be done carefully. dana _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Mon Mar 8 14:40:54 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Thanks-frozen section storage Message-ID: A hearty thanks to all for their suggestions on how to store frozens for future fat stains. FYI, I cut duplicates. Stored half in the fridge and half in the freezer, over the weekend. I went first with the ones stored in the fridge. They turned out fine. Thanks again, Fred From nena.dimaano <@t> stryker.com Mon Mar 8 15:47:47 2004 From: nena.dimaano <@t> stryker.com (Dimaano, Nena) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Glycerol Message-ID: <20BE9059B6FC2D4B9E8FF2A9E2C4EEBD13F26E@HOS2KEXCHCL.howost.strykercorp.com> Hello One investigator handed me a procedure for decalcification but before decalcification he mentioned something about washing the samples in 0.01 PBS containing increasing concentration of glycerol like (5%,10% and 15%). What is the purpose of increasing concentration of glycerol? thanks, Nena Dimaano, MT/HT(ASCP) Advanced Technology Stryker Orthopaedics Mahwah, NJ 07430 tel: 201-831-5338 fax: 201-831-6224 email: Nena.Dimaano@stryker.com From asuarez <@t> mrl.ubc.ca Mon Mar 8 16:45:44 2004 From: asuarez <@t> mrl.ubc.ca (Agripina Suarez) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] AP substrates Message-ID: Hi Nancy, My favorite substrate for alkaline phosphatase is Vector Red from Vector Laboratories. You can read your results in both brightfield and darkfield. When viewing under darkfield microscopy, use filters for rhodamine. Agripina Agripina C. Suarez McDonald Research Laboratories iCAPTURE Centre, St Paul's Hospital 1081 Burrard St, Vancouver B.C., Canada V6Z 1Y6 Tel no. 604 682 2344 local 62703 Fax no. 604 806 8351 >>> 03/08/04 6:48 AM >>> Hello, Looking for the best (most sensitive) substrate for Alkaline Phoshatase. Heard great things about BCIP/TNBT but tried Chemicon's ready to use liquid, and saw nothing compared to good old BCIP/NBT. I see Abcam has also a ready to use BCIP/TNBT, and so does Moss Inc. Does anyone have any comments, or suggestions. thanks, Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179 fax :(33)561004001 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rfail <@t> toolkitmail.com Tue Mar 9 08:38:54 2004 From: rfail <@t> toolkitmail.com (rfail) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] racemase and Muc-1 Message-ID: <404dd6fe.195.3a6c.1424166930@toolkitmail.com> Good morning, I have had a request to order these 2 antibodies. Is anyone using them and from what companies were they purchased? Rena Fail From ahorstma <@t> waldenu.edu Tue Mar 9 08:39:45 2004 From: ahorstma <@t> waldenu.edu (Anne M. Horstmann) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Job Opening Message-ID: <0HUB005WNCQ5IV@mta9.srv.hcvlny.cv.net> A busy Dermatopathology Laboratory on Long Island, NY has a full time (M-F) Histology Technician job opening from 6:00am to 3:00pm. We welcome all interested to fax (516) 883-2936 or e-mail resume. You must have at least an Associates Degree or higher for eligibility to sit for the HT/HTL exam. Students interested should apply because we are willing to train. Health benefits, paid vacations, and pension all available. Please forward this information on to anyone you know that is interested. Thank you, Anne From gcallis <@t> montana.edu Tue Mar 9 09:15:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Need an item for old AutoTechnicon processor Message-ID: <3.0.6.32.20040309081559.00bc56e8@gemini.msu.montana.edu> I have a gentleman who needs a blank clock/timer disk aka plate, the one that is cut to provide time changes during processing. I am not sure if IMEB has these or if anyone has one stashed in a drawer, please let me know. Thanks, Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jstaruk <@t> masshistology.com Tue Mar 9 09:30:56 2004 From: jstaruk <@t> masshistology.com (Mass Histology Service) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Need an item for old AutoTechnicon processor In-Reply-To: <3.0.6.32.20040309081559.00bc56e8@gemini.msu.montana.edu> Message-ID: We've got a few. Have him contact us. Jim ____________________ James Staruk, HT(ASCP) Mass Histology Service www.masshistology.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Tuesday, March 09, 2004 10:16 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Need an item for old AutoTechnicon processor I have a gentleman who needs a blank clock/timer disk aka plate, the one that is cut to provide time changes during processing. I am not sure if IMEB has these or if anyone has one stashed in a drawer, please let me know. Thanks, Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Mar 9 09:42:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Alkaline phosphatase chromogens or chromogens in general Message-ID: <3.0.6.32.20040309084256.00bc56e8@gemini.msu.montana.edu> Chris van der Loos, in his book on Multiple Staining and also his course for mutiple staining, has a wonderful chromogen chart that tells which chromogen is more sensitive than another. If the chromogen is less sensitive, i.e. Vector Red as compared to one of the fuchsin chromogens, then you must increase the concentration of your primary antibody to compensate. The chart is extremely useful when you are setting up IHC, particularly for double staining. This applies to both peroxidase and alkaline phosphatase immunostaining methods. NBT/BCIP is as sensitive as DAB. DAB+ from DAKO combined with DAB enhancer, can make a weakly stained antigen must more apparent and crisp. These two chromogens are on the more sensitive as compared to Vector red. We are able to do murine biotinylated CD4 diluted 1:15,000 and more (0.5mg/ml) with Strepavidin-HRP, followed by DAB+ and DAB enhancer. However when we do this antibody with SA-AP, followed by Vector Red the dilution of the CD4 falls to 1:250 or in that range. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From rlane <@t> ualberta.ca Tue Mar 9 10:12:21 2004 From: rlane <@t> ualberta.ca (rlane) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] source for paraaldehyde Message-ID: <404E7FB3@webmail.ualberta.ca> We can no longer purchase paraldehyde from our pharmacy for the making of Gomori's aldehyde fuchsin. We know of a source in Germany but am wondering if anyone knows of a source in Canada or USA? Thankyou Rosalind Lane Instructor Histotechnology From jbrod <@t> tvmdl.tamu.edu Tue Mar 9 10:08:16 2004 From: jbrod <@t> tvmdl.tamu.edu (Jordan Brod) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Speed Up Luxol Fast Blue Stain? Message-ID: I have a doctor who needs a LFB stain today and the procedure I use requires that slides be left in the Luxol fast blue overnight. Is there a way to speed that step up? Have a great day!!!! ******************************************************************** Jordan W. Brod, HTL (ASCP) Diagnostic Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) jbrod@tvmdl.tamu.edu From sa.drew <@t> hosp.wisc.edu Tue Mar 9 10:13:54 2004 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] racemase and Muc-1 Message-ID: We currently use racemase from BioCare Medical and are happy with it. It's being used on our Ventana instruments, and my co-worker has worked out a lovely dual stain of P504S and HWMCK... We've not done anything with Muc-1. -----Original Message----- From: rfail [mailto:rfail@toolkitmail.com] Sent: Tuesday, March 09, 2004 8:39 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] racemase and Muc-1 Good morning, I have had a request to order these 2 antibodies. Is anyone using them and from what companies were they purchased? Rena Fail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mcauliff <@t> umdnj.edu Tue Mar 9 13:30:29 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] source for paraaldehyde In-Reply-To: <404E7FB3@webmail.ualberta.ca> References: <404E7FB3@webmail.ualberta.ca> Message-ID: <404E1B55.4080506@umdnj.edu> You can substitute acetaldehyde but you need to use 3 times as much. See Buehner et al., J. Histochem. Cytochem. 27:782-787, 1979 for details. Geoff rlane wrote: >We can no longer purchase paraldehyde from our pharmacy for the making of >Gomori's aldehyde fuchsin. We know of a source in Germany but am wondering if >anyone knows of a source in Canada or USA? > >Thankyou >Rosalind Lane >Instructor >Histotechnology > > > >_______________________________________________ >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 joseph-galbraith <@t> uiowa.edu Tue Mar 9 10:55:11 2004 From: joseph-galbraith <@t> uiowa.edu (Galbraith, Joe) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Speed Up Luxol Fast Blue Stain? Message-ID: <5D03ED7B9391D4119D9B0008C76B7B2403005B19@uihc-mail1.uihc.uiowa.edu> Jordan: Modified Kluveer-Barrera Method for LFB Cut 5 micron sections, dry, bake at 60 degrees C for 30 minutes. Deparaffinize and hydrate to 95% ETOH (note not to DI H2O). Place slides in a dish with 400 ml of 0.1% LFB (diluted in 95% ETOH). Microwave on high power for 2 minutes (liquid should be near boiling) (Alternatively you can put slides in a coplin jar with 50 ml of LFB and microwave for 30-40 seconds.) Allow slides to cool in LFB at room temp for 20 minutes. Rinse slides briefly in 95% ETOH to remove excess stain. Rinse slides thoroughly in DI H2O. Differentiate slides in 0.1% Lithium Carbonate for 30 seconds. Three rinses to complete differentiation with 70% ETOH. Three rinses with DI H2O. Check microscopically for bright blue myelin fibers while vessel interiors are nearly colorless, RBC's should be colorless, gray matter should be almost colorless. Repeat differentiation steps (95% ETOH and Lithium Carbonate) if further differentiation is needed. Counterstain with routine H+E stain or Nuclear Fast Red. Good luck! Joe Galbraith -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jordan Brod Sent: Tuesday, March 09, 2004 10:08 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Speed Up Luxol Fast Blue Stain? I have a doctor who needs a LFB stain today and the procedure I use requires that slides be left in the Luxol fast blue overnight. Is there a way to speed that step up? Have a great day!!!! ******************************************************************** Jordan W. Brod, HTL (ASCP) Diagnostic Supervisor - Pathology Texas Veterinary Medical Diagnostic Laboratory (TVMDL) jbrod@tvmdl.tamu.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From siksik03 <@t> comcast.net Tue Mar 9 11:01:10 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Speed Up Luxol Fast Blue Stain? In-Reply-To: References: Message-ID: Dear HistoNetters Jorddan, if you have access to a laboratory microwave, you can preheat the Luxol fast blue solution to 60?C, then place your slides in the preheated solution and microwave at 60?C for 2 hours. Use stirring or agitation. best regards, Steven Slap Microwave Consultant At 10:08 AM -0600 3/9/04, Jordan Brod wrote: >I have a doctor who needs a LFB stain today and >the procedure I use requires that slides be left >in the Luxol fast blue overnight. Is there a >way to speed that step up? > >Have a great day!!!! > >******************************************************************** >Jordan W. Brod, HTL (ASCP) >Diagnostic Supervisor - Pathology >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) >jbrod@tvmdl.tamu.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Mar 9 11:33:59 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] source for paraaldehyde In-Reply-To: <404E7FB3@webmail.ualberta.ca> Message-ID: <3.0.6.32.20040309103359.00bbfc28@gemini.msu.montana.edu> Because paraldehyde is a controlled substance, you can substitute acetaldehyde. Peggy Wenk did this and may respond to your inquiry. The equivalent is 2.5 ml acetaldehyde instead of 1.5 ml paraldehyde. Peggy had a Letter to Editor in J of Histotechnology, Dec 1996 on page 353 discussing this. Acetaldehyde is not controlled nor it is very expensive. Any reliable chemical distributor should have it. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Tue Mar 9 11:49:06 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Faster luxol fast blue In-Reply-To: Message-ID: <3.0.6.32.20040309104906.00bbfc28@gemini.msu.montana.edu> Go to Sakura Finetek Histologic website, and search for Diane Sterchi's Modified Kluver-Barrera for Melin and Nerve Cells, unfortunately, I do not have the date nor issue. Sections at 10 um thick. She did it in 3 hours following a preheating of Luxol fast blue to 60C, stain for 3 hrs in this hot LFB, rinse in 95% ETOH to remove excess stain, rinse in distilled water, begin differentiation in 0.05% lithium carbonate, continue this step in 70% ethanol, until gray and white mater are distinguished. This step is FAST! rinse well in distilled water, counterstain. Good luck. At 10:08 AM 3/9/2004 -0600, you wrote: >I have a doctor who needs a LFB stain today and the procedure I use requires that slides be left in the Luxol fast blue overnight. Is there a way to speed that step up? > >Have a great day!!!! > >******************************************************************** >Jordan W. Brod, HTL (ASCP) >Diagnostic Supervisor - Pathology >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) >jbrod@tvmdl.tamu.edu > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From AFeatherstone <@t> KaleidaHealth.Org Tue Mar 9 11:53:49 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] FAST LFB Message-ID: We routinely stain our LFB's for 2 hours in a 60 degree oven in Luxol Fast Blue and then do the usual differentiation steps. Always works beautifully. Annette Featherstone HT/MLT Kaleida Health Neuropathology 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 portera203 <@t> yahoo.com Tue Mar 9 12:06:10 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] source for paraaldehyde In-Reply-To: <404E7FB3@webmail.ualberta.ca> Message-ID: <20040309180610.48946.qmail@web40913.mail.yahoo.com> We just purchased some from Sigma Chemical - had to have DEA # for controlled substance. rlane wrote:We can no longer purchase paraldehyde from our pharmacy for the making of Gomori's aldehyde fuchsin. We know of a source in Germany but am wondering if anyone knows of a source in Canada or USA? Thankyou Rosalind Lane Instructor Histotechnology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com --------------------------------- Do you Yahoo!? Yahoo! Finance: Get your refund fast by filing online From jo-ann <@t> lan1.molonc.mcgill.ca Tue Mar 9 13:08:57 2004 From: jo-ann <@t> lan1.molonc.mcgill.ca (jo-ann) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Xylene substitute Message-ID: Hi to all, I am setting up a small histology lab, but since the renovations will probably take awhile I still would like to set up my tissue processor and stainer anyway. There is no hood or ventilation canopy in the room at the moment so0 xylene fume will be a major problem. I would appreciate advice from anyone on what xylene substitute is the all around best; taking into consideration the immunohistochemistry may be done in the future. Thanks in advance. Jo-Ann From stancelb <@t> msn.com Tue Mar 9 14:09:51 2004 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Faster luxol fast blue Message-ID: We preheat 45 ml of LFB for 20 seconds at full power in our microwave, place the slides in the stain and place in a 68C oven for 30 minutes. The rest of the stain is as printed in most manuals. Always works. Pathologists happy. Histologically yours, Barbara Eastern Laboratory, Pathology Athens, Georgia 30604 >From: Gayle Callis >To: "Jordan Brod" , Histonet@lists.utsouthwestern.edu >Subject: [Histonet] Faster luxol fast blue >Date: Tue, 09 Mar 2004 10:49:06 -0700 > >Go to Sakura Finetek Histologic website, and search for Diane Sterchi's >Modified Kluver-Barrera for Melin and Nerve Cells, unfortunately, I do not >have the date nor issue. Sections at 10 um thick. > >She did it in 3 hours following a preheating of Luxol fast blue to 60C, >stain for 3 hrs in this hot LFB, rinse in 95% ETOH to remove excess stain, >rinse in distilled water, begin differentiation in 0.05% lithium carbonate, >continue this step in 70% ethanol, until gray and white mater are >distinguished. This step is FAST! rinse well in distilled water, >counterstain. > >Good luck. > >At 10:08 AM 3/9/2004 -0600, you wrote: > >I have a doctor who needs a LFB stain today and the procedure I use >requires that slides be left in the Luxol fast blue overnight. Is there a >way to speed that step up? > > > >Have a great day!!!! > > > >******************************************************************** > >Jordan W. Brod, HTL (ASCP) > >Diagnostic Supervisor - Pathology > >Texas Veterinary Medical Diagnostic Laboratory (TVMDL) > >jbrod@tvmdl.tamu.edu > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > > > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Get a FREE online computer virus scan from McAfee when you click here. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From Deborah.Wallin <@t> WTH.org Tue Mar 9 14:20:45 2004 From: Deborah.Wallin <@t> WTH.org (Wallin, Deborah) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Steiner for Lyme disease Message-ID: I just wanted to clarify a stain. It is recommended to do a Warthin Starry for Lyme disease. Can we perform a Steiner instead? Thank you, D. Wallin From gcallis <@t> montana.edu Tue Mar 9 14:22:23 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Xylene substitute In-Reply-To: Message-ID: <3.0.6.32.20040309132223.00bd8ff0@gemini.msu.montana.edu> We use Richard Allan Clearite 3 without problems, even for IHC. Gayle Callis Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 At 02:08 PM 3/9/2004 -0500, you wrote: >Hi to all, > >I am setting up a small histology lab, but since the renovations will >probably take awhile I still would like to set up my tissue processor and >stainer anyway. There is no hood or ventilation canopy in the room at the >moment so0 xylene fume will be a major problem. I would appreciate advice >from anyone on what xylene substitute is the all around best; taking into >consideration the immunohistochemistry may be done in the future. > >Thanks in advance. > >Jo-Ann > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From SJones <@t> cvm.tamu.edu Tue Mar 9 14:56:45 2004 From: SJones <@t> cvm.tamu.edu (Sarah Jones) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Steiner for Lyme disease Message-ID: Hi Deborah, The Steiners can be done for Lyme disease as well. Sarah Jones HT(ASCP) Dept. of Vet. Anatomy & Public Health Histology Lab Texas A&M University College Station, TX 77843-4458 phone: 979-845-3177 fax: 979-458-3499 >>> "Wallin, Deborah" 3/9/2004 2:20:45 PM >>> I just wanted to clarify a stain. It is recommended to do a Warthin Starry for Lyme disease. Can we perform a Steiner instead? Thank you, D. Wallin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Tue Mar 9 15:10:54 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:39 2005 Subject: Fwd: [Histonet] re: cryostat sectioning Message-ID: <6.0.1.1.0.20040309150912.02597870@mailhost.vetmed.auburn.edu> Please pardon my ignorance, but what kind of frozen tissues are you sectioning on the sliding microtome, what brand & type sliding microtome are you using, and how are you keeping your samples frozen for sectioning on it? Thanks, Atoska >X-Mailer: Novell GroupWise Internet Agent 6.5.1 >Date: Mon, 08 Mar 2004 13:36:00 -0500 >From: "Mollie Lange" >To: >X-Scan-Signature: 6e25c54ecfa6545cca04a3e1c9f5b689 >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >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: 05628b3a735271e1629bc35bb8abda07 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] re: cryostat sectioning >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=6.5 tests=none autolearn=no version=2.63 >X-SA-Exim-Version: 3.1 (built Tue Oct 14 16:21:02 CDT 2003) >X-SA-Exim-Scanned: Yes > >Hi Kathleen, > >I feel your pain. I've been cutting cryostat sections for almost 30 >years, and sometimes it can be completely maddening for no discernible >reason. > >It could possibly be static as one Histonetter suggested. A Japanese >fellow in our lab showed us that spraying Cytocool on the knife and roll >plate really works. It's the ONLY thing that consistently works. > >Are your tissue blocks cryoprotected in sucrose? Have you tried a >freshly made solution of your cryoprotectant in case the original was >not made proplerly? > >We are a research lab, and we always cut our fixed frozen sections for >immuno on a sliding microtome. We find this method much easier than the >cryostat for fixed frozen. Do you have a sliding microtome available and >have you tried it? > >Good luck, >Mollie Lange >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From gentras <@t> vetmed.auburn.edu Tue Mar 9 15:35:31 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:39 2005 Subject: Fwd: [Histonet] Need an item for old Autotechnicon processor Message-ID: <6.0.1.1.0.20040309152859.025982e0@mailhost.vetmed.auburn.edu> G & G Instrument Corp. of Ardsley, NY probably has one for sale if you're unable to get one donated. Their PH # is: 1-800-882-2288. Best wishes, Atoska >X-Sender: gcallis@gemini.msu.montana.edu >X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) >Date: Tue, 09 Mar 2004 08:15:59 -0700 >To: Histonet@lists.utsouthwestern.edu >From: Gayle Callis >X-Scan-Signature: 6e3e8a9b2a8472a697c9c9c003db8af6 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >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: ff0f5448b767a158ef77d41af1533689 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Need an item for old AutoTechnicon processor >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=6.5 tests=none autolearn=no version=2.63 >X-SA-Exim-Version: 3.1 (built Tue Oct 14 16:21:02 CDT 2003) >X-SA-Exim-Scanned: Yes > >I have a gentleman who needs a blank clock/timer disk aka plate, the one >that is cut to provide time changes during processing. I am not sure if >IMEB has these or if anyone has one stashed in a drawer, please let me know. > >Thanks, > > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From Pathologyarts <@t> aol.com Tue Mar 9 16:47:09 2004 From: Pathologyarts <@t> aol.com (Pathologyarts@aol.com) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Distilled Water Message-ID: <197.26a05821.2d7fa36d@aol.com> Does anyone know where I might be able to purchase distilled water for routine histo work? We don't have a distiller really don't want to purchase one. Thank you in advance. From asmith <@t> mail.barry.edu Tue Mar 9 17:49:40 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Distilled Water Message-ID: <494304423C63E246A5CF87A3AEEB577011EE0F@bumail01.barrynet.barry.edu> Most supermarkets sell gallon bottles of distilled water. It is usually right next to the bottled spring water. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pathologyarts@aol.com Sent: Tuesday, March 09, 2004 5:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Distilled Water Does anyone know where I might be able to purchase distilled water for routine histo work? We don't have a distiller really don't want to purchase one. Thank you in advance. _______________________________________________ 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 CrochiereSteve <@t> aol.com Tue Mar 9 20:20:13 2004 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Xylene substitute Message-ID: <104.41047024.2d7fd55d@aol.com> We use Shandon Histosolve. It works well and is easy to recycle with the CBG recycler. Steven M. Crochiere, HT (ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew St. Springfield, MA 01104 From doscwk <@t> nus.edu.sg Tue Mar 9 20:46:02 2004 From: doscwk <@t> nus.edu.sg (Chan Wai Kam) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Toluene substitute Message-ID: Hi Histonetters, Our lab uses toluene in the tissue processor and as we are unable to have an overhead hood for this equipment, I'm wondering if there is any available toluene substitute we could use for specimen clearing. Another problem with leaving the solutions (alcohol and solvent) in the processor(carousel type) is the rapid rate of evaporation from the buckets. Would appreciate any advice. Thanks Julee Chan Musculoskeletal Tissue Lab Orthopaedic Surgery National University of Singapore From DonnaWillis <@t> texashealth.org Tue Mar 9 21:17:18 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Texas Society for Histotechnology Convention Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B201A32F91@ftwex01.txhealth.org> TSH will be holding it's annual convention April 15-18, 2004 at the Holiday Inn Park Plaza in Lubbock, Tx. Workshops to be presented are as follows: Thursday April 15, 2004 Golf Tournament Friday April 16, 2004 House of Delegate Meeting and President's Reception Saturday April 17, 2004 AM Workshops Workshop #1 "HT(ASCP) Examination Readiness Workshop- Part 1", Glenda Hoye speaker Workshop #2 "The Continuum from Antibodies to Personalized Medicine", Mary Cheles speaker Workshop #3 "Preparing for a CAP Inspection", Hector Hernandez Jr and Joe Nocito speakers Workshop #4 "Microwave: The Whole Enchilada", Jan Minshew and Donna Willis speakers Symposium A "Tissue Transfer and Tissue Protection Techniques", Patty Kubier speaker Symposium B "Technical Approaches to Research Histology", Jeffery Stark speaker PM Workshops Workshop #5 "HT(ASCP) Examination Readiness Workshop- Part 2", Glenda Hoye Speaker Workshop #6 "The Effects of Fixation and Epitope Retrieval on Immunohistochemical Staining of Paraffin Sections", Jerry Fredenburgh and Russell Myers speakers Workshop #7 "Microtomy: The Melding of Art and Science", Jan Minshew speaker Workshop #8 "Achieving Excellence with Routine H&E Staining and Immunochemical Staining of Bone Specimens and Bone Marrow Biopsies Following Decalcification", James Biesecker speaker Symposium C "Let Alcohol Clear your Brain" Pamela Marcum speaker Symposium D "Usining Your Tissue Processor as a Problem-Solver" Pamela Marcum speaker Saturday night Casino Bash Sunday April 18, 2004 AM Workshops Workshop #9 "MonKEY Business-the KEY to Delegation", Jan Gardner and Judi Stasko speakers Workshop #10 "New Method and Techniques for Frozen Section", Stephen Peters speaker Workshop #11 "Mystery Theater II Troubleshooting Special Stains", Joan Vesey speaker Workshop #12 " Immunohistochemistry 2004", Ethel Macrea speaker Symposium E "Cancer: A Collection of Diseases" Hazel Dalton, Gabriel Ayala, Patricia Ammon, Manuel Garcia, Shashi Reddy, Olga Rodriquez speakers Symposium F "Special Histologic Techniques", Hazel Dalton, Martin Dominquez, Elizabeth Pancake, Victoria Socin speakers For further information you can contact Donna Willis at 817-878-5644 or Sharon Whitley at 210-237-6488. Thanks, Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From Stanley.Stylli <@t> mh.org.au Tue Mar 9 23:28:25 2004 From: Stanley.Stylli <@t> mh.org.au (Stylli, Stanley) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] a search for FFPE rat compatible antibodies Message-ID: <00519F9F41D2844D9C75BEE1F6AC09AF0127BF2F@rmhmail1.ssg.org.au> Dear All, I am after primary antibodies to use on rat FFPE (especially in brain) for the following TNF-alpha Tenascin Capsase-3 PECAM-1 I would preferably like an antibody that doesnt need antigen retrieval but am willing to try a tried and true antigen retieval method if the antibody exists. thanks for your help Stan From jqb7 <@t> cdc.gov Wed Mar 10 05:45:00 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Steiner for Lyme disease Message-ID: Yes, the Mod. Steiner is the stain we use. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Wallin, Deborah Sent: Tuesday, March 09, 2004 3:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Steiner for Lyme disease I just wanted to clarify a stain. It is recommended to do a Warthin Starry for Lyme disease. Can we perform a Steiner instead? Thank you, D. Wallin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Wed Mar 10 06:33:49 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Apoptosis stain Message-ID: <20040310123349.18717.qmail@web10004.mail.yahoo.com> Hi, I need to do staining for apoptosis on FFPE tissue (mouse and rat). Can anyone recommend a good kit? Thanks in advance, Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Search - Find what you’re looking for faster. From histo20 <@t> hotmail.com Wed Mar 10 07:56:35 2004 From: histo20 <@t> hotmail.com (Paula Wilder) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] (no subject) Message-ID: Hi Histonetters - quick question from Cytology - does anyone have a procedure for handling possible CJD spinal fluids in the Cytology Department? Any help would be greatly appreciated!!! Paula Wilder St.Joseph Medical Center Towson, MD 21204 _________________________________________________________________ Store more e-mails with MSN Hotmail Extra Storage – 4 plans to choose from! http://click.atdmt.com/AVE/go/onm00200362ave/direct/01/ From kmerriam2003 <@t> yahoo.com Wed Mar 10 08:59:03 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Apoptosis stain In-Reply-To: Message-ID: <20040310145903.18756.qmail@web10002.mail.yahoo.com> I already have IHC for caspase-3, but the investigator wants something more. I am not familiar with apoptosis, so this is all new to me. Kim Luis Chiriboga wrote: cell signaling caspase (rabbit), works in ffpe mouse and rat with standard biotinylated secondary & SA-HRP detection Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kim Merriam Sent: Wednesday, March 10, 2004 7:34 AM To: Histonet Subject: [Histonet] Apoptosis stain Hi, I need to do staining for apoptosis on FFPE tissue (mouse and rat). Can anyone recommend a good kit? Thanks in advance, Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Search - Find what youre looking for faster. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Search - Find what you’re looking for faster. From stancelb <@t> msn.com Wed Mar 10 09:21:42 2004 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] old equipment manuals Message-ID: Dear Histonetters, While cleaning out the back of my filing cabinet, I came across some old equipment manuals. The equipment is looooong gone, but the packrat in me has been keeping the manuals….for nostalgia’s sake. But I just can’t keep everything. If any one would like these old manuals, please contact me off the HISTONET. First come. First serve. 1 Minotome (microtome-cryostat) Installation, operation and service manual. Damon/IEC 1975 2.Leitz Rotary Microtomes and Cryostat parts list 3. International Model CTD Microtome-Cryostat IEC 4. Autotechnicon MONO/DUO illustrated parts breakdown 5. Autotechnicon MONO/DUO combination manual for the operation of both Technical publication No. TA1-0225-00 Sept 1971 6. Autotechnicon Ultra II System Hardback manual 1978 Please don’t make me trash them. Histologically yours, Barbara _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ From gcallis <@t> montana.edu Wed Mar 10 09:41:49 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] distilled water Message-ID: <3.0.6.32.20040310084149.00bc3b10@gemini.msu.montana.edu> I work with a young lady who worked with water treatment. She indicated you can buy under the sink attachments on your water supply that are Reverse Osmosis devices (RO) water for very little, even from WalMart or another discount store. This will give you excellent water for histo purposes. How about going to the local grocery store/Target/drug store and buy distilled water from them - you would not have to pay shipping costs, the weight alone is on the heavy side and shipping isn't cheap anymore. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Deborah.Wallin <@t> WTH.org Wed Mar 10 10:03:49 2004 From: Deborah.Wallin <@t> WTH.org (Wallin, Deborah) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Steiner for Lyme disease Message-ID: Thanks so much for the information. I was sure that I could use the Steiner but had not done any testing for Lyme's disease in so long I thought that I would check. Thanks Again, Deborah -----Original Message----- From: Bartlett, Jeanine [mailto:jqb7@cdc.gov] Sent: Wednesday, March 10, 2004 5:45 AM To: Wallin, Deborah; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Steiner for Lyme disease Yes, the Mod. Steiner is the stain we use. Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Wallin, Deborah Sent: Tuesday, March 09, 2004 3:21 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Steiner for Lyme disease I just wanted to clarify a stain. It is recommended to do a Warthin Starry for Lyme disease. Can we perform a Steiner instead? Thank you, D. Wallin _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Bauer.Karen <@t> mayo.edu Wed Mar 10 10:30:12 2004 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Monoclonal Calretinin Message-ID: <8C6E05FA69571948B461F1327CBB893E0629F00C@lmmail2.ad.lmmhs.org> Does anyone make a monoclonal Calretinin? Karen Bauer Histology Supervisor Department of Pathology Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. From juan.gutierrez <@t> christushealth.org Wed Mar 10 10:49:14 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Monoclonal Calretinin Message-ID: Accurate Chemical has one and USBiological has two. Accurate 1-800-645-6264 USBiological 1-800-520-3011 I have not used any of them, but they claim to work on ffpe tissue. Good luck, Juan -----Original Message----- From: Bauer, Karen [mailto:Bauer.Karen@mayo.edu] Sent: Wed 3/10/2004 10:30 AM To: Histonet (E-mail) Cc: Subject: [Histonet] Monoclonal Calretinin Does anyone make a monoclonal Calretinin? Karen Bauer Histology Supervisor Department of Pathology Luther Hospital 715-838-3205 ********************Confidentiality Notice******************** This message is intended for the sole use of the individual and entity to whom it is addressed, and may contain information that is privileged, confidential and exempt from disclosure under applicable law. Any unauthorized review, use, disclosure or distribution of this email message, including any attachment, is prohibited. If you are not the intended recipient, please advise the sender by reply email and destroy all copies of the original message. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mark.lewis <@t> thermo.com Wed Mar 10 11:07:56 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Processing "plastics" / resins Message-ID: Good morning everyone ! Excuse my ignorance on this issue,but I have never processed or stained tissues with resins such as Glycolmethacrylate or Methylmethacrylate. Do you normally run the tissues through a clearing agent for processing and staining? I didn't think you needed to except for the end of the staining process. If you do use a clearant in any step of the process, can you use a Xylene substitute like Shandon Xylene substitute ? Thanks in advance for your replies ! Mark From nmuvarak <@t> facstaff.wisc.edu Wed Mar 10 11:11:11 2004 From: nmuvarak <@t> facstaff.wisc.edu (NIDAL E MUVARAK) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Tissue Damage Marker??? Message-ID: <4ba7f4a734.4a7344ba7f@wiscmail.wisc.edu> Hello everyone. My PI asked me to look for a marker that would show if there's any damage in the tissue. Basically, we're using this ventilation-perfustion system for mice lungs. I've section and stained them hematoxylin, and didn't see any damage... well, at least I couldn't tell. So is there such a thing out there that would mark tissue damage, edema, and such? Thank you. Nidal E Muvarak Associate Research Specialist Vascular Tissue Biomechanics Laboratory Department of Biomedical Engineering University of Wisconsin-Madison 1550 Engineering Dr.; Rm. 2158 Madison, WI 53706-1609 Lab: (608) 265-8921; Office: (608) 265-4205; Home: (608) 256-7934; Cell: (608) 332-6068 http://vtb.bme.wisc.edu From dingj <@t> nicemice.cn Wed Mar 10 11:33:02 2004 From: dingj <@t> nicemice.cn (dingjun) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Use Glycol methacrylate to embed mouse eye Message-ID: <006e01c406c5$bd6e4160$ef64a8c0@djxmg> SGkgRmVsbG93cw0KSGF2ZSBhbnlvbmUgdXNlZCBHTUEgYXMgdGhlIGVtYmVkZGluZyBtZWRpdW0g Zm9yIG1vdXNlIGV5ZSBzZWN0aW9uPw0KSGVyZSBpcyBteSBwcmVzY3JpcHRpb246DQpBIHNvbHV0 aW9uIDogR01BIDk1bWwNCiAgICAgICAgICAgICBQRUctNDAwIDhtbA0KICAgICAgICAgICAgIEJl bnpveWwgcGVyb3hpZGUgMC43Zw0KQiBzb2x1dGlvbiA6IFBFRy00MDAgMjBtbA0KICAgICAgICAg ICAgIE4sTiBkaW1ldGh5bGFuaWxpbmUgMW1sDQpXaGVuIHBvbHltZXJpemF0aW9uICBtaXggQSB3 aXRoIEIgYXQgMTAwOjMNCg0KQ2FuIHNvbWVvbmUgZ2l2ZSBtZSBzb21lIGRpcmVjdGlvbj8NCg0K VGhhbmtzIGluIGFkdmFuY2UuDQoNCkNoZWVycw0KDQpKdW4gDQpNb2RlbCBBbmltYWwgUmVzZWFy Y2ggQ2VudGVyKE1BUkMpDQpOYW5qaW5nIFVuaXZlcnNpdHkNCk5hbkppbmcgUC5SLkNoaW5hDQpQ b3N0YWwgQ29kZTogMjEwMDkzDQpQaG9uZTogODYtMjUtODY0MTUxMQ0KRmF4OiA4Ni0yNS04NjQx NTAwDQpodHRwOi8vd3d3Lm5pY2VtaWNlLmNuL2VuX2luZGV4Lmh0bQ0KDQogDQo= From cmather <@t> origentherapeutics.com Wed Mar 10 11:47:00 2004 From: cmather <@t> origentherapeutics.com (Christine Mather) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] PAS on 24 hour chicken embryo Message-ID: Has anyone any advice on how to avoid non-specific staining of PAS in 24 hour chicken embryos? The yolky tissue seems to be very PAS positive. We use whole mounts or 10um cryosections of 4% para fixed whole embryos, then 0.5% PAS for 5 mins at RT, 2 mins washes in d water, Schiff's for 5 mins at 4C, 2x2mins rinses in H20/sodium metabisulfite/1N HCL, 10 mins wash in cool tap water. From azdudley <@t> hotmail.com Wed Mar 10 12:00:56 2004 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Xylene substitute Message-ID: jo-ann, we use richard allans clear-rite 3 with good results, even with our ihc. we also recycle iit. anita dudley providence hosp. mobile ala >From: jo-ann >To: >Subject: [Histonet] Xylene substitute >Date: Tue, 09 Mar 2004 14:08:57 -0500 > >Hi to all, > >I am setting up a small histology lab, but since the renovations will >probably take awhile I still would like to set up my tissue processor and >stainer anyway. There is no hood or ventilation canopy in the room at the >moment so0 xylene fume will be a major problem. I would appreciate advice >from anyone on what xylene substitute is the all around best; taking into >consideration the immunohistochemistry may be done in the future. > >Thanks in advance. > >Jo-Ann > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Frustrated with dial-up? Lightning-fast Internet access for as low as $29.95/month. http://click.atdmt.com/AVE/go/onm00200360ave/direct/01/ From jryan <@t> sleh.com Wed Mar 10 12:04:36 2004 From: jryan <@t> sleh.com (John Ryan) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] I'm sorry I can not reply immediately because I am out of the office, returning on Monday March 15, Message-ID: I'm sorry I can not reply immediately because I am out of the office, returning on Monday March 15, 2004. +++++CONFIDENTIALITY NOTICE+++++ The information in this e-mail may be confidential and/or privileged. If you are not the intended recipient or an authorized representative of the intended recipient, you are hereby notified that any review, dissemination or copying of this e-mail and its attachments, if any, or the information contained herein is prohibited. If you have received this e-mail in error, please immediately notify the sender by return e-mail and delete this e-mail from your computer system. Thank you. From katherine-walters <@t> uiowa.edu Wed Mar 10 12:03:58 2004 From: katherine-walters <@t> uiowa.edu (katherine-walters@uiowa.edu) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Protein A or G?? Message-ID: <1078941838.404f588e434a5@webmail3.its.uiowa.edu> Hi everyone, Does anyone know of a reference that states which animal sources of primary antibody works with Protein A and which animal sources of antibody works with protein G. I remember having read it back in the day, but now can't seem to lay my hands on it. Thanks for your help, Kathy From jo-ann <@t> lan1.molonc.mcgill.ca Wed Mar 10 12:40:45 2004 From: jo-ann <@t> lan1.molonc.mcgill.ca (jo-ann) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Thank you Message-ID: Thank you to everyone who has sent information on xylene substitutes. I have downloaded the product information and will be going through it all with our Health and Safety Officer. I am sure that my problem is solved. Thanks again. Jo-Ann Bader -- Director's Assistant Molecular Oncology Group McGill University Health Centre 687 Pine Ave. W. Room H5-21 Montreal, QC, Canada, H3A-1A1 Tel: 514-843-1479 Fax; 514-843-1478 E-mail: jo-ann@molonc.mcgill.ca From cgin <@t> pen.eiu.edu Wed Mar 10 14:52:47 2004 From: cgin <@t> pen.eiu.edu (ikenwosu) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Xylene substitute Message-ID: <404f801f.208.1c60.4573@pen.eiu.edu> hi Gayle, I use hito-clear from national diagnostics. order# hs-200. Ike Thedore Nwosu Grad. Student Dept. of Biological Sciences Eastern Illinois University 600 Linclon Ave Charleston, IL 61938 From caroline.stott <@t> anatomy.otago.ac.nz Wed Mar 10 16:13:06 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Processing "plastics" / resins Message-ID: <5.2.1.1.0.20040311110509.02a28490@anatomy.otago.ac.nz> Hi Mark, Heres what we do: After fixation place into 70% a few hrs 95% alc 10 min-1 hr Absolute alc 10 min-1hr Glycolmethacrylate (GMA) 1st change 30 mins-4 hrs GMA 2 30 mins-overnight GMA 3 30 mins-24 hrs GMA 4 30 mins-24 hrs Prepare new monomer 88ml 2-hydroxyethylmethacrylate 12ml 2 butoxyethanol 0.45g Benzoyl peroxide Then add promoter to monomer as 1:50. 8 ml polyethylene glycol 400 1 ml NN Dimethylaniline There is no clearing necessary. You can dip straight into your dye with out hydrating the slides. Hope that helps Caroline Good morning everyone ! Excuse my ignorance on this issue,but I have never processed or stained tissues with resins such as Glycolmethacrylate or Methylmethacrylate. Do you normally run the tissues through a clearing agent for processing and staining? I didn't think you needed to except for the end of the staining process. If you do use a clearant in any step of the process, can you use a Xylene substitute like Shandon Xylene substitute ? Thanks in advance for your replies ! Mark Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From LuckG <@t> empirehealth.org Wed Mar 10 16:34:50 2004 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] "High Iron Diamine" stain Message-ID: Hello, I'm looking for some assistance with doing a procedure called a "High Iron Diamine" stain. I do not have a written procedure and have never done one myself. Any help would be greatly appreciated. Thanks in advance, Greg Greg Luck Anatomic Pathology Supervisor Deaconess Medical Center 800 W. 5th Ave Spokane, WA 99204 509.473.7077 luckg@empirehealth.org From KHays <@t> mbhs.org Wed Mar 10 16:33:28 2004 From: KHays <@t> mbhs.org (KHays@mbhs.org) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] to do or not to do egfr Message-ID: Kathy Tedford-Hays HT (ASCP) Technical Specialist, Histology Dept (601)-968-3070 ext 7398 Baptist Medical Center 1225 North State Street Jackson, MS 39202 we are currently getting several requests for the test egfr as many of you. my questions are: is this another immunostain or is this something more? we were told that if the patients are positive then they are candidates for a new chemotherapy for colon cancer. if this is not another immunostain then what labs are doing the egfr for this new chemo? any information would be appreciated. thank you. The information contained in this email is confidential and is intended for the recipient only. Do not forward this email without permission from its originator. From IKirbis <@t> onko-i.si Thu Mar 11 00:25:06 2004 From: IKirbis <@t> onko-i.si (=?WINDOWS-1250?Q?Kirbi=9A_Srebotnik_Irena?=) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Apoptosis stain Message-ID: you could try with antibody M30 CytoDeath (Roche)detecting CK18 cleaved = by caspases Irena Kirbi=9A -----Original Message----- From: Kim Merriam [mailto:kmerriam2003@yahoo.com] Sent: Wednesday, March 10, 2004 3:59 PM To: Luis Chiriboga; Histonet Subject: RE: [Histonet] Apoptosis stain I already have IHC for caspase-3, but the investigator wants something = more. I am not familiar with apoptosis, so this is all new to me. =20 Kim =20 Luis Chiriboga wrote: cell signaling caspase (rabbit), works in ffpe mouse and rat with = standard biotinylated secondary & SA-HRP detection Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Kim Merriam Sent: Wednesday, March 10, 2004 7:34 AM To: Histonet Subject: [Histonet] Apoptosis stain Hi, I need to do staining for apoptosis on FFPE tissue (mouse and rat). Can anyone recommend a good kit? Thanks in advance, Kim Merriam Novartis Cambridge, MA --------------------------------- Do you Yahoo!? Yahoo! Search - Find what you=12re looking for faster. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Do you Yahoo!? Yahoo! Search - Find what you=92re looking for faster. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpwenk <@t> covad.net Thu Mar 11 03:45:00 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] "High Iron Diamine" stain References: Message-ID: <003f01c4074d$862d2bc0$0520d445@domainnotset.invalid> Try the site from Queen's Hospital, Nottingham, England - home of John Bancroft (of the Bancroft-Gamble histotechnology book fame)> http://www.nottingham.ac.uk/pathology/pathprot.html Click on "Methods", then click on "High Iron Diamine". Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Luck, Greg D." To: Sent: Wednesday, March 10, 2004 5:34 PM Subject: [Histonet] "High Iron Diamine" stain > Hello, > > I'm looking for some assistance with doing a procedure called a "High Iron > Diamine" stain. I do not have a written procedure and have never done one > myself. Any help would be greatly appreciated. Thanks in advance, Greg > > Greg Luck > Anatomic Pathology Supervisor > Deaconess Medical Center > 800 W. 5th Ave > Spokane, WA 99204 > 509.473.7077 > luckg@empirehealth.org > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From RFail <@t> Charleston.net Thu Mar 11 03:36:52 2004 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] MEL 5 Message-ID: <000b01c4074c$635dff30$e211a6a5@rena> Hello, Is this antibody available for paraffin sections? Rena Fail IHC/SS Medical University of SC 165 Ashley Ave. Charleston, SC 29425 From DDDeltour <@t> sig.med.navy.mil Thu Mar 11 05:10:36 2004 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Slides and Food/Drink Message-ID: Hello, The hospital safety officer came through today and said that there should be no eating or drinking while the cytologists screen slides. This is just a screening room. Does anyone know the reference or instruction that states this? Thanks HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. From stancelb <@t> msn.com Thu Mar 11 06:24:25 2004 From: stancelb <@t> msn.com (Barbara Stancel) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] My old equipment manuals Thank you Message-ID: My old equipment manuals thank every one who replied to adopt them. They have all been claimed. I will e-mail each of you who responded. Thank you again, Barbara >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] old equipment manuals >Date: Wed, 10 Mar 2004 15:21:42 +0000 > >Dear Histonetters, > >While cleaning out the back of my filing cabinet, I came across some old >equipment manuals. The equipment is looooong gone, but the packrat in me >has been keeping the manuals….for nostalgia’s sake. But I just can’t keep >everything. > >If any one would like these old manuals, please contact me off the >HISTONET. First come. First serve. > >1 Minotome (microtome-cryostat) Installation, operation and service manual. >Damon/IEC 1975 >2.Leitz Rotary Microtomes and Cryostat parts list >3. International Model CTD Microtome-Cryostat IEC >4. Autotechnicon MONO/DUO illustrated parts breakdown >5. Autotechnicon MONO/DUO combination manual for the operation of both > Technical publication No. TA1-0225-00 Sept 1971 >6. Autotechnicon Ultra II System Hardback manual 1978 > >Please don’t make me trash them. > >Histologically yours, > >Barbara _________________________________________________________________ Learn how to help protect your privacy and prevent fraud online at Tech Hacks & Scams. http://special.msn.com/msnbc/techsafety.armx From lefortb <@t> crhsc.umontreal.ca Thu Mar 11 07:01:05 2004 From: lefortb <@t> crhsc.umontreal.ca (Bertrand Lefort) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] RT-PCR in situ In-Reply-To: <20031201180001.12076.85479.Mailman@swlx167.swmed.edu> Message-ID: Hello, I am trying to developpe RT-PCR in situ in the lab. Is there someone in the group who is using this technology. I have problems to find a real positiv signal and need to discuss about many points in my protocols. You can contact me directly by mail at lefortb@crhsc.umontreal.ca Thanks Bertrand _______________________________________ Bertrand Lefort Research Assistant Laboratory of Neuro-immunology of Asthma Research Center, Room J3155 5400, Blv. Gouin West Montreal, QC H4J 1C5 T?l : 514 338 2222 est. 3622 Web site : http://www.crhsc.umontreal.ca/maghni From ASelf <@t> gmhsc.com Thu Mar 11 08:09:54 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] NexES APK wash bottle Message-ID: Does anyone have an extra APK wash bottle for the Ventana NexES that they would like to get rid rid of. We have a hole in ours and we would have to purchase the whole assembly kit to get the bottle. They do have the bottles available, but they are now new and improved and will not work without getting a whole new assembly kit. Thanks in advance, Amy Amy Self Georgetown Memorial Hospital 527-7179 (843) Note: 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 pengbw <@t> sjtu.edu.cn Thu Mar 11 08:17:00 2004 From: pengbw <@t> sjtu.edu.cn (Baowei Peng) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] block DC migration Message-ID: <20040311141700.822DF101D8A3@sjtu.edu.cn> Hello to all, My question maybe a little apart from IHC. If anyone here can tell me a regeant used to block DC migration to LN? I\'m going to look for a role of cells other than DCs played in antigen presentation after vacination. Any suggestion will be appreciated. Baowei Peng Shanghai Jiaotong University, Shanghai, China From DonnaWillis <@t> texashealth.org Thu Mar 11 08:57:13 2004 From: DonnaWillis <@t> texashealth.org (Willis, Donna) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] to do or not to do egfr Message-ID: <5C6CBCCEB04B894C8BD15B312487F7B201A32F95@ftwex01.txhealth.org> Kathy, Just to let you know. We have been using the DAKO Clone H11 on both the Dako and Ventana instruments for a long time. When we got a request from on of our Oncologist in Feb for EGFR on archived colon cases we thought no problem. We used the H11 clone and everything was negative. The oncologist questioned the clone we were using and asked us to get the new EGFR pharmDx clone 2-18C9 and retest the cases. We did and they were positive. Just beware that if the patient is being tested for the Erbitux drug we all might have to use the 2-18C9 clone. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of KHays@mbhs.org Sent: Wednesday, March 10, 2004 4:33 PM To: histonet@pathology.swmed.edu Subject: [Histonet] to do or not to do egfr Kathy Tedford-Hays HT (ASCP) Technical Specialist, Histology Dept (601)-968-3070 ext 7398 Baptist Medical Center 1225 North State Street Jackson, MS 39202 we are currently getting several requests for the test egfr as many of you. my questions are: is this another immunostain or is this something more? we were told that if the patients are positive then they are candidates for a new chemotherapy for colon cancer. if this is not another immunostain then what labs are doing the egfr for this new chemo? any information would be appreciated. thank you. The information contained in this email is confidential and is intended for the recipient only. Do not forward this email without permission from its originator. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. From JWEEMS <@t> sjha.org Thu Mar 11 09:09:33 2004 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] to do or not to do egfr Message-ID: <8C54DF3F71F6E34A80B1DE628C7EF4DD027F943D@exch2.sjha.org> Don't we need to be sure that the FDA approved reagents are used? If not a patient can be denied clinical trials. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Willis, Donna Sent: Thursday, March 11, 2004 9:57 AM To: 'KHays@mbhs.org'; histonet@pathology.swmed.edu Subject: RE: [Histonet] to do or not to do egfr Kathy, Just to let you know. We have been using the DAKO Clone H11 on both the Dako and Ventana instruments for a long time. When we got a request from on of our Oncologist in Feb for EGFR on archived colon cases we thought no problem. We used the H11 clone and everything was negative. The oncologist questioned the clone we were using and asked us to get the new EGFR pharmDx clone 2-18C9 and retest the cases. We did and they were positive. Just beware that if the patient is being tested for the Erbitux drug we all might have to use the 2-18C9 clone. Donna Willis Histology Lab Manager Harris Methodist Fort Worth, Tx -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of KHays@mbhs.org Sent: Wednesday, March 10, 2004 4:33 PM To: histonet@pathology.swmed.edu Subject: [Histonet] to do or not to do egfr Kathy Tedford-Hays HT (ASCP) Technical Specialist, Histology Dept (601)-968-3070 ext 7398 Baptist Medical Center 1225 North State Street Jackson, MS 39202 we are currently getting several requests for the test egfr as many of you. my questions are: is this another immunostain or is this something more? we were told that if the patients are positive then they are candidates for a new chemotherapy for colon cancer. if this is not another immunostain then what labs are doing the egfr for this new chemo? any information would be appreciated. thank you. The information contained in this email is confidential and is intended for the recipient only. Do not forward this email without permission from its originator. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message and any attachments is intended only for the use of the individual or entity to which it is addressed, and may contain information that is PRIVILEGED, CONFIDENTIAL, and exempt from disclosure under applicable law. If you are not the intended recipient, you are prohibited from copying, distributing, or using the information. Please contact the sender immediately by return e-mail and delete the original message from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Josephs Health System, Inc. From peoshel <@t> wisc.edu Thu Mar 11 10:13:17 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] Brucella staining Message-ID: Histomavens, I've got a project coming in to look at bacteria in the SEM. No problem, I do that routinely. But, it seems to me that this group would be better off sectioning their samples and staining the bacteria. Except they claim the bugs won't stain. This is for Brucella melitensis in mouse tails. Anyone have a good stain for these bacteria? The stain would be done on sections. Calcified or decalified better? I don't think that matters to the study, but it might to the staining. Thanks! Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From a.hannamitchell <@t> ucc.ie Thu Mar 11 12:58:10 2004 From: a.hannamitchell <@t> ucc.ie (Hanna-Mitchell, Ann) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] A question on setting up of organotypic cultures , on the use of heparin in perfusion fluid (non-fixative) and finally , sucrose and ECF Message-ID: <9FBB394A25826C46B2C6F0EBDAD42755059C256A@xch2.ucc.ie> Good Evening from a very wet and windy Ireland! I am setting up spinal cord slice cultures . Does anyone have any particular opinion on the use of the anti-coagulant heparin in perfusion buffer ( non-fixative)prior to removal of 'wet' tissue ( in my case spinal cord) to be processed for culture. It is used by some researchers in my Department but in this instance they are pre-perfusing with 'buffer' prior to perfusing with fixative for other purposes. As I want to maintain the tissue in as healthy a condition as possible, I worry that heparin in my protocol might have unwanted effects on it . At the moment I have stopped using heparin. As regards sucrose, how does pressure perfusion cause sucrose molecules to move from the intravascular to the extravascular compartment in the brain? Lastly,I should really appreciate some advice on the following: having sliced the spinal cord segment on a tissue chopper (slice thickness: 400?m),and having separated out single slices , how does one get these slices onto a millicell insert? Thank you for reading this . I should very much appreciate any assistance offered. Yours Sincerely, Ann T. Hanna-Mitchell PhD Dr. Ann T. Hanna-Mitchell Department of Anatomy BioSciences Institute University College Cork Ireland. Phone +353 21 4902246/+353 21 4901309 E mail a.hannamitchell@ucc.ie/athm@ucc.ie From BRIAN.CHELACK <@t> usask.ca Thu Mar 11 13:44:56 2004 From: BRIAN.CHELACK <@t> usask.ca (Brian Chelack) Date: Fri Sep 16 15:22:39 2005 Subject: [Histonet] regarding Protein A, G, L etc binding affinities Message-ID: <4050C1B8.5B2CA5B3@sask.usask.ca> Try looking on the biocan website: WWW.Biocan.com/frames/acti_f.htm or for the less computer literate: go to Biocan.com click on products; immunology; actigen; rprotein L rprotein LA; scroll down the page for a real nice chart that I've found to be accurate in my experiences. regards Brian Chelack Prairie Diagnostic Services From Jennifer_Harvey <@t> URMC.Rochester.edu Thu Mar 11 13:38:42 2004 From: Jennifer_Harvey <@t> URMC.Rochester.edu (Harvey, Jennifer) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] custom antibodies Message-ID: Hi, We are interested in getting a custom antibody made, including the peptide. Has anyone in histoland had this done? Do you recommend a company you had luck with? I saw that Zymed makes both the peptide and the antibodies. Has anyone ever used them? Thanks for you help. Jennifer Harvey Center for Cellular and Molecular Cardiology University of Rochester Medical Center 601 Elmwood Avenue, Box 679-CCMC Rochester, NY 14642 Telephone: (585) 273-5161 From csr <@t> meyerinst.com Thu Mar 11 13:46:33 2004 From: csr <@t> meyerinst.com (Cheryl Rehfeld - Meyer Instruments, Inc) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] cryosectioning drilling cores Message-ID: <001001c407a2$371f21a0$7800a8c0@CSRLaptop> Hi all, I have a customer that needs to cut drilling mud cores in the cryostat. The materials are extremely hard such as quartz and silica. The problem we have encountered is that the section falls apart. He does not want to use the tape system since when we tried packing tape and it did not hold the sample together. We are using a tungsten carbide knife which is working fine and we section the material at 30 microns thick. I have thought about trying to infiltrate the specimen with plastic resin and section it on a microtome instead. The problem is that the cores are 3 inches in diameter and it could prove difficult getting all the water out out of the sample. He is doing image analysis studies on the sample. I am hoping that some of the materials or hard sample people on the histonet may have some ideas to try. Thank you for your help. Cheryl Rehfeld Meyer Instruments, Inc. 281-579-0342 e-mail csr@meyerinst.com From rocan <@t> mac.com Thu Mar 11 14:20:34 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] custom antibodies In-Reply-To: References: Message-ID: <8D09E9C3-7399-11D8-9341-000A9589219E@mac.com> Jennifer, We had very good service from ProSci. Here is their web site address. http://www.prosci-inc.com/Antibody/Contact-Info.html If you are going invest in peptides I recommend immunizing each rabbit (or mouse) with at least three different peptides from your protein. This will increase your chances of a good antibody and lower the cost. You would need to analyze the protein sequence on a hydrophobicity?plot to find good candidates for antigenicity. I am sure ProSci (for a fee), can do this analysis as well. Good luck, Rocio On Mar 11, 2004, at 11:38 AM, Harvey, Jennifer wrote: > > Hi, > We are interested in getting a custom antibody made, including the > peptide. > Has anyone in histoland had this done? Do you recommend a company you > had > luck with? I saw that Zymed makes both the peptide and the antibodies. > Has > anyone ever used them? > > Thanks for you help. > > Jennifer Harvey > Center for Cellular and Molecular Cardiology > University of Rochester Medical Center > 601 Elmwood Avenue, Box 679-CCMC > Rochester, NY 14642 > Telephone: (585) 273-5161 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Mar 11 14:33:11 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] cryosectioning drilling cores In-Reply-To: <001001c407a2$371f21a0$7800a8c0@CSRLaptop> Message-ID: <3.0.6.32.20040311133311.00bc27d8@gemini.msu.montana.edu> Cheryl, I think he is going to need to go to metallurgical/geology cutting methods. For one thing, the quartz and silica are NOT doing your TC knife any favors, but mainly you cannot cut muddy water!! You could dehydrate and infiltrate the sample with methylmethacrylate, and then cut with the TC knife. I suggest you look into doing methylmethacrylate methods used for undecalcified bone, and do LONG dehydrations on the thick cores. You may be able to use an acetone gradient instead of alcohol, since you are NOT maintaining tissue proteins with acetone being more efficient for water removal in the end. Then embed in MMA and cut the cores with diamond embedded saws, grind them thinner - you can grind them down to 30 um after mounting on a clear plastic slide (glass will break during grinding/polishing). Buehler has all the equipment for this, and it will NOT be cheap. Buehler Isomet saws would cut thin 100 um slabs easily. Most geology/metallurgical labs have this type of equipment available. I am sure there are geology labs who do this kind of thing all the time, a search?? and the methods may be published. Paleoarcheologists deal with similar problems, silicalized bone or silica filled bone - they have methods - mud may be the difference, but they also deal with muddy specimens. I know they sometimes do their sample preps differently than bonehead types with metallurgical resins, etc. Give Buehler a call, they may be able to help you too. Personally, I think he needs to be using geology/metallurgy expertise rather than histotechnics - he should contact a geology dept to see what they do or is he already in a geology dept? (oh dear!) Want to bet oil companies in the business of exploration and sample analysis have the answers also. Maybe a literature search would help - gee I think I gave you more to do than you wanted. Good luck on a muddy subject! At 01:46 PM 3/11/2004 -0600, you wrote: >Hi all, > >I have a customer that needs to cut drilling mud cores in the cryostat. The >materials are extremely hard such as quartz and silica. The problem we have >encountered is that the section falls apart. He does not want to use the >tape system since when we tried packing tape and it did not hold the sample >together. We are using a tungsten carbide knife which is working fine and >we section the material at 30 microns thick. > >I have thought about trying to infiltrate the specimen with plastic resin >and section it on a microtome instead. The problem is that the cores are 3 >inches in diameter and it could prove difficult getting all the water out >out of the sample. He is doing image analysis studies on the sample. > >I am hoping that some of the materials or hard sample people on the histonet >may have some ideas to try. Thank you for your help. >Cheryl Rehfeld >Meyer Instruments, Inc. >281-579-0342 >e-mail csr@meyerinst.com > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gsennello <@t> osip.com Thu Mar 11 14:34:40 2004 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] custom antibodies Message-ID: We have used Bethyl Labs in Texas, 1-800-338-9579 Gina Sennello Associate Scientist/ Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 -----Original Message----- From: Harvey, Jennifer [mailto:Jennifer_Harvey@URMC.Rochester.edu] Sent: Thursday, March 11, 2004 12:39 PM To: Histonet (E-mail) Subject: [Histonet] custom antibodies Hi, We are interested in getting a custom antibody made, including the peptide. Has anyone in histoland had this done? Do you recommend a company you had luck with? I saw that Zymed makes both the peptide and the antibodies. Has anyone ever used them? Thanks for you help. Jennifer Harvey Center for Cellular and Molecular Cardiology University of Rochester Medical Center 601 Elmwood Avenue, Box 679-CCMC Rochester, NY 14642 Telephone: (585) 273-5161 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Zorbutt <@t> aol.com Thu Mar 11 16:07:49 2004 From: Zorbutt <@t> aol.com (Zorbutt@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] The role of Human Papilloma Virus in patients with melanoma. Message-ID: Hi, I would be interested to know if anyone has information linking HPV and melanoma, and which strain of HPV is of interest. Thank you Zohra Butt zorbutt@aol.com From laurie.reilly <@t> jcu.edu.au Thu Mar 11 16:38:49 2004 From: laurie.reilly <@t> jcu.edu.au (Laurie Reilly) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Brucella staining In-Reply-To: Message-ID: <5.1.0.14.0.20040312083243.00a34630@mail.jcu.edu.au> Phil and all, The Putt Modification of the Zeihl-Neelsen Stain is reputed to stain bovine Brucella. The method is a cold ZN utilising New Fuchsin at room temperature and then treating with Saturated Aqueous Lithium Carbonate. The method is in "Animal Tissue Techniques" by Gretchen L. Humason. Regards, Laurie. At 10:13 AM 03/11/04 -0600, Philip Oshel wrote: >Histomavens, > >I've got a project coming in to look at bacteria in the SEM. No problem, I >do that routinely. But, it seems to me that this group would be better off >sectioning their samples and staining the bacteria. >Except they claim the bugs won't stain. >This is for Brucella melitensis in mouse tails. >Anyone have a good stain for these bacteria? The stain would be done on >sections. >Calcified or decalified better? I don't think that matters to the study, >but it might to the staining. >Thanks! > >Phil >-- >Philip Oshel >Supervisor, BBPIC microscopy facility >Department of Animal Sciences >University of Wisconsin >1675 Observatory Drive >Madison, WI 53706 - 1284 >voice: (608) 263-4162 >fax: (608) 262-5157 (dept. fax) > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From bryand <@t> netbistro.com Thu Mar 11 17:10:26 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Identifying staining methods Message-ID: <00b101c407be$207a4080$a870c2cf@yourlk4rlmsu41> Hi all, I received the following communication in November last year, and am posting it with permission. My response is below it, after the line of asterisks. I have never heard the subject of standard identification of staining methods raised before. Could I get you all to read the letter and my reply, then comment on either the specifics in the e-mail, or the subject of identifying specific staining methods from the jumble of names by which they go. Would there be any benefit to a naming scheme to identify specific staining methods and their variants, or do you think it would be a completely unattainable goal, or a pointless exercise, or something else?. If you want to know what LOINC is go to :http://www.loinc.org Awaiting with bated breath, Bryan Llewellyn ----- Original Message ----- From: "Jeremiah H Sable" To: "Bryan D. Llewellyn" Sent: Tuesday, November 25, 2003 9:15 AM Subject: Questions about stains Hi, I've been helping develop a public health information system for the Centers for Disease Control in Atlanta, GA. It uses the LOINC coding system for laboratory test names, and I've found some ambiguous and redundant names for stain methods. I've done a lot of web searching and found the Stains File to be the most organized and helpful site. I'm hoping to get a little more "un-confused" before I send my suggestions on to the LOINC developers. My questions concern three groups of stains: Kinyoun, Gomori, and Silver stains. 1. Kinyoun stain: LOINC has a stain method called "Kinyoun stain." It also has the following methods: "Kinyoun hematoxylin stain" "Kinyoun iron hematoxylin stain" "Kinyoun acid fast stain" "Modified Kinyoun acid fast stain" My question: Is the term "Kinyoun stain" meaningful without further qualification? Or would it be better to delete "Kinyoun stain" from the list of LOINC methods? 2. Gomori stains: LOINC has a stain method called "Gomori stain". This seems ambiguous because "Gomori stain" can mean a Methenamine silver stain or a Trichrome stain. I've also seen references to "Gomori reticulin stain," and a "Gomori stain for pancreatic island cells." (Also collagen and cartillage stains, which might be the same as the Gomori reticulin stain.) Can you help clarify this? How many types of Gomori stains are there? Are there unambiguous names for the methods referred to as Gomori stains? 3. Silver stains: LOINC has nine silver stains: Silver impregnation stain, Dieterle Silver stain Silver stain, Grimelius Silver stain, Fontana-Masson Silver stain, Warthin-Starry Methenamine silver nitrate stain Methenamine silver stain, Grocott Methenamine silver stain, Jones Silver nitrate stain My impressions about this list are: 1. "Methenamine silver nitrate stain" should be renamed "Methenamine silver stain." (I think all Methenemain silver stains have silver nitrate in the recipe.) 2. "Silver nitrate stain" is redundant and actually means "Methenamine silver stain." Is this correct? Is Grocott's Methenamine silver stain a modification of the Gomori Silver Methenamine stain (which isn't on the list)? Should "Silver methenamine stain, Gomori" be added to the list? Also, Is "Silver impregnation stain" just another way of saying "Silver stain?" Should the "Silver impregnation stain, Dieterle" be renamed "Silver stain, Dieterle"? These LOINC names for silver stains are very confusing to me and I'd appreciate any information that might help. Much appreciated, JS ------------------------------------------- Jerry Sable, NEDSS Vocabulary ********************************** Hello, You raise an interesting subject. As far as I know there is no standard way to designate specific staining methods, so it tends to be at the whim of the person referencing them. Most people use the name of the person who published the technique, but not always. Usually the reference includes the tissue component to be stained, but again, not always. Some are referenced colloquially due to their very common use, since nearly all histotechnologists would know what method is meant. I have never seen this subject raised before, and I would like to refer it to a histology discussion group (Histonet), composed of many histotechnologists with some pathologists and others. may I have your permission to do this with your query and my response below? Incidentally, I am not familiar with LOINC. What is it? Kinyoun's hematoxylin and Kinyoun's iron hematoxylin could refer to the same solution, depending on whether Kinyoun ever published a hematoxylin using a mordant other than iron. If one was published (I don't know) then it could refer to two different solutions. The second designation (Kinyoun's iron hematoxylin) is preferable as an identification. Kinyoun's acid fast stain and Modified Kinyoun acid fast stain would likely refer to two slightly different solutions (or procedures, since "stain" is sometimes used to refer to the solution and sometimes to the technique in which the solution is used), the modified one being slightly changed from the original by someone else, nnamed. My opinion is that a designation must identify, if it does not, then it is useless. The term "Kinyoun's stain", with no further identification to help in choosing between alternatives, is useless and can cause confusion. I would remove it. Gomori published several staining methods, many completely unrelated to each other. The term "Gomori's stain" is useless in identifying any of his techniques. To identify the particular method recommended it should be followed by some further identification, either the tissue element being targeted, or some reference to the underpinnings to the technique, i.e. Gomori's silver impregnation for reticulin, Gomori's aldehyde fuchsin for pituitary cells and so on. This should make it clear. Silver stains are very common. Strictly speaking they are not stains but impregnations, although most histotechnologists refer to them as stains, I am sure. Still, formal documents should perhaps use the word impregnation. The term "Silver stain" simply means that a source of silver has been used in a method, nothing more. When it is followed by a name it refers to a specific method. Again, using the name of the published author is very common. When the term "methenamine silver" is included it refers to those methods which incorporate methenamine into the silver solution. Not all silver stains are the same. There are those that use silver nitrate alone, those that use silver nitrate and ammonia, and those that use silver nitrate and methenamine. Not all silver solutions contain methenamine, so your assumption that "methenamine silver nitrate stain" and "methenamine silver stain" mean the same thing is correct. Your assumption that "silver nitrate stain" means the same as "methenamine silver stain" is not correct. Gomori's and Grocott's methenamine silver methods are very similar, however, rather than refer to methenamine silver stains by author's name, perhaps you should consider listing by target, i.e. "Methenamine silver impregnation for fungi, basement membranes and carbohydrates". This covers the three common uses for these techniques by any variant, and they are fairly similar anyway. Dieterle's method demonstrates microorganisms (spirochetes and others). Perhaps "Silver impregnation for microrganisms, Dieterle", or "Silver impregnation for spirochetes, Dieterle", would be clearer. If you would like some more clarification, please ask. Bryan Llewellyn From billions <@t> public1.sz.js.cn Thu Mar 11 20:06:54 2004 From: billions <@t> public1.sz.js.cn (Sinoera Tech) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Re: Substrate for alkaline phosphatase Message-ID: <000a01c407d6$b3b81d00$a001a8c0@ming> Dear Histoneters, We are supplying following compounds. Sulfobromophthalein sodium hydrate Thymolphthalein monophosphate magnesium salt Thymolphthalein monophosphate disodium salt O-cresolphthalein monophosphoric acid and salts phenolphthalein monophosphate bis(cyclohexylamine)salt Other phthalein monophosphoric acid and their salts We would like to have your inquiries. Kind Regards. - Minggeng Wang, Ph.D / President SUZHOU SINOERA CHEM CO., LTD. No.5508, 125 Binhe Road Suzhou New & Hi-Tech District 215011 China Fax: +86 512 68224995 Tel: +86 512 68246939 E-Mail: billions@public1.sz.js.cn From suselore <@t> ihug.co.nz Fri Mar 12 01:49:28 2004 From: suselore <@t> ihug.co.nz (Susie Warwick) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions for small project. Message-ID: <6.0.1.1.0.20040312204517.01b8e7b8@pop.ihug.co.nz> My manager has suggested I do a small research project to help further my knowledge. I am a junior lab worker and am familiar with most aspects of Histology excluding immunos. I would like to do a small project involving special stains, But I need help brainstorming! I enjoy special stains and would like to do something involving them. Does anyone have any suggestions? Thanks Susie From barbara.bublava <@t> meduniwien.ac.at Fri Mar 12 01:39:17 2004 From: barbara.bublava <@t> meduniwien.ac.at (Ing. Barbara Bublava) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Thanks for Replys to "cryosectioning of the lung" Message-ID: <001701c40805$20182b10$9865fea9@GERICHTS9XOZZ8> I wish to thank all people who responded to my mail. The suggested procedures (OTC 1:1, saccarose) did work fine - at least I decided to go on with the saccarose for the fixed lung and a drop of OTC on the surface for not fixed lung. Barbara Bublava From RBARNHART <@t> summithealth.org Fri Mar 12 07:17:15 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Slides and Food/Drink Message-ID: Our cytotech and pathologist both eat and drink while they are screening slides. Nothing has been said to this point. Our pathologist's office is not in the main lab, but the cytotech's office is the first office in the main lab (but she has a door that closes and no other specimens every go in either office, just coverslipped slides). Please keep us informed of any information that you gather from this. >>> "Deltour, Douglas D.(HM2)" 03/11/04 06:10AM >>> Hello, The hospital safety officer came through today and said that there should be no eating or drinking while the cytologists screen slides. This is just a screening room. Does anyone know the reference or instruction that states this? Thanks HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ssq5977 <@t> yahoo.com.cn Fri Mar 12 07:19:30 2004 From: ssq5977 <@t> yahoo.com.cn (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Help! Message-ID: <20040312131930.8872.qmail@web15211.mail.bjs.yahoo.com> Dear Histonetters: ¡¡I found the presence of IgA along capillary loop in renal biopsies by employing fluorescence technique was too common in our institute. Would you please give me some explanations and share your protocol for IF on kidney biopsies? Thanks. Shuqiong Shen Reseach Institute of Nephropathy Jingling Hospital,Nanjing,China --------------------------------- Do You Yahoo!? ÍêÈ«Ãâ·ÑµÄÑÅ»¢µçÓÊ£¬ÂíÉÏ×¢²á»ñÔù¶îÍâ60Õ×ÍøÂç´æ´¢¿Õ¼ä From Sarah.Blachford <@t> ngh.nhs.uk Fri Mar 12 07:47:35 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Pemphigoid/pemphigus Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58E3@NGH_EXCHANGE1> Dear all, Does anyone know of, or use any other staining methods for pemphigoid/pemphigus other than the traditional immunofluorescent techniques. Thanks Sarah Blachford (Sarah.Blachford@ngh.nhs.uk) Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From ssq5977 <@t> yahoo.com.cn Fri Mar 12 08:06:41 2004 From: ssq5977 <@t> yahoo.com.cn (=?gb2312?q?=CA=E7=C7=ED=20=C9=F2?=) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] help! Message-ID: <20040312140641.41446.qmail@web15208.mail.bjs.yahoo.com> Dear Histonetters: ¡¡I found the presence of IgA along capillary loop in renal biopsies by employing fluorescence technique was too common in our institute. Would you please give me some explanations and share your protocol for IF on kidney biopsies? Thanks. Shuqiong Shen Reseach Institute of Nephropathy Jingling Hospital,Nanjing,China --------------------------------- Do You Yahoo!? ÍêÈ«Ãâ·ÑµÄÑÅ»¢µçÓÊ£¬ÂíÉÏ×¢²á»ñÔù¶îÍâ60Õ×ÍøÂç´æ´¢¿Õ¼ä From peoshel <@t> wisc.edu Fri Mar 12 08:49:33 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Brucella staining In-Reply-To: <5.1.0.14.0.20040312083243.00a34630@mail.jcu.edu.au> References: <5.1.0.14.0.20040312083243.00a34630@mail.jcu.edu.au> Message-ID: List, Searching Amazon.com and other internet booksellers, I find the 5th, 1997 edition of "Humason's Animal Tissue Techniques" to be out of print, with no new edition indicated. Is there a 6th edition in the works? Thanks. Phil Phil and all, The Putt Modification of the Zeihl-Neelsen Stain is reputed to stain bovine Brucella. The method is a cold ZN utilising New Fuchsin at room temperature and then treating with Saturated Aqueous Lithium Carbonate. The method is in "Animal Tissue Techniques" by Gretchen L. Humason. Regards, Laurie. At 10:13 AM 03/11/04 -0600, Philip Oshel wrote: Histomavens, I've got a project coming in to look at bacteria in the SEM. No problem, I do that routinely. But, it seems to me that this group would be better off sectioning their samples and staining the bacteria. Except they claim the bugs won't stain. This is for Brucella melitensis in mouse tails. Anyone have a good stain for these bacteria? The stain would be done on sections. Calcified or decalified better? I don't think that matters to the study, but it might to the staining. Thanks! Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Mr.Laurie Reilly Ph 07 4781 4468 Physiology & Pharmacology Fax 07 4779 1526 Aust.Inst.of Tropical Vet.& Animal Sc. James Cook University Townsville Qld. 4811 laurie.reilly@jcu.edu.au Australia. From p.vandeplas <@t> aurion.nl Fri Mar 12 09:08:20 2004 From: p.vandeplas <@t> aurion.nl (Peter van de Plas) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Protein A or G?? In-Reply-To: <1078941838.404f588e434a5@webmail3.its.uiowa.edu> Message-ID: <1917A15C-7437-11D8-8BAE-003065816A84@aurion.nl> Dear Kathy, Reference in literature: Protein G-gold complex: comparative evaluation with protein A-gold for high-resolution immunocytochemistry. Bendayan M and Garzon S.J. Histochem Cytochem. 1988 Jun;36(6):597-607. Info can also be found on page 7 of our catalog. Can send you a pdf.file upon request. Hope this is of help. Peter On Wednesday, March 10, 2004, at 07:03 PM, katherine-walters@uiowa.edu wrote: > > Hi everyone, > > Does anyone know of a reference that states which animal sources of > primary > antibody works with Protein A and which animal sources of antibody > works with > protein G. I remember having read it back in the day, but now can't > seem to > lay my hands on it. > > Thanks for your help, > Kathy > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ----------- Peter van de Plas Aurion Costerweg 5 6702 AA Wageningen The Netherlands Phone: +31 317 497676 Fax: +31 317 415955 From tpmorken <@t> labvision.com Fri Mar 12 10:49:00 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Slides and Food/Drink Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA21773C3@usca0082k08.labvision.apogent.com> The places I have worked have had screening areas separate from the lab and were officially designated "clean" areas, so eating and drinking was ok. In each case only a door separated these areas from the lab. Tim Morken -----Original Message----- From: Rebecca Barnhart [mailto:RBARNHART@summithealth.org] Sent: Friday, March 12, 2004 5:17 AM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Slides and Food/Drink Our cytotech and pathologist both eat and drink while they are screening slides. Nothing has been said to this point. Our pathologist's office is not in the main lab, but the cytotech's office is the first office in the main lab (but she has a door that closes and no other specimens every go in either office, just coverslipped slides). Please keep us informed of any information that you gather from this. >>> "Deltour, Douglas D.(HM2)" 03/11/04 06:10AM >>> Hello, The hospital safety officer came through today and said that there should be no eating or drinking while the cytologists screen slides. This is just a screening room. Does anyone know the reference or instruction that states this? Thanks HM2(FMF) Douglas D. Deltour Naval Hospital Sigonella Italy Anatomic Pathology, Histology Supervisor (HT) FROM US: 01139095564862 DSN: 624-4862 FAX FROM US: 01139095564680 DSN: 624-4680 This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. _______________________________________________ Histonet mailing list Histonet@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 AJMixon21 <@t> aol.com Thu Mar 11 19:16:06 2004 From: AJMixon21 <@t> aol.com (AJMixon21@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] I have a new email address at BellSouth.net. Message-ID: <0CA7173D.42091CE3.01F469A9@aol.com> Hello, I have a new email address at ajmixon2@bellsouth.net. From lance.erickson <@t> ihc.com Fri Mar 12 11:07:51 2004 From: lance.erickson <@t> ihc.com (Lance Erickson) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] New Utah Society for Histotechnology website Message-ID: We're excited to announce the launch of our new website at www.utahhisto.org . With the generosity of Tim Morken and the people at Lab Vision Corporation we are now up and running with a new and hopefully useful resource of information for our members and anyone else visiting us. The Exhibitor Registration Packet for our annual meeting is available for download and payment for fees is also available through the website. Take a look. Any comments/suggestions/opinions are welcome. Lance Erickson President - Utah Society for Histotechnology Anatomic Pathology Supervisor Primary Children's Medical Center 100 N Medical Dr Salt Lake City, UT 84113-1100 (801) 588-3110 fax (801) 588-3169 From Jackie.O'Connor <@t> abbott.com Thu Mar 11 16:43:48 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Osteoblast marker Message-ID: Does anyone have a lead for an osteoblast ab for mice? Jackie O'Connor HT(ASCP) Abbott Laboratories Discovery Chemotheraputics Jackie.O'Connor@abbott.com From mward <@t> wfubmc.edu Fri Mar 12 13:22:42 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Autopsy question Message-ID: <61135F0455D33347B5AAE209B903A304076A4DE3@EXCHVS2.medctr.ad.wfubmc.edu> We are currently in Autopsy negotiations between the school and the hospital. We pay the school (Pathologists) for hospitals autopsies performed and they pay the hospital for services and supplies for Medical Examiner cases. Could you share how you handle this scenario and what the dollar split is for your institution? This would be greatly appreciated! Martha Ward Wake Forest University Baptist Medical Center From lizchlipala <@t> premierhistology.com Fri Mar 12 18:00:38 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Immunohistochemistry discussion group Message-ID: <000001c4088e$3b37afe0$74d48a80@LIZ> Hello all At one time I was registered on this great immunohistochemistry discussion group. It was associated with a web page that had images, protocols, etc. I found out about it when I posted an immuno question. Someone had e-mailed me back about this web page. I have lost the bookmark and I have been unable to find the web page again. Can anyone help me out with this? Thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP) Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ 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 daniel.eberhard <@t> uni-bielefeld.de Sat Mar 13 02:57:36 2004 From: daniel.eberhard <@t> uni-bielefeld.de (Daniel Eberhard) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Quality of tissues: freeze - thaw - freeze Message-ID: <4052CD00.9060500@uni-bielefeld.de> Dear Listers, some month ago I prepared some mouse tissues for cryo-sectioning without fixing the tissues before. Now I have found out that usual fixing of sections is not efficient for our purposes, and whole mount fixation (e.g. with pFA) would have been much better. Thus my question: Have some of you ever thawed unfixed tissue blocks, fixed the tissues and re-frozen them? I?m afraid the integrity of the tissue might be spoiled and of course: some of the tissues are valuable and unique. Any hints are welcome thanks Daniel. -- (-)-(-) --------------------------- \"/ --- Dr. Daniel Eberhard =V= Developmental Biology & Molecular Pathology Graduate Programe on Pattern Formation University of Bielefeld D 33501 Bielefeld/Germany FAX: xx49(0)521-106-5654 (-)-(-) --------------------------- \"/ --- =V= From gudrun.lang <@t> aon.at Sat Mar 13 04:42:44 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Identifying staining methods References: <00b101c407be$207a4080$a870c2cf@yourlk4rlmsu41> Message-ID: <001401c408e7$eb1b9ee0$eeeea8c0@SERVER> I like the idea of standardized names of staining methods. And something like a genealogical tree of stains would give advantages in the jungle of methods. What method is the origin, and what is the modification? For a beginner in histological staining the names are often confusing and in the lab we use often abbreviations, that have nothing in common with the original name. If you are'nt familiar to the history and the histo-investigators the names have no meaning. And a standardized system would help with international comunication. Our german "BerlinerBlau" or "Gitterfaser" would say nothing to an English speaker and the other way round. My thoughts about this issue, from Austria Gudrun Lang ----- Original Message ----- From: "Bryan Llewellyn" To: "Histonet" Sent: Friday, March 12, 2004 12:10 AM Subject: [Histonet] Identifying staining methods > Hi all, > I received the following communication in November last year, and am posting > it with permission. My response is below it, after the line of asterisks. > I have never heard the subject of standard identification of staining > methods raised before. Could I get you all to read the letter and my reply, > then comment on either the specifics in the e-mail, or the subject of > identifying specific staining methods from the jumble of names by which they > go. Would there be any benefit to a naming scheme to identify specific > staining methods and their variants, or do you think it would be a > completely unattainable goal, or a pointless exercise, or something else?. > > If you want to know what LOINC is go to :http://www.loinc.org > > Awaiting with bated breath, > > Bryan Llewellyn > > > > > ----- Original Message ----- > From: "Jeremiah H Sable" > To: "Bryan D. Llewellyn" > Sent: Tuesday, November 25, 2003 9:15 AM > Subject: Questions about stains > > > Hi, I've been helping develop a public health information system for the > Centers for Disease Control in Atlanta, GA. It uses the LOINC coding system > for laboratory test names, and I've found some ambiguous and redundant names > for stain methods. I've done a lot of web searching and found the Stains > File to be the most organized and helpful site. > > I'm hoping to get a little more "un-confused" before I send my suggestions > on to the LOINC developers. My questions concern three groups of stains: > Kinyoun, Gomori, and Silver stains. > > 1. Kinyoun stain: LOINC has a stain method called "Kinyoun stain." It also > has the following methods: > "Kinyoun hematoxylin stain" > "Kinyoun iron hematoxylin stain" > "Kinyoun acid fast stain" > "Modified Kinyoun acid fast stain" > > My question: Is the term "Kinyoun stain" meaningful without further > qualification? Or would it be better to delete "Kinyoun stain" from the > list of LOINC methods? > > 2. Gomori stains: LOINC has a stain method called "Gomori stain". This > seems ambiguous because "Gomori stain" can mean a Methenamine silver stain > or a Trichrome stain. I've also seen references to "Gomori reticulin > stain," and a "Gomori stain for pancreatic island cells." (Also collagen > and cartillage stains, which might be the same as the Gomori reticulin > stain.) > > Can you help clarify this? How many types of Gomori stains are there? Are > there unambiguous names for the methods referred to as Gomori stains? > > 3. Silver stains: LOINC has nine silver stains: > Silver impregnation stain, Dieterle > Silver stain > Silver stain, Grimelius > Silver stain, Fontana-Masson > Silver stain, Warthin-Starry > Methenamine silver nitrate stain > Methenamine silver stain, Grocott > Methenamine silver stain, Jones > Silver nitrate stain > > My impressions about this list are: > > 1. "Methenamine silver nitrate stain" should be renamed "Methenamine silver > stain." (I think all Methenemain silver stains have silver nitrate in the > recipe.) > 2. "Silver nitrate stain" is redundant and actually means "Methenamine > silver stain." > > Is this correct? > > Is Grocott's Methenamine silver stain a modification of the Gomori Silver > Methenamine stain (which isn't on the list)? Should "Silver methenamine > stain, Gomori" be added to the list? > > Also, Is "Silver impregnation stain" just another way of saying "Silver > stain?" Should the "Silver impregnation stain, Dieterle" be renamed "Silver > stain, Dieterle"? > > These LOINC names for silver stains are very confusing to me and I'd > appreciate any information that might help. > > Much appreciated, > > JS > ------------------------------------------- > Jerry Sable, NEDSS Vocabulary > > > ********************************** > > > > Hello, > You raise an interesting subject. As far as I know there is no standard way > to designate specific staining methods, so it tends to be at the whim of the > person referencing them. Most people use the name of the person who > published the technique, but not always. Usually the reference includes the > tissue component to be stained, but again, not always. Some are referenced > colloquially due to their very common use, since nearly all > histotechnologists would know what method is meant. > > I have never seen this subject raised before, and I would like to refer it > to a histology discussion group (Histonet), composed of many > histotechnologists with some pathologists and others. may I have your > permission to do this with your query and my response below? > > Incidentally, I am not familiar with LOINC. What is it? > > Kinyoun's hematoxylin and Kinyoun's iron hematoxylin could refer to the same > solution, depending on whether Kinyoun ever published a hematoxylin using a > mordant other than iron. If one was published (I don't know) then it could > refer to two different solutions. The second designation (Kinyoun's iron > hematoxylin) is preferable as an identification. > > Kinyoun's acid fast stain and Modified Kinyoun acid fast stain would likely > refer to two slightly different solutions (or procedures, since "stain" is > sometimes used to refer to the solution and sometimes to the technique in > which the solution is used), the modified one being slightly changed from > the original by someone else, nnamed. > > My opinion is that a designation must identify, if it does not, then it is > useless. The term "Kinyoun's stain", with no further identification to help > in choosing between alternatives, is useless and can cause confusion. I > would remove it. > > Gomori published several staining methods, many completely unrelated to each > other. The term "Gomori's stain" is useless in identifying any of his > techniques. To identify the particular method recommended it should be > followed by some further identification, either the tissue element being > targeted, or some reference to the underpinnings to the technique, i.e. > Gomori's silver impregnation for reticulin, Gomori's aldehyde fuchsin for > pituitary cells and so on. This should make it clear. > > Silver stains are very common. Strictly speaking they are not stains but > impregnations, although most histotechnologists refer to them as stains, I > am sure. Still, formal documents should perhaps use the word impregnation. > The term "Silver stain" simply means that a source of silver has been used > in a method, nothing more. When it is followed by a name it refers to a > specific method. Again, using the name of the published author is very > common. When the term "methenamine silver" is included it refers to those > methods which incorporate methenamine into the silver solution. > > Not all silver stains are the same. There are those that use silver nitrate > alone, those that use silver nitrate and ammonia, and those that use silver > nitrate and methenamine. Not all silver solutions contain methenamine, so > your assumption that "methenamine silver nitrate stain" and "methenamine > silver stain" mean the same thing is correct. Your assumption that "silver > nitrate stain" means the same as "methenamine silver stain" is not correct. > > Gomori's and Grocott's methenamine silver methods are very similar, however, > rather than refer to methenamine silver stains by author's name, perhaps you > should consider listing by target, i.e. "Methenamine silver impregnation for > fungi, basement membranes and carbohydrates". This covers the three common > uses for these techniques by any variant, and they are fairly similar > anyway. > > Dieterle's method demonstrates microorganisms (spirochetes and others). > Perhaps "Silver impregnation for microrganisms, Dieterle", or "Silver > impregnation for spirochetes, Dieterle", would be clearer. > > If you would like some more clarification, please ask. > > Bryan Llewellyn > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ThisisAnn <@t> aol.com Sat Mar 13 07:52:32 2004 From: ThisisAnn <@t> aol.com (ThisisAnn@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Forcep Sterilization Message-ID: We are having a problem with carry-over at the grossing station. We do not have a sink available and are presently dipping our forceps in ethanol. Is there another solution we can use besides ethanol? Does anyone have any other ideas as how to avoid this contamination issue we are experiencing? Thank you Ann Angelo From bill501 <@t> mindspring.com Sat Mar 13 08:47:50 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Forcep Sterilization In-Reply-To: References: Message-ID: At 8:52 AM -0500 3/13/04, ThisisAnn@aol.com wrote: >We are having a problem with carry-over at the grossing station. 1. Get a sink 2. Get a sink 3. Identify those with sloppy technique and refresh them 4. Consider other sources of contamination besides the grossing station -- _______________ Bill Blank, MD Heartland Lab, Inc From goinghisto <@t> hotmail.com Sat Mar 13 11:46:44 2004 From: goinghisto <@t> hotmail.com (Kathleen Crawford) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] uterus tissue for HT exam Message-ID: Looking for uterus tissue, 2cm x 2cm. Requirements: must have endometrium completely across one side and must include myometrium. Serosa not necessary. Must mail tissue by end of March. Am desperate! Will reimburse you for next day delivery charges. Thank you in advance!!!!! Kathy Crawford 5 Wingate Court Bangor, Maine 04401 _________________________________________________________________ Get a FREE online computer virus scan from McAfee when you click here. http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From lizchlipala <@t> premierhistology.com Sat Mar 13 18:49:31 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] immunohistochemistry discussion group Message-ID: <000b01c4095e$3975be10$74d48a80@LIZ> Hello all I have located the web address to the immunohistochemistry discussion group. Several individuals also wanted the web page it is: http://home.no.net/immuno/ Liz Elizabeth A. Chlipala, BS, HTL(ASCP) Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ 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 arename <@t> hotmail.com Sun Mar 14 02:11:49 2004 From: arename <@t> hotmail.com (hanim shueb) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] IHC for dysfunction neurons Message-ID: Hi there, I was wondering if anyone out there knows whether is it possible to detect dysfunction neurons (not neuronal death; just neurons which are not capable of performing normal functions because infected by virus) by IHC methods and what markers should be used. advance thanking, Hanim Shueb Australia _________________________________________________________________ Are you in love? Find a date on MSN Personals http://match.msn.com.my/ From Rcartun <@t> harthosp.org Sun Mar 14 11:57:16 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] EGFR - My thoughts Message-ID: Approximately two years ago, I said to my colleagues that EGFr testing would make HER-2/neu testing look like "a walk in the park" (i.e, very simple). We tested DAKO's EGFR kit back then and compared it to the monoclonal that we were using at the time (clone 31G7 from Zymed). Although the results were similar (but not identical), the mAb from Zymed consistently produced stronger immunoreactivity which was easier to evaluate. Like you, we have received many requests during the past 3 weeks from our clinical colleagues to test their patients' colorectal tumors for EGFR so that they can be considered eligible for treatment with "Erbitux" (the anti-EGFR mAb drug). Although I started with the Zymed mAb, I am now convinced that this testing must be performed with DakoCytomation's "EGFR pharmDx" kit (primarily for reimbursement issues). Although the kit works well, I find the resulting immunoreactivity very difficult to score (this is true with the Zymed mAb as well). Most tumors have shown what I call "wishy-washing" staining that, if it were HER-2, I would call it negative. However, we are instructed to score it 1+ "Positive". I find it difficult to believe that "Erbitux" will help a patient whose tumor shows rare to focal immunoreactivity and often less intense than normal cells present in the same tissue section; however, I am not familiar with the clinical trials and patient outcomes. I have also noticed EGFR variability from block to block in the same patient when we have tested multiple tissue blocks. In addition, in my experience, it is very unusual to find a tumor that shows the classic "3+" staining pattern that we see with c-erbB-2 (HER-2/neu protein) in breast CA's with obvious gene amplification. It will be interesting to see where we are 6 months to a year from now with EGFR testing. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT From DPALLP <@t> aol.com Sun Mar 14 14:17:19 2004 From: DPALLP <@t> aol.com (DPALLP@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] ergonomics policy Message-ID: <179616FF.15E2C1B4.00042A59@aol.com> Does anyone have an ergonomics policy that they would like to share? Thanks. Susie Duhon, HT(ASCP)QIHC Diagnostic Pathology Associates, LLP Beaumont, Texas From escott8 <@t> houston.rr.com Sun Mar 14 17:53:21 2004 From: escott8 <@t> houston.rr.com (Edward Scott) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Georgia Society for Histotechnology Info. Message-ID: <000001c40a1f$8b0ff800$c5e2f218@thescotts> Hello to all in histoland. I would like for someone from the Georgia society to contact me. I would like an application. The information that I have is outdated. You can email me or fax me. My fax number is 281-540-2106. Thanks in advance. Allison Scott. From mariatere <@t> infovia.com.ar Sun Mar 14 18:23:32 2004 From: mariatere <@t> infovia.com.ar (Teresa Dominguez) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] AgNOR Message-ID: <002101c40a23$c1a27880$537f46c8@FAMILIA> From: "Teresa Dominguez" To: "prasad sbr prasad" Sent: Friday, March 12, 2004 9:38 PM Subject: Re: [Histonet] AgNOR > Sorry If I'm late for this, but at least I tried to help someone. > This is the AgNOR method I had in my notes. > Solutions: > A) Ag. Nitrate Sol. > AgNO3................50 Gr. > Destilled Water...100 ml > > B) Jelly > Dest. Water........100 ml. > Jelly........................2 Gr > > C) Formic Acid > Formic Acid ...........1 ml. > Dest. Water.........100 ml. > Method: 1- Desparaffinize and hydrate to destilled water > 2-Wash in dest. water (2 changes) > 3-Place in this solution for 30 minutes, room temperature: > One part of 2% Jelly , > One part of 1% Formic Acid, > Two part of 50%Ag NO3 > 4- Wash in hot dest. water > 5- Rinse in dest. water > 6- Counterstain, if desire with 0,5% Light Green > 7- Deshydrate in95% Alcohol, Absolute Alcohol,and clear in > Xylene, two > changes each. > 8- Mount with Permount > > Results: > NORs, M's G1 period: Dark Brown or black > Cytoplasms: Green or light brown > > Good Luck!!! > > > H.T Maria T Dominguez > Anatomy Pathology Service > Hospital Regional R?o Grande, > R?o Grande, Tierra del Fuego, Argentina > 54- 02964-422086/88 Ext. 143 > ----- Original Message ----- > From: "prasad sbr prasad" > To: > Sent: Saturday, March 06, 2004 12:26 AM > Subject: [Histonet] AgNOR > > > > > > > > Dear List, > > > > I shall be thankful to you if you send me the detaails of AgNOR staining > and scoring methods. > > > > with regards, > > Prasad. > > Dr.CSBR.Prasad,MD., > > Al Hakeem Polyclinic, > > PO.BOX: 34985, > > Riyadh-11478, > > Saudi Arabia. > > ************************************************ > > ************************************************ > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > From Sven.Terclavers <@t> med.kuleuven.ac.be Mon Mar 15 04:53:47 2004 From: Sven.Terclavers <@t> med.kuleuven.ac.be (Sven Terclavers) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] MicroFil Message-ID: Dear listers, I'm looking for a solution to perfuse bloodvessels which hardens after cooling down or after a certain timepoint of being liquid to fill not only larger bloodvessels but also cappillars. On the internet I found MicroFil, now I was wondering if some of you already tried this. I would like to know the following: -What is the smallest cappillar that will be filled? -Can sections be made without the MicroFil falling out of the cappillars? -Can this MicroFil solution be mixed with e.g. FITC to perform epi-fluorescence/CLS-microscopy? Thanks in advance, Sven Terclavers From Nancy.Walker <@t> sanofi-synthelabo.com Mon Mar 15 06:32:34 2004 From: Nancy.Walker <@t> sanofi-synthelabo.com (Nancy.Walker@sanofi-synthelabo.com) Date: Fri Sep 16 15:22:40 2005 Subject: =?iso-8859-1?Q?R=E9f=2E_=3A_[Histonet]_organotypic_cultures?= Message-ID: Don't understand why you want to perfuse and after do organo culture. Never heard of this! We dissect as quick as possible, use a tissue chopper to get those 400? slice, get them off the chopper support with flush of culture medium into a 25ml Corning tube. Swirl them around a little because this helps separate slices that stick together. Then under a disecting microscope, pick out slices that seem nicest then scoop them up with the flat side of two "paddle pastette" (Alpha laboratories cat n? LW4295 address 40 Parham drive, Eastleigh, Hampshire SO50 4NU UK tel. 44-1703483000). We sterilize the paddles in ETOH. Make sure you preincubate the millicell inserts with culture medium a couple of hours before use. This seems to increase health and survival. good luck, Nancy Walker Molecular Pharmacology Sanofi-Synthelbo Research B.P. 37 Lab?ge Innopole 31676 LABEGE CEDEX FRANCE nancy.walker@sanofi-synthelabo.com tel : (33)561004179? fax :(33)561004001 From leopold <@t> mnsi.net Tue Mar 16 06:38:36 2004 From: leopold <@t> mnsi.net (Derek & Lynda Leopold) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Steiner probs/H. pylori on Nexes Message-ID: <000801c40b53$a5ff9c80$3643fea9@leopold> Hi Netters, I've been searching the Histonet archives for some ideas on how to fix our pesky Steiner stain for H. pylori. I suspect our problems are typical: 1) some slides work and some don't, even in the same batch 2) lots of non-specific silver ppte. 3) we use the microwave method, and often fry the specimens. Ok, so while I don't want to take up the list's time by revisiting this old problem, anyone who feels like writing off-list with some tips is more than welcome. My other question is: Can an IHC for H. pylori be done on the Ventana Nexes? If so, does anyone have any opinions as to cost comparison with Steiner? We do about 15 slides a day with microwave Steiner, BUT we usually have to recut and redo about half of those. I argue that the techs could be doing something else, and the Ventana could be doing our stains for us. Please reply off list, because I know this is old news to most of you. Thanks! Lynda Leopold From GAshton <@t> PICR.man.ac.uk Mon Mar 15 06:17:22 2004 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] XIAP Message-ID: Dear all, I have been asked to do some immunostaining for a protein called XIAP. The tissue used will initially be 10%NBF fixed human cell lines, pelleted into agarose and processed the usual way to wax. Once a method is seen to be working I intend to use human tissue. Does anybody have any experience / methodology for using a XIAP antibody, as there seems to be several available looking thro the literature. Many thanks in advance. Garry PICR UK -------------------------------------------------------- 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 and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. 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. From Barry.R.Rittman <@t> uth.tmc.edu Mon Mar 15 08:24:31 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] MicroFil Message-ID: <566FB0B522443D43AF02D2ADBE35A6F06359AE@UTHEVS3.mail.uthouston.edu> Sven I used Microfil latex several years ago to demonstrate blood vessels in ginigiva, mostly using dogs for soemone elses project. The following is from memory as I am at work. The Microfil solution is a little viscous but can be perfused into even the smallest capillaries. We used a peristaltic pump first rinsing out the blood using saline (with heparin added) until the lips blanched followed by the final Microfil mixture. All solutions were at 37 degrees C. The Microfil was prpeared only 15 minutes before use and bubbles removed using an ultrasonic bath so that they would not become trapped in small vessels and block them. Care must be taken not to use too much presure with the pump or else some vesels will be ruptured. Once the latex had set we then immersed blocks in buffered formalin. Some blocks were dehydrated and cleared in methyl salicylate to visualize the blood vessels. Tissue with bone and teeth were demineralized using 1% formic acid before dehydrating and clearing. Other tissue were processed to paraffin wax for evaluation. The Microfil survived the processing in all procedures. In the paraffin wax however the Microfil tended to shrink somewhat in diameter so that the diameter of some vessels appeared initially to be smaller. We used a combination of blocks and paraffin sections to evaluate blood vessels in the gingiva. Never carried out immunofluorescence on these samples but see no logical reason why this could not be done. Hope that this helps Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sven Terclavers Sent: Monday, March 15, 2004 4:54 AM To: Mailinglist_Histology Subject: [Histonet] MicroFil Dear listers, I'm looking for a solution to perfuse bloodvessels which hardens after cooling down or after a certain timepoint of being liquid to fill not only larger bloodvessels but also cappillars. On the internet I found MicroFil, now I was wondering if some of you already tried this. I would like to know the following: -What is the smallest cappillar that will be filled? -Can sections be made without the MicroFil falling out of the cappillars? -Can this MicroFil solution be mixed with e.g. FITC to perform epi-fluorescence/CLS-microscopy? Thanks in advance, Sven Terclavers _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Joyce.Smith <@t> FLHOSP.ORG Mon Mar 15 09:14:48 2004 From: Joyce.Smith <@t> FLHOSP.ORG (Smith, Joyce (FH)) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] IHC Message-ID: Hi List, Our IHC person is looking for a protocol for MLH-1 - they are having a problem in getting it to work. The power setting on their microwave is not set up like the procedure and they are using colon as the suggested control, but don't know if it is working. Please let us know what you would suggest we try. Thanks Joyce Florida Hospital The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From jen.jones <@t> tufts.edu Mon Mar 15 09:20:34 2004 From: jen.jones <@t> tufts.edu (Jennifer A Jones) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] smooth muscle alpha actin antibody for mouse cells and mouse tissue? Message-ID: <4055C9C2.90705@tufts.edu> Hi everyone - I want to stain mouse tissues and smooth muscle cells in culture for alpha-actin. Apparently clone 1A4 (mouse monoclonal) is quite popular, but my PI wants me to find an antibody from a different species (rabbit, for example) to prevent excessive background staining. Any recommendations? And what dilutions do you use? Thank you!! - Jen From escott8 <@t> houston.rr.com Mon Mar 15 09:45:12 2004 From: escott8 <@t> houston.rr.com (Edward Scott) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Georgia Society App Message-ID: <000301c40aa4$83c979b0$c5e2f218@thescotts> I just missed a fax from Mercer Univ. Please fax me again. Thanks Allison Scott. From gcallis <@t> montana.edu Mon Mar 15 09:52:32 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] smooth muscle alpha actin antibody for mouse cells andmouse tissue? In-Reply-To: <4055C9C2.90705@tufts.edu> Message-ID: <3.0.6.32.20040315085232.00bc2ff0@gemini.msu.montana.edu> With mouse on mouse IHC staining, have you tried the DAKO ARK kit? We also had less background using the Chemicon Mouse on Mouse kit, they will supply you with a free sample kit, so be sure to ask for one. Nice when a company does that for us. At 10:20 AM 3/15/2004 -0500, you wrote: >Hi everyone - I want to stain mouse tissues and smooth muscle cells in >culture for alpha-actin. Apparently clone 1A4 (mouse monoclonal) is >quite popular, but my PI wants me to find an antibody from a different >species (rabbit, for example) to prevent excessive background staining. >Any recommendations? And what dilutions do you use? Thank you!! - Jen Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Stephen.Spencer <@t> intervet.com Mon Mar 15 10:00:08 2004 From: Stephen.Spencer <@t> intervet.com (Spencer, S (Stephen)) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] UK histotechnician Message-ID: <6BE0EFB134CAD611B0A400408102ECD14E1CFD@mksn71.mks.intra> Dear list, I would like to post an advertisement for a vacant histopathology technician position in the UK. Could you please advise on which journal(s) or web sites carry such situations vacant and are likely to be read by potential candidates? Thank you, _________________________ Stephen Spencer Research Veterinarian BSD - Intervet UK Milton Keynes United Kingdom -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- From Sarah.Blachford <@t> ngh.nhs.uk Mon Mar 15 09:04:04 2004 From: Sarah.Blachford <@t> ngh.nhs.uk (Blachford, Sarah - Histopath Main) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] pemphigoid Message-ID: <854FA06D8BE2D611ABA30030052D1DF6FA58E4@NGH_EXCHANGE1> Dear anyone, I want to look at pemphigoid/ pemphigus but do not want to use immunofluorescence. Can anyone help. Sarah Blachford Northampton General Hospital Billing Road Northants NN1 5BD England Sarah.Blachford@ngh.nhs.uk Northampton General Hospital NHS Trust Cliftonville, Northampton NN1 5BD This e-mail may contain confidential information and/or copyright material and is intended for the use of the addressee only. Any unauthorised use may be unlawful. The contents of this e-mail may be subject to public disclosure under the NHS Code of Openness or the Freedom of Information Act 2000. Unless legally exempt, the confidentiality of the message and your reply cannot be guaranteed. If you receive this e-mail by mistake, please advise the sender immediately. Thank you. From nick.kirk3 <@t> btopenworld.com Mon Mar 15 11:26:07 2004 From: nick.kirk3 <@t> btopenworld.com (=?iso-8859-1?q?NICK=20KIRK?=) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] UK histotechnician In-Reply-To: <6BE0EFB134CAD611B0A400408102ECD14E1CFD@mksn71.mks.intra> Message-ID: <20040315172607.41630.qmail@web86310.mail.ukl.yahoo.com> The one most BMS's (histology technicians) read is the Biomedical Scientist. Contact details are: Alison Jones The Biomedical Scientist Step Publishing Ltd Step House North farm Road Tunbridge Wells Kent TN2 3DR Tel: 01892 518877 Fax: 01892 616177 Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England "Spencer, S (Stephen)" wrote: Dear list, I would like to post an advertisement for a vacant histopathology technician position in the UK. Could you please advise on which journal(s) or web sites carry such situations vacant and are likely to be read by potential candidates? Thank you, _________________________ Stephen Spencer Research Veterinarian BSD - Intervet UK Milton Keynes United Kingdom -------------------------------------- This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. -------------------------------------- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From meligroc <@t> zgi.com Mon Mar 15 11:34:54 2004 From: meligroc <@t> zgi.com (CRME (Criss Meligro)) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] flat bed scanners Message-ID: Wish to know the most used flat bed scanner for gross photography of small samples (mouse kidneys). I played a little with my home HP photo scanner and we seem pleased but in purchasing one for the lab I would like to know what people are using or if they are.... thanks! Criss From funderwood <@t> mcohio.org Mon Mar 15 11:42:37 2004 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Forcep Sterilization Message-ID: A toothbrush or test tube brush works well to clean out the grooves of the forceps. Have a bucket with soapy water handy and change it as frequently as possible. 4 out of 5 dentists recommend not re-using the toothbrush on your teeth, however. >>> 03/13/04 08:52AM >>> We are having a problem with carry-over at the grossing station. We do not have a sink available and are presently dipping our forceps in ethanol. Is there another solution we can use besides ethanol? Does anyone have any other ideas as how to avoid this contamination issue we are experiencing? Thank you Ann Angelo _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Mon Mar 15 12:24:18 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] EGFR - My thoughts In-Reply-To: Message-ID: Rich, Thank you for your insight on EGFR. Indeed it will be interesting to see how this all comes out in a year or so. I hope that we all continue to share our experiences so that we can do the best we can for the patients involved. I am so pleased that the DAKO kit does not require HIER and uses enzyme digestion instead. HIER for her2 has been a night mare for those of us at altitude. Best regards, Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Richard Cartun Sent: Sunday, March 14, 2004 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EGFR - My thoughts Approximately two years ago, I said to my colleagues that EGFr testing would make HER-2/neu testing look like "a walk in the park" (i.e, very simple). We tested DAKO's EGFR kit back then and compared it to the monoclonal that we were using at the time (clone 31G7 from Zymed). Although the results were similar (but not identical), the mAb from Zymed consistently produced stronger immunoreactivity which was easier to evaluate. Like you, we have received many requests during the past 3 weeks from our clinical colleagues to test their patients' colorectal tumors for EGFR so that they can be considered eligible for treatment with "Erbitux" (the anti-EGFR mAb drug). Although I started with the Zymed mAb, I am now convinced that this testing must be performed with DakoCytomation's "EGFR pharmDx" kit (primarily for reimbursement issues). Although the kit works well, I find the resulting immunoreactivity very difficult to score (this is true with the Zymed mAb as well). Most tumors have shown what I call "wishy-washing" staining that, if it were HER-2, I would call it negative. However, we are instructed to score it 1+ "Positive". I find it difficult to believe that "Erbitux" will help a patient whose tumor shows rare to focal immunoreactivity and often less intense than normal cells present in the same tissue section; however, I am not familiar with the clinical trials and patient outcomes. I have also noticed EGFR variability from block to block in the same patient when we have tested multiple tissue blocks. In addition, in my experience, it is very unusual to find a tumor that shows the classic "3+" staining pattern that we see with c-erbB-2 (HER-2/neu protein) in breast CA's with obvious gene amplification. It will be interesting to see where we are 6 months to a year from now with EGFR testing. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From EMarquez <@t> crch.hawaii.edu Mon Mar 15 14:19:06 2004 From: EMarquez <@t> crch.hawaii.edu (Eddie Marquez) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] xylene substitute Message-ID: Aloha from Hawaii, Just needed some info.-Has anybody used xylene substitute with a linear stainer; for standard H&E's? If so, how good are the results? May I also have the brand name. Thank you, Eddie from the Cancer Research From ramona.tolliver <@t> yale.edu Mon Mar 15 14:51:33 2004 From: ramona.tolliver <@t> yale.edu (Ramona Tolliver) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Yale University - Opening Message-ID: <6.0.1.1.2.20040315154144.01d4cee8@email.med.yale.edu> Good afternoon, Yale University has a Histology Manager (1st shift) position available from 7:00 a.m. - 3:00 p.m. If you know of anyone who may be interested in an outstanding growth opportunity in a department committed to excellence in patient care, teaching, and discovery, please forward this information on to them. Salary is commensurate with experience. Relocation assistance is available. Inquiries should be sent to ramona.tolliver@yale.edu. Thank you for your time! Ramona Tolliver Ramona E. Tolliver Human Resource Manager Yale University School of Medicine Department of Pathology 310 Cedar Street New Haven, CT 06520-8023 Telephone: (203) 785-6689 Fax: (203) 785-7303 From Diane.Gladney <@t> se.amedd.army.mil Mon Mar 15 15:00:52 2004 From: Diane.Gladney <@t> se.amedd.army.mil (Gladney, Diane C Ms MACH) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] xylene substitute Message-ID: <9D41AB7C56F8304F98537ABD87B249F66261A4@dasmthgbz001.amedd.army.mil> Eddy, I have used Thermo Electron (formerly Shandon) Xylene Substitute in my auto-stainer for years without any problems. BUT I only use it in the deparaffinization at the beginning of the staining series. I use Xylene in the last three stations of the stainer. Our mounting medium has xylene as the solvent so the substitute is not compatible with the mounting medium. I have tried using the xylene substitute mounting media and I do not like the results. Therefore, we stay with a good old tried and true mounting medium. At least we have considerably cut our use of Xylene as we also use the same xylene substitute in our processors. If you have any other questions, please feel free to contact me. Happy Cutting, Diane Diane C. Gladney, HT (ASCP) Histology /Cytology Supervisor Moncrief Army Community Hospital P.O. BOX 484 4500 Stuart Ave. FT. Jackson, SC 29207 (803) 751-2530 DSN 734-2530 EMAIL: diane.gladney@se.amedd.army.mil OR dcgx1@aol.com -----Original Message----- From: Eddie Marquez [mailto:EMarquez@crch.hawaii.edu] Sent: Monday, March 15, 2004 3:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene substitute Aloha from Hawaii, Just needed some info.-Has anybody used xylene substitute with a linear stainer; for standard H&E's? If so, how good are the results? May I also have the brand name. Thank you, Eddie from the Cancer Research _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kbowden <@t> ucsd.edu Mon Mar 15 15:01:17 2004 From: kbowden <@t> ucsd.edu (K. Bowden) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] flat bed scanners References: Message-ID: <4056199D.2030301@ucsd.edu> Agfa Arcus and Agfa Duoscan T1200. We the like the software as well and the quility of the scanner. -- Karen Bowden Staff Research Associate II University of CA, San Diego Department of Orthopedics 9500 Gilman Dr. 0630 La Jolla, CA 92093-0630 858-277-4970 CONFIDENTIALITY NOTICE: THE INFORMATION TRANSMITTED IN THIS E-MAIL IS INTENDED ONLY FOR THE PERSON OR ENTITY TO WHICH IT IS ADDRESSED AND MAY CONTAIN CONFIDENTIAL AND/OR PRIVILEGED MATERIAL. ANY REVIEW, RETRANSMISSION, DISSEMINATION OR OTHER USE OF OR TAKING OF ANY ACTION IN RELIANCE UPON THIS INFORMATION BY PERSONS OR ENTITIES OTHER THAN THE INTENDED RECIPIENT IS PROHIBITED. IF YOU RECEIVED THIS E-MAIL IN ERROR, PLEASE CONTACT THE SENDER AND DELETE THE MATERIAL FROM ANY COMPUTER. CRME (Criss Meligro) wrote: > Wish to know the most used flat bed scanner for gross photography of small samples (mouse kidneys). I played a little with my home HP photo scanner and we seem pleased but in purchasing one for the lab I would like to know what people are using or if they are.... thanks! Criss > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From zandra <@t> gwu.edu Mon Mar 15 15:03:10 2004 From: zandra <@t> gwu.edu (Alexandra de Sousa) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] imidizole? Message-ID: <1ab8d121ab6a61.1ab6a611ab8d12@gwu.edu> I have been using imidizole in cytochrome oxidase reactions and have been ordering from Sigma-Aldrich, catalog #I0250. They are backordered for the quantity I need until June so I was wondering whether anyone know of another supplier I could order from. Thanks! Alexandra From TJasper <@t> smdc.org Mon Mar 15 15:36:30 2004 From: TJasper <@t> smdc.org (Jasper, Thomas) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Lower Hutt, New Zealand Message-ID: <1DB04B57B04C5747B87C3B39F3E605AA02E44E53@harrier> I'm quite certain I've seen postings from Lower Hutt, New Zealand on this server before. Would someone from there please contact me. I apologize in advance if I've incorrectly referenced the community or region. A reply would be much appreciated. Thank you! Thomas G. Jasper Anatomic Pathology Coordinator SMDC Duluth, MN 55805 (218) 786-4510 tjasper@smdc.org From gsennello <@t> osip.com Mon Mar 15 16:25:42 2004 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] EGFR - Message-ID: I was wondering if those of you who use the ACIS by ChromaVision to score your Hercept slides are planning to use it to score the EGFR PharmDx? If so I would be interested in knowing what criteria will be used to. We do not do Hercept but are looking at EGFR PharmDx. Thanks, Gina Gina Sennello Associate Scientist/ Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Sunday, March 14, 2004 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EGFR - My thoughts Approximately two years ago, I said to my colleagues that EGFr testing would make HER-2/neu testing look like "a walk in the park" (i.e, very simple). We tested DAKO's EGFR kit back then and compared it to the monoclonal that we were using at the time (clone 31G7 from Zymed). Although the results were similar (but not identical), the mAb from Zymed consistently produced stronger immunoreactivity which was easier to evaluate. Like you, we have received many requests during the past 3 weeks from our clinical colleagues to test their patients' colorectal tumors for EGFR so that they can be considered eligible for treatment with "Erbitux" (the anti-EGFR mAb drug). Although I started with the Zymed mAb, I am now convinced that this testing must be performed with DakoCytomation's "EGFR pharmDx" kit (primarily for reimbursement issues). Although the kit works well, I find the resulting immunoreactivity very difficult to score (this is true with the Zymed mAb as well). Most tumors have shown what I call "wishy-washing" staining that, if it were HER-2, I would call it negative. However, we are instructed to score it 1+ "Positive". I find it difficult to believe that "Erbitux" will help a patient whose tumor shows rare to focal immunoreactivity and often less intense than normal cells present in the same tissue section; however, I am not familiar with the clinical trials and patient outcomes. I have also noticed EGFR variability from block to block in the same patient when we have tested multiple tissue blocks. In addition, in my experience, it is very unusual to find a tumor that shows the classic "3+" staining pattern that we see with c-erbB-2 (HER-2/neu protein) in breast CA's with obvious gene amplification. It will be interesting to see where we are 6 months to a year from now with EGFR testing. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Mon Mar 15 18:13:56 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] EGFR - Message-ID: I don't think you need a machine to score the EGFR immunoreactivity. Richard Cartun >>> "Sennello, Gina" 03/15/04 05:25PM >>> I was wondering if those of you who use the ACIS by ChromaVision to score your Hercept slides are planning to use it to score the EGFR PharmDx? If so I would be interested in knowing what criteria will be used to. We do not do Hercept but are looking at EGFR PharmDx. Thanks, Gina Gina Sennello Associate Scientist/ Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 -----Original Message----- From: Richard Cartun [mailto:Rcartun@harthosp.org] Sent: Sunday, March 14, 2004 10:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] EGFR - My thoughts Approximately two years ago, I said to my colleagues that EGFr testing would make HER-2/neu testing look like "a walk in the park" (i.e, very simple). We tested DAKO's EGFR kit back then and compared it to the monoclonal that we were using at the time (clone 31G7 from Zymed). Although the results were similar (but not identical), the mAb from Zymed consistently produced stronger immunoreactivity which was easier to evaluate. Like you, we have received many requests during the past 3 weeks from our clinical colleagues to test their patients' colorectal tumors for EGFR so that they can be considered eligible for treatment with "Erbitux" (the anti-EGFR mAb drug). Although I started with the Zymed mAb, I am now convinced that this testing must be performed with DakoCytomation's "EGFR pharmDx" kit (primarily for reimbursement issues). Although the kit works well, I find the resulting immunoreactivity very difficult to score (this is true with the Zymed mAb as well). Most tumors have shown what I call "wishy-washing" staining that, if it were HER-2, I would call it negative. However, we are instructed to score it 1+ "Positive". I find it difficult to believe that "Erbitux" will help a patient whose tumor shows rare to focal immunoreactivity and often less intense than normal cells present in the same tissue section; however, I am not familiar with the clinical trials and patient outcomes. I have also noticed EGFR variability from block to block in the same patient when we have tested multiple tissue blocks. In addition, in my experience, it is very unusual to find a tumor that shows the classic "3+" staining pattern that we see with c-erbB-2 (HER-2/neu protein) in breast CA's with obvious gene amplification. It will be interesting to see where we are 6 months to a year from now with EGFR testing. Richard Cartun Director, Immunopathology Hartford Hospital Hartford, CT _______________________________________________ Histonet mailing list Histonet@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 CrochiereSteve <@t> aol.com Mon Mar 15 19:03:34 2004 From: CrochiereSteve <@t> aol.com (CrochiereSteve@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions for small project. Message-ID: <104.4167f727.2d87ac66@aol.com> Sue, I once did a little project at AFIP involving a Slim-Jim and a bunch of different special stains to demonstrate what was actually in those tasty little meat snax. Steven M. Crochiere, HT (ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew St. Springfield, MA 01104 From lpwenk <@t> covad.net Mon Mar 15 20:06:04 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Georgia Society for Histotechnology Info. References: <000001c40a1f$8b0ff800$c5e2f218@thescotts> Message-ID: <005f01c40afb$3d6deda0$acd3d445@domainnotset.invalid> For a list of all the states with histology societies, along with the presidents' names, phone numbers and email addresses (in case anyone else wants to join their state society) (hint, hint), go to the National Society for Histotechnology (NSH) web site, under state membership: http://www.nsh.org/membership/constituentsoc.html If you want information about joining the NSH (hint, hint, nudge, nudge), go to: http://www.nsh.org/membership/ and click on forms. You do NOT have to be a certified HT or HTL to join NSH or any histology state society. Just have to be someone interested in the field of histology. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Edward Scott" To: Sent: Sunday, March 14, 2004 6:53 PM Subject: [Histonet] Georgia Society for Histotechnology Info. > Hello to all in histoland. I would like for someone from the Georgia > society to contact me. I would like an application. The information > that I have is outdated. You can email me or fax me. My fax number is > 281-540-2106. Thanks in advance. > Allison Scott. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From peptolab <@t> hamptons.com Mon Mar 15 19:52:03 2004 From: peptolab <@t> hamptons.com (peptolab) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Ergonomics Policy Message-ID: <000a01c40af9$4888dd00$6e7dbd18@JEFF> I recently underwent CAP inspection and had to write a lab safety ergonomics policy- I think it was eventually used by all hopitals in our large multihospital system, after tweaking for institutional varitions. Sorry folks, but I'm not going to just post the finished product but here's how I approached it: I did a google search for "laboratory ergonomics". Numerous governmental, industrial, and educational websites came up with a wide array of ergonomics information broken down for for various lab tasks as well as general approaches for monitoring and investigating complaints. I shamelessly cut and pasted sections from all these websites (including the sites in the references for the final policy) until I had covered precautions and concerns for every activity in our lab. That was a lot of fun actually. Then I asked the employee health nurse and lab management about the specific steps to be followed by employees who want to evalutae a percieved problem and included these in the policy. Voila. Jeff Silverman HT HTL QIHC ASCP Pathologists' Assistant-Lab Safety Officer Southside Hospital North Shore Long Island Jewish Hospital System Bay Shore New York USA From Histopatty <@t> aol.com Mon Mar 15 20:37:33 2004 From: Histopatty <@t> aol.com (Histopatty@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Alport's Message-ID: <17.43cf6c2d.2d87c26d@aol.com> I am looking for an antibody or kit for type IV collagen in Alport's Syndrome. The kit we currently use is expired and it is from Sweden, and to reorder would be a huge deal to say the least. Any help would be appreciated. Thank you Patty Eneff OU MEDICAL CENTER Patricia.Eneff@hcahealthcare.com From DDDeltour <@t> sig.med.navy.mil Tue Mar 16 00:08:14 2004 From: DDDeltour <@t> sig.med.navy.mil (Deltour, Douglas D.(HM2)) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] xylene substitute Message-ID: Eddie, I use Histo-Clear made by national diagnostics. It is the best substitute that I have found. http://www.nationaldiagnostics.com/. I spent three years in Hawaii. Lived in Kailua. I miss it. Take care Doug -----Original Message----- From: Eddie Marquez [mailto:EMarquez@crch.hawaii.edu] Sent: Monday, March 15, 2004 9:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] xylene substitute Aloha from Hawaii, Just needed some info.-Has anybody used xylene substitute with a linear stainer; for standard H&E's? If so, how good are the results? May I also have the brand name. Thank you, Eddie from the Cancer Research _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This document may contain information covered under the privacy Act, 5 USC 552(a), and/or the Health Insurance Portability and Accountability Act (PL 104-191) and its various implementing regulations and must be protected in accordance with those provisions. Healthcare information is personal and sensitive and must be treated accordingly. If this correspondence contains healthcare information it is being provided to you after applying the appropriate security controls and authorization from the patient, or under circumstances that don't require patient authorization. You, the recipient, are obligated to maintain it in a safe, secure and confidential manner. Redisclosure without additional patient consent or as permitted by law is prohibited. Unauthorized redisclosure or failure to maintain confidentiality subjects you to application of appropriate sanction. If you have received this correspondence in error, please notify the sender and the command Privacy Officer at privacy@sig.med.navy.mil at once and destroy any copies you have made. From carl.hobbs <@t> kcl.ac.uk Tue Mar 16 02:15:14 2004 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] re smooth muscle actin Message-ID: <001901c40b2e$cf6ffa80$e8345c9f@Carlos> I use DakoM851 and Sigma A 2547 on mouse FFPWP tissues with std StreptABCpx DAB system with no problems with b'ground . So, cells shouldn't present with any problems. Embyronic skeletal muscle will also be +ve with Dako reagent if using HIER. From siksik03 <@t> comcast.net Tue Mar 16 06:31:21 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] IHC In-Reply-To: References: Message-ID: Dear HistoNetters It would help, Joyce, to know exactly what the procedure calls for, and how your microwave is set up. If you can provide this information, someone might be able to help. best regards, Steven Slap Microwave Consultant At 10:14 AM -0500 3/15/04, Smith, Joyce (FH) wrote: >From: "Smith, Joyce (FH)" >To: "'Histonet@pathology.swmed.edu'" >Cc: >Subject: [Histonet] IHC > >Hi List, >Our IHC person is looking for a protocol for MLH-1 - they are having a >problem in getting it to work. >The power setting on their microwave is not set up like the procedure and >they are using colon as the suggested control, but don't know if it is >working. >Please let us know what you would suggest we try. >Thanks >Joyce >Florida Hospital From siksik03 <@t> comcast.net Tue Mar 16 06:38:36 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] UK histotechnician In-Reply-To: <6BE0EFB134CAD611B0A400408102ECD14E1CFD@mksn71.mks.intra> References: <6BE0EFB134CAD611B0A400408102ECD14E1CFD@mksn71.mks.intra> Message-ID: Dear HistoNetters "The Histotech's Home Page" accepts free job postings for its "Jobs in Histology" page (http://www.histology.to/jobs.html). The last figures I have indicate that the site is visited by approximately 300,000 histotechs a month, of which about 8000 are from the UK. best regards, Steven Slap co-webmaster > "Spencer, S (Stephen)" >To: "'histonet@lists.utsouthwestern.edu'" >Subject: [Histonet] UK histotechnician > >Dear list, > >I would like to post an advertisement for a vacant histopathology technician >position in the UK. Could you please advise on which journal(s) or web sites >carry such situations vacant and are likely to be read by potential >candidates? > >Thank you, > >_________________________ >Stephen Spencer >Research Veterinarian >BSD - Intervet UK >Milton Keynes >United Kingdom From bill501 <@t> mindspring.com Tue Mar 16 07:30:11 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] flat bed scanners In-Reply-To: References: Message-ID: For 3D objects CCD scanners are much better than the cheaper CMOS scanners, for depth of field reasons. There are a number out there, but you have to look at the specs. I use UMAX Powerlook IIIs. Bill At 9:34 AM -0800 3/15/04, CRME (Criss Meligro) wrote: >Wish to know the most used flat bed scanner for gross photography of >small samples (mouse kidneys). -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) http://www.druidry/org http://www.druidry.org/board From Terry.Marshall <@t> rothgen.nhs.uk Tue Mar 16 08:08:38 2004 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] flat bed scanners Message-ID: Bill, This is fascinating information, but we could do with knowing a bit more. What are CCD and CMOS? How do you tell what scanner is/does/uses what? Look at the specification for what? What is good and what is bad? This is a completely new one to me, and I'm sure we would all like to here a fuller story. (See what you've done now - let yourself in for it :-) ) Dr Terry L Marshall, B.A.(Law), M.B.,Ch.B.,F.R.C.Path Consultant Pathologist Rotherham General Hospital South Yorkshire England terry.marshall@rothgen.nhs.uk -----Original Message----- From: Bill Blank [mailto:bill501@mindspring.com] Sent: 16 March 2004 13:30 To: CRME (Criss Meligro); Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] flat bed scanners For 3D objects CCD scanners are much better than the cheaper CMOS scanners, for depth of field reasons. There are a number out there, but you have to look at the specs. I use UMAX Powerlook IIIs. Bill At 9:34 AM -0800 3/15/04, CRME (Criss Meligro) wrote: >Wish to know the most used flat bed scanner for gross photography of >small samples (mouse kidneys). -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) http://www.druidry/org http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mark.lewis <@t> thermo.com Tue Mar 16 08:44:58 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions for small project. Message-ID: So Steve, What did you find ?? I'm curious...I never eat them, but I'm still curious.. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com CrochiereSteve@aol.com Sent by: To: suselore@ihug.co.nz, histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: Re: [Histonet] Junior Histo worker seeks suggestions for small project. 03/15/2004 08:03 PM Sue, I once did a little project at AFIP involving a Slim-Jim and a bunch of different special stains to demonstrate what was actually in those tasty little meat snax. Steven M. Crochiere, HT (ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew St. Springfield, MA 01104 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BlazekL <@t> childrensdayton.org Tue Mar 16 08:51:07 2004 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions forsmall project. Message-ID: If you ever saw what was in them you would never eat them again! Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center 1 Children's Plaza Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org >>> 03/16/04 09:44AM >>> So Steve, What did you find ?? I'm curious...I never eat them, but I'm still curious.. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com CrochiereSteve@aol.com Sent by: To: suselore@ihug.co.nz, histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: Re: [Histonet] Junior Histo worker seeks suggestions for small project. 03/15/2004 08:03 PM Sue, I once did a little project at AFIP involving a Slim-Jim and a bunch of different special stains to demonstrate what was actually in those tasty little meat snax. Steven M. Crochiere, HT (ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew St. Springfield, MA 01104 _______________________________________________ Histonet mailing list Histonet@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 vherrington <@t> sumterregional.org Tue Mar 16 08:53:25 2004 From: vherrington <@t> sumterregional.org (Herrington, Vicki) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Leica RM 2025 Knife Holder Message-ID: <3E09D94FAC8AD411A04900D0B77E645002815520@SRHNTMAIL> I am trying to locate a Leica RM 2025 knife holder for low-profile blades. This is the one that has screws on the front plate for tightening and a lever on the right to release the blade. If you have an old one for sale, or can tell me where I might find used microtome parts, I'd would appreciate it! Thanks, Vicki Herrington, M.S. AP Department Director Sumter Regional Hospital Americus, GA 31709 PH: 229-931-1150 Note: The information contained in this message and any attachments 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 reading, dissemination, distribution, copying, or other use of this communication or any of its attachments is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting this message, any attachments, and all copies and backups from your computer. If you have received this communication in error and are unable to reply to this message, please notify the sender immediately by telephone at (229) 931-1380. Thank you. Sumter Regional Hospital From Barry.R.Rittman <@t> uth.tmc.edu Tue Mar 16 09:00:41 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions forsmall project. Message-ID: <566FB0B522443D43AF02D2ADBE35A6F06359B1@UTHEVS3.mail.uthouston.edu> As far as I understand, the industry making hot dogs etc. usually have a requirement that the grinding process is such that tissue within a section can only be identified above a certain magnification. This is I think chiefly so that the public will not be put off by finding easily identifiable items in the meat. Thus the need for histology of these meats - in fact the sections are really interesting. We should have a competition at conventions to ask attendees to identify individual components although this might decrease the consumption of these products. Perhaps there should be a subdivision of histopathology, a title of wiernerologist, patho-frankologist, patho-bolognologist spring to mind. Now that the seed has been sown perhaps we shall get some interesting suggestions to take our minds of the more serious problems? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mark.lewis@thermo.com Sent: Tuesday, March 16, 2004 8:45 AM To: CrochiereSteve@aol.com Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Junior Histo worker seeks suggestions forsmall project. So Steve, What did you find ?? I'm curious...I never eat them, but I'm still curious.. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com CrochiereSteve@aol.com Sent by: To: suselore@ihug.co.nz, histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: Re: [Histonet] Junior Histo worker seeks suggestions for small project. 03/15/2004 08:03 PM Sue, I once did a little project at AFIP involving a Slim-Jim and a bunch of different special stains to demonstrate what was actually in those tasty little meat snax. Steven M. Crochiere, HT (ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew St. Springfield, MA 01104 _______________________________________________ Histonet mailing list Histonet@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 rjr6 <@t> psu.edu Tue Mar 16 09:08:20 2004 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] A little lab safety humor Message-ID: <001901c40b68$85055500$8861ba92@padlspsu.psu.edu> The following is an excerpt from the university's environmental health and safety compliance document. According to this I am not compliant. 1. Inspect storage areas and inventory stored materials immediately and annually to ensure that they are safety and secure and not accessible to authorized personnel. Roberta Horner HT/HTL Penn State University Animal Diagnostic Lab From Jackie.O'Connor <@t> abbott.com Tue Mar 16 09:12:51 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions forsmall project. Message-ID: Let's not forget the patho-bratologist - I once sectioned a bratwurst for a German pathologist (Hans Dolz) - he never ate bratwurst again. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics "Barry R Rittman" Sent by: histonet-bounces@lists.utsouthwestern.edu 03/16/2004 09:00 AM To: cc: Subject: RE: [Histonet] Junior Histo worker seeks suggestions forsmall project. As far as I understand, the industry making hot dogs etc. usually have a requirement that the grinding process is such that tissue within a section can only be identified above a certain magnification. This is I think chiefly so that the public will not be put off by finding easily identifiable items in the meat. Thus the need for histology of these meats - in fact the sections are really interesting. We should have a competition at conventions to ask attendees to identify individual components although this might decrease the consumption of these products. Perhaps there should be a subdivision of histopathology, a title of wiernerologist, patho-frankologist, patho-bolognologist spring to mind. Now that the seed has been sown perhaps we shall get some interesting suggestions to take our minds of the more serious problems? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mark.lewis@thermo.com Sent: Tuesday, March 16, 2004 8:45 AM To: CrochiereSteve@aol.com Cc: histonet@lists.utsouthwestern.edu; histonet-bounces@lists.utsouthwestern.edu Subject: Re: [Histonet] Junior Histo worker seeks suggestions forsmall project. So Steve, What did you find ?? I'm curious...I never eat them, but I'm still curious.. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com CrochiereSteve@aol.com Sent by: To: suselore@ihug.co.nz, histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: Re: [Histonet] Junior Histo worker seeks suggestions for small project. 03/15/2004 08:03 PM Sue, I once did a little project at AFIP involving a Slim-Jim and a bunch of different special stains to demonstrate what was actually in those tasty little meat snax. Steven M. Crochiere, HT (ASCP) Histology Supervisor LifePath Partners @ Mercy Medical Center 299 Carew St. Springfield, MA 01104 _______________________________________________ Histonet mailing list Histonet@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 peoshel <@t> wisc.edu Tue Mar 16 09:13:27 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions forsmall project. In-Reply-To: <566FB0B522443D43AF02D2ADBE35A6F06359B1@UTHEVS3.mail.uthouston.edu> References: <566FB0B522443D43AF02D2ADBE35A6F06359B1@UTHEVS3.mail.uthouston.edu> Message-ID: Given the fat content of brats, should they only be cryosectioned? Phil >As far as I understand, the industry making hot dogs etc. usually have >a requirement that the grinding process is such that tissue within a >section can only be identified above a certain magnification. This is I >think chiefly so that the public will not be put off by finding easily >identifiable items in the meat. Thus the need for histology of these >meats - in fact the sections are really interesting. We should have a >competition at conventions to ask attendees to identify individual >components although this might decrease the consumption of these >products. >Perhaps there should be a subdivision of histopathology, a title of >wiernerologist, patho-frankologist, patho-bolognologist spring to mind. >Now that the seed has been sown perhaps we shall get some interesting >suggestions to take our minds of the more serious problems? >Barry -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From escott8 <@t> houston.rr.com Tue Mar 16 09:22:33 2004 From: escott8 <@t> houston.rr.com (Edward Scott) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] CAP Check List Message-ID: <000001c40b6a$846a3b90$c5e2f218@thescotts> I just received my Cap self check list and I'm having a problem with the question ANP.23075 : Is there evidence of ongoing evaluation of results of instrument maintenance and functions for all devices. Would having maintenance charts and a policy of how often instruments are checked by Bio Med and or the company from which the instrument was purchased be enough. I also have all of the PM/ service records on all instruments that require annual checks. Any help with this would be appreciated. Thanks in advance. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas From mcauliff <@t> umdnj.edu Tue Mar 16 12:41:31 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Re: Microscopy of food/was Junior Histo worker seeks suggestions forsmall project. In-Reply-To: References: <566FB0B522443D43AF02D2ADBE35A6F06359B1@UTHEVS3.mail.uthouston.edu> Message-ID: <40574A5B.5030800@umdnj.edu> I have a copy of a parody/satire/vallstein (inside joke for Seinfeld fans), "Fine Structural studies of common meat and dairy products using the Electron Microscopy I. Liverwurst." that I got from one of my mentors about 30 years ago. Pokes fun at EM and the critics of EM. Very funny. I have no idea where it was originally published. Geoff Philip Oshel wrote: > Given the fat content of brats, should they only be cryosectioned? > Phil > >> As far as I understand, the industry making hot dogs etc. usually have >> a requirement that the grinding process is such that tissue within a >> section can only be identified above a certain magnification. This is I >> think chiefly so that the public will not be put off by finding easily >> identifiable items in the meat. Thus the need for histology of these >> meats - in fact the sections are really interesting. We should have a >> competition at conventions to ask attendees to identify individual >> components although this might decrease the consumption of these >> products. >> Perhaps there should be a subdivision of histopathology, a title of >> wiernerologist, patho-frankologist, patho-bolognologist spring to mind. >> Now that the seed has been sown perhaps we shall get some interesting >> suggestions to take our minds of the more serious problems? >> Barry > > -- -- ********************************************** 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 aostrand <@t> seattlecca.org Tue Mar 16 09:44:17 2004 From: aostrand <@t> seattlecca.org (Ostrander, Anita B) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Shandon Varistain Gemini Message-ID: <5E6BFDF4F0AB2C4DA69CF4473FC7B94823B50D@ldap.seattlecca.org> Hi, Our Laboratory is currently in the process of purchasing an automated H&E stainer. We are testing out the Shandon Varistain Gemini. I would greatly appreciate any feedback (positive, negative or indifferent) that anyone could give me. If you are more comfortable sending me feedback directly; my e-mail is aostrand@seattlecca.org. Thank you! Anita Anita Ostrander Pathology Manager Seattle Cancer Care Alliance Office:206-288-1347 Cell Phone: 206-226-0785 aostrand@seattlecca.org This electronic message transmission contains information which may be confidential or privileged. The information is intended to be for the use of the individual or entity named above. If you are not the intended recipient, be aware that any disclosure, copying, distribution or use of the contents of this information is prohibited. If you have received this electronic transmission in error, please leave a message via telephone at (206) 288-6266, notify me by electronic reply, and delete this message. Opinions and ideas in this message that do not relate to official business are understood as neither given nor endorsed by the Seattle Cancer Care Alliance. To view our complete Notice of Privacy Practices, visit our web site at www.seattlecca.org. From Djemge <@t> aol.com Tue Mar 16 10:04:37 2004 From: Djemge <@t> aol.com (Djemge@aol.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] AO parts and Histology equipment manuals Message-ID: Hi all, this is my first time adding a message. Does anyone know where I can find AO microtome and cryostat parts? Also is there a place on-line to obtain manuals for some of the AO and other histology equipment? I specifically need a copy of a manual for an old AO 855 cryostat. The one I will soon be using needs some oiling, greasing, and overall attention. Thank you for your help, Donna From STapper <@t> slhduluth.com Tue Mar 16 10:47:28 2004 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Case differentiation Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D6716C9@slhw2smail01.slhdomain.com> Is anyone out there doing anything extraordinary to identify their cases? One of my pathologists went to a meeting last week - (Don't you just hate that!) And saw a poster about how one lab was using a round robin of six different colors of blocks that were then cut and mounted on matching colored slides - all in an effort to prevent a case mix-up. I know that come places will put their decal tissue in an alternate color cassette, endoscopy in another etc., but I am looking for other ideas. Any thoughts would be appreciated. Sheila Tapper HT(ASCP) St. Lukes' Hospital Duluth, MN stapper@slhduluth.com CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. From STapper <@t> slhduluth.com Tue Mar 16 10:51:15 2004 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] CPT questions Message-ID: <3BFBBD68413CB443A7125A63EC0ACD1D6716CA@slhw2smail01.slhdomain.com> 1. Does anyone use the 22 modifier when billing for the Hercept test? It was recommended that we do this - but our billing person is entirely opposed. 2. When billing an 88342 - do you bill different amounts based on the actual cost of the test, or do you bill one amount for the CPT code? Your input is greatly appreciated. Sheila Tapper HT(ASCP) St. Luke's Hospital Duluth, MN stapper@slhduluth.com CONFIDENTIAL: The confidential information accompanying this transmission may contain protected health information and is legally privileged under state and federal law. This information is intended only for the use of the individual to which it is addressed. The recipient or person responsible for delivering this information is prohibited by law from disclosing this information without proper authorization to any other party, unless required to do so by law or regulation. If you are not the intended recipient, you are hereby notified that any review, dissemination, distribution, or copying of this message is strictly prohibited. If you have received this communication in error, please destroy and delete this message from any computer and contact us immediately by return e-mail. No response indicates that the information was received by the appropriate authorized party. Warning: Although St. Luke's has taken reasonable precautions to ensure no viruses are present in this email, St. Luke's cannot accept responsibility for any loss or damage arising from the use of this email or attachments. From jcline <@t> wchsys.org Tue Mar 16 11:51:45 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Edward Scott/ Cap ? Message-ID: <003e01c40b7f$5aa60b80$1d2a14ac@wchsys.org> I have been an inspector and from your answer as to what you keep track of, should suffice. From STEGTM <@t> samcstl.org Tue Mar 16 12:45:04 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Immunostainers Message-ID: We're about to purchase an immunostainer for our lab. We've sort of narrowed the choices down between the Ventana and Dako autostainer models. Since both companies are competetive for pricing to us, we would like to hear any feedback good/bad/otherwise about these stainers from folks using them. Any comments/advice you can offer will be appreciated. Peace, Terre From Don.Birgerson <@t> leica-microsystems.com Tue Mar 16 14:00:37 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] AO parts and Histology equipment manuals Message-ID: Hi Donna, The AO, American Optical, Reichert, Leitz, B&L, Wild and Jung are all companies that are now known as Leica-Microsystems. The "855" is the freezing cabinet of the 975C Cryostat from our old AO company. This cryostat was manufactured from 1981 through 1989 and will need 14970000 Cryo Lubricant and 14021608292 Knife 120mm (old #942 round back knife). I can supply you with a copy of the operators manual if you will contact me with a phone number and address. Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 Djemge@aol.com Sent by: To: histonet@lists.utsouthwestern.edu histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] AO parts and Histology equipment manuals 03/16/2004 10:04 AM Hi all, this is my first time adding a message. Does anyone know where I can find AO microtome and cryostat parts? Also is there a place on-line to obtain manuals for some of the AO and other histology equipment? I specifically need a copy of a manual for an old AO 855 cryostat. The one I will soon be using needs some oiling, greasing, and overall attention. Thank you for your help, Donna _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From jcline <@t> wchsys.org Tue Mar 16 13:45:20 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] therersa stegall Message-ID: <001a01c40b8f$378e0660$1d2a14ac@wchsys.org> Immunostainers: I currently use Ventana, I am satisfied with the staining quality. I am considering getting another module to increase the amount of slides I can stain at one time. We do not have the volume that the Dako can handle. But I have talked with other techs and they are satisfied with the Dako because they have a large volume of immuno's. From dmnelson <@t> iastate.edu Tue Mar 16 13:46:32 2004 From: dmnelson <@t> iastate.edu (Diane Gerjets) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] slim jims and more Message-ID: <6.0.1.1.2.20040316133347.01b247e8@dmnelson.mail.iastate.edu> Hi Steve, One year for April Fool's Day, we put a piece of hot dog through the grossing area as a joke for the pathologists. We were somewhat surprised to find what they were made of. Mainly of fat, muscle and some sort of vegetation. We also had a pathologist who read the results of a research project done with spam. It looked like they wasted no body parts. It was disgusting. However, I was brought up eating hot dogs and spam, so I will continue to eat them. It hasn't hurt me yet. Diane Gerjets Iowa State University Collage of Veterinary Medicine Ames, Iowa From dmnelson <@t> iastate.edu Tue Mar 16 13:54:19 2004 From: dmnelson <@t> iastate.edu (Diane Gerjets) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] colored cassettes Message-ID: <6.0.1.1.2.20040316134823.01b7bd50@dmnelson.mail.iastate.edu> Sheila, We use white cassettes for research, aqua for biopsy, necropsy ( we do change these colors yearly so it's not as hard to find the right block or slide for the right year ), we use red cassettes for anything that is cut at 3 microns. This system seems to work fine for us. Diane Gerjets Iowa State University Ames, Iowa From HETZER <@t> surgery.wisc.edu Tue Mar 16 14:03:45 2004 From: HETZER <@t> surgery.wisc.edu (Michael Hetzer) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] xylene substitutes Message-ID: We have used Propar clearent from Anatech in our linear stainer for ten years now. We have it in the last tank where the slides can sit till they are coverslipped. There is no odor and we find it to be quite tolerant of high humidity conditions and it doesn't dry out the tissue. It is not the fastest paraffin remover but then, we are only processing frozen sections. Buzz:{) Mohs Surgery Madison, WI From bwhitaker <@t> brownpathology.com Tue Mar 16 14:50:47 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] another cytology question Message-ID: <000001c40b98$5b985160$3601a8c0@brownpathology.net> Hi All, The new laboratory that we will be opening soon (hopefully), is anticipating about 20-30 cytospins/month. I have quotes on a demo unit Cytospin IV and a refurbished Cytospin III. The IV was quoted to us at around $5300 and the refurbished III at around $4000. For our low volume, do any of you have strong feelings one way or the other? I don't have any idea how much goes wrong with the Cytospins, and I don't know the difference between a III and a IV at all. Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From juan.gutierrez <@t> christushealth.org Tue Mar 16 14:51:37 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Immunostainers Message-ID: I have both machines in my lab. Which one do I use the most? The Benchmark from Ventana. It is far easier to use with less prep time and in my and my pathologists opinion, a lot better staining. It is also my personal opinion that Ventana offers much better customer support than DAKO does. Ultimately it comes down to what your needs are. If you are running regular antibodies with regular detection kits then Ventana is the one for you. If you are using some difficult primaries with some obscure detection mix then you should try the DAKO, or contact Ventana and ask them about their Discovery system. DAKO might give you more flexibility with your detection systems, but on the other hand Ventana gives you more ease of use. All you do is cut your section, dry it, slap a bar-code on it and run it. On the DAKO you have to cut, dry, depar., hydrate, do epitope retrieval and then load on the machine. If you are short of staff like most of us are, the choice should be easy. By the way the only reason I have both machines is because tha DAKO was already here, but right now I.m trying to trade it for the new BMK XT. That baby does IHC, CISH AND FISH on the same run. Sorry to rattle on. Good Luck making your choice, you can call me if you want to know more about it. Juan C. Gutierrez,HT(ASCP) Histology Supervisor Christus Santa Rosa Healthcare (210)704-2533 -----Original Message----- From: Therersa Stegall [mailto:STEGTM@samcstl.org] Sent: Tue 3/16/2004 12:45 PM To: Histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] Immunostainers We're about to purchase an immunostainer for our lab. We've sort of narrowed the choices down between the Ventana and Dako autostainer models. Since both companies are competetive for pricing to us, we would like to hear any feedback good/bad/otherwise about these stainers from folks using them. Any comments/advice you can offer will be appreciated. Peace, Terre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kennedya <@t> email.cs.nsw.gov.au Tue Mar 16 15:01:07 2004 From: kennedya <@t> email.cs.nsw.gov.au (Andrew Kennedy) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] "Meat" Histology Message-ID: <20040317075973.SM01292@crgcsls813> Hi Histonetters, You would be freaked out if you looked at a section of cabanosi under the microscope - cartilage fragments everywhere, lots of artery and vein and even pancreas and bowel mucosa....still tastes good with cheese and water crackers though. My brother was studying food technology and I recall him doing some DNA testing on minced meats and sausages some year ago - beef mince and sausages were found to have traces of horse meat in it and some supposed "halal" and "kosher" meats actually had pork in them! As an aside, some of the engineering/trades people that I know in our hospital call myself and another colleague, "the butchers" due to the bits of meat that are on the benches when they visit... Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Sydney, Australia ? "noli illegitimi carborundum" ? "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." From amosbrooks <@t> earthlink.net Tue Mar 16 16:30:57 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Junior Histo worker seeks suggestions for small project. In-Reply-To: References: Message-ID: <6.0.0.22.1.20040316162713.01b1edd0@pop.earthlink.net> Hi, Hot dogs are just as groody and not as dehydrated. Interesting project in tissue identification (cartilage, adipose, skin, I think there was some brain tx. too, not nearly as much muscle as one would expect from an attempt at a meat). Amos Brooks I wish I was an Osar Meyer Weiner ... At 09:41 AM 3/16/2004, you wrote: >Message: 10 >Date: Mon, 15 Mar 2004 20:03:34 EST >From: CrochiereSteve@aol.com >Subject: Re: [Histonet] Junior Histo worker seeks suggestions for > small project. >To: suselore@ihug.co.nz, histonet@lists.utsouthwestern.edu >Message-ID: <104.4167f727.2d87ac66@aol.com> >Content-Type: text/plain; charset="US-ASCII" > >Sue, >I once did a little project at AFIP involving a Slim-Jim and a bunch of >different special stains to demonstrate what was actually in those tasty >little >meat snax. > > >Steven M. Crochiere, HT (ASCP) >Histology Supervisor >LifePath Partners @ Mercy Medical Center >299 Carew St. >Springfield, MA 01104 From mward <@t> wfubmc.edu Tue Mar 16 15:32:29 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] Autopsy Survey Message-ID: <61135F0455D33347B5AAE209B903A304076A4DE9@EXCHVS2.medctr.ad.wfubmc.edu> I am posting this brief survey at the request of my department manager. Thank you in advance for your replies. 1. How many Pathologists perform autopsies in your institution? ______ 2. Does your institution perform Medical Examinar autopsies? Y N volume_____ 3. Does your institution perform hospital autopsies? Y N volume_____ 4. What are your fees for a private autopsy? _____ 5. Does your hospital pay the Pathologist for performing hospital cases? Y N What is the fee? 6. Does the Department of Pathology (professional) receive state reimbursement for Medical Examinar cases? Y N What is the fee? ______ 7. Does the Pathology Department reimburse the hospital for technical fees (diener, supplies, histology ) Y N What is the fee?______ If you have another scenario please explain. You may reply to the histonet or to me privately. Thanks, Martha Ward Wake Forest University Baptist Medical Center From Jackie.O'Connor <@t> abbott.com Tue Mar 16 15:36:22 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:40 2005 Subject: [Histonet] "Meat" Histology Message-ID: Exactly why I like to refer to the path lab as the "parts department". Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics "Andrew Kennedy" Sent by: histonet-bounces@lists.utsouthwestern.edu 03/16/2004 03:01 PM To: "Histonet" cc: Subject: [Histonet] "Meat" Histology Hi Histonetters, You would be freaked out if you looked at a section of cabanosi under the microscope - cartilage fragments everywhere, lots of artery and vein and even pancreas and bowel mucosa....still tastes good with cheese and water crackers though. My brother was studying food technology and I recall him doing some DNA testing on minced meats and sausages some year ago - beef mince and sausages were found to have traces of horse meat in it and some supposed "halal" and "kosher" meats actually had pork in them! As an aside, some of the engineering/trades people that I know in our hospital call myself and another colleague, "the butchers" due to the bits of meat that are on the benches when they visit... Andrew Kennedy Senior Science Officer Anatomical Pathology Concord Repatriation General Hospital Sydney, Australia ? "noli illegitimi carborundum" ? "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Tue Mar 16 15:14:11 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Tissue samples Message-ID: Can someone recommend a good vendor for normal human as well as human tumor tissue samples? I'm constructing my own tissue microarrays, and do not have access to normal human tissues. I'd like to get snap frozen samples so I can fix them myself. It used to be so easy! Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Fax 847.938.3266 From portera203 <@t> yahoo.com Tue Mar 16 15:51:51 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mouse on mouse immuno help!!! Message-ID: <20040316215151.43097.qmail@web40909.mail.yahoo.com> Hopefully someone out there can give me some assistance. I am trying to stain mouse tissue with a Monoclonal anti - BRDU. We are using a biotinylated rabbit anti-mouse F(ab')2 secondary at 1:500 in a commercial diluent with normal rabbit serum added to it. Our negative system control (no primary) is coming out almost perfectly clean - however the slides with primary applied are a whole different story. I am seeing what i believe to be good positive signal - with lots of noise. Does anyone have any suggestions that might not be working with commercial kits that are out there. My lab is research and we don't usually work with kits. Just questioning things like - should I avidin/biotin block all mouse tissue?, should i use low or high percentages of normal serum in my primary and secondary?..... I am so close to having this thing worked out any help would be appreciated. Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam From gsennello <@t> osip.com Tue Mar 16 16:05:57 2004 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Tissue samples Message-ID: We have used and been pleased with: Bio-options, Inc 323-933-9775 Ask for Dr. Blocher Gina Sennello Associate Scientist/ Histotechnologist OSIP 2860 Wilderness Place Boulder, CO 80301 phone 303-546-7739 fax 303-444-0672 -----Original Message----- From: Jackie.O'Connor@abbott.com [mailto:Jackie.O'Connor@abbott.com] Sent: Tuesday, March 16, 2004 2:14 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Tissue samples Can someone recommend a good vendor for normal human as well as human tumor tissue samples? I'm constructing my own tissue microarrays, and do not have access to normal human tissues. I'd like to get snap frozen samples so I can fix them myself. It used to be so easy! Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics 847.938.4919 Fax 847.938.3266 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lizchlipala <@t> premierhistology.com Tue Mar 16 16:19:18 2004 From: lizchlipala <@t> premierhistology.com (Elizabeth Chlipala) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mouse on mouse immuno help!!! In-Reply-To: <20040316215151.43097.qmail@web40909.mail.yahoo.com> Message-ID: <000001c40ba4$bc8a9f80$74d48a80@LIZ> Amy If we are performing BrdU on mouse tissue we use a primary from Accurate Chemical and Scientific (800) 645-6264, Clone BUI/75 product number MAS 250. It's a rat anti-BrdU. Then we use a secondary from Dako - rabbit anti-rat (mouse absorbed) Catalog Number: E 0468. I will only use mouse on mouse antibodies when absolutely necessary, and if I have to do that I'll use either the Dako ARK kit or the Vector MOM Kit. Both will work fine. I can't send attachments to the histonet so I'll e-mail you separately with our protocol. Hope this helps Liz Elizabeth A. Chlipala, BS, HTL(ASCP) Premier Histology Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 lizchlipala@premierhistology.com www.premierhistology.com Ship to Address: Premier Histology Laboratory University of Colorado MCBD, Room A3B40 Boulder, Colorado 80309 _________________________________ 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. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Amy Porter Sent: Tuesday, March 16, 2004 2:52 PM To: Histonet Subject: [Histonet] mouse on mouse immuno help!!! Hopefully someone out there can give me some assistance. I am trying to stain mouse tissue with a Monoclonal anti - BRDU. We are using a biotinylated rabbit anti-mouse F(ab')2 secondary at 1:500 in a commercial diluent with normal rabbit serum added to it. Our negative system control (no primary) is coming out almost perfectly clean - however the slides with primary applied are a whole different story. I am seeing what i believe to be good positive signal - with lots of noise. Does anyone have any suggestions that might not be working with commercial kits that are out there. My lab is research and we don't usually work with kits. Just questioning things like - should I avidin/biotin block all mouse tissue?, should i use low or high percentages of normal serum in my primary and secondary?..... I am so close to having this thing worked out any help would be appreciated. Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Mar 16 17:10:45 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mouse on mouse immuno help!!! In-Reply-To: <20040316215151.43097.qmail@web40909.mail.yahoo.com> Message-ID: <3.0.6.32.20040316161045.00bc3e70@gemini.msu.montana.edu> AMy, YOu are doing mouse on mouse immunostaining and you should use a kit designed for that use. Our research lab normally does not work with kits until it comes to frustrating mouse on mouse staining. That is when we go commercial. The DAKO ARK kit should work, Chemicon has a new kit that works well, there are others on the market. The kicker with these kits is the special blockers they supply in order to reduce the mouse to mouse detection, aka high background. Another kit that we have used is Scytek out of Logan Utah. I was surprised (I am a terrible doubtful type when it comes to ms on ms staining!) at how clean the new Chemicon kit was, simple and easy to use. At 01:51 PM 3/16/2004 -0800, you wrote: >Hopefully someone out there can give me some assistance. I am trying to stain mouse tissue with a Monoclonal anti - BRDU. We are using a biotinylated rabbit anti-mouse F(ab')2 secondary at 1:500 in a commercial diluent with normal rabbit serum added to it. Our negative system control (no primary) is coming out almost perfectly clean - however the slides with primary applied are a whole different story. I am seeing what i believe to be good positive signal - with lots of noise. Does anyone have any suggestions that might not be working with commercial kits that are out there. My lab is research and we don't usually work with kits. Just questioning things like - should I avidin/biotin block all mouse tissue?, should i use low or high percentages of normal serum in my primary and secondary?..... I am so close to having this thing worked out any help would be appreciated. > > >Amy S.Porter, HT(ASCP) >Michigan State University >Department of Physiology >Division of Human Pathology >College of Human Medicine >portera203@yahoo.com > > >Do you Yahoo!? >Yahoo! Mail - More reliable, more storage, less spam >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From mariatere <@t> infovia.com.ar Tue Mar 16 19:55:31 2004 From: mariatere <@t> infovia.com.ar (Teresa Dominguez) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] AgNOR References: <000001c40a39$ed95d610$094dbad0@hppav> Message-ID: <00af01c40bc2$f21d6470$217f46c8@FAMILIA> George Cole, I'm really sorry if you have missunderstood something I wrote in my message. The problem was I should translate from my notes in Spanish to english, so the "jelly" I mensionated is the same jelly you will know flavoured or not. That "jelly" of course is without "any" flavour. Maybe you know it as "gelatine", "gelatin" too, and synonims like that. Next point, the NOR is, Nucleolar Organization Releaser; at least this is the information I have. Related with the" wacky silver preps that were very fickle"... I know something about that, thru, you are right. In my 10 years experience as a histotechnician I only have performed Jone's Methenamine for kidney, Ag Reticuline, and Grocott's Method. I have never failed in them. Maybe I am a "lucky woman", or my boss is a Magician, but ... About the Formic acid, if you like to take a look at this, formic acid - a colorless pungent fuming vesicatory liquid acid HCOOH found naturally in ants and many plants or made catalytically from carbon monoxide and steam; used in finishing textiles and paper and in the manufacture of insecticides and fumigants , furthermore you can read this, If you are interested in anything more, this site is interesting http://msds.pdc.cornell.edu/msds/msdsdod/a96/m47919.htm Now, you wonder to know what's happens If I use just room temperature water. Ok, in my opinion I think the warm or hot water (not boiled) is useful to acelerate the reactions; so if you rather you can use Just room temperature water, you choose. Well, I hope all you doubts have been solved, at least I tried to give you "a light". My idea was Just to help to someone, I am not owner of the thru, I do mistakes of course, that's why I choose this list to be updated everyday. If I can tell you something, even a "little thing" of my experience, ok. I am satisfied. I like to learn with discussions too. Thanks for your e-mail. H.T Maria T Dominguez Anatomy Pathology Service Hospital Regional R?o Grande, R?o Grande, Tierra del Fuego, Argentina 54- 02964-422086/88 Ext. 143 ----- Original Message ----- From: "George Cole" To: "'Teresa Dominguez'" Sent: Monday, March 15, 2004 12:02 AM Subject: RE: [Histonet] AgNOR M.Teresa; I was fascinated by your procedure. I spent a life time as a histotech doing silvers on many, many brain sections and on even more nerve sections. I confess I don't know what NOR is. I also wonder just what the jelly is---I can't imagine a blackberry or strawberry flavored silver prep. I see that after a century of wacky silver preps that were very fickle---a sometimes thing---your 16% Silver Nitrate comes close to the 20% that brought silvers into steady and fairly reliable usage. I can't guess what the Formic Acid is for. I thought it might darken the silver from brown to black, but I see your results still indicate brown silver deposits. I wonder what happens if you don't use the hot water rinse, just room temperature water. Thanks for another recipe for the fabled Silver Nitrate. georgecole@ev1.net From azdudley <@t> hotmail.com Tue Mar 16 20:54:43 2004 From: azdudley <@t> hotmail.com (anita dudley) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] xylene substitute Message-ID: eddie, we use richard allans clearrite on our H&E stainer with very good results. anita providence hosp. mobile al >From: "Eddie Marquez" >To: >Subject: [Histonet] xylene substitute >Date: Mon, 15 Mar 2004 10:19:06 -1000 > > >Aloha from Hawaii, > >Just needed some info.-Has anybody used xylene substitute with a linear >stainer; for standard H&E's? If so, how good are the results? May I also >have the brand name. > >Thank you, > >Eddie from the Cancer Research > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ MSN Toolbar provides one-click access to Hotmail from any Web page – FREE download! http://clk.atdmt.com/AVE/go/onm00200413ave/direct/01/ From cmalc <@t> unimelb.edu.au Wed Mar 17 00:26:10 2004 From: cmalc <@t> unimelb.edu.au (Cathy Malcontenti-Wilson) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] TNF alpha and MARCO immunostaining Message-ID: <5.2.1.1.2.20040317171253.050a1c10@mail.staff.unimelb.edu.au> Dear all. Does anyone have any experience with immunostaining for TNFalpha and MARCO (activated macrophage marker) in paraffin embedded, formalin fixed mouse tissues? We routinely use a MOM kit for our primary antibodies which work fine. Any help with antigen retrieval or other tips will be greatly appreciated. Cathy From alexander.nader <@t> wgkk.sozvers.at Wed Mar 17 01:24:40 2004 From: alexander.nader <@t> wgkk.sozvers.at (Nader, Alexander) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] flat bed scanners Message-ID: <4FE843CBBAC1D211A8150008C70952DAAE7D1D@hk01nt05.hkh.wgkk.sozvers.at> > Bill, > This is a completely new one to me, and I'm sure we would all > like to here a fuller story. > (See what you've done now - let yourself in for it :-) ) > >> For 3D objects CCD scanners are much better than the cheaper CMOS >> scanners, for depth of field reasons. There are a number out there, >< but you have to look at the specs. I use UMAX Powerlook IIIs. >> Bill Blank >> http://kernunnos.com (Celtic studies and numismatics) >> http://www.druidry/org http://www.druidry.org/board Dear Terry, dear Bill, one of the answers is already given on the homepage of Bill: the differences between the three scanners (UMAX) are striking and really very interesting, not only for numismatics but also forsimple minded pathologists. I remember a similar article about depth of fields in a German journal for computers (c't) a couple of years ago. Maybe this article http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.o rg.uk/mag/artapr01/dwscanner.html is also interesting to you too. Alexander Nader MD Vienna, Austria From GAshton <@t> PICR.man.ac.uk Wed Mar 17 02:42:15 2004 From: GAshton <@t> PICR.man.ac.uk (Garry Ashton) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] histogel Message-ID: Dear all, I am looking for a supplier of "histogel" a substance used to embed cell pellets before processing. I believe a company called Richard Allan Scientific supply it. Does anybody know of a company that supply histogel or a similar product in the UK. Many thanks. Garry PICR UK -------------------------------------------------------- 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 and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. 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. From settembr <@t> umdnj.edu Wed Mar 17 06:40:12 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Immunostainers Message-ID: Hello Therersa, I have a Dako and am very happy with it. I am sure that you've gone through the demos so you already know how easy it is to use. So I'll tell you about their great technical services dept. Whenever I have any questions, they are fantastic. That dept alone is a reason to lean towards the Dako. If you haven't gone through the demo, call them. We purchased one about 8 years ago and then as our volume increased and we saw the need for another, Dako suggested that we look into their, "Reagent Acquisition" plan. We were purchasing enough of their reagents on a monthly basis, that we were able to get a second virtually for free. Also, please know that you are NOT required to use their products either. The Dako autostainer can use any reagents from any company. Hope this helps. Dana Settembre Immunohistochemistry Lab University Hospital - UMDNJ- Newark, NJ >>> Therersa Stegall 3/16/2004 10:45:04 AM >>> We're about to purchase an immunostainer for our lab. We've sort of narrowed the choices down between the Ventana and Dako autostainer models. Since both companies are competetive for pricing to us, we would like to hear any feedback good/bad/otherwise about these stainers from folks using them. Any comments/advice you can offer will be appreciated. Peace, Terre _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Wed Mar 17 07:24:45 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] CAP Check List In-Reply-To: <000001c40b6a$846a3b90$c5e2f218@thescotts> Message-ID: Allison, That's the way I'm setting my lab up. I think this question pertains to automatic immuno and special stainers. If you have PM performed let's say on a Benchmark, are the staining results the same as before the PM was performed? That's how I'm interpreting the question. I wish CAP would be more specific with some of their questions, Joe Nocito, BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX 78205 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Edward Scott Sent: Tuesday, March 16, 2004 9:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CAP Check List I just received my Cap self check list and I'm having a problem with the question ANP.23075 : Is there evidence of ongoing evaluation of results of instrument maintenance and functions for all devices. Would having maintenance charts and a policy of how often instruments are checked by Bio Med and or the company from which the instrument was purchased be enough. I also have all of the PM/ service records on all instruments that require annual checks. Any help with this would be appreciated. Thanks in advance. Allison Scott HT(ASCP) Histology Supervisor LBJ Hospital Houston, Texas _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************Notice******************************************** This e-mail, including attachments, contains information that is confidential and may be legally privileged. This e-mail, including atachments, constitutes non-public information inteneded to be conveyed only to the designated recipient(s). If you are not an intended recipient,please delete this e-mail, including attachments, and notify me. The unauthorized use, dissemination, distribution or reproduction of this e-mail, including attachments, is prohibited and may be unlawful. ***************************************************************************************** From dmcaloose <@t> wcs.org Wed Mar 17 07:27:18 2004 From: dmcaloose <@t> wcs.org (McAloose, Dee) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] job listing Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A0778A982@WOLF.wcs.org> I would like to post the following job advertisement. Can you please tell me what the process is, how long the ad can run, and whether or not there is a fee for posting the listing. Thanks. Regards. D McAloose, VMD, Diplomate ACVP Head, Department of Pathology Wildlife Conservation Society 2300 Southern Blvd Bronx, NY 10460 phone: (718) 220-7105 fax: (718) 220-7126 email: dmcaloose@wcs.org Wanted - Histotechnician The Department of Pathology of the Wildlife Conservation Society has an immediate opening for a full-time histotechnician. The laboratory provides diagnostic services to one of the largest zoological collections in the country. The successful candidate will be responsible for routine processing, embedding, cutting, staining and cover slipping of slides for H&E and special staining; excellent microtome sectioning skills are essential. In addition, the position requires that you are able to produce high quality slides at high volume and operate and perform routine maintenance of laboratory equipment. Applications will be accepted until May 15, 2004 or until the position is filled. Address all correspondence to: Ms. Tawanda Williams, Human Resources, Wildlife Conservation Society, 2300 Southern Blvd., Bronx, NY 10460. email: twilliams@wcs.org . EOE. From kmerriam2003 <@t> yahoo.com Wed Mar 17 07:53:33 2004 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] another cytology question In-Reply-To: <000001c40b98$5b985160$3601a8c0@brownpathology.net> Message-ID: <20040317135333.18080.qmail@web10001.mail.yahoo.com> Statspin makes a small cytospin machine (can only do 4 samples at a time), but it is cheap (under 2K). It is in the Fisher catalog. Regards, Kim Merriam Novartis Cambridge, MA Bonnie Whitaker wrote: Hi All, The new laboratory that we will be opening soon (hopefully), is anticipating about 20-30 cytospins/month. I have quotes on a demo unit Cytospin IV and a refurbished Cytospin III. The IV was quoted to us at around $5300 and the refurbished III at around $4000. For our low volume, do any of you have strong feelings one way or the other? I don't have any idea how much goes wrong with the Cytospins, and I don't know the difference between a III and a IV at all. Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam From vherrington <@t> sumterregional.org Wed Mar 17 08:14:27 2004 From: vherrington <@t> sumterregional.org (Herrington, Vicki) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] histogel Message-ID: <3E09D94FAC8AD411A04900D0B77E645002815529@SRHNTMAIL> We use Nutrient Agar Slants from Becton & Dickinson (cat # 220971 - 100 tubes for approximately $40). They work just as well and are really cheap. Of course, you have to keep them refrigerated, and if you don't use all the tube you have to be conscious of the need to use good lab techniques and refrigerate the remainder quickly since it is a bacterial culture medium. Vicki -----Original Message----- From: Garry Ashton [mailto:GAshton@PICR.man.ac.uk] Sent: Wednesday, March 17, 2004 3:42 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] histogel Dear all, I am looking for a supplier of "histogel" a substance used to embed cell pellets before processing. I believe a company called Richard Allan Scientific supply it. Does anybody know of a company that supply histogel or a similar product in the UK. Many thanks. Garry PICR UK -------------------------------------------------------- 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 and do not necessarily represent those of the Paterson Institute for Cancer Research or the Christie Hospital NHS Trust. 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Note: The information contained in this message and any attachments 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 reading, dissemination, distribution, copying, or other use of this communication or any of its attachments is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting this message, any attachments, and all copies and backups from your computer. If you have received this communication in error and are unable to reply to this message, please notify the sender immediately by telephone at (229) 931-1380. Thank you. Sumter Regional Hospital From mbryhan <@t> NORTHERNHEALTH.ORG Wed Mar 17 08:51:22 2004 From: mbryhan <@t> NORTHERNHEALTH.ORG (Mary Bryhan) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Rental Cryostat Message-ID: Hi all, just wondering if anyone knows of a short term cryostat rental service? Mary Bryhan HT (ASCP) Northern Michigan Hospital Petoskey, MI From portera203 <@t> yahoo.com Wed Mar 17 09:46:42 2004 From: portera203 <@t> yahoo.com (Amy Porter) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Thanks for mouse on mouse Message-ID: <20040317154642.57633.qmail@web40914.mail.yahoo.com> As always histonet is a fabulous resource, thank you all for you time and help with my dilema. Amy S.Porter, HT(ASCP) Michigan State University Department of Physiology Division of Human Pathology College of Human Medicine portera203@yahoo.com Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam From hemo_de_2000 <@t> yahoo.com Wed Mar 17 09:47:16 2004 From: hemo_de_2000 <@t> yahoo.com (Kevin Oakley) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] xylene substitute In-Reply-To: Message-ID: <20040317154716.23739.qmail@web60710.mail.yahoo.com> Dear Eddie, Hi, I am with Scientific Safety Solvents and saw your post. We make a xylene substitute, called Hemo-De. If you are intrested we can fax you a brochure and our MSDS sheet about our product. If you would like to talk to us direct you can e-mail me back at hemo_de_2000@yahoo.com, or call 1-800-355-3689. Hope you have a wonderful day. Sincerely, Caroline Gruslin anita dudley wrote:eddie, we use richard allans clearrite on our H&E stainer with very good results. anita providence hosp. mobile al >From: "Eddie Marquez" >To: >Subject: [Histonet] xylene substitute >Date: Mon, 15 Mar 2004 10:19:06 -1000 > > >Aloha from Hawaii, > >Just needed some info.-Has anybody used xylene substitute with a linear >stainer; for standard H&E's? If so, how good are the results? May I also >have the brand name. > >Thank you, > >Eddie from the Cancer Research > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ MSN Toolbar provides one-click access to Hotmail from any Web page – FREE download! http://clk.atdmt.com/AVE/go/onm00200413ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam From dmccaig <@t> ckha.on.ca Wed Mar 17 09:47:06 2004 From: dmccaig <@t> ckha.on.ca (Diana McCaig) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Time slides are submitted to pathologist Message-ID: <3E5A3F039F0BD8118B4700C00D0020240430F2@CKHA9> The OLA Checklist for Lab License mandates on item V.C.1 that a record should be kept of the time of delivery of the slides to the pathologist. How are these being recorded? Is there master list that techs indicate the time with each tray, is it indicated on the requisition, or on a worksheet. We are not automated so computer entry is not an option. Diana McCaig, R.T. Charge Tech, Histology Chatham Kent Health Alliance 519-352-6401 (3347) From carl.hobbs <@t> KCL.AC.UK Wed Mar 17 10:38:03 2004 From: carl.hobbs <@t> KCL.AC.UK (Carl Hobbs) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] re mouse on mouse Message-ID: <00d401c40c3e$37bbd9a0$e8345c9f@Carlos> I use Dako mouse anti BrdU( and many other mouse monoclonals) often on pwax mouse tissue, with no problems; lungs, brain, skin and gut. Std goat anti mouse biotin secondary with streptavidin- biotin px DAB system( Dako again, no kits). I personally use BSA as protein additive and ignore serum as a blocker. IMHO, it's probably the way you try to disclose the BrdU that's causing the background( obviously, if it's endog peroxidase, too concentrated primary or biotin... that's different). We've tried many ways to disclose and many give b'ground. Only clean, specific results we got was after protease digestion, followed by HCL hydrolysis pretreatments. From Robert.Lott <@t> bhsala.com Wed Mar 17 11:12:12 2004 From: Robert.Lott <@t> bhsala.com (Lott, Robert) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] IHC ASR List Message-ID: <35B6C610DD1DD311B1FA0008C791400407F3009D@gobexchm3.bhsala.com> Would you wonderful people out in histoland mind sharing with me your list of Immunohistochemistry Analyte Specific Reagents (ASR) in use in your lab by: 1)antibody name 2) Vendor Thanks!! Robert L. Lott, HTL(ASCP) Manager, Anatomic Pathology Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388 phone 205-592-5646 fax robert.lott@bhsala.com From lwhite <@t> lakeridgehealth.on.ca Wed Mar 17 11:22:34 2004 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mercury free fixatives Message-ID: I am trying to find a replacement for B5 fixative for lymph nodes and bone marrow. I am currently testing B-Plus fixative but am not completely happy with the results so far. Has anybody had good results with B-Plus or another mercury free alternative? Any feedback would be greatly appreciated. Lori White, B.Sc., MLT Charge Technologist, Histopathology Lakeridge Health Corporation Tel 905 576-8711 ext. 4358 Fax 905 905 721-4757 From pruegg <@t> colobio.com Wed Mar 17 11:02:37 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] CD98 and PLGF Message-ID: I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160 and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies work in ffpe tissue? Patsy From BoozerKA <@t> pa1.ah.org Wed Mar 17 11:12:59 2004 From: BoozerKA <@t> pa1.ah.org (Kathleen Boozer) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Web site for Cytology Message-ID: Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! From tpmorken <@t> labvision.com Wed Mar 17 11:08:40 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] flat bed scanners (and 3D objects) Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA21773EE@usca0082k08.labvision.apogent.com> Bill Wrote: > >> For 3D objects CCD scanners are much better than the cheaper CMOS >> scanners, for depth of field reasons. There are a number out there, >< but you have to look at the specs. I use UMAX Powerlook IIIs. >> Bill Blank I'm not clear to me how, as Bill suggests, the detector of a scanner could have an effect of depth of field. Isn't the detector essentially like film - a flat surface? It seems to me it is the optics that determines the depth of field and that maybe Bill's CCD scanner simply has different optics than the cmos scanner he mentions. Tim Morken Dear Terry, dear Bill, one of the answers is already given on the homepage of Bill: the differences between the three scanners (UMAX) are striking and really very interesting, not only for numismatics but also forsimple minded pathologists. I remember a similar article about depth of fields in a German journal for computers (c't) a couple of years ago. Maybe this article http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.o rg.uk/mag/artapr01/dwscanner.html is also interesting to you too. Alexander Nader MD Vienna, Austria >> For 3D objects CCD scanners are much better than the cheaper CMOS >> scanners, for depth of field reasons. There are a number out there, >< but you have to look at the specs. I use UMAX Powerlook IIIs. >> Bill Blank From garygill <@t> dcla.com Wed Mar 17 11:38:26 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Web site for Cytology Message-ID: Cytopathnet, cytopathnet-l@cytopathnet.us, is closest available. It's open to the public at no charge. Send an email to their address with Subscribe as the Subject. Cytopathnet used to be a robust vigorous exchange, but it's much less so today after a series of virus infections. Gary Gill -----Original Message----- From: Kathleen Boozer [mailto:BoozerKA@pa1.ah.org] Sent: Wednesday, March 17, 2004 12:13 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Web site for Cytology Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kosmicdog <@t> hotmail.com Wed Mar 17 11:29:09 2004 From: kosmicdog <@t> hotmail.com (jason madore) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] staining frozen sections Message-ID: We are having problems with staining our frozen sections. Slides which once gave excellent morphology soon begin to loose their nuclei. The nucleus looks empty or washed away leaving white holes where the nucleus was. We are cutting the slides at -20. Storing at -80 wrapped in tin foil with dessicant. Warm to RT for 30seconds and fix in cold acetone\EtOH for 5 minutes, wash in dH2O for 1 minute, and then stain with hematoxilyn, dehydrate, clear in xylenes, then mount. Does anyone know what could be causing this? Thanks. for any help. Jason. _________________________________________________________________ MSN Premium with Virus Guard and Firewall* from McAfee® Security : 2 months FREE* http://join.msn.com/?pgmarket=en-ca&page=byoa/prem&xAPID=1994&DI=1034&SU=http://hotmail.com/enca&HL=Market_MSNIS_Taglines From garygill <@t> dcla.com Wed Mar 17 11:38:26 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Web site for Cytology Message-ID: Cytopathnet, cytopathnet-l@cytopathnet.us, is closest available. It's open to the public at no charge. Send an email to their address with Subscribe as the Subject. Cytopathnet used to be a robust vigorous exchange, but it's much less so today after a series of virus infections. Gary Gill -----Original Message----- From: Kathleen Boozer [mailto:BoozerKA@pa1.ah.org] Sent: Wednesday, March 17, 2004 12:13 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Web site for Cytology Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Wed Mar 17 13:04:51 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:41 2005 Subject: Martha Ward Re: [Histonet] Autopsy Survey References: <61135F0455D33347B5AAE209B903A304076A4DE9@EXCHVS2.medctr.ad.wfubmc.edu> Message-ID: <001901c40c52$ba7b2260$1d2a14ac@wchsys.org> ----- Original Message ----- From: "Martha Ward" To: Sent: Tuesday, March 16, 2004 4:32 PM Subject: [Histonet] Autopsy Survey I am posting this brief survey at the request of my department manager. Thank you in advance for your replies. 1. How many Pathologists perform autopsies in your institution? __5 2. Does your institution perform Medical Examinar autopsies? Y N volume___N__ 3. Does your institution perform hospital autopsies? Y N volume___Y__ 4. What are your fees for a private autopsy? $2,000_ 5. Does your hospital pay the Pathologist for performing hospital cases? N What is the fee? 6. Does the Department of Pathology (professional) receive state reimbursement for Medical Examinar cases? N/A What is the fee? ______ 7. Does the Pathology Department reimburse the hospital for technical fees (diener, supplies, histology ) N What is the fee?______ If you have another scenario please explain. You may reply to the histonet or to me privately. Thanks, Martha Ward Wake Forest University Baptist Medical Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RBARNHART <@t> summithealth.org Wed Mar 17 13:22:15 2004 From: RBARNHART <@t> summithealth.org (Rebecca Barnhart) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Shadon Gemini stainer Message-ID: There was a message a couple days ago about a lab thinking about getting a Shandon Gemini. We are also thinking of getting one. We visited another local hospital that has one and like it. But we were told by a vendor that we deal with (not one that we would be buying a stainer from anyway) that they heard that there was software issue with the Gemini. Any input is appreciated. From settembr <@t> umdnj.edu Wed Mar 17 13:45:08 2004 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] IHC ASR List Message-ID: I am still compiling my list but there are not that many. Dako C-kit (CD117) Dana Settembre University Hospital - UMDNJ Newark, NJ >>> "Lott, Robert" 3/17/2004 9:12:12 AM >>> Would you wonderful people out in histoland mind sharing with me your list of Immunohistochemistry Analyte Specific Reagents (ASR) in use in your lab by: 1)antibody name 2) Vendor Thanks!! Robert L. Lott, HTL(ASCP) Manager, Anatomic Pathology Baptist Health System 800 Montclair Road Birmingham, AL 35213 205-592-5388 phone 205-592-5646 fax robert.lott@bhsala.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gudrun.lang <@t> aon.at Wed Mar 17 14:18:19 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] tissueprocessing-xylol Message-ID: <000c01c40c5c$fd6e3800$eeeea8c0@SERVER> Hi I am reading about tissueprocessing. There are many various clearents and I wonder, if everybody uses xylol in the tissueprocessors like we do. Is it really the mostly used reagens? Gudrun Lang From Stephen.J.Scholz <@t> osfhealthcare.org Wed Mar 17 14:49:07 2004 From: Stephen.J.Scholz <@t> osfhealthcare.org (Scholz, Stephen J.) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Job Opening Message-ID: <7F1312711CA7474A89B3DF8BA0BA54D0F5F0E2@pmc-rfd-mx01.intranet.osfnet.org> Hello all, We have a position open for a HT/HTL here in Rockford. The position is full time days working with a great staff and a wonderful Pathologist group in a routine Histology lab. Please contact me if you are interested and I can give you all the details. Thank you, Stephen J. Scholz HT(ASCP) Histology Coordinator OSF St. Anthony Medical Center Rockford IL Phone: 815-395-5410 Fax: 815-395-5364 e-mail: sjscholz@osfhealthcare.org From weneng <@t> hotmail.com Wed Mar 17 14:28:12 2004 From: weneng <@t> hotmail.com (Wendy England) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] iron stain on bone marrow Message-ID: Thanks again to everybody who gave me the information about this stain! Here comes another question: I couldn't see much iron on my slide, only about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 min, then in freshly mixed stain solution(equal part of 20% HCl and 10% potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin after wash. Then wash, air dry then mount. I was told there should be much more iron. I feel bad now. Could any body give me advice or could I ask if it's possible due to the bone marrow preparation since we indeed saw some spots. I didn't prepare the sample. Thanks and look forward to hear from you there! Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar – get it now! http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ From Luis.Chiriboga <@t> med.nyu.edu Wed Mar 17 14:04:45 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Last Call For Nominations (NYSHS Members Only) Message-ID: TO: All New York State Histotechnology Society Members RE: Award nominations for 2004 The NYSHS Awards committee is asking for nominations from its membership for this years awards cycle. In addition to recognition by the society, recipients will also be awarded a small financial grant to further their knowledge and education in the discipline of histotechnology. Applications are due by April 1st 2004. There are 3 awards awaiting nominations. 1.Thermo Electron Student Scholarship Award: Is awarded to a histology student or a histotech who wishes to attend a professional meeting. This award is sponsored by thermo Electron inc. and must be used to defray educational expenses.Please send us: A letter from you, showing evidence of your commitment to continuing education. Two letters of recommendation from supervisor, pathologists, or histotechnologist.. Name and address of your current employer or school, and your current address. 2. Dominic Europa Award: Awarded to a long standing NYSHS member (for use towards an educational meeting) who serves as an inspiration to others in the field. Candidate can be a bench tech or a supervisor (preferably not in the limelight). Please submit a nomination and recommendation letter, detailing the nominees contributions. 3.Biogenex Excellence in Education Award: Awarded to any member in good standing, to be used to fund an educational endeavor. This endeavor could be tuition for a class, educational materials or payment for a meeting that is not funded by an employer. To apply for this award, please send a letter outlining how you would benefit educationally from this award, and how it will help you to better serve the profession. This award is sponsored by Biogenex. We encourage e-mail submission of applications and letters. Please submit applications to: Luis Chiriboga NYSHS Awards Chairperson 318 East 15th Street New York, NY 10003 or vial E-mail Luis.chiriboga@med.nyu.edu From mark.lewis <@t> thermo.com Wed Mar 17 16:24:30 2004 From: mark.lewis <@t> thermo.com (mark.lewis@thermo.com) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mercury free fixatives Message-ID: Lori, Thermo sells Zinc Formal Fixx. That works fairly well for a B-5 substitute. We can send you a sample if you'd like. Let me know. Best regards, Mark Mark Lewis Product Specialist Anatomical Pathology Clinical Diagnostics Thermo Electron Corporation (412) 747-4013 (412) 788-1097 E-mail: mark.lewis@thermo.com "White, Lori" To: "Histonet (histonet@lists.utsouthwestern.edu)" Sent by: histonet-bounces@lists.utsouth cc: western.edu Subject: [Histonet] mercury free fixatives 03/17/2004 12:22 PM I am trying to find a replacement for B5 fixative for lymph nodes and bone marrow. I am currently testing B-Plus fixative but am not completely happy with the results so far. Has anybody had good results with B-Plus or another mercury free alternative? Any feedback would be greatly appreciated. Lori White, B.Sc., MLT Charge Technologist, Histopathology Lakeridge Health Corporation Tel 905 576-8711 ext. 4358 Fax 905 905 721-4757 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LBlack <@t> carilion.com Wed Mar 17 16:52:00 2004 From: LBlack <@t> carilion.com (Lisa Black) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Shadon Gemini stainer Message-ID: My lab upgraded from the older model of the Shandon stainer to the new one. We are a busy laboratory and this stainer is able to stain many more H&E's in half the time as the older model. We have had no software problems. Best Wishes. Lisa Black CCL Histology Roanoke, VA >>> "Rebecca Barnhart" 3/17/04 2:22:15 PM >>> There was a message a couple days ago about a lab thinking about getting a Shandon Gemini. We are also thinking of getting one. We visited another local hospital that has one and like it. But we were told by a vendor that we deal with (not one that we would be buying a stainer from anyway) that they heard that there was software issue with the Gemini. Any input is appreciated. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bill501 <@t> mindspring.com Wed Mar 17 17:03:47 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] flat bed scanners (and 3D objects) In-Reply-To: <0556BE8AC5551E4E8AF6BB9E42509BA21773EE@usca0082k08.labvision.apogent.com> References: <0556BE8AC5551E4E8AF6BB9E42509BA21773EE@usca0082k08.labvision.apogent.com> Message-ID: At 9:08 AM -0800 3/17/04, Morken, Tim - Labvision wrote: >I'm not clear to me how, as Bill suggests, the detector of a scanner could >have an effect of depth of field. Isn't the detector essentially like film - >a flat surface? It seems to me it is the optics that determines the depth of >field and that maybe Bill's CCD scanner simply has different optics than the >cmos scanner he mentions. My Experience is completely empirical. I have tried reading the photonics and optical reasons, but have not really understood them. There was a recent article (in the last year) about CMOS detectors in Photonics magazine which I no longer have. What I remember is the big advantage of CMOS is that it is cheaper. The dynamic color range and something called the fill factor are still inferior to CCD, but getting closer. I did a quick search on Google this AM, but really only found things about digital cameras where the optic effect is more obvious (at least to me when one can vary the aperture of a lens) The CCD scanners tend to be the higher end scanners and it may be, as Tim said, that the optics are different. With my PL III's I can get 3D objects almost an inch thick in focus. With one of the CMOS scanners I got free with something, I couldn't get an entire high relief coin in focus (say 1/8th inch). I gay that scanner away. besides, it was USB 1 and too slow. -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) http://www.druidry/org http://www.druidry.org/board From histo <@t> sdspathology.com.au Wed Mar 17 18:42:40 2004 From: histo <@t> sdspathology.com.au (Histology) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing Message-ID: <001801c40c81$eabac640$c6c710ac@sdspathology.com.au> Hi There Does anyone use Isopropanol in routine overnight parraffin processing replacing xylol?? If you are willing could you please supply any protocols you may use. Thanks for your help. Frances SDS Pathology Sydney, NSW,Australia From Lizbeth_Kelly <@t> hgsi.com Wed Mar 17 14:08:49 2004 From: Lizbeth_Kelly <@t> hgsi.com (Lizbeth_Kelly@hgsi.com) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] NSH Region II Symposium (June 3,4, & 5th) Message-ID: The Maryland Society of Histotechnologists (MSH) is sponsoring the Region II Symposium June 3, 4, and 1/2 day on the 5th at the Holiday Inn Select, North-Baltimore, Maryland. For information, you may contact Terri DeCarli at 410-787-4546 or Renate Jaacks at 410-879-9012. Lizbeth Kelly, HT (ASCP), QIHC Board Member, MSH From lynzjackson <@t> hotmail.com Thu Mar 18 02:55:14 2004 From: lynzjackson <@t> hotmail.com (lynsay jackson) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Chromavision Message-ID: Has anyone had success using the ACIS chromavision platform to measure EGFR, activated EFGR or HER2? What criteria have you used. Do you think there is some bias in quantification methods for IHC? I would be grateful of some feedback, and any other successful automated quatification methods or general opinions on Chromavison image analysis. Lynsay Jackson _________________________________________________________________ Stay in touch with absent friends - get MSN Messenger http://www.msn.co.uk/messenger From lpwenk <@t> covad.net Thu Mar 18 03:46:53 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mercury free fixatives References: Message-ID: <005f01c40ccd$f1ea2720$5223d445@domainnotset.invalid> We recently tested several, all at different time intervals, from 1 to 24 hours. In general, we found that zinc formalin take about 1.5 times as long to fix as B5 (e.g., if you were fixing bone marrows for 2 hours, the zinc formalin needed 3 hours; if lymph nodes required 4 hours in B5, it now needed 6 hours in zinc formalin). We also found that different tissues required different formulations of zinc formalin. We liked one formulation for bone marrow biopsies, and a different one for solid tissues. I think the difference was because the bone marrow biopsy had to be decalcified, and one of the formulations held up better against the acid decalcification. So you need to test out several companies brands, all at different time intervals, to find out what works best for your tissue and your processing. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "White, Lori" To: Sent: Wednesday, March 17, 2004 12:22 PM Subject: [Histonet] mercury free fixatives > I am trying to find a replacement for B5 fixative for lymph nodes and bone > marrow. I am currently testing B-Plus fixative but am not completely happy > with the results so far. Has anybody had good results with B-Plus or > another mercury free alternative? Any feedback would be greatly > appreciated. > > Lori White, B.Sc., MLT > Charge Technologist, Histopathology > Lakeridge Health Corporation > Tel 905 576-8711 ext. 4358 > Fax 905 905 721-4757 > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Christl <@t> chempath.uct.ac.za Thu Mar 18 03:15:37 2004 From: Christl <@t> chempath.uct.ac.za (Christl) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] ENaC-alpha Antibody Message-ID: <405968B8.4858DBFB@chempath.uct.ac.za> Hi all Histonetters I would like to know if there is anyone out there who has used the polyclonal Anti-Rat ENaC-alpha antibody from Alpha Diagnostics in the UK? We are trying to get it to work but there seem to be no literature on this. We are using normal rat kidney as a positive control. I have to confess, we've only tried it at three different dilutions and three incubation times with pressure cooking as the Ag retrieval method of choice at the moment. However, seeing as we have very little antibody, I don't want to let ANY go to waste unnecessarily! Any suggestions would be helpful! Thanks Christl From barbara.bublava <@t> meduniwien.ac.at Thu Mar 18 04:18:56 2004 From: barbara.bublava <@t> meduniwien.ac.at (Ing. Barbara Bublava) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Re: cryosectioning of the lung References: Message-ID: <001601c40cd2$6bd4f340$9865fea9@GERICHTS9XOZZ8> Hi Dan, here are the collected answers Curtis D. King: I soak the tung tissue in OCT and sailine 1:1 if at 4-8C overnight. This helps to fill the spaces in the lung tissue. Richard Edwards Inflate lung with proprietary embedding compound..... Vinnie Della Speranza The next issue of HistoLogic will include a comprehensive article by Gayle Callis on the handling of tissues intended for cryosectioning. Gayle may respond to this query herself but have you considered perfusing the lung with OCT before freezing? The OCT can, in this manner, occupy the spaces filled with air that will provide the support the tissue needs during sectioning. Joost Bruijntjes Do you have the opportunity to inflate this lung tissue with OCT? If not, it will really help you. Douglas Gregg DVM PhD Try this, it works for me. The problem is no OCT in the alveolae. Put a drop of OCT on the face of the block and quickly press the face against the freezing metal block in the bottom of the crystat. This will force some OCT into the tissue and you should be able to get at least a few sections beofre having to repeat this procedure. Let me know if it works for you. and one answer, sorry - can?t find the mail right now suggested to soak overnight in 20% sucrose. I found a lot of answers in the archive about the cryoprotecting role of sucrose. I did not know, that it makes the bubbles go away. Thanks for the help. My lung sections are now real pretty ones :o) Barbara ----- Original Message ----- From: "Dan Luchtel" To: Sent: Thursday, March 18, 2004 2:05 AM Subject: cryosectioning of the lung > Would it be possible for you to send to me (or to the list) an e-mail > summary of the responses as apparently, the responses were sent > directly to you and not to the listserv? > > Many thanks, Dan Luchtel > > > > Message: 13 > Date: Fri, 12 Mar 2004 08:39:17 +0100 > From: "Ing. Barbara Bublava" > Subject: [Histonet] Thanks for Replys to "cryosectioning of the lung" > To: > Message-ID: <001701c40805$20182b10$9865fea9@GERICHTS9XOZZ8> > Content-Type: text/plain; charset="iso-8859-1" > > I wish to thank all people who responded to my mail. > The suggested procedures (OTC 1:1, saccarose) did work fine - at least I > decided to go on with the saccarose for the fixed lung and a drop of OTC on > the surface for not fixed lung. > > Barbara Bublava > > > -- > From siksik03 <@t> comcast.net Thu Mar 18 07:58:53 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing In-Reply-To: <001801c40c81$eabac640$c6c710ac@sdspathology.com.au> References: <001801c40c81$eabac640$c6c710ac@sdspathology.com.au> Message-ID: Dear HistoNetters For isopropanol to work as an intermedium, it has to be evaporated out of the paraffin. It is not, technically, a clearing agent. It works in microwave processing because the paraffin is heated above the boiling temperature of isopropanol (82?C at room temperature). I wouldn't think one would want to heat paraffin like this in a routine overnight processor. best regards, Steven Slap Microwave Consultant At 11:42 AM +1100 3/18/04, Histology wrote: > >Hi There > >Does anyone use Isopropanol in routine overnight >parraffin processing replacing xylol?? If you >are willing could you please supply any >protocols you may use. >Thanks for your help. > >Frances >SDS Pathology >Sydney, NSW,Australia From yichaowu <@t> hotmail.com Thu Mar 18 07:58:37 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Please help with me--routine immunofluorescent staining on kidney specimens Message-ID: Dear Dr. Callis, Dr. Li, and everyone here, Hello! It is a long time no seeing.I am the person who inquired how to prepare frozen sections with LN2 last year in June. Later we successfully isolated RNA from acetone/methanol-fixed paraffin-embedded sections and then we do not pay much attention to frozen sections. (Ref: http://www.histosearch.com/histonet/Jun03A/Aboutquotbetterfrozensect.html) But now we have encourtered with a most enormous difficulty.Ours are kidney biopsy specimens from patients. And We found that immunofluorescent stainings of IgG IgA and IgM on acetone/methanol-fixed paraffin-embedded sections are not accurate. There are somewhat pseudo-positive results in them.Therefore we go on to transform all routine immunofluorescent stainings from paraffin-embedded sections to snap-frozen sections. But how is the most popular standard protocol now in the immunofluorescent stainings in renal specimens? Could you kindly give us some suggestions? The antibodies we used are from DAKO, including rabbit anti-human primary antibodies and swine anti-rabbit secondary antibodies.The washing buffer is PBS. Another questions to Dr. Callis, you have mentioned to me that, "A cryomold is what you embed tissue in - to form a block.The chuck is what holds the block when cutting in cryostat.DO NOT USE THE CHUCK FOR SNAP FREEZING..." I just wonder then, how do you TRANSFER the block from cryomold onto the chuck? The second question is,could I store the frozened biopsy specimens in liquid N2 till the sectioning? Will it increase the possibility of empty spaces in the specimens? Thank you very much for your kindest help! Yichao WU,Ph.D candidate Jinling Hospital Nanjing 210002 P.R.China _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From histomjans <@t> yahoo.com Thu Mar 18 08:12:19 2004 From: histomjans <@t> yahoo.com (Melissa Jans) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Help!! millipore and biopsy orientation Message-ID: <20040318141219.69443.qmail@web14909.mail.yahoo.com> We currently receive all our GI biopsies and cervical biopsies oriented on millipore paper. This millipore paper is made of nitrocellulose. It withstands routine processing and does not effect cutting or staining. The biopsies adhere to the millipore paper very well which allows us to maintain orientation of the biopsy throughout grossing, embedding, cutting and so forth. (Current vendor Millipore MF disc Mixed Cellulose Esters hydrophilic 8.0 um 47 mm white plain). Here is my problem: We are changing the way we currently process our biopsies. The millipore we currently use will not survive this processing and therefore we lose orientation of these biopsies. We have tried two other types of millipore but the biopsies do not stick throughout the entire process. (Millipore Mitex disc PTFE hydophobic 5.0 um 47mm white plain and Millipore Mitex PTFE hydrophobic 10 um 47mm white plain). I need some help here. Are there any other institutes using millipore or something else comparable that I could give a try. I am open to any and all suggestions. I am particularly interested in those that use millipore in the microwave successfully. Any help will be greatly appreciated. Melissa Jans University of Iowa Hospitals and Clinics Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam From DOOLEEO <@t> shands.ufl.edu Thu Mar 18 08:29:25 2004 From: DOOLEEO <@t> shands.ufl.edu (Elaine Dooley) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Folate binding protein Message-ID: Dear Histonetters, Vendors, Does any one know of a source for an antibody to Folate binding protein, Folate receptor or Folic Acid that works in formalin fixed paraffin embedded sections. Thanks in Advance Elaine Dooley 352-265-0111 ext 72117 From flemons <@t> bhset.org Thu Mar 18 08:13:03 2004 From: flemons <@t> bhset.org (Fran Lemons) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Washed out spots Message-ID: Hello fellow 'netters! I am trying to troubleshoot a problem with some of our sections, perhaps you can give me some input. Some of our sections, mainly the biopsies, have areas of "washed out" stain. That is to say, there are "spots" where the tissue just didn't seem to take up the hematoxylin. We use the Gemini Varistain autostainer, and the reagents are fresh & in order as they should be. Our biopsies are processed on a 4 hour run separate from large pieces, and all of the reagents on the processor (VIP) are as they should be. I even considered perhaps something in the waterbath that is getting on the sections and "blocking" the hematoxylin, such as a cleaning agent residual; but I am assured by my techs that they use Paragard on the microtomes and hot water only on the waterbaths. Biopsies are cut before the regular run, maybe there is residual on the 'tomes? Dunno. Didn't mean to write a book, but I wanted to provide as much info as possible in case someone else out there is experiencing the same problems. Any suggestions would be helpful. Thanks in advance, Fran Walker BHET Knoxville From katherine-walters <@t> uiowa.edu Thu Mar 18 08:49:17 2004 From: katherine-walters <@t> uiowa.edu (katherine-walters@uiowa.edu) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing In-Reply-To: References: <001801c40c81$eabac640$c6c710ac@sdspathology.com.au> Message-ID: <1079621357.4059b6ed6e007@webmail1.its.uiowa.edu> Hi Frances, We use it routinely in our processor. We have protocols from biopsy to large tissues. I think the times are pretty similar to xylene times, depending of course on the tissues. If you want specifics I can email you our protocols. Kathy Quoting "Steven E. Slap" : > Dear HistoNetters > > For isopropanol to work as an intermedium, it has > to be evaporated out of the paraffin. It is not, > technically, a clearing agent. It works in > microwave processing because the paraffin is > heated above the boiling temperature of > isopropanol (82?C at room temperature). I > wouldn't think one would want to heat paraffin > like this in a routine overnight processor. > > best regards, > Steven Slap > Microwave Consultant > > At 11:42 AM +1100 3/18/04, Histology wrote: > > > >Hi There > > > >Does anyone use Isopropanol in routine overnight > >parraffin processing replacing xylol?? If you > >are willing could you please supply any > >protocols you may use. > >Thanks for your help. > > > >Frances > >SDS Pathology > >Sydney, NSW,Australia > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From gudrun.lang <@t> aon.at Thu Mar 18 09:05:15 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Washed out spots References: Message-ID: <003501c40cfa$6bc4bf70$eeeea8c0@SERVER> Maybe the deparaffination-step before staining is insufficient. On spots of residual paraffin the hematoxylin will not work well. We deparaffinise in two baths of xylen-substitute for at least 10 minutes in summary. Gudrun Lang ----- Original Message ----- From: "Fran Lemons" To: Sent: Thursday, March 18, 2004 3:13 PM Subject: [Histonet] Washed out spots Hello fellow 'netters! I am trying to troubleshoot a problem with some of our sections, perhaps you can give me some input. Some of our sections, mainly the biopsies, have areas of "washed out" stain. That is to say, there are "spots" where the tissue just didn't seem to take up the hematoxylin. We use the Gemini Varistain autostainer, and the reagents are fresh & in order as they should be. Our biopsies are processed on a 4 hour run separate from large pieces, and all of the reagents on the processor (VIP) are as they should be. I even considered perhaps something in the waterbath that is getting on the sections and "blocking" the hematoxylin, such as a cleaning agent residual; but I am assured by my techs that they use Paragard on the microtomes and hot water only on the waterbaths. Biopsies are cut before the regular run, maybe there is residual on the 'tomes? Dunno. Didn't mean to write a book, but I wanted to provide as much info as possible in case someone else out there is experiencing the same problems. Any suggestions would be helpful. Thanks in advance, Fran Walker BHET Knoxville _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From yichaowu <@t> hotmail.com Thu Mar 18 09:44:26 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Please help with me--routine immunofluorescent staining on kidney specimens Message-ID: Dear Dr. Yaskovich, Thank you for your suggestion.But now we prefer to do renal immunofluorescent stainings of IgG IgA IgM C3 C4 C1q on frozen sections. I have realized that frozen sections are the most frequently applied in other labs for renal immunofluorescent stainings of those immunoglobulins. The difficulty of ours is the pseudo-positive results on acetone-fixed paraffin-embedded sections.That is to say,sometime we have positive result of IgA on such sections,but on the frozen section of the same patient, it is negative! As for other kind of fixatives,formalin is also not suitable for immunofluorescent stainings of those immunoglobulins. The reason for such pseudo-positive results may involve multi-factors,I imagine. 1) Maybe the process of acetone-fixation and paraffin-embedding cause auto-fluorescence. 2) Maybe the antibodies we used could not react with those immunoglobulins specifically? But what we used is from DAKO. 3) Another information is, on acetone-fixed paraffin-embedded sections,we have compared, a) Direct staining of IgA with FITC conjugated rabbit-anti-human IgA antibody b) Indirect staining of IgA with rabbit anti-human IgA as primary antibody first,then FITC-conjugated swine anti-rabbit secondary antibody. The results are just different!And frequently all positive in glomerular capillaries for different degree! And in some cases the frozen sections of the same patient is negative. Could anyone kindly give me suggestions? Now we want to do immunofluorescent staining of such immunoglobulins totally on frozen sections.But we do not know the most popular protocol of it in renal pathology in details. Any suggestion would be helpful to us! And we just could not stand pseudo-positive results anymore. Yichao WU Jinling Hospital Nanjing 210002 P.R.China >From: "Yaskovich, Ruth A (NIH/NIDCR)" >To: 'yichao wu' >Subject: RE: [Histonet] Please help with me--routine immunofluorescent >staining on kidney specimens >Date: Thu, 18 Mar 2004 09:47:38 -0500 > >Can you do it the opposite cut the Snap frozen section first then melt down >for paraffin much better sections? Acetone/Methonol is not a good fixative >for Paraffin. >Ruth Yaskovich >Neuronal Gene Expression Section >National Institute of Dental and Crainiofacial Research >N.I.H. >-----Original Message----- >From: yichao wu [mailto:yichaowu@hotmail.com] >Sent: Thursday, March 18, 2004 8:59 AM >To: histonet@lists.utsouthwestern.edu >Cc: tli1@flowcity.bsd.uchicago.edu >Subject: [Histonet] Please help with me--routine immunofluorescent >staining on kidney specimens > > >Dear Dr. Callis, Dr. Li, and everyone here, Hello! > >It is a long time no seeing.I am the person who inquired how to prepare >frozen sections with LN2 last year in June. Later we successfully isolated >RNA from acetone/methanol-fixed paraffin-embedded sections and then we do >not pay much attention to frozen sections. > >(Ref: >http://www.histosearch.com/histonet/Jun03A/Aboutquotbetterfrozensect.html) > >But now we have encourtered with a most enormous difficulty.Ours are kidney >biopsy specimens from patients. And We found that immunofluorescent >stainings of IgG IgA and IgM on acetone/methanol-fixed paraffin-embedded >sections are not accurate. There are somewhat pseudo-positive results in >them.Therefore we go on to transform all routine immunofluorescent >stainings > >from paraffin-embedded sections to snap-frozen sections. > >But how is the most popular standard protocol now in the immunofluorescent >stainings in renal specimens? Could you kindly give us some suggestions? >The > >antibodies we used are from DAKO, including rabbit anti-human primary >antibodies and swine anti-rabbit secondary antibodies.The washing buffer is >PBS. > >Another questions to Dr. Callis, you have mentioned to me that, >"A cryomold is what you embed tissue in - to form a block.The chuck is what >holds the block when cutting in cryostat.DO NOT USE THE CHUCK FOR SNAP >FREEZING..." > >I just wonder then, how do you TRANSFER the block from cryomold onto the >chuck? > >The second question is,could I store the frozened biopsy specimens in >liquid > >N2 till the sectioning? Will it increase the possibility of empty spaces in >the specimens? > >Thank you very much for your kindest help! > >Yichao WU,Ph.D candidate > >Jinling Hospital >Nanjing 210002 >P.R.China > >_________________________________________________________________ >Add photos to your e-mail with MSN 8. Get 2 months FREE*. >http://join.msn.com/?page=features/featuredemail > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail From histo <@t> bthosp.com Thu Mar 18 09:39:30 2004 From: histo <@t> bthosp.com (O'Brien, Sue) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Oil Red O Stain on Fluid Message-ID: <3653750027C5D6119D0600508BB89A9A0ABFFF@MAIL_SERVER> I was wondering how other places handled staining fluids for Oil Red O (or if they did that stain, or another, such as Sudan Black). If so, how do you do the staining (from prep of specimen to final mounting)? Thank-you, Sue O'Brien, Histology Supervisor Burdette Tomlin Memorial Hospital Cape May Court House, NJ 08210 From Dixon.Leslie <@t> mayo.edu Thu Mar 18 09:58:43 2004 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] active-Caspase Message-ID: Hello all, We are having issues with non-specific background staining. I am interested in anyone's protocol for anti-active Caspase-3 on mouse livers. Any help would be greatly appreciated. Thanks, Leslie From gcallis <@t> montana.edu Thu Mar 18 10:01:13 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing In-Reply-To: <001801c40c81$eabac640$c6c710ac@sdspathology.com.au> Message-ID: <3.0.6.32.20040318090113.00bd6990@gemini.msu.montana.edu> We used isopropanol was used in place of ethyl alcohol as a dehydrant, but never replaced the xylene step just before paraffin with it. I don't think isopropyl alcohol is miscible with paraffin. My Merck index says isopropyl is miscible with water, alcohol, ether and chloroform (carcinogenic) but nothing said about paraffin. After isopropyl gradient we cleared with xylene. You will still need a clearant, ie. xylene or one of the xylene substitutes to remove alcohol before infiltration with paraffin. Maybe a better solution is to find a better clearant substitute before paraffin. At 11:42 AM 3/18/2004 +1100, you wrote: >Hi There > >Does anyone use Isopropanol in routine overnight parraffin processing replacing xylol?? If you are willing could you please supply any protocols you may use. >Thanks for your help. > >Frances >SDS Pathology >Sydney, NSW,Australia >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From jcline <@t> wchsys.org Wed Mar 17 13:13:45 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Lori White/ B plus Message-ID: <000001c40d03$25c5cec0$1d2a14ac@wchsys.org> We fix our nodes & BM's in B Plus for a minimum of 2 hours. Then our cores are decaled with HCL/Formalin for one hour and process overnight. We have no problem with the results. From RossS <@t> BaylorHealth.edu Thu Mar 18 10:10:45 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing Message-ID: I think the company that sells the Bond Immunostainer (Vision Biosystems) has developed a process that makes replacing Xylene with Isopropyl Alcohol possible. It has something to do with the heat I think. I was talking with their rep James about this. I admit I still have a lot of questions about the process, but apparently it is possible. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Thursday, March 18, 2004 10:01 AM To: Histology; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Isopropanol tissue processing We used isopropanol was used in place of ethyl alcohol as a dehydrant, but never replaced the xylene step just before paraffin with it. I don't think isopropyl alcohol is miscible with paraffin. My Merck index says isopropyl is miscible with water, alcohol, ether and chloroform (carcinogenic) but nothing said about paraffin. After isopropyl gradient we cleared with xylene. You will still need a clearant, ie. xylene or one of the xylene substitutes to remove alcohol before infiltration with paraffin. Maybe a better solution is to find a better clearant substitute before paraffin. At 11:42 AM 3/18/2004 +1100, you wrote: >Hi There > >Does anyone use Isopropanol in routine overnight parraffin processing replacing xylol?? If you are willing could you please supply any protocols you may use. >Thanks for your help. > >Frances >SDS Pathology >Sydney, NSW,Australia _______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From Barry.R.Rittman <@t> uth.tmc.edu Thu Mar 18 10:16:35 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing Message-ID: <566FB0B522443D43AF02D2ADBE35A6F06359BA@UTHEVS3.mail.uthouston.edu> Isopropyl alcohol is miscible with paraffin wax but is only slowly replaced by it. This technique has been used in the past for delicate tissues and for insects to prevent excessive hardening. Isopropyl alcohol is a very gentle intermediate agent but while I have not used it in tissue processors, I would suspect that it is not suitable for these as it requires a much longer time for its replacement by the wax. The best method to use for hand processing is to make a "slush" of isopropyl alcohol and paraffin wax and allow to sit at room temperature. Sufficient paraffin wax mixes with the alcohol to allow some to penetrate into the tissues. Best to leave several hours to overnight if possible. This will cut down the time in molten paraffin wax. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gayle Callis Sent: Thu 3/18/2004 10:01 AM To: Histology; Histonet@lists.utsouthwestern.edu Cc: Subject: Re: [Histonet] Isopropanol tissue processing We used isopropanol was used in place of ethyl alcohol as a dehydrant, but never replaced the xylene step just before paraffin with it. I don't think isopropyl alcohol is miscible with paraffin. My Merck index says isopropyl is miscible with water, alcohol, ether and chloroform (carcinogenic) but nothing said about paraffin. After isopropyl gradient we cleared with xylene. You will still need a clearant, ie. xylene or one of the xylene substitutes to remove alcohol before infiltration with paraffin. Maybe a better solution is to find a better clearant substitute before paraffin. At 11:42 AM 3/18/2004 +1100, you wrote: >Hi There > >Does anyone use Isopropanol in routine overnight parraffin processing replacing xylol?? If you are willing could you please supply any protocols you may use. >Thanks for your help. > >Frances >SDS Pathology >Sydney, NSW,Australia >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PDavala <@t> sjh-nh.org Thu Mar 18 10:52:25 2004 From: PDavala <@t> sjh-nh.org (Davala, Pam) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Bouin's Substitute Message-ID: To all out there... Is there a reliable substitute for Bouin's Mordant in the Masson Trichrome. We had a lab accident with Bouin's Mordant from the microwave.... and I would like to get rid of this if possible... Inhaled it is lethel. If someone has an answer... I would really appreciate it. thanks From yichaowu <@t> hotmail.com Thu Mar 18 10:52:12 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Re: Please help with me--routine immunofluorescent staining on kidney specimens Message-ID: Dear Dr. Callis, Thank you very much! And could you kindly suggest me that what kind of FITC-labelled primary antibodies to IgG and IgA etc should be applied on renal frozen sections? Are F(ab'2) antibodies preferred than IgG fraction? Is methanol caused the autofluorescence or unproper fluorescence in glomerular capillaries? Thank you. Yichao WU >From: Gayle Callis >To: "yichao wu" >Subject: Re: Please help with me--routine immunofluorescent staining on >kidney specimens >Date: Thu, 18 Mar 2004 09:15:08 -0700 > >Get rid of methanol, use cold pure acetone fixation ONLY!! > > >Another questions to Dr. Callis, you have mentioned to me that, > >"A cryomold is what you embed tissue in - to form a block.The chuck is >what > >holds the block when cutting in cryostat.DO NOT USE THE CHUCK FOR SNAP > >FREEZING..." > > > >I just wonder then, how do you TRANSFER the block from cryomold onto the > >chuck? > >Pop the block out of mold and freeze block onto little metal chuck that >holds blocks during sectioning. Do this with OCT. Use your fingers and >work quickly to not melt parts of the block where tissue is located. > > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > _________________________________________________________________ The new MSN 8: advanced junk mail protection and 2 months FREE* http://join.msn.com/?page=features/junkmail From la.sebree <@t> hosp.wisc.edu Thu Mar 18 10:43:19 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Please Help with Cresyl Violet Acetate Message-ID: Good Morning Histoneters, I am trying to work-up a CEVstain for nissel substance and I am having difficulties producing a magenta effect on the nissel. I have tried several powders with the CI # of 10510-54-0 along with different staining procedures. I would appreciate any help or suggestions anyone has to offer. Angela Baker 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 pedro.louro <@t> spcorp.com Thu Mar 18 11:18:42 2004 From: pedro.louro <@t> spcorp.com (Louro, Pedro) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Fekete's fixative Message-ID: <4508920F80C0D411B90200508BF9A9F4062B45FB@LAFMSG30.us.schp.com> I need Histo. assistance: I'm looking for a recipe or protocol for Fekete's Fixative. please help! Thanks ********************************************************************* 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 David.Edmondson <@t> christie-tr.nwest.nhs.uk Thu Mar 18 11:34:12 2004 From: David.Edmondson <@t> christie-tr.nwest.nhs.uk (Edmondson David (RBV) NHS Christie Tr) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Immunostainers Message-ID: We are using a DAKO Techmate 500 Plus, Daily runs of upto 240 slides at the moment but there seems no reason not to introduce more runs as the need arises. It is very reliable. Touch wood. The reagent and hardware costs are not insignificant but the manpower savings justify them. We use DAKO's Envision which takes a little off the run time. Having the pretreatments off the machine makes it easier to maximise the run time too. Company support has been good, both machinery and consumables. David Manchester UK -----Original Message----- From: Dana Settembre [mailto:settembr@umdnj.edu] Sent: 17 March 2004 12:40 To: Histonet@lists.utsouthwestern.edu; STEGTM@samcstl.org Subject: Re: [Histonet] Immunostainers Hello Therersa, I have a Dako and am very happy with it. I am sure that you've gone through the demos so you already know how easy it is to use. So I'll tell you about their great technical services dept. Whenever I have any questions, they are fantastic. That dept alone is a reason to lean towards the Dako. If you haven't gone through the demo, call them. We purchased one about 8 years ago and then as our volume increased and we saw the need for another, Dako suggested that we look into their, "Reagent Acquisition" plan. We were purchasing enough of their reagents on a monthly basis, that we were able to get a second virtually for free. Also, please know that you are NOT required to use their products either. The Dako autostainer can use any reagents from any company. Hope this helps. Dana Settembre Immunohistochemistry Lab University Hospital - UMDNJ- Newark, NJ >>> Therersa Stegall 3/16/2004 10:45:04 AM >>> We're about to purchase an immunostainer for our lab. We've sort of narrowed the choices down between the Ventana and Dako autostainer models. Since both companies are competetive for pricing to us, we would like to hear any feedback good/bad/otherwise about these stainers from folks using them. Any comments/advice you can offer will be appreciated. Peace, Terre _______________________________________________ Histonet mailing list Histonet@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 STEGTM <@t> samcstl.org Thu Mar 18 11:42:29 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Great response! Message-ID: Thank you to everyone that responded to our question on Immunostainers. I've got about 70 good responses to consider and filter for information, and several sales pitches. Thanks for the "net", a place where we can receive such help. TS From mward <@t> wfubmc.edu Thu Mar 18 11:50:31 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Autopsy survey Message-ID: <61135F0455D33347B5AAE209B903A304076A4DEB@EXCHVS2.medctr.ad.wfubmc.edu> I would like to thank everyone who responded to our survey. My manager and administrator appreciate everyone who took the time to share information with us. Thank you again! Martha Ward Wake Forest University Baptist Medical Center From DEllenburg2 <@t> stfrancishealth.org Thu Mar 18 12:02:55 2004 From: DEllenburg2 <@t> stfrancishealth.org (DEllenburg2@stfrancishealth.org) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Decontamination of Cryostat Message-ID: <6661F19BB774D711942900A0C905F2533D8AFD@GVL01MSX> Dear Histonetters, I need help with the following question on the CAP checklist. Question ANP.24250 --- Is there a documented procedure for the routine decontamination of the cryostat at defined intervals, and are decontamination records evident? Does anyone know how often "defined intervals" is referring to? Your help would be greatly appreciated. Thanks, Deborah Ellenburg, HT (ASCP) Histology Supervisor BonSecours St. Francis Hospital Greenville, SC The information in this communication is intended to be confidential to the Individual(s) and/or Entity to whom it is addressed. It may contain information of a Privileged and/or Confidential nature, which is subject to Federal and/or State privacy regulations. In the event that you are not the intended recipient or the agent of the intended recipient, do not copy or use the information contained within this communication, or allow it to be read, copied or utilized in any manner, by any other person(s). Should this communication be received in error, please notify the sender immediately either by response e-mail or by phone at (864) 255-1000, and permanently delete the original e-mail, attachment(s), and any copies. From gcallis <@t> montana.edu Thu Mar 18 12:02:57 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Oil Red O Stain on Fluid In-Reply-To: <3653750027C5D6119D0600508BB89A9A0ABFFF@MAIL_SERVER> Message-ID: <3.0.6.32.20040318110257.00bbf090@gemini.msu.montana.edu> You cannot fix cells containing fat/lipids with alcohol or acetone or the lipids are removed by these solvents. For cytospins and cell cultures - fix with NBF or paraformaldehyde, rinse and do the Churukian Oil Red O stain, coverslip with aqueous mounting media to retain Oil Red O staining. Oil Red O/Dextrin, Churukian method: Fresh tissue frozen sections fixed post cutting with NBF can be rinsed and stained immediately, or fixed frozen sections (cryoprotected), mounted on Plus Charge slides. Air dry NBF fixed frozen sections 30 min to 1 hour, or longer to insure they stay on slide. Cells can be cytospun, cultured, and fixed with NBF. Protocol: 1. Immerse dry slides directly into filtered 0.5% Oil Red O in Dextrin , stain 20 minutes 2. Rinse VERY GENTLY in running tap water 3. Counterstain with Gill II hematoxylin for 20 - 30 seconds 4. Rinse gently with water, blue in bluing solution, (NOT AMMONIA WATER), rinse gently, and coverslip with aqueous mounting media Reagents: Dissolve 0.5 gm Oil Red O in absolute isopropyl alcohol, allow to stir overnight. Dissolve 1 gm dextrin (bacteriological grade or TYPE III (Sigma) from corn in 100 ml distilled water Working solution is 60 mls stock Oil Red O and 40 ml 1% dextrin solution Stable for months, and reported to work on paraffin sections. This is a very clean stain, and not MESSY! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From AFeatherstone <@t> KaleidaHealth.Org Thu Mar 18 12:01:52 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Please Help with Cresyl Violet Acetate Message-ID: Our Cresyl Echt Violet is CEV Stock Sol. CI-51010 --5gm D. H2O----80 ml Absolute--20 ml Warm the distilled, add the CEV mix, then add alcohol. Working CEV solution 45 ml of stock Acetic Acid, glacial--15 drops Deparaffinize, stain in working sol for 8 minutes, dehydrate, clear. Nissl substance and nuclei----Blue-purple Background--------------------Colorless Annette Featherstone HT/MLT -----Original Message----- From: Sebree Linda A. [mailto:la.sebree@hosp.wisc.edu] Sent: Thursday, March 18, 2004 11:43 To: Histonet (E-mail) Subject: [Histonet] Please Help with Cresyl Violet Acetate Good Morning Histoneters, I am trying to work-up a CEVstain for nissel substance and I am having difficulties producing a magenta effect on the nissel. I have tried several powders with the CI # of 10510-54-0 along with different staining procedures. I would appreciate any help or suggestions anyone has to offer. Angela Baker 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 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 stevemachinuk <@t> yahoo.co.uk Thu Mar 18 11:50:21 2004 From: stevemachinuk <@t> yahoo.co.uk (=?iso-8859-1?q?Steve=20Machin=20UK?=) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Oil Red O Stain on Fluid In-Reply-To: <3653750027C5D6119D0600508BB89A9A0ABFFF@MAIL_SERVER> Message-ID: <20040318175021.28930.qmail@web25108.mail.ukl.yahoo.com> We use a Cytospin centrifuge on a slow speed to avoid splattering the lipid, fix in conc formalin vapour, then stain in Oil Red O etc. --- "O'Brien, Sue" wrote: > I was wondering how other places handled staining fluids for Oil > Red O (or > if they did that stain, or another, such as Sudan Black). If so, > how do you > do the staining (from prep of specimen to final mounting)? > Thank-you, > Sue O'Brien, Histology Supervisor > Burdette Tomlin Memorial Hospital > Cape May Court House, NJ 08210 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___________________________________________________________ Yahoo! Messenger - Communicate instantly..."Ping" your friends today! Download Messenger Now http://uk.messenger.yahoo.com/download/index.html From Janet.Bonner <@t> FLHOSP.ORG Thu Mar 18 11:56:04 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] iron stain on bone marrow-SOLUTIONS Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB3F9A@fh2k093.fhmis.net> The 20% Hydrochloric acid solution should be added to the 10% Potassium ferrocyanide solution - (not 10% potassium ferrocyanide to 20% Hydrochloric acid solution) when making the 1:1 mixture. -----Original Message----- From: Wendy England [mailto:weneng@hotmail.com] Sent: Wednesday, March 17, 2004 3:28 PM To: histonet@pathology.swmed.edu Subject: [Histonet] iron stain on bone marrow Thanks again to everybody who gave me the information about this stain! Here comes another question: I couldn't see much iron on my slide, only about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 min, then in freshly mixed stain solution(equal part of 20% HCl and 10% potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin after wash. Then wash, air dry then mount. I was told there should be much more iron. I feel bad now. Could any body give me advice or could I ask if it's possible due to the bone marrow preparation since we indeed saw some spots. I didn't prepare the sample. Thanks and look forward to hear from you there! Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar - get it now! http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From ldunikoski <@t> rbmc.org Thu Mar 18 12:35:06 2004 From: ldunikoski <@t> rbmc.org (Dunikoski, Leonard PhD) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] iron stain on bone marrow-SOLUTIONS Message-ID: "Remember the order: acid to water" -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: Thursday, March 18, 2004 12:56 PM To: 'Wendy England'; histonet@pathology.swmed.edu Subject: RE: [Histonet] iron stain on bone marrow-SOLUTIONS The 20% Hydrochloric acid solution should be added to the 10% Potassium ferrocyanide solution - (not 10% potassium ferrocyanide to 20% Hydrochloric acid solution) when making the 1:1 mixture. -----Original Message----- From: Wendy England [mailto:weneng@hotmail.com] Sent: Wednesday, March 17, 2004 3:28 PM To: histonet@pathology.swmed.edu Subject: [Histonet] iron stain on bone marrow Thanks again to everybody who gave me the information about this stain! Here comes another question: I couldn't see much iron on my slide, only about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 min, then in freshly mixed stain solution(equal part of 20% HCl and 10% potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin after wash. Then wash, air dry then mount. I was told there should be much more iron. I feel bad now. Could any body give me advice or could I ask if it's possible due to the bone marrow preparation since we indeed saw some spots. I didn't prepare the sample. Thanks and look forward to hear from you there! Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar - get it now! http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet NOTICE: The information included in this email contains confidential information belonging to the sender. This information is intended only for the use of the individual or entity named above. If you are not the intended recipient, you are hereby notified that any disclosure, copying, distribution or action taken in reliance on the contents hereof is strictly prohibited. If you have received this email in error, please notify the sender immediately. From brett_connolly <@t> merck.com Thu Mar 18 13:02:30 2004 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Iron stain on frozen sections Message-ID: Along with the bone marrow thread..... Has anyone had experience with iron stains on frozen sections? What fixative? Procedure? Thx, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ------------------------------------------------------------------------------ Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ------------------------------------------------------------------------------ From pruegg <@t> colobio.com Thu Mar 18 12:50:38 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] iron stain on bone marrow In-Reply-To: Message-ID: Actually we used to get our best iron stains on smears since processing can leach out iron in the tissue. Perhaps a shorter fixation in methanol would help, alcohol can also leach out the hemosiderin. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Wendy England Sent: Wednesday, March 17, 2004 1:28 PM To: histonet@pathology.swmed.edu Subject: [Histonet] iron stain on bone marrow Thanks again to everybody who gave me the information about this stain! Here comes another question: I couldn't see much iron on my slide, only about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 min, then in freshly mixed stain solution(equal part of 20% HCl and 10% potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin after wash. Then wash, air dry then mount. I was told there should be much more iron. I feel bad now. Could any body give me advice or could I ask if it's possible due to the bone marrow preparation since we indeed saw some spots. I didn't prepare the sample. Thanks and look forward to hear from you there! Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar ? get it now! http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From victor <@t> pathology.washington.edu Thu Mar 18 12:11:43 2004 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Fekete's fixative References: <4508920F80C0D411B90200508BF9A9F4062B45FB@LAFMSG30.us.schp.com> Message-ID: <4059E65F.8020909@pathology.washington.edu> Tellyesniczky/Fekete (Telly's) Fixative - [Lillie, 1965] 70%ETOH 100ml Glacial acetic acid 5ml 37-40% formaldehyde conc. 10ml Recommended for the preservation of glycogen. Lyses rbc's. Widely used by botanist's. Transfer to 85% ETOH. Humason,G. (1972) Animal Tissue Techniques. W.H.Freeman and Co., San Francisco. Eric C .Kellar Histology/Immunohistochemistry University of Pittsburgh Medical Center Louro, Pedro wrote: >I need Histo. assistance: > >I'm looking for a recipe or protocol for Fekete's Fixative. > >please help! > >Thanks > > > >********************************************************************* >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 > > -- Victor Tobias Clinical Applications Analyst Dept of Pathology University of Washington Medical Center 206-598-2792 206-598-7659 Fax From nick.kirk3 <@t> btopenworld.com Thu Mar 18 13:13:09 2004 From: nick.kirk3 <@t> btopenworld.com (=?iso-8859-1?q?NICK=20KIRK?=) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] iron stain on bone marrow-SOLUTIONS In-Reply-To: <07AB60D5D7B9754EBF56F360F98D083DEB3F9A@fh2k093.fhmis.net> Message-ID: <20040318191309.39869.qmail@web86308.mail.ukl.yahoo.com> Hi there In my experience it doesn't matter which way round you add the solutions, plus I think the HCl solution you are using is far too strong. The Perl's prussian blue technique I use uses 2% HCl and 2% Pot ferro and works excellently every time even on marrows. We incubate for about 15 minutes. Hope that helps Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England "Bonner, Janet" wrote: The 20% Hydrochloric acid solution should be added to the 10% Potassium ferrocyanide solution - (not 10% potassium ferrocyanide to 20% Hydrochloric acid solution) when making the 1:1 mixture. -----Original Message----- From: Wendy England [mailto:weneng@hotmail.com] Sent: Wednesday, March 17, 2004 3:28 PM To: histonet@pathology.swmed.edu Subject: [Histonet] iron stain on bone marrow Thanks again to everybody who gave me the information about this stain! Here comes another question: I couldn't see much iron on my slide, only about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 min, then in freshly mixed stain solution(equal part of 20% HCl and 10% potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin after wash. Then wash, air dry then mount. I was told there should be much more iron. I feel bad now. Could any body give me advice or could I ask if it's possible due to the bone marrow preparation since we indeed saw some spots. I didn't prepare the sample. Thanks and look forward to hear from you there! Wendy _________________________________________________________________ FREE pop-up blocking with the new MSN Toolbar - get it now! http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From acjanes <@t> bu.edu Thu Mar 18 13:17:54 2004 From: acjanes <@t> bu.edu (Amy Janes) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Storing Tissue Message-ID: Hi, I wrote before because my mouse brain tissue was full of holes and I was told I should flash freeze the tissue, which I did and it seemed to work. However, when I used this technique I cut the tissue immediatley after freezing. Now I am running ICC on tissue that was flash frozen and then stored at -20C. The tissue again looks like it has tiny holes. I am running ICC for a retrograde tracer which is supposed to label cell bodies but instead looks like a smear of color and I cannot see individual cells. I am double labeling for fos and I get no staining (because I think the cells are destroyed). Any suggestions? Is there a better/safer way to store tissue if I won't be able to cut it quickly? Thanks, Amy Janes, MA Boston University Department of Psychology Molecular Neurobiology and Behavior Laboratory From shive003 <@t> umn.edu Thu Mar 18 13:50:55 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] mouse bone osteoblast IHC Message-ID: <005401c40d22$538a53c0$78065486@vdl220FAC> I am posting this for an MD not on the Histonet listserv; he is requesting information on staining mouse osteoblasts using the IHC method. (I have had no experience with osteoblasts myself). Does anyone know of an antibody source for detecting murine osteoblasts (or one made against another species that will cross-react)? What decalcification method is used on the bone samples? Any complete protocols that anyone is willing to share would be greatly appreciated. Thanks... Jan Shivers U of MN Vet Diag Lab From gcallis <@t> montana.edu Thu Mar 18 14:06:44 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] iron stain on bone marrow-SOLUTIONS In-Reply-To: Message-ID: <3.0.6.32.20040318130644.00bea8f0@gemini.msu.montana.edu> Have used this rule with CONCENTRATED acid going into water to prevent spattering plus creating heat. However, mixing 20% HCl (more water than acid) with 10% potassium ferrocyanide didn't make any difference. I am sure I have mixed these both ways, and I never noticed spattering. Histotechnic textbooks do not address the safety issue here nor state any sequence of events. The old poem learned in my college chemistry class Chem 101 back in the dark ages was applied to concentrated stock acids, and still used - goes: "Always do what you oughter, pour the acid in the water!" At 01:35 PM 3/18/2004 -0500, you wrote: >"Remember the order: acid to water" > >-----Original Message----- >From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] >Sent: Thursday, March 18, 2004 12:56 PM >To: 'Wendy England'; histonet@pathology.swmed.edu >Subject: RE: [Histonet] iron stain on bone marrow-SOLUTIONS > > >The 20% Hydrochloric acid solution should be added to the 10% Potassium >ferrocyanide solution - (not 10% potassium ferrocyanide to 20% Hydrochloric >acid solution) when making the 1:1 mixture. > > >-----Original Message----- >From: Wendy England [mailto:weneng@hotmail.com] >Sent: Wednesday, March 17, 2004 3:28 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] iron stain on bone marrow > > >Thanks again to everybody who gave me the information about this stain! Here >comes another question: I couldn't see much iron on my slide, only >about 2~3 spots per rat bone marrow slide. I fixed them in Methanol for 15 >min, then in freshly mixed stain solution(equal part of 20% HCl and 10% >potassium ferrocyanide) for 30 min in 50-60C. Then counterstain in Safranin >after wash. Then wash, air dry then mount. > >I was told there should be much more iron. I feel bad now. Could any body >give me advice or could I ask if it's possible due to the bone marrow >preparation since we indeed saw some spots. I didn't prepare the sample. > >Thanks and look forward to hear from you there! > >Wendy > >_________________________________________________________________ >FREE pop-up blocking with the new MSN Toolbar - get it now! >http://clk.atdmt.com/AVE/go/onm00200415ave/direct/01/ > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >The information contained in this message may be privileged and/or >confidential and protected from disclosure. If the reader of this message >is not the intended recipient or agent responsible for delivering this >message to the intended recipient, you are hereby notified that any >dissemination, distribution or copying of this communication is strictly >prohibited. If you have received this communication in error, please notify >the sender immediately by replying to this message and deleting the material >from any computer. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet >NOTICE: The information included in this email contains confidential >information belonging to the sender. This information is intended only for >the use of the individual or entity named above. If you are not the intended >recipient, you are hereby notified that any disclosure, copying, >distribution or action taken in reliance on the contents hereof is strictly >prohibited. If you have received this email in error, please notify the >sender immediately. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From caroline.stott <@t> anatomy.otago.ac.nz Thu Mar 18 14:19:51 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:41 2005 Subject: [Histonet] Isopropanol tissue processing Message-ID: <5.2.1.1.0.20040319091830.02908710@anatomy.otago.ac.nz> Hi Yeah with microwave processing we go straight from isopropyl alcohol to paraffin. Never had any trouble. Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From mcauliff <@t> umdnj.edu Thu Mar 18 17:07:39 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Storing Tissue In-Reply-To: References: Message-ID: <405A2BBB.1000509@umdnj.edu> Hi Amy: Amy Janes wrote: >Hi, >I wrote before because my mouse brain tissue was full of holes and I was >told I should flash freeze the tissue, which I did and it seemed to work. >However, when I used this technique I cut the tissue immediatley after >freezing. Now I am running ICC on tissue that was flash frozen and then >stored at -20C. > How long was the storage? Did you store sections or the tissue block? Minus 20 may not be cold enough to retain antigenicity, some antigens degrade at minus 80. >The tissue again looks like it has tiny holes. I am >running ICC for a retrograde tracer which is supposed to label cell bodies but instead looks like a smear of color and I cannot see individual cells. > Is this fixed or fresh frozen tissue? >I am double labeling for fos and I get no staining (because I think the >cells are destroyed). Any suggestions? > Have you done each label by itself to see the results? If so, how do they look? How does your known positive control look? >Is there a better/safer way to >store tissue if I won't be able to cut it quickly? > Depends on the antigen and how you are storing the tissue. First off, -20 may not be cold enough but I need to know more (see above). Geoff > >Thanks, > >Amy Janes, MA >Boston University >Department of Psychology >Molecular Neurobiology >and Behavior Laboratory > >_______________________________________________ >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 nyilmaz <@t> mersin.edu.tr Thu Mar 18 22:28:52 2004 From: nyilmaz <@t> mersin.edu.tr (nyilmaz@mersin.edu.tr) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Placental section ihc... Message-ID: We are using placental tissue for immunohistochemistry, and have some problems about background staining especially with serum remnants in the vessels. Does anybody have ideas about this problem. If you inform us we'd be greatly appreciate... Nejat Yilmaz PhD Mersin University Medical School From staceylburton <@t> yahoo.com Thu Mar 18 14:26:06 2004 From: staceylburton <@t> yahoo.com (Stacey Burton) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] fume hood regulation Message-ID: <20040318202606.65831.qmail@web14521.mail.yahoo.com> I am currently installing 12 fume hoods into 12 laboratories. I have been informed, by the installer, of some new fume hood safety standards set by OSHA. I have been told in order to be OSHA compliant, I must have an audible and visual alarm on each fume hood that will react if the fume hood fails to work properly. Have any of you heard of this policy? If so, will you please educate me on these new standards? I want to install the alarms if it is indeed a new rule of compliance; however, I could do without the costly expense if this is only a rumor. Thanks for your input, Stacey Burton, HT (ASCP) Laboratoy Manager Uropath LLC San Antonio TX 210) 521-7700 office 210) 521-7710 fax staceylburton@yahoo.com Do you Yahoo!? Yahoo! Mail - More reliable, more storage, less spam From acjanes <@t> bu.edu Thu Mar 18 14:30:56 2004 From: acjanes <@t> bu.edu (Amy Janes) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Storing Tissue In-Reply-To: <405A2BBB.1000509@umdnj.edu> References: <405A2BBB.1000509@umdnj.edu> Message-ID: > How long was the storage? Less than a week > Did you store sections or the tissue block? tissue block imbedded in OCT and wrapped in foil. > Is this fixed or fresh frozen tissue? Fixed. > Have you done each label by itself to see the results? Yes. I ran the labeling on Brain cut at 30microns and olfactory Bulb cut at 20 microns and labeled them free floating. The fos has lots of background and little to know fos staining. The retrograde label stains in the correct area and the background is fine. There are no individual cells (I should see distinict cell body staining) but just a smear of staining. > they look? How does your known positive control look? I havent run a positive control with this group. I have run positive controls on other sections using the same procedure and didnt think it was necessary. Thanks, Amy From garsha <@t> itg.uiuc.edu Thu Mar 18 14:55:23 2004 From: garsha <@t> itg.uiuc.edu (Karl Garsha) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] digesting soft tissue Message-ID: <405A0CBB.6080202@itg.uiuc.edu> Greetings, I'm trying to find a good cocktail for non-specific digestion of tissue from bone samples--only the mineralized bone needs to be left unscathed. I'm looking for a relatively cheap solution that will yield nice clean bones. I'm aware of a product from Fisher, but it costs about $70/100ml. It seems to me that there must be a simple solution that I'm just not aware of (aside from culturing maggots). Thanks in advance for any suggestions. Regards, Karl G. -- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu From mcauliff <@t> umdnj.edu Thu Mar 18 18:10:24 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] digesting soft tissue In-Reply-To: <405A0CBB.6080202@itg.uiuc.edu> References: <405A0CBB.6080202@itg.uiuc.edu> Message-ID: <405A3A70.6000502@umdnj.edu> Household bleach, 1 part bleach to 4 parts water. or Adolph's meat tenderizer is papain, don't know the concentration needed. You can get papain form Sigma, works well at 37C. I don't know any other details. Karl Garsha wrote: > Greetings, > I'm trying to find a good cocktail for non-specific digestion of > tissue from bone samples--only the mineralized bone needs to be left > unscathed. I'm looking for a relatively cheap solution that will > yield nice clean bones. I'm aware of a product from Fisher, but it > costs about $70/100ml. It seems to me that there must be a simple > solution that I'm just not aware of (aside from culturing maggots). > Thanks in advance for any suggestions. > Regards, > Karl G. > -- -- ********************************************** 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 RossS <@t> BaylorHealth.edu Thu Mar 18 15:27:37 2004 From: RossS <@t> BaylorHealth.edu (Stapf, Ross) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] digesting soft tissue Message-ID: Found these in the archives. I thought I remembered this coming up before. Enjoy. Ross M Stapf Histopathology Manager Baylor University Medical Center 3500 Gaston Ave. Dallas, TX 75246 214-820-2465 214-820-4110 fax RossS@baylorhealth.edu To macerate tissue away, use Biz detergent (local grocery store)- an enzyme detergent. Make a 20% solution and heat bone at 80C until muscle comes, slides off bone surfaces. The enzyme is the kicker here, eats away at protein attachments. It bleachs bones as well, smells better than just boiling bones. After Biz treatment, rinse bones well with running tap water - let them air dry. It is a good idea to fix bones with NBF or 70% alcohol, then rinse well overnight to get rid of formalin - you don't want to inactivate enzyme in detergent. Also, fixed bone holds together better (cartilage is removed by BIZ!) and if bones (or parts of same) separate, glue with Duco Cement, Superglue, or a glue gun after total drying. Nice way to learn bone anatomy by putting it back together. This method came from Journal of Anatomy, many moons ago, 1980's - to prepare skulls for gross specimen study. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology - Marsh Lab Montana State University - Bozeman S. 19th and Lincoln St Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) email: gcallis@montana.edu This is for those who would like something whose odor befits its work. I think Gayle Callis' BIZ might be better, but Gee Whiz folks, we're talking about Gross Anatomy here! These are procedures that one used to be forced to carry out on the roof, or out in an open field, away from civilized folk. So, I thought I would contribute a recipe from an old, VERY reputable, source. "Liquid Soap". Reighard and Jennings, "Anatomy of the Cat" [Oops! Sorry kitty!], 1935, Holt Reinhart Soft water 2000ml ("Soft" is not a scientific term!) Strong ammonia 150ml (for glazed eyes, opaque NICtitating membranes and, of course, alkalinity) KNO3 12g (for suppression of something? John and Abigail?) Hard soap (brown) 75g (We used to save the shavings of the brown soap used in the local Cub Scout carving tutorials. We liked the brown soap, because it WAS harder than Ivory! This soap is still available in a double bar with attractive white wrapping with pink printing. You can't tell a soap by its cover!) Heat the mixture to boiling until homogeneous. Immerse the bone and boil for 40 min Pour off the liquid and renew it. Boil another 30min or until the soft parts come away easily. [Boil too long and the epiphyses will separate.] The bone can be rinsed in water. The bone can be defatted, after drying, in bnenzene/zylene. Cheers (it almost feels like Friday), Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson@wcupa.edu -----Original Message----- From: Kim Merriam [mailto:kmerriam@tktx.com] Sent: Tuesday, December 10, 2002 12:30 PM To: Histonet Subject: Dissolving skeletal muscle from bone Hi all, Does anyone know what solution to use to dissolve skeletal muscle off of bone, so that the bone remains in tact. This is not for a histology study, but I thought that someone out there on the histonet would know how to do this! Thanks in advance, Kim Merriam TKT Cambridge, MA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Geoff McAuliffe Sent: Thursday, March 18, 2004 6:10 PM To: Karl Garsha Cc: histonet@lists.utsouthwestern.edu; Microscopy@MSA.Microscopy.Com Subject: Re: [Histonet] digesting soft tissue Household bleach, 1 part bleach to 4 parts water. or Adolph's meat tenderizer is papain, don't know the concentration needed. You can get papain form Sigma, works well at 37C. I don't know any other details. Karl Garsha wrote: > Greetings, > I'm trying to find a good cocktail for non-specific digestion of > tissue from bone samples--only the mineralized bone needs to be left > unscathed. I'm looking for a relatively cheap solution that will > yield nice clean bones. I'm aware of a product from Fisher, but it > costs about $70/100ml. It seems to me that there must be a simple > solution that I'm just not aware of (aside from culturing maggots). > Thanks in advance for any suggestions. > Regards, > Karl G. > -- -- ********************************************** 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 ********************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet This e-mail, facsimile, or letter and any files or attachments transmitted with it contains information that is confidential and privileged. This information is intended only for the use of the individual(s) and entity(ies) to whom it is addressed. If you are the intended recipient, further disclosures are prohibited without proper authorization. If you are not the intended recipient, any disclosure, copying, printing, or use of this information is strictly prohibited and possibly a violation of federal or state law and regulations. If you have received this information in error, please notify Baylor Health Care System immediately at 1-866-402-1661 or via e-mail at privacy@baylorhealth.edu. Baylor Health Care System, its subsidiaries, and affiliates hereby claim all applicable privileges related to this information. From peoshel <@t> wisc.edu Thu Mar 18 15:09:41 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] digesting soft tissue In-Reply-To: <405A0CBB.6080202@itg.uiuc.edu> References: <405A0CBB.6080202@itg.uiuc.edu> Message-ID: Karl, NaOH or KOH or bleach, I've used all. Check the fish people on campus -- this is a routine matter for them. Mind, using base or bleach creates a pleasant odor your lab mates will be hanging you for. Phil >Greetings, >I'm trying to find a good cocktail for non-specific digestion of >tissue from bone samples--only the mineralized bone needs to be left >unscathed. I'm looking for a relatively cheap solution that will >yield nice clean bones. I'm aware of a product from Fisher, but it >costs about $70/100ml. It seems to me that there must be a simple >solution that I'm just not aware of (aside from culturing maggots). >Thanks in advance for any suggestions. >Regards, >Karl G. > >-- >Karl Garsha >Light Microscopy Specialist >Imaging Technology Group >Beckman Institute for Advanced Science and Technology >University of Illinois at Urbana-Champaign >405 North Mathews Avenue >Urbana, IL 61801 >Office: B650J >Phone: 217.244.6292 >Fax: 217.244.6219 >Mobile: 217.390.1874 >www.itg.uiuc.edu > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From dmcaloose <@t> wcs.org Thu Mar 18 15:45:05 2004 From: dmcaloose <@t> wcs.org (McAloose, Dee) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Job opening - Wildlife Conservation Society Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD6B0@WOLF.wcs.org> Hello, I'd like to announce an immediate opening for a full-time histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity and in becoming part of our team! Thanks for listening and we look forward to hearing from you. D McAloose, VMD, Dipl ACVP Head, Department of Pathology Wildlife Conservation Society Bronx, NY 10464 (phone) 718-220-7105 (fax) 718-220-7126 dmcaloose@wcs.org From ccrowder <@t> mail.vetmed.lsu.edu Thu Mar 18 15:57:46 2004 From: ccrowder <@t> mail.vetmed.lsu.edu (Cheryl Crowder) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Fekete fixative Message-ID: According to Lillie (p. 34) Fekete fixative is an acetic acid-alcohol- formalin fixative. The recipe is: 37-40% formaldehyde 10 ml Glacial acetic acid 5 ml 70% alcohol 100 ml It is just slightly different from several other formulae containing the same ingredients. Cheryl Crowder, BA, HTL(ASCP) Chief Technologist Anatomic Pathology Department of Pathobiological Sciences School of Veterinary Medicine Louisiana State University Skip Bertman Drive Baton Rouge, LA ?70803 225-578-9734 FAX: ?225-578-9720 From pruegg <@t> colobio.com Thu Mar 18 16:20:17 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Placental section ihc... In-Reply-To: Message-ID: I too am staining placenta for IHC and it has lots of problems. There is a tremendous amount of red cells, hemosiderin, etc. and so it is difficult to quench all of the endogenous peroxidase. If you are using a biotin detection system you could have real trouble with endogenous biotin as well. I use a labelled polymer detection to get around that. I also block with serum free protein before the primary antibody and then again before the secondary. If I still have problems I use serum block (serum from the animal species the secondary is made in) again before primary and before secondary. It might be better to use alk. phos. or aec as a chromogen because like melanin in skin the general tissue background is brown so for better contrast some other chromogen besides DAB might be more appealing. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of nyilmaz@mersin.edu.tr Sent: Thursday, March 18, 2004 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Placental section ihc... We are using placental tissue for immunohistochemistry, and have some problems about background staining especially with serum remnants in the vessels. Does anybody have ideas about this problem. If you inform us we'd be greatly appreciate... Nejat Yilmaz PhD Mersin University Medical School _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Thu Mar 18 16:42:16 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] digesting soft tissue In-Reply-To: <405A0CBB.6080202@itg.uiuc.edu> Message-ID: <3.0.6.32.20040318154216.00bc9090@gemini.msu.montana.edu> This is actually a published method in J of Anatomy to produce museum specimens for teaching sometime in early 80's??? Buy BIZ laundry detergent (or raid the family laundry supply!) make a 20% solution, heat to 80C, less than boiling. Suspend bones in this hot solution (or suspend and bring solution to working temperature, don't boil!)using cheesecloth to retain small boney parts that can fall off and handle a slippery, hot specimen easily. BIZ enzyme detergent loosens soft tissue to the point you can push it off with gloved fingers. It will take cartilage off but that was acceptable. Wear a mask when weighing - BIZ dust is very irritating to nasal passages and also skin. Digestion takes several hours if bone is large, so test frequently to see if soft tissue is releasing. I have done formalin fixed bone, rinsed over night in running tap water to remove the formalin (NBF kills the enzyme!), then BIZ. NBF fixed bone stablilizes the soft tissue attachments to make them more resistant to enzyme. Wear gloves, safety glasses and rinse off detergent before handling bone excessively, slippery, and irritates skin. Push soft tissue off, use old tooth brush and a dull knife to do gentle scraping if needed. When finished, rinse bone well, then air dry. The result is a BLEACHED, clean bone that doesn't smell bad - particularly when the digestion process is happening. Fixed is less smelly, unfixed means you are cooking bone while enzyme works - not so pleasant - use a hood. Have done whole bovine calf heads, bovine vertebral columns, bones from hind and forelimbs; rat and mouse heads, rat femurs/tibias. Beware - the enzyme will digest any soft attachments IF you let bones digest too long. Bones will fall apart, then you have to be a good anatomist and glue (thicker superglue works) tiny parts back in their relative, interdigitating, tight positions - a tedious but educational procedure on how our boney parts go together. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From peoshel <@t> wisc.edu Thu Mar 18 16:56:09 2004 From: peoshel <@t> wisc.edu (Philip Oshel) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] digesting soft tissue In-Reply-To: <3.0.6.32.20040318154216.00bc9090@gemini.msu.montana.edu> References: <3.0.6.32.20040318154216.00bc9090@gemini.msu.montana.edu> Message-ID: Gayle, Nice post -- I'd forgotten about Biz. Works a treat. We've used it for high-resolution SEM after digesting away the soft tissue. Phil >This is actually a published method in J of Anatomy to produce museum >specimens for teaching sometime in early 80's??? > >Buy BIZ laundry detergent (or raid the family laundry supply!) make a 20% >solution, heat to 80C, less than boiling. Suspend bones in this hot >solution (or suspend and bring solution to working temperature, don't >boil!)using cheesecloth to retain small boney parts that can fall off and >handle a slippery, hot specimen easily. BIZ enzyme detergent loosens soft >tissue to the point you can push it off with gloved fingers. It will take >cartilage off but that was acceptable. Wear a mask when weighing - BIZ >dust is very irritating to nasal passages and also skin. Digestion takes >several hours if bone is large, so test frequently to see if soft tissue is >releasing. > >I have done formalin fixed bone, rinsed over night in running tap water to >remove the formalin (NBF kills the enzyme!), then BIZ. NBF fixed bone >stablilizes the soft tissue attachments to make them more resistant to >enzyme. Wear gloves, safety glasses and rinse off detergent before handling >bone excessively, slippery, and irritates skin. Push soft tissue off, use >old tooth brush and a dull knife to do gentle scraping if needed. When >finished, rinse bone well, then air dry. The result is a BLEACHED, clean >bone that doesn't smell bad - particularly when the digestion process is >happening. Fixed is less smelly, unfixed means you are cooking bone while >enzyme works - not so pleasant - use a hood. > >Have done whole bovine calf heads, bovine vertebral columns, bones from >hind and forelimbs; rat and mouse heads, rat femurs/tibias. Beware - the >enzyme will digest any soft attachments IF you let bones digest too long. >Bones will fall apart, then you have to be a good anatomist and glue >(thicker superglue works) tiny parts back in their relative, >interdigitating, tight positions - a tedious but educational procedure on >how our boney parts go together. > > > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) From DWilliams <@t> ciit.org Thu Mar 18 17:46:08 2004 From: DWilliams <@t> ciit.org (Delorise Williams) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] NCSHT Spring Meeting Message-ID: <12816CB59E68184F9F5F28063E68B047285DC0@xsrvr.ciit.org> The North Carolina Society OF Histopathology Technologists presents the 2004 Spring Meeting on April 23 &24 at the Hilton Wilmington Riverside in Wilmington NC. Below is a summary of the program: Friday: April 23, 2004 Using Histological Techniques for Protein and mRNA Expression Analysis in Pharmaceutical Research and 8:15-9:15AM Drug Discovery Stephanie Broka, BS GlaxoSmithKline, Research Triangle Park, NC 9:15 - 10:15 AM Basic Immunology and Immunohistochemistry Godfrey Guerzon , MS,MBA, HT, DLM (ASCP) New Hanover Regional Medical Center,Wilmington, NC 11:00 - 12:00 Organize for Success Beth Moore,MT (ASCP) DLM, MBA New Hanover Regional Medical Center, Wilmington, NC WORKSHOP # 1 (1:30-4:30) HT (ASCP) Examination Readiness-The Practical Exam-Robert Lott, HTL(ASCP) Baptist Health System,Birmingham, Al WORKSHOP # 2 (1:30-4:30) Preparation of Tissue for Evaluation of Sexually Dimorphic Nuclei in Rat Brain-Melanie Struve, BS and Renee Thacker,BS, HT(ASCP) CIIT Centers for Health Research, Research Triangle Park, NC Saturday: April 24,2004 WORKSHOP # 3 (8:15-12:00) HT (ASCP) Examination Readiness-The Written Exam-Robert Lott, HTL (ASCP) Baptist Health Systems, Birmingham, Al WORKSHOP #4 (8:15-12:00) Catalyzed Signal Amplification system-Leslie Sells, BS, HTL, Technical Support Representative, DakoCytomation WORKSHOP # 5 (1:00-4:30) Dyes and Tissue: Chemistry Mechanism of Staining-Bert Dotson,MBA, HTL (ASCP) Duke University Medical Center, Durham, NC WORKSHOP # 6 (1:00-4:30) Microwave: Routine Histological Application-Theory to Practice Jim Milios, International Applications Specialist, Milestone Srl., Italy Delorise Williams CIIT Centers for Health Research PO Box 12137 Research Triangle Park, NC 27709 (919) 558-1200 Voice Mail-(919) 558-1252 Fax-(919) 558-1300 From John.Garlits <@t> stjude.org Thu Mar 18 17:51:40 2004 From: John.Garlits <@t> stjude.org (Garlits, John) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] need help with IHC detection of human cells in mouse tissue Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC238ABF@SJMEMXMB04.stjude.sjcrh.local> Hi, I was wondering if anybody could help me out. We want to look for human cells engrafted in mice, particularly in formalin-fixed, acid decalcified, paraffin-embedded tissue. I have so far tried two anti-human beta-2 microglobulin antibodies. The first I tried has been used in human/sheep transplant tissue, and I tried it, but unfortunately it is an IgM kappa, so it failed with the Biogenex mouse-on-mouse IHC kit, which it turns out is not made for IgM antibodies. If anybody is aware of a good way to use a mouse IgM on both human and mouse tissue, I'd love to know. I have looked for a good secondary antibody for this purpose, but have had no luck so far. The second antibody I tried is from Novocastra and distributed by Vector Labs. It is a rabbit polyclonal to human beta-2-microglobulin. The trouble I had was nonspecific binding in control mouse tissue. I was thinking to use some of the Biogenex mouse-on-mouse kit reagents to help block mouse antigens, but I do not know if this would actually work. The second problem with this antibody was that I did not always see positive marking in human tissue (especially osteocytes), which I thought should be pretty well all positive since it is human tissue. Is the expression of beta-2-microglobulin so variable? Has anyone tried any other general human cell marker in other animals? I did a quick search for beta actin, but it seems most of those cross-react with mouse. Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 From Yang.Wang <@t> tufts.edu Thu Mar 18 17:53:28 2004 From: Yang.Wang <@t> tufts.edu (Yang Wang) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] (no subject) Message-ID: <1079654008.405a367856f5c@webmail.tufts.edu> Hi, dear histonet friends: We need help! We have been working on the conditions for LCM for about 7 months, which is still very frustrating:-(.We tried to capture single cells from intestinal tissues. However, the major problem we are facing now is that we always get extra tissues from the capture. (We could pull off a whole villi when we capture a single cellL) .We used the standard protocols provided by ?ARCTURUS? for the tissue section and dehydration. Here is what we have done: 1.We cut 5um frozen sections and do either ?pre? or ?post? fixation in 4% paraformaldehyde. a. ?Pre?: fix 2h on ice before frozen, since we are working on GFP expressing tissue, which required pre-fixation. b. ? Post?: fix 15-30 min after frozen 2.The slides will then be dehydrated by: 75% ethyl 30s, 95% ethy 30s, 100% ethyl 1min and xylene 5min, then air dry 30min. We always pull off extra tissues, which adjacent to the cell interested. We tried treat slides and cap with ?ARCTURUS prep strip?. But it didn?t help. We are wondering whether we need specific treatment with slides (we use fisher superfrost plus)? Or our procedures are not proper? Thanks a lot for help! Yang Wang New England Medical Center Yang.wang@tufts.edu From Yang.Wang <@t> tufts.edu Thu Mar 18 17:56:48 2004 From: Yang.Wang <@t> tufts.edu (Yang Wang) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Extra tissues in LCM capture Message-ID: <1079654208.405a3740c18b2@webmail.tufts.edu> Hi, dear histonet friends: We need help! We have been working on the conditions for LCM for about 7 months, which is still very frustrating:-(.We tried to capture single cells from intestinal tissues. However, the major problem we are facing now is that we always get extra tissues from the capture. (We could pull off a whole villi when we capture a single cellL) .We used the standard protocols provided by ?ARCTURUS? for the tissue section and dehydration. Here is what we have done: 1.We cut 5um frozen sections and do either ?pre? or ?post? fixation in 4% paraformaldehyde. a. ?Pre?: fix 2h on ice before frozen, since we are working on GFP expressing tissue, which required pre-fixation. b. ? Post?: fix 15-30 min after frozen 2.The slides will then be dehydrated by: 75% ethyl 30s, 95% ethy 30s, 100% ethyl 1min and xylene 5min, then air dry 30min. We always pull off extra tissues, which adjacent to the cell interested. We tried to treat slides and cap with ?ARCTURUS prep strip?. But it didn?t help. We are wondering whether we need specific treatment with slides (we use fisher superfrost plus)? Or our procedures are not proper? Thanks a lot for help! Yang Wang New England Medical Center Yang.wang@tufts.edu From AnthonyH <@t> chw.edu.au Thu Mar 18 18:38:27 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Iron stain on frozen sections Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E167@simba.kids> Brett, Following is an excerpt from a paper I published a year or so ago: Henwood, A.F., (2002) "Microwave Perl's Stain for urgent frozen sections" Aust J Med Sc 23(2):68-69). Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html MICROWAVE PERLS' STAIN FOR URGENT FROZEN SECTIONS Anthony F. Henwood Laboratory Manager, Histopathology, The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA Abstract A rapid Perls' Stain for hemosiderin using a microwave oven is described. It takes one minute to perform and is particularly suitable for urgent frozen section diagnosis. Introduction Urgent frozen section diagnosis is usually restricted to the interpretation of haematoxylin and eosin stained sections (1). With the increased utilisation of microwave modifications of routine special stains, it is possible to perform a range of such stains on frozen sections (1). To be applicable to urgent frozen section diagnosis, a special stain needs to be rapid, preferably taking 1-2 minutes to perform, easy to prepare, using stock solutions with few preparation steps and produce results equal to those obtained on paraffin sections. The following microwave Perls' technique for hemosiderin takes about one minute to perform. Method Frozen sections can be fixed in Shoobridge's fixative (10ml concentrated formalin in 30ml absolute alcohol) (2) or 95% alcohol. Kennedy and Forbes (1) have also successfully used Wolman's fixative (5% acetic acid in absolute ethanol). 1. Rinse fixed frozen sections in water and then place in a coplin jar containing 20m1 each of 2% Potassium Ferrocyanide and 2% Hydrochloric acid. Place in the microwave oven and microwave for 20 seconds at 650 watts. Slides can be left in this solution for up to 3 minutes. Remove slides if the solution begins to turn cloudy. 2. Rinse slides in water. 3. Counterstain using agitation, with eosin (as routinely used for the frozen section HE) for 5-7 seconds. Other counterstains such as Nuclear Fast Red (3) or 0.25% basic fuchsin (4) can also be used, though eosin is readily available at the frozen section site and works quite well. Since the frozen block is usually fixed and processed to paraffin, sections of this block should also be stained with the routine Perls' as a quality control procedure. Discussion Iron in the body is stored in the forms of hemosiderin (ferric hydroxide polymer) or ferritin (a ferrous iron-protein complex) (5). Iron in tissues occurs mainly in the ferric state (6,7). Microscopically, hemosiderin appears similar to other yellow to brown pigments, such as melanin and fine carbon dust. Macrophages can contain any of these pigments. Heavily pigmented macrophages, often masking the cell nucleus, can be confused with malignant melanoma. The identification of the pigment as being hemosiderin, especially at the time of frozen section, is diagnostically useful. The rapid Perls' stain described above uses the same solution as used by Schaffner (4). This solution is easier to prepare than others reported in the literature (3). The stain takes approximately one minute to perform and gives results equivalent to those obtained in paraffin sections. It is especially applicable to frozen section diagnosis. References: 1. Kennedy, A., Foulis, A.K. (1989) "Use of microwave oven improves morphological and staining of cryostat sections", J.Clin.Pathol. 42:101-105. 2. Shoobridge, M.P.K., (1978) "Improving frozen sections by wet fixation", (Abstract) Pathology 10:195. 3. Brian, N.T., (1983) "Rapid Metallic Histological staining using the microwave oven", J.Histotechnol. 6(3): 125-129. 4. Schaffner, R., (1986) "The Perls' Iron staining procedure for use in the microwave oven using a temperature probe", J.Histotechnol. 9(2): 107-108. 5. Barka, T., Anderson, P.J., (1963) "Histochemistry: Theory, practice and bibliography" Harper & Row Publishers Inc, New York, p172-174. 6. Davenport, H.A., (1961) "Histological and Histochemical Technics" W.B. Saunders Co., Philadelphia, 280-284. 7. Gabe, M., (1976) "Histological Techniques" Masson, Paris, p311-317. -----Original Message----- From: Connolly, Brett M [mailto:brett_connolly@merck.com] Sent: Friday, 19 March 2004 6:03 AM To: 'HISTONET' (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Iron stain on frozen sections Along with the bone marrow thread..... Has anyone had experience with iron stains on frozen sections? What fixative? Procedure? Thx, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ---------------------------------------------------------------------------- -- Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ---------------------------------------------------------------------------- -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From RFail <@t> Charleston.net Thu Mar 18 21:09:52 2004 From: RFail <@t> Charleston.net (Rena Fail) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Placental section ihc... In-Reply-To: Message-ID: <010501c40d5f$cde15f50$dc10a6a5@rena> We are on occasion asked to stain placenta tissue for IHC. We use non-fat milk as a protein block. No problems with background Rena Fail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of nyilmaz@mersin.edu.tr Sent: Thursday, March 18, 2004 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Placental section ihc... We are using placental tissue for immunohistochemistry, and have some problems about background staining especially with serum remnants in the vessels. Does anybody have ideas about this problem. If you inform us we'd be greatly appreciate... Nejat Yilmaz PhD Mersin University Medical School _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JNocito <@t> Pathreflab.com Fri Mar 19 06:51:05 2004 From: JNocito <@t> Pathreflab.com (Joe Nocito) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Using H & E slides on the Benchmark XT Message-ID: Morning Histoland, We have acquired the Ventana Benchmark XT and I really need some help. When we were performing the slides manually, we were able to take an H&E slide, take off the coverslip and run an immuno on it. The doctors fell in love with this since we processes a lot of cervical biopsies and sometimes the lesion is not there on recuts. We have had a problem with H&E slides not picking up the immuno staining. I set up different programs using the wet slide protocol, but I'm having problems still. Would anyone be willing to share their protocol with me? Thanks Joe Nocito, BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX 78205 ***************************************Notice******************************************** This e-mail, including attachments, contains information that is confidential and may be legally privileged. This e-mail, including atachments, constitutes non-public information inteneded to be conveyed only to the designated recipient(s). If you are not an intended recipient,please delete this e-mail, including attachments, and notify me. The unauthorized use, dissemination, distribution or reproduction of this e-mail, including attachments, is prohibited and may be unlawful. ***************************************************************************************** From subratab <@t> bdonline.com Fri Mar 19 08:20:32 2004 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] DAB-Mn histochemistry for superoxide: help to remove background Message-ID: <200403191425.i2JEPrGC023459@mailout.proshikanet.com> Dear histo-experts I am trying to stain superoxide producing cells on frozen tissue sections of rat kidney. I am using Brigg?s (and Karnovsky) cytochemical method in unfixed tissue. The reaction medium contains 1 mg/ml DAB, 1 mM sodium azide and 0.5 mM MnCl2 in 100mM Tris-HCl (pH 7.2). Incubation time 30 minutes at 37 degree C. (The principle of the reaction is that intracellular superoxide will oxidize Mn++ to Mn+++; then Mn+++ will in turn cause DAB oxidation and precipitaion). MY PROBLEM: I am getting profuse DAB staining throughout the tubulo-interstitial area (glomerulus is clean). Most probably its from endogenous peroxidase activity. So I tried to block endo perox by H2O2 (and also by sodium azide, and H2O2+azide) before incubation with reaction medium. This way I have removed tubulo-interstitial DAB staining. But I am not getting any positive stain. I suspect that my blocking step is somehow damaging (blocking) superoxide producing enzymes as well. At this stage I need some expert opinion. How can I specifically block endogenous peroxidase keeping other enzymes intact? Hope to get some effective ways from histo-experts. Thanks in advance Subrata Biswas MD Lab de Fisiopatologia Renal Uni of Campinas, SP, Brazil. From angela.mcnabola.b <@t> bayer.com Fri Mar 19 08:31:13 2004 From: angela.mcnabola.b <@t> bayer.com (Angela McNabola) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] "Animal" person seeking clinical folks help! Message-ID: Hi all, I soliciting protocols that any of you may be willing to share on how to make slides suitable for IHC using FNA's. I am looking for how to best make slide smears (I guess!). Clinical sites will be collecting samples from patients and sending the slides to my lab for staining/evaluation. We are fairly new to this, even new to processing human samples, so any help you can provide would be greatly appreciated. Also, keep in mind that we potentially may be receiving samples from all over the world, so the easier the better since we will be asking seeral sites to do it all the same way thanks in advance! Angela McNabola, MS, HT(ACSP)QIHC, SLS, RLATG Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 angela.mcnabola.b@bayer.com From nick.kirk3 <@t> btopenworld.com Fri Mar 19 09:25:09 2004 From: nick.kirk3 <@t> btopenworld.com (=?iso-8859-1?q?NICK=20KIRK?=) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] "Animal" person seeking clinical folks help! In-Reply-To: Message-ID: <20040319152509.87494.qmail@web86308.mail.ukl.yahoo.com> Angela Here's a useful recipe/technique for cytological material. The immuno results are very good, plus you have the bonus of being able to keep any residual material for a longer period. RECIPE for PEG/IMS fixative 3% Polyethylene glycol in 50% Industrial methylated spirit (IMS) {=99% ethanol}. --------------------------------- Method 1. Spin cells down 2. Perform blood lysis (if required) with Ammonium chloride solution 3. Re-suspend in PEG/IMS fixative 4. Leave to fix for 2 hours 5. Make cytospins 6. Air dry completely 7. Place in 95% I.M.S. for at least 10 minutes to remove PEG 8. Immunostain Any left over material can be stored in the PEG/IMS fixative for prolonged periods of time. Hope this is of some help Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England Angela McNabola wrote: Hi all, I soliciting protocols that any of you may be willing to share on how to make slides suitable for IHC using FNA's. I am looking for how to best make slide smears (I guess!). Clinical sites will be collecting samples from patients and sending the slides to my lab for staining/evaluation. We are fairly new to this, even new to processing human samples, so any help you can provide would be greatly appreciated. Also, keep in mind that we potentially may be receiving samples from all over the world, so the easier the better since we will be asking seeral sites to do it all the same way thanks in advance! Angela McNabola, MS, HT(ACSP)QIHC, SLS, RLATG Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 angela.mcnabola.b@bayer.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DSTULTS <@t> fremontmemorial.org Fri Mar 19 12:48:02 2004 From: DSTULTS <@t> fremontmemorial.org (Deb Stults) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] help with elastic staining Message-ID: We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen From pruegg <@t> colobio.com Fri Mar 19 12:58:04 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] CD98 and PLGF In-Reply-To: Message-ID: I guess no one is using cd98 and/or Placental Growth Factor on ffpe tissue???? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, March 17, 2004 10:03 AM To: Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: [Histonet] CD98 and PLGF I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160 and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies work in ffpe tissue? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Fri Mar 19 12:59:57 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] FW: Question Message-ID: Message -----Original Message----- From: Garza-Williams, Sara [mailto:Garza-Williams.Sara@tchden.org] Sent: Tuesday, March 16, 2004 1:47 PM To: pruegg@colobio.com Subject: Question Patsy, Can you tell me or do you know someone who can help me find CD52 and CD33 antibodies that work in paraffin? Thanks Sara Williams Anatomic Pathology Supervisor The Children's Hospital Denver, CO 303.861.6177 garza-williams.sara@tchden.org CONFIDENTIALITY NOTICE: The information contained in this message is legally privileged and confidential information intended for the use of the individual or entity named above. If the reader of this message is not the intended recipient, or the employee or agent responsible to deliver it to the intended recipient, you are hereby notified that any release, dissemination, distribution, or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the author immediately by replying to this message and delete the original message. Thank you. From ylee <@t> bccancer.bc.ca Fri Mar 19 13:03:10 2004 From: ylee <@t> bccancer.bc.ca (Yin Ping Lee) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Background staining in EBER ISH Message-ID: <6BAF4D075F07D411B30900508B94CBA0C1F559@SERVER20> Hello, Histonetter: We have done EBER ISH staining using the same protocol for months without problem. However, recently we encounter heavy background staining in * some of the runs (not all the runs) and * sometimes only some of the slides (not all the slides) in a run. We do manual staining and use PNA probe, anti-FITC/HRP and DAB. We checked all the reagents and even measure the pH of the buffer and deionized water but cannot find out what is going wrong. Does anyone have the same experience or know how to fix it? Thank in advance for any help! Yin Ping Lee Histopathology BC Cancer Agency Vancouver, BC From pruegg <@t> colobio.com Fri Mar 19 13:09:55 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] active-Caspase In-Reply-To: Message-ID: if you are using a biotinylated antibody and/or detection system your non-specific staining is probably due to endogenous biotin in liver and in my experience even with the most aggressive ab block you cannot get rid of the biotin in liver. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Dixon, Leslie E. Sent: Thursday, March 18, 2004 8:59 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] active-Caspase Hello all, We are having issues with non-specific background staining. I am interested in anyone's protocol for anti-active Caspase-3 on mouse livers. Any help would be greatly appreciated. Thanks, Leslie _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From pruegg <@t> colobio.com Fri Mar 19 13:18:46 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] help with elastic staining In-Reply-To: Message-ID: I have this problem when I do a Pentachrome stain for elastic fibers, the fibers stain fine when I first do the hematoxylin but as I continue with the rest of the stain the fiber stain is lost, I have remedied this by repeating the hematoxylin again at the end of the staining process to get back the fiber staining. Try it. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Deb Stults Sent: Friday, March 19, 2004 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help with elastic staining We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ROENN <@t> surgery.wisc.edu Fri Mar 19 13:33:07 2004 From: ROENN <@t> surgery.wisc.edu (Drew Allan Roenneburg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] help with elastic staining Message-ID: Deb and Karen Do you make up any of your own solutions? Try combining 30 mL of a 5% alcoholic hematoxylin, 10mL of a 10% aq. ferric chloride, and 10 mL Verhoeff's iodine soln (Potassium iodide and Iodine). Stain for 12 mins. Wash under running tap water. Differentiate by dipping slide in 2% aq ferric chloride, checking microscopically between. Drew From Dodgymatt <@t> aol.com Fri Mar 19 13:49:59 2004 From: Dodgymatt <@t> aol.com (Dodgymatt@aol.com) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] please remove me from your mailing list Message-ID: <5539C29B.5C90FB20.0237CBF1@aol.com> please remove me from your mailing list From gentras <@t> vetmed.auburn.edu Fri Mar 19 14:15:57 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:42 2005 Subject: Fwd: [Histonet] Lori White/ B plus Message-ID: <6.0.1.1.0.20040319141112.0259b480@mailhost.vetmed.auburn.edu> Hello, please what's your source of this B plus and do you know it's components? Thanks, Atoska >From: "Joyce Cline" >To: "Histonet" >Date: Wed, 17 Mar 2004 14:13:45 -0500 >X-Mailer: Microsoft Outlook Express 6.00.2800.1158 >X-Virus-Scanned: by amavisd-new at wchsys.org >X-Scan-Signature: 690e9200c080ca55fd7a8d0fb0193eeb >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >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: 148abf0c5acfe33f338c6ed44fdf9350 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Lori White/ B plus >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.4 required=6.5 tests=DATE_IN_PAST_12_24 > autolearn=no version=2.63 >X-SA-Exim-Version: 3.1 (built Tue Oct 14 16:21:02 CDT 2003) >X-SA-Exim-Scanned: Yes >X-MIME-Autoconverted: from quoted-printable to 8bit by >cvm4.vetmed.auburn.edu id i2IG9dn16152 > > We fix our nodes & BM's in B Plus for a minimum of 2 hours. Then our > cores are decaled with HCL/Formalin for one hour and process overnight. > We have no problem with the results. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From kcboswell <@t> grandecom.net Fri Mar 19 14:29:47 2004 From: kcboswell <@t> grandecom.net (Boswell) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using Zenker's Message-ID: <000e01c40df0$efab0050$e1edbcd8@Boswell> Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain without using Zenker's. We have looked everywhere and are unable to find it. Thank you. From pruegg <@t> colobio.com Fri Mar 19 15:20:16 2004 From: pruegg <@t> colobio.com (Patsy Ruegg) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] mouse bone osteoblast IHC In-Reply-To: <005401c40d22$538a53c0$78065486@vdl220FAC> Message-ID: You can use osteocalcin, osteopontin or osteonectin (osteopontin and nectin can be purchased from U of Iowa Hybridoma Bank, the osteocalcin from another co. I can't think of right now ?something like Southern Biotechnologies???? do a search for the antibody) for osteoblasts, I used 5% formic acid decal (Immunocal from Decal Chemicals comes premade and worked well for me)with zinc formalin fixation and pepsin digestion with labelled polymer (envision) detection. Another way to label osteoblasts is by enzyme histochemical method (alk. phosphatase) but it does not work well in my experience on decalcified ffpe tissue, I used in on cold processed GMA embedded bone without decalcification. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jan Shivers Sent: Thursday, March 18, 2004 12:51 PM To: histonet Subject: [Histonet] mouse bone osteoblast IHC I am posting this for an MD not on the Histonet listserv; he is requesting information on staining mouse osteoblasts using the IHC method. (I have had no experience with osteoblasts myself). Does anyone know of an antibody source for detecting murine osteoblasts (or one made against another species that will cross-react)? What decalcification method is used on the bone samples? Any complete protocols that anyone is willing to share would be greatly appreciated. Thanks... Jan Shivers U of MN Vet Diag Lab _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Mar 19 15:31:24 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without usingZenker's In-Reply-To: <000e01c40df0$efab0050$e1edbcd8@Boswell> Message-ID: <3.0.6.32.20040319143124.00bca308@gemini.msu.montana.edu> There is a variation of this method for CNS tissue. CAVEAT:You get rid of the mercury, however, dichromate solution cannot be dumped down a drain, collect for chemically safe disposal. Gamble and Bancroft, 5th Edition, Theory and Practice of Histological Techniques, 2001. Using naturally oxidized PTAH solution: Dewax, and rehydrate to dist. water Acid dichromate solution for 30 min 10% HCl in absolute ethanol 12 ml 3% aqueous potassium dichromate 36 ml Chromate solutions must be collected, cannot be dumped down sink. Wash in tap water Acid permangante solution for 1 min 0.5% potassium permanganate 50 ml 3% sulfuric acid 2.5ml Wash in tap water Bleach in 1% oxalic acid Wash in tap water Mallory's PTAH stain overnight dehydrate starting in 95% alcohol, if sections are too blue, some of the stain will be removed during dehydration. Dehydration should be rapid to prevent water and alcohol from removing stain. IF tissue has been fixed in a chromate based fixative, you can eliminate dichromate step i.e Zenkers fixation. If you do not have naturally oxidized PTAH, PTAH can be made with chemical oxidation by potassium permanganate, but this solution continues to oxidize and has a short shelf life. A. hematoxylin, 0.5g -dissolve in 100 ml distilled water B. 10 g phosphotungstic acid in 400 mls distilled water C. Add A to B, then add 25 ml of 0.25% potassium permanganate. Stain can be used next day with optimal (they said peak) staining is at 7 days. In past, we used permanganate oxidized PTAH with success, not sure of others experience. If you have trouble, you should buy a naturally oxidized PTAH from a reliable commercial source, but check with them to make sure it is not chemically oxidized version. PTAH, natural variety, can last for years. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From pam <@t> ategra.com Fri Mar 19 19:19:59 2004 From: pam <@t> ategra.com (Pam Barker (extension 234)) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Job Opportunities in Histology Latest Update 03/19/04 Message-ID: Hi Connie, Here is the latest update on opportunities with some of my best clients throughout the US who are seeking Histology Supervisors, Histo Technologists and Histo Technicians. These are positions as direct employees of our client. There are fulltime 40 hour per week positions.. As a direct employee of one of our clients you will be provided with full benefits including Health Insurance, Vacation, Sick Pay, Relocation money and a lucrative sign-on bonus. I have part time and temporary positions as well. I have supervisory, team lead and bench positions. These positions require HTL or HT certification or registry eligibility. Here are some of my HOTTEST Histology Supevisory positions: 1. Colorado - Histology Manager 2. Connecticut - Histology Supervisor 3. Oregon - Lead Histo Tech Here are some of my HOTTEST Histo Tech bench positions: 1. Florida - Histo Tech (full time and part time positions) 2. Florida - MOHS Tech 3. Virginia - Histo Tech 4. Pennsylvania - Histo Tech 5. Oregon - Lead Histo Tech 6. Massachusetts - Histo Tech 8. Massachusetts - MOHS Tech 9. Minnesota - Histo Tech 10. Alabama - Histo Tech 11. Wisconsin - Histo Tech 12. Ohio - Histo Tech 13. California - Histo Tech (multiple positions in research and clinical) 14. Nevada - Histo Tech 15. Nebraska - Histo Tech If you are interested in these jobs, please CALL ME ASAP at 800 466 9919 x234. To speed things up, please also send me a copy of your resume, (if you haven't already done so). If you are interested in jobs outside the above-mentioned areas, please send me your resume as well. I have clients throughout the US. I will keep your resume confidential and will not release it to anyone without your permission (This is Ategra policy as well as my own). My services are at no charge to you. Of course, you may be happy in your present job, but it never hurts to to keep an eye open. Also, if you have friends/peers who do not have an email address, if you could pass my query & name on to them I'd be very grateful. I don't want to be a bother - I was told that you were a hands-on Histo Tech or a Lab Supervisor. If you are no longer working in a lab please send me an email and I will remove you from my list of people to contact. However, if you are interested in any of the jobs above, please call me. Thank You !! Pam - 800 466 9919 ext 234 --------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing Learn More About Ategra: Pam Barker Senior Lab Recruiter Ategra Systems Inc Specialists in Permanent & Contract Staffing 7085 University Blvd. Winter Park, FL 32792 VOICE: 407-671-5800 ext 234 TOLLFREE: 800-466-9919 ext 234 EMAIL: pam@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 lpwenk <@t> covad.net Fri Mar 19 21:23:42 2004 From: lpwenk <@t> covad.net (Lee & Peggy Wenk) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without usingZenker's References: <000e01c40df0$efab0050$e1edbcd8@Boswell> Message-ID: <002b01c40e2a$bf467ea0$e83dd445@domainnotset.invalid> We've postmordanted in Bouins in 60 degree C. oven for 1 hour. Wash in running water until yellow is gone. Then stain as usual. The striated muscle fibers seemed less "splotchy" to me with this method than with postmordanting in Zenker or saturated mecuric chloride. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 ----- Original Message ----- From: "Boswell" To: Sent: Friday, March 19, 2004 3:29 PM Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without usingZenker's Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain without using Zenker's. We have looked everywhere and are unable to find it. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Gldnsthisto <@t> aol.com Fri Mar 19 21:21:52 2004 From: Gldnsthisto <@t> aol.com (Gldnsthisto@aol.com) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Memorial to Jim Becker Message-ID: <2b.52d3feb9.2d8d12d0@aol.com> I was informed today that a dear friend and colleague in Histology field passed on after a long illness. Jim Becker and I met over twenty years ago at a California State Histology meeting in San Jose. He befriended me as I was still learning the Histology field. He was the owner of Marketing International, a supply company of which we all have bought something from at one time or another. He was a quality person with a smile on his face along with his pipe. He liked everyone. Although I haven't seen him for a number of years, I would see his advertisements in most of the news letters of each state society. I wish my deepest sympathy to his wife and family. We will miss him. California Larry Fields From yichaowu <@t> hotmail.com Sat Mar 20 03:25:27 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen sections---High background! Message-ID: Dear Dr.Callis,Dr.Yaskovich,,Dr. Li,and all histonetters, Thank you very much for all of your reply.The frozen sections we make now have better mophology with Liquid N2 freezing. Still three questions with regarding to kidney immunofluorescent staining. 1) We stain human IgG on kidney frozen sections with antibody from DAKO (F0202),and dilution is 1:50,but the background is always strong. Other antibodies to IgA(F0204 from DAKO),IgM(F0203 DAKO),C3(F0201 DAKO),C4(F0169 DAKO)&C1q (F0254) all produce dark background under fluorescent microscope,which is highly preferred. Why IgG is so strange? Any problem with the antibody? Or the unspecific staining for IgG is special? 2) What kind of O.C.T is the best? Thank you very much! Yichao WU Research Institute of Nephrology Jinling Hospital Nanjing 210002 P.R.China >From: Terry Li >To: "yichao wu" >Subject: Re: Please help with me--routine immunofluorescent staining on >kidney specimens >Date: Thu, 18 Mar 2004 11:35:36 -0600 > >Yichao, > >Autofluoresence is a big disadvantage from paraffin tissues. Chosing >fixatives is also important depend on what protein you are dealing with. > >My answer for your first question: It's very easy to get the block out of >the plastic mold. Use your finger to warm the bottom of the block, then >you can get it out easily. Add OCT on the tissue holder (chuck) and put >the tissue block on top of it. They will stick together in cryostat and >your tissue is ready to be cut. > >Second question: Store the frozen samples in -80oC freezer until to use. I >am working on a hand book about laboratory methods with English versus >Chinese. Here is part of it. I hope it may be helpful for you. "Feed >back" is very welcome. > >Immunohistochemistry on Frozen Tissue >The best method for antigen preservation in immunohistochemistry is >freezing. Freezing is a very suitable method for preserving antigens which >lose their immunoreactivity. > >1. Preparation of Frozen Tissues for sectioning > Materials: > 2-methylbutane; > Dry ice; > Plastic molders; > Frozen tissue matrix (O.C.T. compound, 4583 from SAKURA); > Long forceps; > Metal or plastic tall cylinder. > >Methods: >1) Pour 2-methylbutane into a metal or plastic tall cylinder. Keep >adding small pieces of dry ice into the solution to make the temperature >cool down to -40-50o C and keep it for certain long time as desired. >2) Label plastic models and fill the bottle with O.C.T.. >3) Collect desired tissues. Preserve the best tissue integrity is the >key to have high quality of immunohistochemistry result. Have fine tools >and use fine technique. >4) Place tissue into the labeled plastic model and fill the molder with >more O.C.T. until the tissue get fully covered. Arrange tissue in the >matrix near the bottom so tissue is easily exposed when sections are cut, >and adjust the orientation if desire. Place molder with tissue into the >cylinder of cold 2-methylbutane. Allow the tissue matrix to solidify from >the bottle of the molder to the top to avoid bubbles and leave it in the >solution for 5-10 minutes. >5) Store blocks in the -80oC freezer until ready sectioning. > >Note: >1. Get rid of extra aqueous from tissue specimen with paper towel, do >not wash tissues before frozen. >2. Some tissues need to be fixed first. Tissue samples can be treated >with 10%-30% sucrose-PBS after fixation, and then frozen as above. > >2. Sectioning of Frozen Tissues > >Materials: >Frozen tissue block; >Superfrost plus slides (12-550-15 from Fisher Scientific); >Blade (low or high-profile disposable blades or non-disposable blade); >Frozen Tissue Matrix (O.C.T.). > >Instrumentation: >Cryostat (Leica, Microm or other brands). > >Methods: >1) Before cutting sections, allow the temperature of the frozen tissue >block to equilibrate to the temperature of the cryostat (-20oC in general). >2) Use O.C.T. to freeze tissue block on the specimen disk and fix the >disk on the machine. Adjust the positioning of the block to align the >block with the knife blade. Cut the tissue until the desired tissue is >exposed and start collection. Put sections on super plus slides and air >dry over night. The thickness of sections is various depend upon your >research. 3-6 um sections are commonly used. >3) Fix the sections by immersion in cold acetone/methanol for 20 min, >air dry and start the staining procedure or store the slides at sealed >slide box in -80oC until you are ready. Or store the slides without any >fixation. >4) Use O.C.T. to cover exposed tissue and store it in -80oC. > > >Shihong Li >Immunohistochemistry Faciltiy >University of Chicago > > > > >>_________________________________________________________________ >>Add photos to your e-mail with MSN 8. Get 2 months FREE*. >>http://join.msn.com/?page=features/featuredemail _________________________________________________________________ MSN 8 helps eliminate e-mail viruses. Get 2 months FREE*. http://join.msn.com/?page=features/virus From gudrun.lang <@t> aon.at Sat Mar 20 06:03:42 2004 From: gudrun.lang <@t> aon.at (Gudrun Lang) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] help with elastic staining References: Message-ID: <00c101c40e73$638699d0$eeeea8c0@SERVER> As far as I know, Weigert stains the nuclei. So if your nuclei are pale-black Weigert has worked. Elastic fibers are stained by Resorcin Fuchsin. In our procedure there is a "washing-out-step" afterwards. Perhaps it was too long? There is a good description on the following website: http://www-medlib.med.utah.edu/WebPath/HISTHTML/MANUALS/WEIGERTS.PDF greetings Gudrun Lang ----- Original Message ----- From: "Deb Stults" To: Sent: Friday, March 19, 2004 7:48 PM Subject: [Histonet] help with elastic staining We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scoop <@t> mail.nih.gov Sat Mar 20 10:43:04 2004 From: scoop <@t> mail.nih.gov (Sharon Cooperman) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] ED1 Ab from Serotec Message-ID: Hi Inga (and everyone else), Does you know if the ED1 antibody from Serotec (anti-CD68) stains all (or at least most) macrophages or only activated macrophages? Also, in the brain, can you use it to distinguish between activated and non-activated microglia? I have used it on non-activated macrophages in various tissues and it seems to work great and be specific for macrophages. Also, the data sheet that comes with it says it works for most macrophages. However, I've been told that it only stains activated microglia. Do you know the answer? Thanks, Sharon -- Sharon Cooperman NIH, NICHD, CBMB 301.435-7735 Building 18T, room 101 301.402-0078 fax Bethesda, MD 20892 From RSRICHMOND <@t> aol.com Sat Mar 20 13:27:58 2004 From: RSRICHMOND <@t> aol.com (RSRICHMOND@aol.com) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using Zenker's Message-ID: Omnipotente Ptah! - sings Aida in Verdi's famous opera. I think the PTAH stain has declined a lot since then. I haven't seen one since I was a resident around 1970. I think it's been replaced by various immunohistochemical stains. A good batch of PTAH had to age like a fine wine - mine improved with age - I might still have it, but I think it got lost in a move. Certainly I never had any luck with PTAH unless the tissue had been first fixed in Zenker's (Helly's) fixative, with both mercury and chromium as well as formaldehyde. Them days is gone forever! But it sure was a great stain for reactive or neoplastic astrocytes - at its best, almost as good as the Cajal gold technique - now my age is really showing. The old joke was that, since the stain took overnight to do, that the PTAH stain gave the pathologist time to think about the diagnosis for an unusual tumor! Bob Richmond Knoxville TN and Gastonia NC From cwscouten <@t> myneurolab.com Sat Mar 20 18:59:22 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Forcep Sterilization Message-ID: And after that stick in a hot glass bead bed sterilizer to sterilize, if the subject line is correct. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=415001&catdesc=Surgical+Equipment&CatThreeID=51&CatOneID=2&subcatdesc=Sterilizers&idsubcategory=10 Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Monday, March 15, 2004 11:43 AM To: histonet@lists.utsouthwestern.edu Cc: ann_angelo@labcorp.com; RogerA@Dianon.com Subject: Re: [Histonet] Forcep Sterilization A toothbrush or test tube brush works well to clean out the grooves of the forceps. Have a bucket with soapy water handy and change it as frequently as possible. 4 out of 5 dentists recommend not re-using the toothbrush on your teeth, however. >>> 03/13/04 08:52AM >>> We are having a problem with carry-over at the grossing station. We do not have a sink available and are presently dipping our forceps in ethanol. Is there another solution we can use besides ethanol? Does anyone have any other ideas as how to avoid this contamination issue we are experiencing? Thank you Ann Angelo _______________________________________________ Histonet mailing list Histonet@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 yichaowu <@t> hotmail.com Sun Mar 21 05:35:55 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen Message-ID: Thank you for your reply!It is very helpful to me.I have once thought of the problem too. Since the background of IgA and IgM is very low and they are serum immunoglobulins too,it seems strange for the strong background of IgG. Mr.George Cole advised me that the concentration (1:50 in dilution) for the anti-human IgG antibody may be too high.However, our tech told me that further dilution would prevent those positive signals from being present. Hence I just wonder how other departments of renal diseases handle the immunofluorescent staining in biopsy specimens? Please kindly give me some hints since it is terrible for the diagnosis of patients with bad immunofluorescent staining results.It really hurts :-( Thank you very much in advance! Yichao WU,Ph.D Candidate Research Institute of Nephrology Jinling Hospital 305 East Zhongshan Road Nanjing 210002 P.R.China >From: andreah@imclone.com >To: yichaowu@hotmail.com >Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney frozen >Date: Sat, 20 Mar 2004 13:07:18 -0500 > > > > > >I expect it is not background but real signal from all the IgG in the >tissue. IgG is normally at high concentration in mouse tissue and will give >staining especially in a blood filled organ like the kidney. > >----- Message from "yichao wu" on Sat, 20 Mar 2004 >17:25:27 +0800 ----- > >To: histonet@lists.utsouthwestern.edu, tli1@flowcity.bsd.uchicago.edu >Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen >sections---High background! > >Dear Dr.Callis,Dr.Yaskovich,,Dr. Li,and all histonetters, > >Thank you very much for all of your reply.The frozen sections we make now >have better mophology with Liquid N2 freezing. > >Still three questions with regarding to kidney immunofluorescent staining. > >1) We stain human IgG on kidney frozen sections with antibody from DAKO >(F0202),and dilution is 1:50,but the background is always strong. > >Other antibodies to IgA(F0204 from DAKO),IgM(F0203 DAKO),C3(F0201 >DAKO),C4(F0169 DAKO)&C1q (F0254) all produce dark background under >fluorescent microscope,which is highly preferred. > >Why IgG is so strange? Any problem with the antibody? Or the unspecific >staining for IgG is special? > >2) What kind of O.C.T is the best? > >Thank you very much! > >Yichao WU >Research Institute of Nephrology >Jinling Hospital >Nanjing 210002 >P.R.China > _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus From jizv66 <@t> hotmail.com Sun Mar 21 06:59:35 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] CD marker in zinc fixed mouse liver Message-ID: hello everybody. I am new in immunohisotology and I have trying to stain for CD4,CD8 and CD45R but i had problems to get good staining. I am using Pharmingen protocol (rat primary antibody, biotinylated goat anti rat secondary antibody, streptavidina-HRP). I have tried H2O2 as endogenous peroxidase blocking solution (3% in Metanol) but eritrocytes are still brown after DAB. I am blocking with white egg and skim mil, for endogenous biotin. May one help me? May I have any good protocol? Thank you very much. JORGE IVAN ZAPATA Universidad del Valle Cali, Colombia _________________________________________________________________ Protect your PC - get McAfee.com VirusScan Online http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 From subratab <@t> bdonline.com Sun Mar 21 09:00:29 2004 From: subratab <@t> bdonline.com (Subratab) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Re: ED1 Message-ID: <200403211506.i2LF6IGC026236@mailout.proshikanet.com> Hi Sharon Here are few points about ED1. >Does you know if the ED1 antibody from Serotec (anti-CD68) stains all (or at least most) macrophages or only activated macrophages? mAb ED1 is a pan-macrophage marker in rats that recognizes majority of macrophage populations, as well as monocytes in the peripheral blood. Its not only for the detection of activated macrophages. But actively fagocytosing macrophages show strong positivity. This Ab is a rat version of anti-CD68 since it binds with an antigen homologous to human CD68 and mouse macrosialin. >Also, in the brain, can you use it to distinguish between activated and non-activated microglia? I think ED1 cannot be used to distinguish activated from non-activated microglia. You can read the following article for detail: Dijkstra CD : Rat macrophage lysosomal membrane antigen recognized by mAb ED1. Immunology 83: 140-7, 1994. Subrata Biswas University of Campinas SP, Brazil From John.Garlits <@t> stjude.org Sun Mar 21 13:38:19 2004 From: John.Garlits <@t> stjude.org (Garlits, John) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] need help with IHC detection of human cells in mouse tissue Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC238AC6@SJMEMXMB04.stjude.sjcrh.local> Dear Andrea, Thank you for your helpful suggestions! I will try some of the things you mentioned very soon. We use Regular Cal Immuno from BBC Biochemical, and it has not been a problem at least for the GFP staining we do. I would like to try some other decal methods, but unfortunately I am quite short on time. Do you mind if I ask, what primary antibodies have you used to detect human cells in mouse? Beta-2-microglobulin is commonly used, which is why we started with that. Are you aware of any others, ideally proteins with high uniform expression regardless of cell type? I'm a little worried about the microglobulin, because I don't know if it will be expressed in osteocytes inside the bone in particular. I had the thought of trying beta-actin, but so far I have not seen any antibody that would be specific to human vs mouse. Maybe a nuclear protein, such as histones? Thanks again, John -----Original Message----- From: andreah@imclone.com [mailto:andreah@imclone.com] Sent: Saturday, March 20, 2004 12:01 PM To: Garlits, John Subject: RE: [Histonet] need help with IHC detection of human cells in mouse tissue Dear John, I routintely stain for human antigens on human xenografts in mice so I have tried to address your concerns as best as I udnerstood your problems. Let me know if I am missing something and I will try to help. For routine antogens I have no problems as I often try to specifically use rabbit pAbs to avoid mouse-on-mouse issues. When this is not an option, we try to use directly labeled mouse primaries with biotin etc so we do not have to use secondaries against mouse. When that is also not an option, we use special blocking methods or kits from places such as DAKO etc (their ARK kit is decent). I do not routinely use decalcified tissue. When we have had to do decalc tissue it caused problems as many of the decalc reagents obliterate antigens (as they are HCl acid based). We now use gentler decalc methods such as formic acid decalcifiers or EDTA decalc for such situations. This might explain why you are not seeing the intensity or amount of stain even on the human tissue as the antigens are being destroyed? Companies such as Zymed and KPL sell anti-IgM antibodies absorbed against mouse and human IgG which you could use probably directly in mouse tissue without too much background. The amount of IgM should not that much in any given mouse certainly compared to IgG (especially if you are working with nude or SCID immunocompromised mice for the xenografts) so I would expect that you don't even have to use a mouse on mouse kit ... For the rabbit polyclonal which you are having issues with I would make sure your secondary is absorbed against mouse serum proteins. Otherwise you will get background. Jackson ImmunoResearch sells excellent secondaries for this purpose and this is what I use. Buy a donkey anti-rabbit min cross to mouse, human etc. The other option for staining mouse tissue with rabbit antibodies is to use DAKO's Rabbit EnVision+ kit. This is an expensive kit but is EXCELLENT and gives great intensity of stain with not that much background and you can reduce your primary concentrations way down! I will be glad to help as much as I can. Cheers! Andrea ----- Message from "Garlits, John" on Thu, 18 Mar 2004 17:51:40 -0600 ----- To: Subject: [Histonet] need help with IHC detection of human cells in mouse tissue Hi, I was wondering if anybody could help me out. We want to look for human cells engrafted in mice, particularly in formalin-fixed, acid decalcified, paraffin-embedded tissue. I have so far tried two anti-human beta-2 microglobulin antibodies. The first I tried has been used in human/sheep transplant tissue, and I tried it, but unfortunately it is an IgM kappa, so it failed with the Biogenex mouse-on-mouse IHC kit, which it turns out is not made for IgM antibodies. If anybody is aware of a good way to use a mouse IgM on both human and mouse tissue, I'd love to know. I have looked for a good secondary antibody for this purpose, but have had no luck so far. The second antibody I tried is from Novocastra and distributed by Vector Labs. It is a rabbit polyclonal to human beta-2-microglobulin. The trouble I had was nonspecific binding in control mouse tissue. I was thinking to use some of the Biogenex mouse-on-mouse kit reagents to help block mouse antigens, but I do not know if this would actually work. The second problem with this antibody was that I did not always see positive marking in human tissue (especially osteocytes), which I thought should be pretty well all positive since it is human tissue. Is the expression of beta-2-microglobulin so variable? Has anyone tried any other general human cell marker in other animals? I did a quick search for beta actin, but it seems most of those cross-react with mouse. Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 From kwuny <@t> email.cs.nsw.gov.au Sun Mar 21 16:37:12 2004 From: kwuny <@t> email.cs.nsw.gov.au (Young Kwun) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen Message-ID: <01C40FF1.418BC330.kwuny@email.cs.nsw.gov.au> Hi, We use IgG F(ab)2 (Dako F0315) instead of Ig fraction you are using. We follow manufacturer's suggested dilution which is 1:10 without any problem with the background staining. We used to use another IgG Fc in 1:50 dilution from a company called DiaSorin in USA. It was a very good antibody. But we were unable to get their antibodies anymore for some reason. I hope this helps. Young Kwun, PhD Senior Hospital Scientist Dept. of Anatomical Pathology Concord Hospital Concord NSW 2139 Australia Tel)61-2-9767-6075 Fax)61-2-9767-8427 kwuny@email.cs.nsw.gov.au The information contained in this message is intended for the named addressee only, and is confidential to the sender and intended recipient. If you are not the named addressee please do not copy, distribute, take any action reliant on, or disclose anything in this E-mail message to any other person or organisation. If you have received this message in error please delete the email and notify me immediately. Views expressed in this message are those of the individual sender and are not necessarily the views of Central Sydney Area Health Service. -----Original Message----- From: yichao wu [SMTP:yichaowu@hotmail.com] Sent: Sunday, 21 March 2004 22:36 To: andreah@imclone.com Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney frozen Thank you for your reply!It is very helpful to me.I have once thought of the problem too. Since the background of IgA and IgM is very low and they are serum immunoglobulins too,it seems strange for the strong background of IgG. Mr.George Cole advised me that the concentration (1:50 in dilution) for the anti-human IgG antibody may be too high.However, our tech told me that further dilution would prevent those positive signals from being present. Hence I just wonder how other departments of renal diseases handle the immunofluorescent staining in biopsy specimens? Please kindly give me some hints since it is terrible for the diagnosis of patients with bad immunofluorescent staining results.It really hurts :-( Thank you very much in advance! Yichao WU,Ph.D Candidate Research Institute of Nephrology Jinling Hospital 305 East Zhongshan Road Nanjing 210002 P.R.China >From: andreah@imclone.com >To: yichaowu@hotmail.com >Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney frozen >Date: Sat, 20 Mar 2004 13:07:18 -0500 > > > > > >I expect it is not background but real signal from all the IgG in the >tissue. IgG is normally at high concentration in mouse tissue and will give >staining especially in a blood filled organ like the kidney. > >----- Message from "yichao wu" on Sat, 20 Mar 2004 >17:25:27 +0800 ----- > >To: histonet@lists.utsouthwestern.edu, tli1@flowcity.bsd.uchicago.edu >Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen >sections---High background! > >Dear Dr.Callis,Dr.Yaskovich,,Dr. Li,and all histonetters, > >Thank you very much for all of your reply.The frozen sections we make now >have better mophology with Liquid N2 freezing. > >Still three questions with regarding to kidney immunofluorescent staining. > >1) We stain human IgG on kidney frozen sections with antibody from DAKO >(F0202),and dilution is 1:50,but the background is always strong. > >Other antibodies to IgA(F0204 from DAKO),IgM(F0203 DAKO),C3(F0201 >DAKO),C4(F0169 DAKO)&C1q (F0254) all produce dark background under >fluorescent microscope,which is highly preferred. > >Why IgG is so strange? Any problem with the antibody? Or the unspecific >staining for IgG is special? > >2) What kind of O.C.T is the best? > >Thank you very much! > >Yichao WU >Research Institute of Nephrology >Jinling Hospital >Nanjing 210002 >P.R.China > _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet "This message is intended for the addressee named and may contain confidential information. If you are not the intended recipient, please destroy it and notify the sender. Views expressed in this message are those of the individual sender, and are not necessarily the views of the Central Sydney Area Health Service." From Jason.PALMER <@t> svhm.org.au Sun Mar 21 18:47:42 2004 From: Jason.PALMER <@t> svhm.org.au (PALMER Jason (SVHM)) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] RE: need help with IHC detection of human cells in mouse tissue Message-ID: I have used a mouse anti human mitochondria antibody from Calbiochem before to successfully identify a variety of human cells in mouse background in paraffin embedded tissue. I am pretty sure, however, that they don't manufacture this one anymore, but I wouldn't be surprised if other companies are producing other human-specific mitochondrial Abs. 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 Hi, I was wondering if anybody could help me out. We want to look for human cells engrafted in mice, particularly in formalin-fixed, acid decalcified, paraffin-embedded tissue. I have so far tried two anti-human beta-2 microglobulin antibodies. The first I tried has been used in human/sheep transplant tissue, and I tried it, but unfortunately it is an IgM kappa, so it failed with the Biogenex mouse-on-mouse IHC kit, which it turns out is not made for IgM antibodies. If anybody is aware of a good way to use a mouse IgM on both human and mouse tissue, I'd love to know. I have looked for a good secondary antibody for this purpose, but have had no luck so far. The second antibody I tried is from Novocastra and distributed by Vector Labs. It is a rabbit polyclonal to human beta-2-microglobulin. The trouble I had was nonspecific binding in control mouse tissue. I was thinking to use some of the Biogenex mouse-on-mouse kit reagents to help block mouse antigens, but I do not know if this would actually work. The second problem with this antibody was that I did not always see positive marking in human tissue (especially osteocytes), which I thought should be pretty well all positive since it is human tissue. Is the expression of beta-2-microglobulin so variable? Has anyone tried any other general human cell marker in other animals? I did a quick search for beta actin, but it seems most of those cross-react with mouse. Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 -------------- next part -------------- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From kegraves <@t> utmb.edu Sun Mar 21 20:03:02 2004 From: kegraves <@t> utmb.edu (Graves, Kerry) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] CD marker in zinc fixed mouse liver Message-ID: <71C7F1D937BE6947BB95FA30532BFF9BFA8DA2@EXCHANGE2K2.utmb.edu> -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Jorge Ivan Zapata Valencia Sent: Sun 3/21/2004 6:59 AM To: histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] CD marker in zinc fixed mouse liver hello everybody. I am new in immunohisotology and I have trying to stain for CD4,CD8 and CD45R but i had problems to get good staining. I am using Pharmingen protocol (rat primary antibody, biotinylated goat anti rat secondary antibody, streptavidina-HRP). I have tried H2O2 as endogenous peroxidase blocking solution (3% in Metanol) but eritrocytes are still brown after DAB. I am blocking with white egg and skim mil, for endogenous biotin. May one help me? May I have any good protocol? Thank you very much. JORGE IVAN ZAPATA Universidad del Valle Cali, Colombia _________________________________________________________________ Protect your PC - get McAfee.com VirusScan Online http://clinic.mcafee.com/clinic/ibuy/campaign.asp?cid=3963 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Sun Mar 21 21:46:27 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] help with elastic staining Message-ID: The iron hematoxylin used for an elastic stain is usually Verhoeff. This is alcoholic hematoxylin (5%), ferric chloride (10%) and Lugol iodine. This is prepared fresh and the solutions are added in the order given. The differentiating solution is 2% ferric chloride. Jennifer MacDonald From JMacDonald <@t> mtsac.edu Sun Mar 21 22:12:59 2004 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] amphoteric dyes Message-ID: Can anyone help clarify amphoteric dyes for me? I have read conflicting information. A couple of sources state that amphoteric dyes are cationic if the pH is below the IEP and anionic if the pH is above the IEP. Other sources state the opposite. Also, what is the purpose of the potassium metabisulfite after the gold chloride toning step in the Gomori Reticulin? Jennifer MacDonald From Janet.Bonner <@t> FLHOSP.ORG Mon Mar 22 09:35:21 2004 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Lori White/ B plus Message-ID: <07AB60D5D7B9754EBF56F360F98D083DEB3F9C@fh2k093.fhmis.net> We get our B+ from BBC Chemical (1-800-635-4477). Solution contains water, 37% formaldehyde, zinc chloride and selected buffers. No iodine clearing is necessary and all 14 of our Pathologists are happy with the results. -----Original Message----- From: Atoska S. Gentry [mailto:gentras@vetmed.auburn.edu] Sent: Friday, March 19, 2004 3:16 PM To: Histonet Subject: Fwd: [Histonet] Lori White/ B plus Hello, please what's your source of this B plus and do you know it's components? Thanks, Atoska >From: "Joyce Cline" >To: "Histonet" >Date: Wed, 17 Mar 2004 14:13:45 -0500 >X-Mailer: Microsoft Outlook Express 6.00.2800.1158 >X-Virus-Scanned: by amavisd-new at wchsys.org >X-Scan-Signature: 690e9200c080ca55fd7a8d0fb0193eeb >X-Content-Filtered-By: Mailman/MimeDel 2.1.3 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >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: 148abf0c5acfe33f338c6ed44fdf9350 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: [Histonet] Lori White/ B plus >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.4 required=6.5 tests=DATE_IN_PAST_12_24 > autolearn=no version=2.63 >X-SA-Exim-Version: 3.1 (built Tue Oct 14 16:21:02 CDT 2003) >X-SA-Exim-Scanned: Yes >X-MIME-Autoconverted: from quoted-printable to 8bit by >cvm4.vetmed.auburn.edu id i2IG9dn16152 > > We fix our nodes & BM's in B Plus for a minimum of 2 hours. Then our > cores are decaled with HCL/Formalin for one hour and process overnight. > We have no problem with the results. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet The information contained in this message may be privileged and/or confidential and protected from disclosure. If the reader of this message is not the intended recipient or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify the sender immediately by replying to this message and deleting the material from any computer. From Ian.Bernard <@t> LACKLAND.AF.MIL Mon Mar 22 09:36:58 2004 From: Ian.Bernard <@t> LACKLAND.AF.MIL (Bernard Ian R SSgt 59 CRES/MSROP) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Looking for a lab shaker with platform. Message-ID: Need this to place specimen containers with formalin for shaking just before grossing. Need price make and recommendations Thanks SSgt B From ASelf <@t> gmhsc.com Mon Mar 22 09:51:49 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] CPT charge code Message-ID: Histonetters, I need some help about CPT charge codes.... We have been in several discussions about the CPT code 88108.The AMA coding book states the 88108 is Cytopathology, concentration technique, smears and interpretation. What does concentration technique mean? I was told if the smears were prepared from one of the following methods that is concentration technique; cytospin, autocyte, or thin prep. And should be coded 88108. Now we spin our fluids down by centrifuge, pour off the supernatent and pipette a drop of the sediment that collects in the bottom of the tube onto a slide and smear with a sister slide. I was told that this was not concentration technique and therefore should be coded as 88104. I have been using 88104 until this morning - I was told to start using code 88108. Which code should I really use for these cytopathology cases? Thanks, Amy(confused) Note: 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 yichaowu <@t> hotmail.com Mon Mar 22 10:01:00 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen sections Message-ID: Dear Young (What a nice name!:-)) Thank you so much for your kindest help and the important information!And our director has urged me to order such kind of F(ab')2 antibody for our immunostaining,when she heard of your suggestions today. Yes,the usage of IgA F0316 (vs F0204), IgM F0317 (vs F0203) as well as IgG F031 (vs F0315) is very reasonable for lower background. I agree this entirely after reading the production explanations at DAKO website. I just wonder, since F(ab')2 antibodies are inclined to produce lower background if there is any,should we change all antibodies we used now to this kind of antibodies if possible? And I just wonder what is the reason of its low background for the F(ab')2 antibody?:-) Would the Fc part of the primary antibody react with the Fc receptors on human cells in the sections? However, the primary antibody is from a different species, would the Fc part of this antibody still react with Fc receptor on human cells? Another possibility may be, the completements on the human sections may fix the primary antibody with Fc part.Still is the different species problem.Maybe there is a kind of reaction accross different species? (Is my description clear? Sorry for any confusion) Besides,could you also advise me how the frozen sections should be blocked? Now we used 10% Bovine Calf Serum in PBS. And before the mounting of the frozen sections for inspection under fluorescent microscope, should the sections be rinsed in some kind of solution like CuSO4 or NaBO4 to decrease autofluorescence if there is any? I am afraid that these two kind of solutions would decrease the intensity of the "real positive"signals. Thank you very much for all of your help! I know a friend from Australian Sydney,a very very nice lady. Connie Yichao WU Research Institute of Nephrology Jinling Hospital 305 East Zhongshan Road Nanjing 210002 P.R.China >From: Young Kwun >To: 'yichao wu' >Subject: RE:Renal IF >Date: Mon, 22 Mar 2004 15:50:54 +1100 > >Dear Yichao, >Next time please call me just Young. That's the Australian way (In most >cases). >Other FITC antibodies we use Fc as well. IgA( All Dako's, F0316), M >(F0317), C3 (F0201), CIq(F0254), >Fibrinogen (F0111) Kappa(F0340) and Lambda(F0341). I normally follow their >suggested upper dilution. If the dilution they suggest is 1:10 - 1:20, then >1:20 would be a quite safe bet generally. >Another thing is that you do not need to fix in aceton or methanol at all >for renal or skin IF. If you have a know positive case, try to do staining >without fixation. Unless we are busy, we normally dry the slides about 30 >mins at room temp. >Feel free to write to me if you have further questions. > >Regards, > >Young > > > >-----Original Message----- >From: yichao wu [SMTP:yichaowu@hotmail.com] >Sent: Monday, 22 March 2004 14:59 >To: kwuny@email.cs.nsw.gov.au >Subject: Thank you very much > >Dear Dr. Kwun, > >Thank you very much for your kindest help! You are the first one who shares >this experience with me.Thank you, really. > >Connie > >Yichao WU >Nanjing China > > > >From: Young Kwun > >To: 'yichao wu' ,"Histonet (E-mail)" > > > >Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney >frozen > >Date: Mon, 22 Mar 2004 09:37:12 +1100 > > > >Hi, > >We use IgG F(ab)2 (Dako F0315) instead of Ig fraction you are using. We > >follow manufacturer's suggested dilution which is 1:10 without any >problem > >with the background staining. > >We used to use another IgG Fc in 1:50 dilution from a company called > >DiaSorin in USA. It was a very good antibody. But we were unable to get > >their antibodies anymore for some reason. > >I hope this helps. > > > > > > > > > > > > > >Young Kwun, PhD > >Senior Hospital Scientist > >Dept. of Anatomical Pathology > >Concord Hospital > >Concord NSW 2139 Australia > >Tel)61-2-9767-6075 > >Fax)61-2-9767-8427 > >kwuny@email.cs.nsw.gov.au > > > >The information contained in this message is intended for the named > >addressee only, and is confidential to the sender and intended recipient. > >If you are not the named addressee please do not copy, distribute, take >any > >action reliant on, or disclose anything in this E-mail message to any >other > >person or organisation. If you have received this message in error please > >delete the email and notify me immediately. Views expressed in this >message > >are those of the individual sender and are not necessarily the views of > >Central Sydney Area Health Service. > > > > > > > >-----Original Message----- > >From: yichao wu [SMTP:yichaowu@hotmail.com] > >Sent: Sunday, 21 March 2004 22:36 > >To: andreah@imclone.com > >Cc: histonet@lists.utsouthwestern.edu > >Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney >frozen > > > >Thank you for your reply!It is very helpful to me.I have once thought of > >the > >problem too. > > > >Since the background of IgA and IgM is very low and they are serum > >immunoglobulins too,it seems strange for the strong background of IgG. > > > >Mr.George Cole advised me that the concentration (1:50 in dilution) for >the > >anti-human IgG antibody may be too high.However, our tech told me that > >further dilution would prevent those positive signals from being present. > > > >Hence I just wonder how other departments of renal diseases handle the > >immunofluorescent staining in biopsy specimens? Please kindly give me >some > >hints since it is terrible for the diagnosis of patients with bad > >immunofluorescent staining results.It really hurts :-( > > > >Thank you very much in advance! > > > >Yichao WU,Ph.D Candidate > >Research Institute of Nephrology > >Jinling Hospital > >305 East Zhongshan Road > >Nanjing 210002 > >P.R.China > > > > >From: andreah@imclone.com > > >To: yichaowu@hotmail.com > > >Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney > >frozen > > >Date: Sat, 20 Mar 2004 13:07:18 -0500 > > > > > > > > > > > > > > > > > >I expect it is not background but real signal from all the IgG in the > > >tissue. IgG is normally at high concentration in mouse tissue and will > >give > > >staining especially in a blood filled organ like the kidney. > > > > > >----- Message from "yichao wu" on Sat, 20 Mar >2004 > > >17:25:27 +0800 ----- > > > > > >To: histonet@lists.utsouthwestern.edu, tli1@flowcity.bsd.uchicago.edu > > >Subject: [Histonet] Immunofluorescent staining of IgG on kidney >frozen > > >sections---High background! > > > > > >Dear Dr.Callis,Dr.Yaskovich,,Dr. Li,and all histonetters, > > > > > >Thank you very much for all of your reply.The frozen sections we make >now > > >have better mophology with Liquid N2 freezing. > > > > > >Still three questions with regarding to kidney immunofluorescent > >staining. > > > > > >1) We stain human IgG on kidney frozen sections with antibody from DAKO > > >(F0202),and dilution is 1:50,but the background is always strong. > > > > > >Other antibodies to IgA(F0204 from DAKO),IgM(F0203 DAKO),C3(F0201 > > >DAKO),C4(F0169 DAKO)&C1q (F0254) all produce dark background under > > >fluorescent microscope,which is highly preferred. > > > > > >Why IgG is so strange? Any problem with the antibody? Or the unspecific > > >staining for IgG is special? > > > > > >2) What kind of O.C.T is the best? > > > > > >Thank you very much! > > > > > >Yichao WU > > >Research Institute of Nephrology > > >Jinling Hospital > > >Nanjing 210002 > > >P.R.China > > > > > > >_________________________________________________________________ > >MSN 8 with e-mail virus protection service: 2 months FREE* > >http://join.msn.com/?page=features/virus > > > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > >"This message is intended for the addressee named and may contain > >confidential information. If you are not the intended recipient, please > >destroy it and notify the sender. Views expressed in this message are >those > >of the individual sender, and are not necessarily the views of the >Central > >Sydney Area Health Service." > > > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >_________________________________________________________________ >MSN 8 helps eliminate e-mail viruses. Get 2 months FREE*. >http://join.msn.com/?page=features/virus > > >"This message is intended for the addressee named and may contain >confidential information. If you are not the intended recipient, please >destroy it and notify the sender. Views expressed in this message are those >of the individual sender, and are not necessarily the views of the Central >Sydney Area Health Service." _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail From medjnw <@t> emory.edu Mon Mar 22 10:19:51 2004 From: medjnw <@t> emory.edu (Josiah N. Wilcox, Ph.D.) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Histopathology positions at Medtronic Vascular Message-ID: There are currently two openings at Medtronic Vascular in Santa Rosa, California for a histopathologist (senior manager, PhD, DVM or equivalent) and histology technician/histotechnologist (BS or MS with experience) to help establish a new histopathology laboratory to work on drug eluting stents and other vascular projects. Individuals with experience in plastic embedding and sectioning or previous experience in processing stents for histopathology and morphometric analysis are encouraged to apply. Josiah N. Wilcox, Ph.D. Visiting Scholar Medtronic Vascular 3576 Unocal Place SS-48, FBMC Santa Rosa, CA 95403 Professor of Hematology/Oncology and Internal Medicine (on sabbatical) The Winship Cancer Institute Dept. of Hematology/Oncology 1635-B Clifton Rd., Suite B5100 Atlanta, GA 30322 Email: medjnw@emory.edu WebPage: www.emory.edu/WILCOX/ Medtronic is one of the world?s leading medical technology companies dedicated to providing lifelong solutions to people with chronic disease. Medtronic is dedicated to the total well being of its employees and has been recognized as a Fortune ?Best 100 Companies to Work for in America.? Histology Technician Responsibilities: Tissue Processing, including assignment of accession numbers to tissue specimens, monitoring of the automatic tissue processors, and embedding of tissues in frozen, plastic and paraffin media. Slide Preparation and sectioning, including frozen, paraffin and plastic tissue blocks, operating microtomes, and slide review for quality control of tissue sections. Specialized staining of histology slides including immunohistochemistry. Complete various histomorphometric measurements using computer image analysis following specific requirement in a study protocol. Assist with the generation of reports and various interpretive gross and histopathologic evaluations of highly complex host-biomaterial interactions. Collaborate with the generation, review and implementation of applicable SOPs. Other tasks may include maintenance of files of scientific histology publications; instrument and equipment maintenance; preparation of records of quality control checks on stains prepared. Requirements/ skills: BS required; MS preferred. 2-5 years relevant histopathology experience in a medical laboratory technology program or equivalent. ASCP registration preferred. Prior experience with computer based histomorphometry and plastic tissue processing/sectioning helpful. Excellent computer skills, including word processing, spreadsheets, and database management. Team Player, dependability; good written, communication and interpersonal skills; logical problem solving ability. Ability to take initiative in team environment and find creative solutions to problems. Organizational skills, prioritization skills, timely decision-making and time management abilities. With people like you, who have purpose, potential and passion, we supply physicians and their patients around the world with the means to extend life, restore health and alleviate pain. To route your resume directly to the hiring department, visit www.medtronic.com/employment, select Requisition 35672 and apply online using "Apply to this Job" or "Add to Cart". No agencies or phone calls, please. Medtronic is an equal opportunity employer committed to cultural diversity in the workforce. Histopathologist Responsibilities: Setting up and subsequently managing the Medtronic Vascular Histology lab in compliance with GLP requirements (21 CFR, Part 58). The primary responsibility of this role is to develop and manage the new histology laboratory in Medtronic Vascular. Responsible for histopathological analysis of tissues arising from pre-clinical studies, and subsequently, interpreting, analyzing, documenting, and reporting of these results. Additionally, the Histopathologist will collaborate with scientists, regulatory, clinical, and R&D personnel on Drug Eluting Stents pre-clinical studies as well as other biologic programs called out in the Vascular Strategic plan. As an active member of the pre-clinical research team, this individual will be responsible for leading the histopathology component of pre-clinical studies for the Vascular Division. Requirements/ skills: PhD.or Doctor of Veterinary Medicine. Prior experience with computer based histomorphometry. Experience in veterinary histopathology is essential; experience in cardiovascular histopathology is preferred. 6-10 years of relevant histopathology experience including plastic embedding techniques, histochemistry and computer based morphological analysis. Scientific rigor within the context of GLP or similar QA scheme. Good technical writing, communications, prioritization, organizational and computer skills. Ability to take initiative in team matrix environment and find creative solutions to problems. Avid collaborator/ team player. Knowledgeable educator with proven critical thinking skills. With people like you, who have purpose, potential and passion, we supply physicians and their patients around the world with the means to extend life, restore health and alleviate pain. To route your resume directly to the hiring department, visit www.medtronic.com/employment, select Requisition 35673 and apply online using "Apply to this Job" or "Add to Cart". No agencies or phone calls, please. Medtronic is an equal opportunity employer committed to cultural diversity in the workforce. From Jennifer_Harvey <@t> URMC.Rochester.edu Mon Mar 22 10:07:19 2004 From: Jennifer_Harvey <@t> URMC.Rochester.edu (Harvey, Jennifer) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Looking for antibodies for ERK5, BCR, IKB-alpha Message-ID: Histonet, I am looking for antibodies that work on formalin fixed paraffin sections for ERK5, BCR, and IKB- alpha. Is there anyone that has found ones that work? Thanks for your help, Jennifer Harvey Center for Cellular and Molecular Cardiology University of Rochester Medical Center 601 Elmwood Avenue, Box 679-CCMC Rochester, NY 14642 Telephone: (585) 273-5161 From yichaowu <@t> hotmail.com Mon Mar 22 10:33:20 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen Message-ID: Dear LuAnn, Thank you for your suggestions.And George Cole has also suggest that to me.Thank you all. Yichao >From: LuAnn Anderson >To: "yichao wu" >Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney frozen >Date: Mon, 22 Mar 2004 10:21:44 -0600 > >Hello Yichao, >You will need to find a known positive control and titer the IgG. Start at >1:50 and increase 1:100, 1:200, 1:400, 1:800, etc (checkerboard). Then >screen them and determine which titration works best for low background but >positive staining. You could do dilutions in between if you think >necessary. 1:50 sound much too high, which would produce the background you >are seeing. > > > > > >At 07:35 PM 3/21/04 +0800, you wrote: >>Thank you for your reply!It is very helpful to me.I have once thought of >>the problem too. >> >>Since the background of IgA and IgM is very low and they are serum >>immunoglobulins too,it seems strange for the strong background of IgG. >> >>Mr.George Cole advised me that the concentration (1:50 in dilution) for >>the anti-human IgG antibody may be too high.However, our tech told me that >>further dilution would prevent those positive signals from being present. >> >>Hence I just wonder how other departments of renal diseases handle the >>immunofluorescent staining in biopsy specimens? Please kindly give me some >>hints since it is terrible for the diagnosis of patients with bad >>immunofluorescent staining results.It really hurts :-( >> >>Thank you very much in advance! >> >>Yichao WU,Ph.D Candidate >>Research Institute of Nephrology >>Jinling Hospital >>305 East Zhongshan Road >>Nanjing 210002 >>P.R.China >> >>>From: andreah@imclone.com >>>To: yichaowu@hotmail.com >>>Subject: RE: [Histonet] Immunofluorescent staining of IgG on kidney >>>frozen >>>Date: Sat, 20 Mar 2004 13:07:18 -0500 >>> >>> >>> >>> >>> >>>I expect it is not background but real signal from all the IgG in the >>>tissue. IgG is normally at high concentration in mouse tissue and will >>>give >>>staining especially in a blood filled organ like the kidney. >>> >>>----- Message from "yichao wu" on Sat, 20 Mar 2004 >>>17:25:27 +0800 ----- >>> >>>To: histonet@lists.utsouthwestern.edu, tli1@flowcity.bsd.uchicago.edu >>>Subject: [Histonet] Immunofluorescent staining of IgG on kidney frozen >>>sections---High background! >>> >>>Dear Dr.Callis,Dr.Yaskovich,,Dr. Li,and all histonetters, >>> >>>Thank you very much for all of your reply.The frozen sections we make now >>>have better mophology with Liquid N2 freezing. >>> >>>Still three questions with regarding to kidney immunofluorescent >>>staining. >>> >>>1) We stain human IgG on kidney frozen sections with antibody from DAKO >>>(F0202),and dilution is 1:50,but the background is always strong. >>> >>>Other antibodies to IgA(F0204 from DAKO),IgM(F0203 DAKO),C3(F0201 >>>DAKO),C4(F0169 DAKO)&C1q (F0254) all produce dark background under >>>fluorescent microscope,which is highly preferred. >>> >>>Why IgG is so strange? Any problem with the antibody? Or the unspecific >>>staining for IgG is special? >>> >>>2) What kind of O.C.T is the best? >>> >>>Thank you very much! >>> >>>Yichao WU >>>Research Institute of Nephrology >>>Jinling Hospital >>>Nanjing 210002 >>>P.R.China >> >>_________________________________________________________________ >>MSN 8 with e-mail virus protection service: 2 months FREE* >>http://join.msn.com/?page=features/virus >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________________________________________ MSN 8 with e-mail virus protection service: 2 months FREE* http://join.msn.com/?page=features/virus From STEGTM <@t> samcstl.org Mon Mar 22 10:46:21 2004 From: STEGTM <@t> samcstl.org (Therersa Stegall) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Dako Artisan Message-ID: Hello. Again, thank you to everyone who responded to our inquiries re:immunostainers. I am now asking for input regarding the combination unit from Dako, called the "Artisan". Any/all remarks, comments, kudos, jeers are welcome. Peace, TS From mrsgbd2001 <@t> yahoo.com Mon Mar 22 11:44:25 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Cryosectioning Rat eyes Message-ID: <20040322174425.38438.qmail@web13310.mail.yahoo.com> HI all, Wondering if I could get some opinions on the best way to embed rat eyes in OCT. I usually see people cut the eye in half, or in a cup shape (removing the lense), we are interested in the retinal area. Any ideals? Ms.Gareth Davis Research Assistant Tennessee State University Do you Yahoo!? Yahoo! Finance Tax Center - File online. File on time. From jcline <@t> wchsys.org Mon Mar 22 12:15:40 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Atoska Gentry/ B Plus Message-ID: <006901c41039$af661660$1d2a14ac@wchsys.org> Hi Atoska I get my B Plus from Market Lab, order # ML0407, Phone # 800-237-3604. I do not know it's components, other than formaldehyde. The company that makes it is Advanced Biomedical Reagents & Technologies. From AFeatherstone <@t> KaleidaHealth.Org Mon Mar 22 12:47:13 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Ventana staining Avidin Biotin Block Message-ID: hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 muololi <@t> umdnj.edu Mon Mar 22 13:21:12 2004 From: muololi <@t> umdnj.edu (Lilia Muolo) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Ventana staining Avidin Biotin Block Message-ID: Hi, our lab only uses the A /B blocking on the Ventana if necessary but usually we don't need to use it. We use the Discovery here but I do remember using the A/B blocking more often when we had the Nexes. I would imagine that the Benchmark would be like the Discovery. Lil Muolo Cancer Institute of New Jersey >>> "Featherstone, Annette" 3/22/04 1:47:13 PM >>> hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From juan.gutierrez <@t> christushealth.org Mon Mar 22 13:21:59 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Ventana staining Avidin Biotin Block Message-ID: We use it on everything. It's a lot easier than having a different protocol for aech type of tissue. Juan -----Original Message----- From: Featherstone, Annette [mailto:AFeatherstone@KaleidaHealth.Org] Sent: Mon 3/22/2004 12:47 PM To: Histonet (E-mail) Cc: Subject: [Histonet] Ventana staining Avidin Biotin Block hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jcline <@t> wchsys.org Mon Mar 22 14:56:30 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Wendy England/ iron smears Message-ID: <001f01c41050$277dcb00$1d2a14ac@wchsys.org> I use an old fashioned way of fixing the bone marrow smears. We had trouble with no reaction on our smears. Now we use a coplin jar with a small piece of paper towel in the bottom, drop 10% buffered formalin to saturate the paper towel but not to reach the blood on the slides. Leave the slides in for 10 minutes and stain normally. We have not had any trouble with an iron reaction since using this method. But do not leave the slides in more than 15 minutes max. From tpmorken <@t> labvision.com Mon Mar 22 16:30:56 2004 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] IHC position in San Francisco bay area. Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA217741F@usca0082k08.labvision.apogent.com> Lab Vision, a fast-growing biotech company located in Fremont, California, just southeast of San Francisco, has an opening for an Immunohistochemistry Technologist in the Quality Control laboratory. The position entails testing of manufactured lots of antibody and detection reagents as well as prospective reagents. The applicant must have extensive experience in immunohistochemistry, especially in development of new antibodies and other IHC reagents. Advancement is limited only by personal initiative. Certification by ASCP as HT or HTL and/or QIHC is preferred. Salary is negotiable. Lab Vision (www.labvision.com) is a world-leading manufacturer of IHC-automation instruments and immunohistochemistry reagents for the pathology laboratory and biological research. Lab Vision is an Apogent company (www.apogent.com) If interested please send resume by email to: Tim Morken Product Development Lab Vision / Neomarkers 47790 Westinghouse Dr. Fremont, CA 94539 USA PH: 510-991-2840 FAX: 510-991-2826 email: tpmorken@labvision.com www.labvision.com From gcallis <@t> montana.edu Mon Mar 22 17:12:28 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Small Animal atlas (Volumes 1 and 2) are back in print! Message-ID: <3.0.6.32.20040322161228.00bcc478@gemini.msu.montana.edu> Dear All, For those of you who work with or dissect laboratory animals and are in need of an atlas to guide the way. After many years of frustration and sadness when these atlases went out of print, they are back on the market. I saw the new atlas today, and couldn't wait to order it. A huge silent hooray! There are two color atlases (volume 1 is for rabbit and guinea pig and Volume 2 is rat, mouse, and Golden hamster. Pricing for Vol 1 and Vol 2 are $139 per atlas, and that is a deal. ISBN:0702027030 Title:Colour Atlas of Anatomy of Small Laboratory Animals, Volume 2, mouse, rat, hamster Price:$139.00 Go to Elsevier website to order, it is very user friendly, excellent. I just gave/ordered myself an early birthday gift! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From AnthonyH <@t> chw.edu.au Mon Mar 22 18:22:18 2004 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E16A@simba.kids> Dear Boswell, The following Cherukian's modification works very well. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html Document Procedure: Phosphotungstic Acid Haematoxylin Principle: Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Use brain sections and section of muscle. A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined. Reagents: 1. Eosin: - Warning: Flammable - see MSDS Eosin Y, water soluble (CI 145380) 0.5g Distilled Water 10ml 80% Ethanol 190ml Working: 10ml stock and Before use add 50ul glacial acetic acid. 2. 1% Periodic Acid 3. PTAH solution Haematoxylin (CI 75290) 0.5g Phosphotungstic Acid 10g Distilled water 500ml Dissolve solid ingredients in separate portions of the water. Use gentle heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium permanganate to ripen. The stain is ready to use. Procedure: 1. Dewax and hydrate sections to 80% alcohol. 2. Place slides in eosin for 30 seconds. 3. Wash slides in distilled water for a few seconds. 4. Place slides in 1% periodic acid for 20 minutes. 5. Wash slides in water for 3 minutes. 6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30 minutes on. 7. Dehydrate, clear and mount. Results: Dendrites, nuclei, fibrin, platelets and muscle - blue Red blood cells and collagen - red. Notes: Reference: 1. Cherukian, C.J., Histologic. 8(4); 105, (1977). 2. Luna, L., Histologic. 5(2); 66, (1975). -----Original Message----- From: Boswell [mailto:kcboswell@grandecom.net] Sent: Saturday, 20 March 2004 7:30 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using Zenker's Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain without using Zenker's. We have looked everywhere and are unable to find it. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From bozz <@t> icm.csic.es Tue Mar 23 03:14:40 2004 From: bozz <@t> icm.csic.es (Anna Bozzano) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] please remove me from your mailing list Message-ID: <3.0.1.32.20040323101440.0120fd88@cucafera.icm.csic.es> please remove me from your mailing list From kemlo <@t> tiscali.co.uk Tue Mar 23 03:21:37 2004 From: kemlo <@t> tiscali.co.uk (Kemlo Rogerson) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Web site for Cytology In-Reply-To: Message-ID: <000001c410b8$3f376b80$48362850@KEMLOS> Cytopathnet.org I think. It sort of comes and goes depending on the variety of virus it becomes infected with. I'm always amused that it becomes regularly infected by viruses; sort of gives you a feeling of its authenticity. I think it is negative for the oncogenic variety but hope it has regular tests including a silicone biopsy. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Boozer Sent: 17 March 2004 17:13 To: histonet@pathology.swmed.edu Subject: [Histonet] Web site for Cytology Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vanann702 <@t> skmc.gov.ae Tue Mar 23 02:59:07 2004 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] subscribe Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E01F18A95@SKMCEMAIL.skmc.gov.ae> From StarkusL <@t> ummhc.org Tue Mar 23 06:56:38 2004 From: StarkusL <@t> ummhc.org (Starkus, Laurie) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Mouse Lung Message-ID: I've recently started working with mouse lung and was given a protocol for processing. The protocol calls for 0.85% NaCl for 60 minutes at 4 degrees celsius. Then a 1:1 0.85% NaCl to Absolute ethanol for 30 minutes at room temp. Then the processing looks rather normal. Since my experience is with human tissue, I've never seen a sodium chloride step in processing. What does it do? And, is it really necessary? This is mouse lung infected with TB or cryptococcus. Thanks in advance. From AFeatherstone <@t> KaleidaHealth.Org Tue Mar 23 07:34:14 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] help with elastic staining Message-ID: When doing the pentachrome stain do not over differentiate the elastic too much in the ferric chloride, keep it kind of dark. The acid solutions that follow continue to lighten the elastic fibers. Annette FeatherstoneHT/MLT Kaleida Health -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 14:19 To: Deb Stults; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] help with elastic staining I have this problem when I do a Pentachrome stain for elastic fibers, the fibers stain fine when I first do the hematoxylin but as I continue with the rest of the stain the fiber stain is lost, I have remedied this by repeating the hematoxylin again at the end of the staining process to get back the fiber staining. Try it. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Deb Stults Sent: Friday, March 19, 2004 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help with elastic staining We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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 komo <@t> bu.edu Tue Mar 23 08:48:31 2004 From: komo <@t> bu.edu (komo@bu.edu) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Problems with c-fos immediate early gene labelling Message-ID: <1080053311.40604e3f585a5@www.bu.edu> I'm hoping I could get some helpful advice on what might cause my c-fos IHC labelling to work well in one lab, but not my own. I am running the protocol on rat brains using a hydrogen peroxide/methanol step for endogenous peroxidase blocking, a normal goat serum blocking step, a c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated secondary, a streptavidin horeradish peroxidase, and a nickle enhanced DAB reaction. Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. Does anyone has any suggestions as to why this might be? One person at the other lab suggested that possibly the wells that we run the tissue in were not being cleaned properly (we soak them in bleach while the other lab uses just pex). Could this have any effect? Thanks for any advice you can provide!! Rob Komorowski From AFeatherstone <@t> KaleidaHealth.Org Tue Mar 23 08:50:21 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] CD98 and PLGF Message-ID: YES!!I am! Or was... I used a pepsin pretreatment @37* for 15 minutes, Block with a protein block, primary antibody 1:100 for 1hr, Goat secondary 30 min rt, Alk phos 30 minutes rt, and whatever chromgen you use for alk phos, rt 40 min. My Plgf is from Santa Cruz. Annette Featherstone HT/MLT -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 13:58 To: Patsy Ruegg; Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: RE: [Histonet] CD98 and PLGF I guess no one is using cd98 and/or Placental Growth Factor on ffpe tissue???? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, March 17, 2004 10:03 AM To: Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: [Histonet] CD98 and PLGF I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160 and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies work in ffpe tissue? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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 mcauliff <@t> umdnj.edu Tue Mar 23 12:24:38 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Problems with c-fos immediate early gene labelling In-Reply-To: <1080053311.40604e3f585a5@www.bu.edu> References: <1080053311.40604e3f585a5@www.bu.edu> Message-ID: <406080E6.2080605@umdnj.edu> Hi Ron: komo@bu.edu wrote: >I'm hoping I could get some helpful advice on what might cause my c-fos >IHC labelling to work well in one lab, but not my own. I am running the >protocol on rat brains using a hydrogen peroxide/methanol step for >endogenous peroxidase blocking, a normal goat serum blocking step, a >c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated >secondary, a streptavidin horeradish peroxidase, and a nickle enhanced >DAB reaction. > >Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. > When you say you are using the "exact same antibodies, chemicals, and ..... " do you mean the same bottle of reagent? You are carrying the bottle or vial of reagent from one lab to another? If not, look at the batch of antibody, your peroxide, your DAB, etc. If you are storing antibody in a freezer at home it is being subjected to freeze-thaw cycles (unless you have a very old freezer that does not defrost itself on a timer). If your bottle of peroxide is more than a few months old it may be in the process of turning into water, peroxide does that. I have had DAB work one day and fail the next. Since you have found that the brain slices stain well in the "other lab", the problem must lie with your home lab. Since bleach reacts with DAB I would avoid using staining dishes treated with bleach. Geoff >Does anyone has any suggestions as to why this might be? One person >at the other lab suggested that possibly the wells that we run the tissue in >were not being cleaned properly (we soak them in bleach while the other >lab uses just pex). Could this have any effect? > >Thanks for any advice you can provide!! > >Rob Komorowski > > > >_______________________________________________ >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 dmcaloose <@t> wcs.org Tue Mar 23 09:46:32 2004 From: dmcaloose <@t> wcs.org (McAloose, Dee) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] FW: Job opening - Wildlife Conservation Society Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD6D3@WOLF.wcs.org> > Hello, > > I'd like to announce an immediate opening for a full-time histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity and in becoming part of our team! > > Thanks for listening and we look forward to hearing from you. > > D McAloose, VMD, Dipl ACVP > Head, Department of Pathology > Wildlife Conservation Society > Bronx, NY 10464 > (phone) 718-220-7105 > (fax) 718-220-7126 > dmcaloose@wcs.org > > From shive003 <@t> umn.edu Tue Mar 23 10:21:54 2004 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] IHC on blood and cytology smears (ICC) Message-ID: <006301c410f2$f45d1ed0$78065486@vdl220FAC> Could someone please send me a protocol for performing IHC/ICC on smears (I normally only do FFPE slides)? I'd like info mostly on if the slides have to be coated/charged , smears air-dried or fixed (and in what) for how long, and if you do any modification of the immuno protocol, such as blocking endogenous peroxidase later in the procedure (and with what concentration of H2O2), and/or changing incubation times/dilutions. I've done smears on very rare occasions, but have had trouble getting the cells to remain on the uncoated slides that the smears are being made on. Thank you very much in advance. Jan Shivers Univ of Minnesota Vet Diag Lab shive003@tc.umn.edu From Luis.Chiriboga <@t> med.nyu.edu Tue Mar 23 11:02:46 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] NYSHS 2004 Annual Meeting Message-ID: The 2004 New York State Histotechnology Society annual meeting will be held at the Holiday Inn in beautiful Saratoga Springs, New York from Friday April 30th through Saturday May 1st. This years meeting theme is "Histology is no mystery: Let New York State solve your problems". An e-mail mini program is available upon request by replying to this message. For a program and registration packet, please contact Judy LaDuc at 518-897-2247 or jaladuc@capital.net We hope to see you there. From mprice26 <@t> juno.com Tue Mar 23 12:38:43 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Texas Society for Histotechnology Message-ID: <20040323.103935.1731.2431688@webmail06.nyc.untd.com> Hi Histonetters, Can someone send me the info for the TSH meeting in April? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From psanquin <@t> lugo.usc.es Tue Mar 23 13:00:59 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:42 2005 Subject: [Histonet] Fetus fixation In-Reply-To: <20040323.103935.1731.2431688@webmail06.nyc.untd.com> Message-ID: <3.0.6.32.20040323200059.007b7440@pop.lugo.usc.es> Dear listers, I am trying to cut brains from fetal mice (E19) fixed in Paraformaldehyde 4% with the vibratome. I have problems with the perfusion of the mother. The fixative does not reach the fetuses and these even after 12 hours postfixation in the same fixative are too soft to take the brains out. Could you give me some input? Have you tried to cut in vibratome the whole head of the fetus? Is it possible? Thanks in advance Pablo From lwhite <@t> lakeridgehealth.on.ca Tue Mar 23 14:42:10 2004 From: lwhite <@t> lakeridgehealth.on.ca (White, Lori) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] RE: Histonet Ventana staining Avidin Biotin block Message-ID: Hi, We use the avidin/block on the Ventana Nexes as necessary and it works very well. I don't know that it would be advantageous to use for all tissues, considering the extra time that would be built into the run..... Lori -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, March 23, 2004 1:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 4, Issue 27 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. Atoska Gentry/ B Plus (Joyce Cline) 2. Ventana staining Avidin Biotin Block (Featherstone, Annette) 3. Re: Ventana staining Avidin Biotin Block (Lilia Muolo) 4. RE: Ventana staining Avidin Biotin Block (GUTIERREZ, JUAN) 5. Wendy England/ iron smears (Joyce Cline) 6. IHC position in San Francisco bay area. (Morken, Tim - Labvision) 7. Small Animal atlas (Volumes 1 and 2) are back in print! (Gayle Callis) 8. RE: Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's (Tony Henwood) 9. please remove me from your mailing list (Anna Bozzano) 10. RE: Web site for Cytology (Kemlo Rogerson) 11. subscribe (Anne Van Binsbergen) 12. Mouse Lung (Starkus, Laurie) 13. RE: help with elastic staining (Featherstone, Annette) 14. Problems with c-fos immediate early gene labelling (komo@bu.edu) 15. RE: CD98 and PLGF (Featherstone, Annette) 16. Re: Problems with c-fos immediate early gene labelling (Geoff McAuliffe) 17. FW: Job opening - Wildlife Conservation Society (McAloose, Dee) 18. IHC on blood and cytology smears (ICC) (Jan Shivers) 19. NYSHS 2004 Annual Meeting (Luis Chiriboga) ---------------------------------------------------------------------- Message: 1 Date: Mon, 22 Mar 2004 13:15:40 -0500 From: "Joyce Cline" Subject: [Histonet] Atoska Gentry/ B Plus To: "Histonet" Message-ID: <006901c41039$af661660$1d2a14ac@wchsys.org> Content-Type: text/plain; charset="iso-8859-1" Hi Atoska I get my B Plus from Market Lab, order # ML0407, Phone # 800-237-3604. I do not know it's components, other than formaldehyde. The company that makes it is Advanced Biomedical Reagents & Technologies. ------------------------------ Message: 2 Date: Mon, 22 Mar 2004 13:47:13 -0500 From: "Featherstone, Annette" Subject: [Histonet] Ventana staining Avidin Biotin Block To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. ------------------------------ Message: 3 Date: Mon, 22 Mar 2004 14:21:12 -0500 From: Lilia Muolo Subject: Re: [Histonet] Ventana staining Avidin Biotin Block To: AFeatherstone@KaleidaHealth.Org, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hi, our lab only uses the A /B blocking on the Ventana if necessary but usually we don't need to use it. We use the Discovery here but I do remember using the A/B blocking more often when we had the Nexes. I would imagine that the Benchmark would be like the Discovery. Lil Muolo Cancer Institute of New Jersey >>> "Featherstone, Annette" 3/22/04 1:47:13 PM >>> hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 22 Mar 2004 13:21:59 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Ventana staining Avidin Biotin Block To: "Featherstone, Annette" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="utf-8" We use it on everything. It's a lot easier than having a different protocol for aech type of tissue. Juan -----Original Message----- From: Featherstone, Annette [mailto:AFeatherstone@KaleidaHealth.Org] Sent: Mon 3/22/2004 12:47 PM To: Histonet (E-mail) Cc: Subject: [Histonet] Ventana staining Avidin Biotin Block hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 22 Mar 2004 15:56:30 -0500 From: "Joyce Cline" Subject: [Histonet] Wendy England/ iron smears To: "Histonet" Message-ID: <001f01c41050$277dcb00$1d2a14ac@wchsys.org> Content-Type: text/plain; charset="iso-8859-1" I use an old fashioned way of fixing the bone marrow smears. We had trouble with no reaction on our smears. Now we use a coplin jar with a small piece of paper towel in the bottom, drop 10% buffered formalin to saturate the paper towel but not to reach the blood on the slides. Leave the slides in for 10 minutes and stain normally. We have not had any trouble with an iron reaction since using this method. But do not leave the slides in more than 15 minutes max. ------------------------------ Message: 6 Date: Mon, 22 Mar 2004 14:30:56 -0800 From: "Morken, Tim - Labvision" Subject: [Histonet] IHC position in San Francisco bay area. To: "Histology Net List Server (E-Mail)" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA217741F@usca0082k08.labvision.apogent.com> Content-Type: text/plain Lab Vision, a fast-growing biotech company located in Fremont, California, just southeast of San Francisco, has an opening for an Immunohistochemistry Technologist in the Quality Control laboratory. The position entails testing of manufactured lots of antibody and detection reagents as well as prospective reagents. The applicant must have extensive experience in immunohistochemistry, especially in development of new antibodies and other IHC reagents. Advancement is limited only by personal initiative. Certification by ASCP as HT or HTL and/or QIHC is preferred. Salary is negotiable. Lab Vision (www.labvision.com) is a world-leading manufacturer of IHC-automation instruments and immunohistochemistry reagents for the pathology laboratory and biological research. Lab Vision is an Apogent company (www.apogent.com) If interested please send resume by email to: Tim Morken Product Development Lab Vision / Neomarkers 47790 Westinghouse Dr. Fremont, CA 94539 USA PH: 510-991-2840 FAX: 510-991-2826 email: tpmorken@labvision.com www.labvision.com ------------------------------ Message: 7 Date: Mon, 22 Mar 2004 16:12:28 -0700 From: Gayle Callis Subject: [Histonet] Small Animal atlas (Volumes 1 and 2) are back in print! To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040322161228.00bcc478@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Dear All, For those of you who work with or dissect laboratory animals and are in need of an atlas to guide the way. After many years of frustration and sadness when these atlases went out of print, they are back on the market. I saw the new atlas today, and couldn't wait to order it. A huge silent hooray! There are two color atlases (volume 1 is for rabbit and guinea pig and Volume 2 is rat, mouse, and Golden hamster. Pricing for Vol 1 and Vol 2 are $139 per atlas, and that is a deal. ISBN:0702027030 Title:Colour Atlas of Anatomy of Small Laboratory Animals, Volume 2, mouse, rat, hamster Price:$139.00 Go to Elsevier website to order, it is very user friendly, excellent. I just gave/ordered myself an early birthday gift! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 8 Date: Tue, 23 Mar 2004 11:22:18 +1100 From: Tony Henwood Subject: RE: [Histonet] Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's To: "'Boswell'" , Histonet@lists.utsouthwestern.edu Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E16A@simba.kids> Content-Type: text/plain; charset="iso-8859-1" Dear Boswell, The following Cherukian's modification works very well. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html Document Procedure: Phosphotungstic Acid Haematoxylin Principle: Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Use brain sections and section of muscle. A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined. Reagents: 1. Eosin: - Warning: Flammable - see MSDS Eosin Y, water soluble (CI 145380) 0.5g Distilled Water 10ml 80% Ethanol 190ml Working: 10ml stock and Before use add 50ul glacial acetic acid. 2. 1% Periodic Acid 3. PTAH solution Haematoxylin (CI 75290) 0.5g Phosphotungstic Acid 10g Distilled water 500ml Dissolve solid ingredients in separate portions of the water. Use gentle heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium permanganate to ripen. The stain is ready to use. Procedure: 1. Dewax and hydrate sections to 80% alcohol. 2. Place slides in eosin for 30 seconds. 3. Wash slides in distilled water for a few seconds. 4. Place slides in 1% periodic acid for 20 minutes. 5. Wash slides in water for 3 minutes. 6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30 minutes on. 7. Dehydrate, clear and mount. Results: Dendrites, nuclei, fibrin, platelets and muscle - blue Red blood cells and collagen - red. Notes: Reference: 1. Cherukian, C.J., Histologic. 8(4); 105, (1977). 2. Luna, L., Histologic. 5(2); 66, (1975). -----Original Message----- From: Boswell [mailto:kcboswell@grandecom.net] Sent: Saturday, 20 March 2004 7:30 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using Zenker's Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain without using Zenker's. We have looked everywhere and are unable to find it. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 9 Date: Tue, 23 Mar 2004 10:14:40 +0100 From: Anna Bozzano Subject: [Histonet] please remove me from your mailing list To: histonet@lists.utsouthwestern.edu Message-ID: <3.0.1.32.20040323101440.0120fd88@cucafera.icm.csic.es> Content-Type: text/plain; charset="us-ascii" please remove me from your mailing list ------------------------------ Message: 10 Date: Tue, 23 Mar 2004 09:21:37 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Web site for Cytology To: "'Kathleen Boozer'" , Message-ID: <000001c410b8$3f376b80$48362850@KEMLOS> Content-Type: text/plain; charset="us-ascii" Cytopathnet.org I think. It sort of comes and goes depending on the variety of virus it becomes infected with. I'm always amused that it becomes regularly infected by viruses; sort of gives you a feeling of its authenticity. I think it is negative for the oncogenic variety but hope it has regular tests including a silicone biopsy. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Boozer Sent: 17 March 2004 17:13 To: histonet@pathology.swmed.edu Subject: [Histonet] Web site for Cytology Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 23 Mar 2004 12:59:07 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] subscribe To: Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E01F18A95@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="iso-8859-1" ------------------------------ Message: 12 Date: Tue, 23 Mar 2004 07:56:38 -0500 From: "Starkus, Laurie" Subject: [Histonet] Mouse Lung To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="ISO-8859-1" I've recently started working with mouse lung and was given a protocol for processing. The protocol calls for 0.85% NaCl for 60 minutes at 4 degrees celsius. Then a 1:1 0.85% NaCl to Absolute ethanol for 30 minutes at room temp. Then the processing looks rather normal. Since my experience is with human tissue, I've never seen a sodium chloride step in processing. What does it do? And, is it really necessary? This is mouse lung infected with TB or cryptococcus. Thanks in advance. ------------------------------ Message: 13 Date: Tue, 23 Mar 2004 08:34:14 -0500 From: "Featherstone, Annette" Subject: RE: [Histonet] help with elastic staining To: 'Patsy Ruegg' , Deb Stults , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" When doing the pentachrome stain do not over differentiate the elastic too much in the ferric chloride, keep it kind of dark. The acid solutions that follow continue to lighten the elastic fibers. Annette FeatherstoneHT/MLT Kaleida Health -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 14:19 To: Deb Stults; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] help with elastic staining I have this problem when I do a Pentachrome stain for elastic fibers, the fibers stain fine when I first do the hematoxylin but as I continue with the rest of the stain the fiber stain is lost, I have remedied this by repeating the hematoxylin again at the end of the staining process to get back the fiber staining. Try it. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Deb Stults Sent: Friday, March 19, 2004 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help with elastic staining We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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. ------------------------------ Message: 14 Date: Tue, 23 Mar 2004 09:48:31 -0500 From: komo@bu.edu Subject: [Histonet] Problems with c-fos immediate early gene labelling To: histonet@lists.utsouthwestern.edu Message-ID: <1080053311.40604e3f585a5@www.bu.edu> Content-Type: text/plain I'm hoping I could get some helpful advice on what might cause my c-fos IHC labelling to work well in one lab, but not my own. I am running the protocol on rat brains using a hydrogen peroxide/methanol step for endogenous peroxidase blocking, a normal goat serum blocking step, a c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated secondary, a streptavidin horeradish peroxidase, and a nickle enhanced DAB reaction. Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. Does anyone has any suggestions as to why this might be? One person at the other lab suggested that possibly the wells that we run the tissue in were not being cleaned properly (we soak them in bleach while the other lab uses just pex). Could this have any effect? Thanks for any advice you can provide!! Rob Komorowski ------------------------------ Message: 15 Date: Tue, 23 Mar 2004 09:50:21 -0500 From: "Featherstone, Annette" Subject: RE: [Histonet] CD98 and PLGF To: 'Patsy Ruegg' , "Histonet@Pathology. Swmed. Edu" Cc: "Ihcrg@Yahoogroups. Com" Message-ID: Content-Type: text/plain; charset="iso-8859-1" YES!!I am! Or was... I used a pepsin pretreatment @37* for 15 minutes, Block with a protein block, primary antibody 1:100 for 1hr, Goat secondary 30 min rt, Alk phos 30 minutes rt, and whatever chromgen you use for alk phos, rt 40 min. My Plgf is from Santa Cruz. Annette Featherstone HT/MLT -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 13:58 To: Patsy Ruegg; Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: RE: [Histonet] CD98 and PLGF I guess no one is using cd98 and/or Placental Growth Factor on ffpe tissue???? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, March 17, 2004 10:03 AM To: Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: [Histonet] CD98 and PLGF I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160 and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies work in ffpe tissue? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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. ------------------------------ Message: 16 Date: Tue, 23 Mar 2004 10:24:38 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] Problems with c-fos immediate early gene labelling To: komo@bu.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <406080E6.2080605@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Ron: komo@bu.edu wrote: >I'm hoping I could get some helpful advice on what might cause my c-fos >IHC labelling to work well in one lab, but not my own. I am running the >protocol on rat brains using a hydrogen peroxide/methanol step for >endogenous peroxidase blocking, a normal goat serum blocking step, a >c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated >secondary, a streptavidin horeradish peroxidase, and a nickle enhanced >DAB reaction. > >Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. > When you say you are using the "exact same antibodies, chemicals, and ..... " do you mean the same bottle of reagent? You are carrying the bottle or vial of reagent from one lab to another? If not, look at the batch of antibody, your peroxide, your DAB, etc. If you are storing antibody in a freezer at home it is being subjected to freeze-thaw cycles (unless you have a very old freezer that does not defrost itself on a timer). If your bottle of peroxide is more than a few months old it may be in the process of turning into water, peroxide does that. I have had DAB work one day and fail the next. Since you have found that the brain slices stain well in the "other lab", the problem must lie with your home lab. Since bleach reacts with DAB I would avoid using staining dishes treated with bleach. Geoff >Does anyone has any suggestions as to why this might be? One person >at the other lab suggested that possibly the wells that we run the tissue in >were not being cleaned properly (we soak them in bleach while the other >lab uses just pex). Could this have any effect? > >Thanks for any advice you can provide!! > >Rob Komorowski > > > >_______________________________________________ >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 ********************************************** ------------------------------ Message: 17 Date: Tue, 23 Mar 2004 10:46:32 -0500 From: "McAloose, Dee" Subject: [Histonet] FW: Job opening - Wildlife Conservation Society To: Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD6D3@WOLF.wcs.org> Content-Type: text/plain; charset="iso-8859-1" > Hello, > > I'd like to announce an immediate opening for a full-time histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity and in becoming part of our team! > > Thanks for listening and we look forward to hearing from you. > > D McAloose, VMD, Dipl ACVP > Head, Department of Pathology > Wildlife Conservation Society > Bronx, NY 10464 > (phone) 718-220-7105 > (fax) 718-220-7126 > dmcaloose@wcs.org > > ------------------------------ Message: 18 Date: Tue, 23 Mar 2004 10:21:54 -0600 From: "Jan Shivers" Subject: [Histonet] IHC on blood and cytology smears (ICC) To: "histonet" Message-ID: <006301c410f2$f45d1ed0$78065486@vdl220FAC> Content-Type: text/plain; charset="iso-8859-1" Could someone please send me a protocol for performing IHC/ICC on smears (I normally only do FFPE slides)? I'd like info mostly on if the slides have to be coated/charged , smears air-dried or fixed (and in what) for how long, and if you do any modification of the immuno protocol, such as blocking endogenous peroxidase later in the procedure (and with what concentration of H2O2), and/or changing incubation times/dilutions. I've done smears on very rare occasions, but have had trouble getting the cells to remain on the uncoated slides that the smears are being made on. Thank you very much in advance. Jan Shivers Univ of Minnesota Vet Diag Lab shive003@tc.umn.edu ------------------------------ Message: 19 Date: Tue, 23 Mar 2004 12:02:46 -0500 From: Luis Chiriboga Subject: [Histonet] NYSHS 2004 Annual Meeting To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" The 2004 New York State Histotechnology Society annual meeting will be held at the Holiday Inn in beautiful Saratoga Springs, New York from Friday April 30th through Saturday May 1st. This years meeting theme is "Histology is no mystery: Let New York State solve your problems". An e-mail mini program is available upon request by replying to this message. For a program and registration packet, please contact Judy LaDuc at 518-897-2247 or jaladuc@capital.net We hope to see you there. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 4, Issue 27 *************************************** From cwscouten <@t> myneurolab.com Tue Mar 23 14:59:40 2004 From: cwscouten <@t> myneurolab.com (Charles Scouten) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Fetus fixation Message-ID: Just as the brain is protected by the blood brain barrier, the fetal and mother blood are kept separate. Perfusion of the mother might not reach the fetus. The blood brain barrier can be broken by high pressure (300 mm Hg.)to wash out the extracellular fluid in brain and avoid shrinkage when the formaldehyde gets there. I would be very curious to know if this works for fetal tissue as well. If the tissue is soft 12 hours post fixation, it has also deteriorated considerably. Cutting whole head might work, but is not the solution you need. You must get fixative to perfuse the fetus. If high pressure does not work, I wonder if there is a blood vessel you can get to to perfuse the fetus instead of the mother. The Perfusion One apparatus at the following site could provide the pressure perfusion and see if that does it. http://www.myneurolab.com/myneurolab/mnl_products_detail.asp?idproduct=471001&catdesc=Histology+Equipment&CatThreeID=674&CatOneID=4&subcatdesc=Sacrifice+Equipment&idsubcategory=21 Call me to arrange a test. Cordially, Charles W.? Scouten, Ph.D. myNeuroLab.com 5918 Evergreen Blvd. St. Louis, MO 63134 Ph: 314 522 0300? FAX? 314 522 0377 cwscouten@myneurolab.com http://www.myneurolab.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pablo S?nchez Quinteiro Sent: Tuesday, March 23, 2004 1:01 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Fetus fixation Dear listers, I am trying to cut brains from fetal mice (E19) fixed in Paraformaldehyde 4% with the vibratome. I have problems with the perfusion of the mother. The fixative does not reach the fetuses and these even after 12 hours postfixation in the same fixative are too soft to take the brains out. Could you give me some input? Have you tried to cut in vibratome the whole head of the fetus? Is it possible? Thanks in advance Pablo _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mrsgbd2001 <@t> yahoo.com Tue Mar 23 15:20:51 2004 From: mrsgbd2001 <@t> yahoo.com (Gareth Davis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] cryo preserving and sectioning of bone Message-ID: <20040323212051.81956.qmail@web13306.mail.yahoo.com> Hello again, Does anyone have a good protocol for preparing bone (after decal is done) for cryosectioning. I've done the sucrose preservation and also have just gone straight to OCT and immediate freezing with LN2. Both protocols have worked for me. But, I was just wondering what the consensus was. Thanks, Ms. Gareth Davis Research Assistant Tennessee State University Do you Yahoo!? Yahoo! Finance Tax Center - File online. File on time. From spoulos <@t> saa.ars.usda.gov Tue Mar 23 16:46:02 2004 From: spoulos <@t> saa.ars.usda.gov (Sylvia Poulos) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? Message-ID: Hi all, I've got some formalin fixed paraffin embedded mouse tissue. Any idea of something I could do to stain capillaries in these? Someone here mentioned mason's trichrome. Any thoughts are always welcome. Thanks! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) From barbara.wright2 <@t> dnax.org Tue Mar 23 16:59:33 2004 From: barbara.wright2 <@t> dnax.org (Wright, Barbara (SPRI 2)) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? Message-ID: <29B25753F6B1D51196110002A589D4440105C862@PALMSG30.us.schp.com> Try using rat anti-mouse CD31 (MEC13.3) from Pharmingen/BD. -----Original Message----- From: Sylvia Poulos [mailto:spoulos@saa.ars.usda.gov] Sent: Tuesday, March 23, 2004 2:46 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] blood vessel staining? Hi all, I've got some formalin fixed paraffin embedded mouse tissue. Any idea of something I could do to stain capillaries in these? Someone here mentioned mason's trichrome. Any thoughts are always welcome. Thanks! Sylvia Sylvia P. Poulos USDA-ARS-Animal Physiology Research Unit Athens, GA 30605 706-583-8279 706-542-0399 (fax) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete. From rocan <@t> mac.com Tue Mar 23 18:02:07 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? In-Reply-To: References: Message-ID: <7DB55CA2-7D26-11D8-A981-000A9589219E@mac.com> Sylvia, The Pharmigen antibody works great but you may have to do some antigen exposure. We have found that the best staining is on thick frozen sections(20um) and we leave a 1:100 dilution incubating overnight AT ROOM TEMPERATURE! Our secondary is an anti-rat biotinylated (I think this one is from Biomeda). We then use the ABC kit and DAB from vector. Good Luck Rocio On Mar 23, 2004, at 2:46 PM, Sylvia Poulos wrote: > Hi all, > I've got some formalin fixed paraffin embedded mouse tissue. Any idea > of something I could do to stain capillaries in these? Someone here > mentioned mason's trichrome. Any thoughts are always welcome. Thanks! > Sylvia > > Sylvia P. Poulos > USDA-ARS-Animal Physiology Research Unit > Athens, GA 30605 > 706-583-8279 > 706-542-0399 (fax) > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rocan <@t> mac.com Tue Mar 23 22:46:30 2004 From: rocan <@t> mac.com (rocan@mac.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? In-Reply-To: <00519F9F41D2844D9C75BEE1F6AC09AFC89E71@rmhmail1.ssg.org.au> References: <00519F9F41D2844D9C75BEE1F6AC09AFC89E71@rmhmail1.ssg.org.au> Message-ID: <37AE85E2-7D4E-11D8-A981-000A9589219E@mac.com> Stan, I avoid antigen retrieval like the plague. Therefore I would recommend to not use the formalin in the first place if at all possible. I believe that for formalin fixed samples my tech used citrate but I am not sure (long time ago). Also, in general we are having very good luck with the L.A.B. antigen retrieval solution that was mentioned here a few months ago. We normally use use Streck's fixative when we plan to use the pharmigen anti-cd31 antibody on certain samples. We also have used the antibody without antigen retrieval when we fix by intracardiac perfusion with 4% paraformaldehyde. Doing the intracardiac perfusion with 4%PFA actually fixes the vessels before they collapse. I think the key for this antibody is thick sections and long incubation at room temperature. The thicker the sections the more you can actually visualize all intact vessel including capillaries. The branching is clearly distinct, beautiful! Rocio On Mar 23, 2004, at 4:20 PM, Stylli, Stanley wrote: > Rocio > WHat sort of antigen retrieval have you used ? > Has it been successful ? > thanks > Stan > > -----Original Message----- > From: rocan@mac.com [mailto:rocan@mac.com] > Sent: Wednesday, 24 March 2004 11:02 AM > To: 'histonet@pathology.swmed.edu' > Cc: Sylvia Poulos > Subject: Re: [Histonet] blood vessel staining? > > > Sylvia, > The Pharmigen antibody works great but you may have to do some antigen > exposure. We have found that the best staining is on thick frozen > sections(20um) and we leave a 1:100 dilution incubating overnight AT > ROOM TEMPERATURE! > Our secondary is an anti-rat biotinylated (I think this one is from > Biomeda). We then use the ABC kit and DAB from vector. > Good Luck > Rocio > > On Mar 23, 2004, at 2:46 PM, Sylvia Poulos wrote: > >> Hi all, >> I've got some formalin fixed paraffin embedded mouse tissue. Any idea >> of something I could do to stain capillaries in these? Someone here >> mentioned mason's trichrome. Any thoughts are always welcome. Thanks! >> Sylvia >> >> Sylvia P. Poulos >> USDA-ARS-Animal Physiology Research Unit >> Athens, GA 30605 >> 706-583-8279 >> 706-542-0399 (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 Juan.Solon <@t> lshtm.ac.uk Wed Mar 24 03:36:30 2004 From: Juan.Solon <@t> lshtm.ac.uk (Juan Solon) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] CD4/CD25/TGFB Antibodies; Filter choices for Leica Message-ID: Dear All, As part of a study on mucosal immunity in The Gambia, W. Africa, I plan to double stain antral and duodenal specimens for CD4 and CD25 & CD8 and CD25. Also, we would attempt to triple stain for TGF beta on these subsets. We have access to a Leica DMRXE although the filter cubes are all currently long pass suppression filters . I have both formalin-fixed and frozen sections of the antrum and the duodenum. I would like to ask for advise on several points: 1. Can anyone share their experience on antibody combinations known to work for CD4-25, CD8-25 and triple staining for CD4-CD25-TGFB either with vectastain elite (peroxidase) or immunofluorescence. I have compiled a list of CD4, CD8, CD25, TGFB antibodies from Dako, Ancell, Novocastra, BD, and Serotec, but before purchasing any of these, I thought it would be wise to ask advise from the Histonet community. 2. I would probably need to acquire different filter cubes to do simultaneous imaging of fluorescence. There are several suppliers I have encountered on the net (Omega Filters and Chroma Technology) * does anyone have experience on using filters from these companies on Leica microscopes (specifically the DMRXE). Any problems encountered with using third-party filters with Leica microscopes? Thanks. Juan Dr. Juan Antonio Solon MRC Keneba Field Station MRC Laboratories Gambia Atlantic Road, Fajara PO Box 273, Banjul The Gambia Email : juan.solon@lshtm.ac.uk Tel No: (++220) 541021 Fax No: (++220) 541022 / (++220) 496513 From psanquin <@t> lugo.usc.es Wed Mar 24 04:18:22 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? In-Reply-To: Message-ID: <3.0.6.32.20040324111822.007bad10@pop.lugo.usc.es> Hi Sylvia, Lectins as WGA or LCA have been used successfully by intravascular delivery in chicken embryos but this is not possible in your tissue. A marker I have read about is PECAM but I have not experience with it. Regards Pablo At 05:46 p.m. 23/03/04 -0500, you wrote: >Hi all, >I've got some formalin fixed paraffin embedded mouse tissue. Any idea >of something I could do to stain capillaries in these? Someone here >mentioned mason's trichrome. Any thoughts are always welcome. Thanks! >Sylvia From Myri37 <@t> aol.com Wed Mar 24 05:31:35 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] MMA Message-ID: <653D3513.5D808838.0005167B@aol.com> hello histonetters i embedded some samples (thin tissue on titane in MMA resin (MMA, butylmethacrylate, methylbenzoate, PEG400, and N,N toluidine) i already done this, and i could deplastizise sections for immunostainnig, but for these last samples i dont understand why it doesnt totaly deplastizise, the protocol was the same, and i embedded them myself does anyone have any idea on what could avoid deplastization of MMA ? thank you very much in advance myriam baali natural implant From Myri37 <@t> aol.com Wed Mar 24 06:53:47 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Bone embedding Message-ID: <4B40DB0D.62A4CFAA.0005167B@aol.com> hello everyone I would like to embedd bone in both paraffin and MMA, but for the first i must decalcifiy before embedding, so i nedd to parts of this bone The bone is fixed in paraformald?hyde 4% at 4?C since a week, do you know a method to cut it avoiding damaging tissue Thank you very much in advance Myriam natural implant From Jackie.O'Connor <@t> abbott.com Wed Mar 24 07:42:41 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? Message-ID: I routinely perform IHC for microvessle density using Pharmingen's PECAM antibody using Streck fixed tissues in paraffin, 5 or 6 micron sections. Pharmingen includes an SABC protocol with their antibody that works extremely well - I use the antibody at a 1:30 dilution overnight at 4 degrees C with no antigen retrieval. It's the best I've found so far. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotheraputics "rocan@mac.com" cc: "Stylli, Stanley" Subject: Re: [Histonet] blood vessel staining? Stan, I avoid antigen retrieval like the plague. Therefore I would recommend to not use the formalin in the first place if at all possible. I believe that for formalin fixed samples my tech used citrate but I am not sure (long time ago). Also, in general we are having very good luck with the L.A.B. antigen retrieval solution that was mentioned here a few months ago. We normally use use Streck's fixative when we plan to use the pharmigen anti-cd31 antibody on certain samples. We also have used the antibody without antigen retrieval when we fix by intracardiac perfusion with 4% paraformaldehyde. Doing the intracardiac perfusion with 4%PFA actually fixes the vessels before they collapse. I think the key for this antibody is thick sections and long incubation at room temperature. The thicker the sections the more you can actually visualize all intact vessel including capillaries. The branching is clearly distinct, beautiful! Rocio On Mar 23, 2004, at 4:20 PM, Stylli, Stanley wrote: > Rocio > WHat sort of antigen retrieval have you used ? > Has it been successful ? > thanks > Stan > > -----Original Message----- > From: rocan@mac.com [mailto:rocan@mac.com] > Sent: Wednesday, 24 March 2004 11:02 AM > To: 'histonet@pathology.swmed.edu' > Cc: Sylvia Poulos > Subject: Re: [Histonet] blood vessel staining? > > > Sylvia, > The Pharmigen antibody works great but you may have to do some antigen > exposure. We have found that the best staining is on thick frozen > sections(20um) and we leave a 1:100 dilution incubating overnight AT > ROOM TEMPERATURE! > Our secondary is an anti-rat biotinylated (I think this one is from > Biomeda). We then use the ABC kit and DAB from vector. > Good Luck > Rocio > > On Mar 23, 2004, at 2:46 PM, Sylvia Poulos wrote: > >> Hi all, >> I've got some formalin fixed paraffin embedded mouse tissue. Any idea >> of something I could do to stain capillaries in these? Someone here >> mentioned mason's trichrome. Any thoughts are always welcome. Thanks! >> Sylvia >> >> Sylvia P. Poulos >> USDA-ARS-Animal Physiology Research Unit >> Athens, GA 30605 >> 706-583-8279 >> 706-542-0399 (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 frouwke <@t> sci.kun.nl Wed Mar 24 07:51:26 2004 From: frouwke <@t> sci.kun.nl (Frouwke Kuijpers) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Alcian Yellow Message-ID: <003201c411a7$1ba31620$4a8aae83@sci.kun.nl> Can somebody tell me how to make an Alcian Yellow solution PH 2,5? And do you have to filter it before use? I already know that 0,2% is the max. but still I get it not dissolved. And on the histonet archive I read that Alcian Yellow is not made anymore. We still have a lot of it, does it has an expiring date? Thanks Frouwke F.J.Kuijpers-Kwant Dept. Cellular Animal Physiology University of Nijmegen Toernooiveld 1 6525 ED Nijmegen frouwke@sci.kun.nl From Tbarnhart <@t> primecare.org Wed Mar 24 07:57:44 2004 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] EDTA pH 8.0 recipe Message-ID: <1779904B5E82D511914C00D0B793339205BFD7F3@exchangent> Does anyone have a recipe to make EDTA buffer at a pH of 8.0. I would like to make our own in an effort to save some dollars. I have checked the archives and only have come up with one rather difficult procedure. Thanks in advance.... Tammy Barnhart, BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND tbarnhart@primecare.org Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From dobbin <@t> upei.ca Wed Mar 24 08:01:26 2004 From: dobbin <@t> upei.ca (Greg Dobbin) Date: Fri Sep 16 15:22:43 2005 Subject: (Fwd) Re: [Histonet] EDTA pH 8.0 recipe Message-ID: <40616A86.20208.6FA22C@localhost> Hi Tammy, I maintain my own "personal" Histonet Archives, and I found the following post from 1999. Good luck. Greg Date sent: Mon, 27 Dec 1999 11:15:29 -0600 From: "Lott, Robert" Subject: EDTA-NaOH, pH 8.0 retrieval fluid Forwarded to: DOBBIN@acad1.cs.upei.ca To: "'histonet@pathology.swmed.edu'" Stock Solution: 1.86 gm. EDTA (same as used for decalcification) 5.0 liters DI water To the above solution, add 2 to 2.5 ml. 2N NaOH. Adjust solution to pH 8.0 with 1N NaOH or 1N HCl solution. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ From: "Barnhart, Tammy" To: "Histonet (E-mail)" Date sent: Wed, 24 Mar 2004 07:57:44 -0600 Subject: [Histonet] EDTA pH 8.0 recipe > Does anyone have a recipe to make EDTA buffer at a pH of 8.0. I would like > to make our own in an effort to save some dollars. I have checked the > archives and only have come up with one rather difficult procedure. Thanks > in advance.... > > Tammy Barnhart, BS, HTL(ASCP) > Anatomic Pathology Supervisor > St. Alexius Medical Center > Bismarck, ND > tbarnhart@primecare.org > > > > Confidentiality Notice:This e-mail message is for sole use of intended > recipient(s) and may contain confidential and privileged information. Any > unauthorized review, use, disclosure, distribution, or copying is > prohibited. If you are not the intended recipient, please contact the sender > by replying to this e-mail and destroy/delete all copies of this e-mail > message. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Wed Mar 24 09:19:21 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:43 2005 Subject: (Fwd) Re: [Histonet] EDTA pH 8.0 recipe Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FD38@UTHEVS3.mail.uthouston.edu> Tammy and Greg EDTA powder is only slightly soluble in water. I prefer to start with disodium EDTA (that is much more soluble)and then adjust the pH to 8 with NaOH. Can also use this in saline. The 1.86 gms in 5 liters is a very weak solution for demineralization. Normally we use 5% EDTA adjusted to pH 7.3. If cost is a the determining factor then EDTA can be recovered from a used demineralization solution - however very few do this because of the time involved. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Greg Dobbin Sent: Wednesday, March 24, 2004 5:01 AM To: histonet@lists.utsouthwestern.edu Subject: (Fwd) Re: [Histonet] EDTA pH 8.0 recipe Hi Tammy, I maintain my own "personal" Histonet Archives, and I found the following post from 1999. Good luck. Greg Date sent: Mon, 27 Dec 1999 11:15:29 -0600 From: "Lott, Robert" Subject: EDTA-NaOH, pH 8.0 retrieval fluid Forwarded to: DOBBIN@acad1.cs.upei.ca To: "'histonet@pathology.swmed.edu'" Stock Solution: 1.86 gm. EDTA (same as used for decalcification) 5.0 liters DI water To the above solution, add 2 to 2.5 ml. 2N NaOH. Adjust solution to pH 8.0 with 1N NaOH or 1N HCl solution. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ From: "Barnhart, Tammy" To: "Histonet (E-mail)" Date sent: Wed, 24 Mar 2004 07:57:44 -0600 Subject: [Histonet] EDTA pH 8.0 recipe > Does anyone have a recipe to make EDTA buffer at a pH of 8.0. I would > like to make our own in an effort to save some dollars. I have > checked the archives and only have come up with one rather difficult > procedure. Thanks in advance.... > > Tammy Barnhart, BS, HTL(ASCP) > Anatomic Pathology Supervisor > St. Alexius Medical Center > Bismarck, ND > tbarnhart@primecare.org > > > > Confidentiality Notice:This e-mail message is for sole use of intended > recipient(s) and may contain confidential and privileged information. > Any unauthorized review, use, disclosure, distribution, or copying is > prohibited. If you are not the intended recipient, please contact the > sender by replying to this e-mail and destroy/delete all copies of > this e-mail message. _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From medjnw <@t> emory.edu Wed Mar 24 09:29:42 2004 From: medjnw <@t> emory.edu (Josiah N. Wilcox, Ph.D.) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] blood vessel staining? In-Reply-To: <3.0.6.32.20040324111822.007bad10@pop.lugo.usc.es> Message-ID: <12488976-7DA8-11D8-9C2C-000393DC4B88@emory.edu> Just do an alpha actin stain, all blood vessels have a SMC coat even small vasa vessels in the adventitia Josiah N. Wilcox, Ph.D. Professor of Hematology/Oncology and Internal Medicine The Winship Cancer Institute Dept. of Hematology/Oncology 1635-B Clifton Rd., Suite B5100 Atlanta, GA 30322 WebPage: www.emory.edu/WILCOX/ On Wednesday, March 24, 2004, at 02:18 AM, Pablo S?nchez Quinteiro wrote: > Hi Sylvia, > > Lectins as WGA or LCA have been used successfully by intravascular > delivery > in chicken embryos but this is not possible in your tissue. A marker I > have > read about is PECAM but I have not experience with it. > > Regards > > Pablo > > > > At 05:46 p.m. 23/03/04 -0500, you wrote: >> Hi all, >> I've got some formalin fixed paraffin embedded mouse tissue. Any idea >> of something I could do to stain capillaries in these? Someone here >> mentioned mason's trichrome. Any thoughts are always welcome. Thanks! >> Sylvia > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ramona.tolliver <@t> yale.edu Wed Mar 24 10:39:19 2004 From: ramona.tolliver <@t> yale.edu (Ramona Tolliver) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Yale University - Opening Message-ID: <6.0.1.1.2.20040324113917.01e29100@email.med.yale.edu> Good afternoon, Yale University has a Histology Manager (1st shift) position available from 7:00 a.m. - 3:00 p.m. If you know of anyone who may be interested in an outstanding growth opportunity in a department committed to excellence in patient care, teaching, and discovery, please forward this information on to them. Salary is commensurate with experience. Relocation assistance is available. Inquiries should be sent to ramona.tolliver@yale.edu. Thank you for your time! Ramona Tolliver Ramona E. Tolliver Human Resource Manager Yale University School of Medicine Department of Pathology 310 Cedar Street New Haven, CT 06520-8023 Telephone: (203) 785-6689 Fax: (203) 785-7303 From gcallis <@t> montana.edu Wed Mar 24 10:41:51 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] CD4/CD25/TGFB Antibodies; Filter choices for Leica In-Reply-To: Message-ID: <3.0.6.32.20040324094151.00bd5248@gemini.msu.montana.edu> What species are you working with? Mouse? Human? At 09:36 AM 3/24/2004 +0000, you wrote: >Dear All, > >As part of a study on mucosal immunity in The Gambia, W. Africa, I plan to double stain antral and duodenal specimens for CD4 and CD25 & CD8 and CD25. Also, we would attempt to triple stain for TGF beta on these subsets. We have access to a Leica DMRXE although the filter cubes are all currently long pass suppression filters . I have both formalin-fixed and frozen sections of the antrum and the duodenum. > >I would like to ask for advise on several points: > >1. Can anyone share their experience on antibody combinations known to work for CD4-25, CD8-25 and triple staining for CD4-CD25-TGFB either with vectastain elite (peroxidase) or immunofluorescence. I have compiled a list of CD4, CD8, CD25, TGFB antibodies from Dako, Ancell, Novocastra, BD, and Serotec, but before purchasing any of these, I thought it would be wise to ask advise from the Histonet community. > >2. I would probably need to acquire different filter cubes to do simultaneous imaging of fluorescence. There are several suppliers I have encountered on the net (Omega Filters and Chroma Technology) * does anyone have experience on using filters from these companies on Leica microscopes (specifically the DMRXE). Any problems encountered with using third-party filters with Leica microscopes? > >Thanks. >Juan > > >Dr. Juan Antonio Solon >MRC Keneba Field Station >MRC Laboratories Gambia >Atlantic Road, Fajara >PO Box 273, Banjul >The Gambia >Email : juan.solon@lshtm.ac.uk >Tel No: (++220) 541021 >Fax No: (++220) 541022 / (++220) 496513 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Mar 24 10:46:18 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] MMA In-Reply-To: <653D3513.5D808838.0005167B@aol.com> Message-ID: <3.0.6.32.20040324094618.00bd5248@gemini.msu.montana.edu> How did you remove plastic? Neil Hand recommends warm (can be up to 60C) xylene X 3 changes, and how thin are your sections? He removed MMA from 3 um sections. One can even use methylmethacrylate monomer to remove polymerized MMA, but this requires some careful handling, and then removal of the monomer with several changes of absolute ethanol. You do not want to breathe monomer fumes at any time nor get it on you. At 06:31 AM 3/24/2004 -0500, you wrote: >hello histonetters >i embedded some samples (thin tissue on titane in MMA resin (MMA, butylmethacrylate, methylbenzoate, PEG400, and N,N toluidine) >i already done this, and i could deplastizise sections for immunostainnig, but for these last samples i dont understand why it doesnt totaly deplastizise, the protocol was the same, and i embedded them myself >does anyone have any idea on what could avoid deplastization of MMA ? >thank you very much in advance >myriam baali >natural implant > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Mar 24 10:53:53 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] EDTA pH 8.0 recipe In-Reply-To: <1779904B5E82D511914C00D0B793339205BFD7F3@exchangent> Message-ID: <3.0.6.32.20040324095353.00bd5248@gemini.msu.montana.edu> Use EDTA Tetrasodium salt, dissolve in 800 mls distilled water and adjust pH DOWN to pH 8 using pH meter and glacial acetic acid, just titrate it with a pipette, watching the meter. This is an old method from Webb Gee from U of Utah. The pH of tetrasodium EDTA is around 9 to begin with. If you are planning to decalcify bone with pH 8, remember this is rather alkaline and protein linkages sensitive to alkaline conditions can be damaged. Maybe work at pH 7.4 - 7.6, and stay out of higher pH range. You can dissolve as much as 14 gm/100 mls buffer or distilled water. IF the bone is totally fixed to begin with, distilled water can be used to make up the EDTA solution. Tetrasodium EDTA goes into solution much faster than EDTA, or EDTA Disodium salt (they are soluble to around 10% aka 10 g/100 mls). Good luck At 07:57 AM 3/24/2004 -0600, you wrote: >Does anyone have a recipe to make EDTA buffer at a pH of 8.0. I would like >to make our own in an effort to save some dollars. I have checked the >archives and only have come up with one rather difficult procedure. Thanks >in advance.... > >Tammy Barnhart, BS, HTL(ASCP) >Anatomic Pathology Supervisor >St. Alexius Medical Center >Bismarck, ND >tbarnhart@primecare.org > > > >Confidentiality Notice:This e-mail message is for sole use of intended >recipient(s) and may contain confidential and privileged information. Any >unauthorized review, use, disclosure, distribution, or copying is >prohibited. If you are not the intended recipient, please contact the sender >by replying to this e-mail and destroy/delete all copies of this e-mail >message. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Mar 24 10:55:52 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: (Fwd) Re: [Histonet] EDTA pH 8.0 recipe In-Reply-To: <40616A86.20208.6FA22C@localhost> Message-ID: <3.0.6.32.20040324095552.00bd5248@gemini.msu.montana.edu> I would like to point that he did NOT specify with EDTA he was using, presumption is EDTA or EDTA disodium. Unfortunately, this is a pet peeve of mine, one should always say WHICH EDTA is being used, or at least give a formula weight. > >Stock Solution: > 1.86 gm. EDTA (same as used for decalcification) > 5.0 liters DI water > >To the above solution, add 2 to 2.5 ml. 2N NaOH. >Adjust solution to pH 8.0 with 1N NaOH or 1N HCl solution. >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ > >From: "Barnhart, Tammy" >To: "Histonet (E-mail)" >Date sent: Wed, 24 Mar 2004 07:57:44 -0600 >Subject: [Histonet] EDTA pH 8.0 recipe > >> Does anyone have a recipe to make EDTA buffer at a pH of 8.0. I would like >> to make our own in an effort to save some dollars. I have checked the >> archives and only have come up with one rather difficult procedure. Thanks >> in advance.... >> >> Tammy Barnhart, BS, HTL(ASCP) >> Anatomic Pathology Supervisor >> St. Alexius Medical Center >> Bismarck, ND >> tbarnhart@primecare.org >> >> >> >> Confidentiality Notice:This e-mail message is for sole use of intended >> recipient(s) and may contain confidential and privileged information. Any >> unauthorized review, use, disclosure, distribution, or copying is >> prohibited. If you are not the intended recipient, please contact the sender >> by replying to this e-mail and destroy/delete all copies of this e-mail >> message. >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From bwhitaker <@t> brownpathology.com Wed Mar 24 11:24:20 2004 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] EDTA pH 8.0 recipe In-Reply-To: <3.0.6.32.20040324095353.00bd5248@gemini.msu.montana.edu> Message-ID: <000201c411c4$d8860320$3601a8c0@brownpathology.net> Did I miss something? I assumed that she wanted it for HIER.... maybe my prejudice is showing, but that's what I use EDTA at pH 8 for. Bonnie Whitaker -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, March 24, 2004 10:54 AM To: Barnhart, Tammy; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] EDTA pH 8.0 recipe Use EDTA Tetrasodium salt, dissolve in 800 mls distilled water and adjust pH DOWN to pH 8 using pH meter and glacial acetic acid, just titrate it with a pipette, watching the meter. This is an old method from Webb Gee from U of Utah. The pH of tetrasodium EDTA is around 9 to begin with. If you are planning to decalcify bone with pH 8, remember this is rather alkaline and protein linkages sensitive to alkaline conditions can be damaged. Maybe work at pH 7.4 - 7.6, and stay out of higher pH range. You can dissolve as much as 14 gm/100 mls buffer or distilled water. IF the bone is totally fixed to begin with, distilled water can be used to make up the EDTA solution. Tetrasodium EDTA goes into solution much faster than EDTA, or EDTA Disodium salt (they are soluble to around 10% aka 10 g/100 mls). Good luck At 07:57 AM 3/24/2004 -0600, you wrote: >Does anyone have a recipe to make EDTA buffer at a pH of 8.0. I would like >to make our own in an effort to save some dollars. I have checked the >archives and only have come up with one rather difficult procedure. Thanks >in advance.... > >Tammy Barnhart, BS, HTL(ASCP) >Anatomic Pathology Supervisor >St. Alexius Medical Center >Bismarck, ND >tbarnhart@primecare.org > > > >Confidentiality Notice:This e-mail message is for sole use of intended >recipient(s) and may contain confidential and privileged information. Any >unauthorized review, use, disclosure, distribution, or copying is >prohibited. If you are not the intended recipient, please contact the sender >by replying to this e-mail and destroy/delete all copies of this e-mail >message. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cmather <@t> origentherapeutics.com Wed Mar 24 11:19:13 2004 From: cmather <@t> origentherapeutics.com (Christine Mather) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Rossman's fluid Message-ID: I would like to try an old method for fixing early chicken embryos that uses Rossman's fluid. It's not commercially available and uses picric acid, rather nasty stuff. Are there any modern alternatives that will do the same job? Thanks, Christine. From Tbarnhart <@t> primecare.org Wed Mar 24 11:34:44 2004 From: Tbarnhart <@t> primecare.org (Barnhart, Tammy) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] EDTA pH 8.0 recipe Message-ID: <1779904B5E82D511914C00D0B793339205BFD7F4@exchangent> Yes, it is for HIER, not decalcification. This list is great.....so many answers, so quickly. Thank you all!!!! Tammy Barnhart,BS, HTL(ASCP) Anatomic Pathology Supervisor St. Alexius Medical Center Bismarck, ND tbarnhart@primecare.org -----Original Message----- From: Bonnie Whitaker [mailto:bwhitaker@brownpathology.com] Sent: Wednesday, March 24, 2004 11:24 AM To: 'Gayle Callis'; 'Barnhart, Tammy'; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] EDTA pH 8.0 recipe Did I miss something? I assumed that she wanted it for HIER.... maybe my prejudice is showing, but that's what I use EDTA at pH 8 for. Bonnie Whitaker Confidentiality Notice:This e-mail message is for sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, distribution, or copying is prohibited. If you are not the intended recipient, please contact the sender by replying to this e-mail and destroy/delete all copies of this e-mail message. From mcauliff <@t> umdnj.edu Wed Mar 24 15:12:20 2004 From: mcauliff <@t> umdnj.edu (Geoff McAuliffe) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Rossman's fluid In-Reply-To: References: Message-ID: <4061F9B4.1000409@umdnj.edu> Hi Christine: Picric acid is only dangerous if allowed to dry out and then subjected to severe shock or high temperatures (well above the boiling point of water). Much of the picric acid paranoia is spread by unsubstantiated rumors. Still, if you don't want it in your lab, try a formalin+alcohol+acetic acid mixture or Bouin's (I think you can buy Bouin's made up or sat. aqueous picric acid, which minimizes the risks). You could also ask your local lab safety police to store a small quantity of picric acid for you. Geoff Christine Mather wrote: >I would like to try an old method for fixing early chicken embryos that uses >Rossman's fluid. It's not commercially available and uses picric acid, >rather nasty stuff. Are there any modern alternatives that will do the same >job? > >Thanks, > >Christine. > > > > >_______________________________________________ >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 Apeters <@t> chsd.org Wed Mar 24 12:31:18 2004 From: Apeters <@t> chsd.org (Peters, Anne) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] job opening Message-ID: <00D7298B3825D511BF540008C75F8A3C8BB57D@excluster.chsd.org> >We currently have an opening for a full-time transmission electron >microscopist with histology experience in a hospital setting. We primarily >work on renal, liver, muscle, ciliary biopsies and a variety of tumors. We >average approximately 200 full EM cases per year. >Responsibilities include EM preparation from specimen processing to darkroom >photography, general histology, specimen accessioning (Meditech). Our lab >also performs immunohistochemistry, immunofluorescence and routine special >stains. >Bachelor's Degree mimimun requirement. > >Children's Hospital >Department of Pathology >3020 Children's Way Mail Code 5007 >San Diego, CA 92123 > >Attn: A. Peters > Anatomic Pathology Technical Specialist From gcallis <@t> montana.edu Wed Mar 24 13:16:35 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] EDTA pH 8.0 recipe In-Reply-To: <000201c411c4$d8860320$3601a8c0@brownpathology.net> References: <3.0.6.32.20040324095353.00bd5248@gemini.msu.montana.edu> Message-ID: <3.0.6.32.20040324121635.00bd5248@gemini.msu.montana.edu> Good point, I just assumed decalcification was her goal but she mentioned neither immunohistochemistry nor bone! No prejudice showing, just heads up!! You wrote: Did I miss something? I assumed that she wanted it for HIER.... maybe my prejudice is showing, but that's what I use EDTA at pH 8 for. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Rcartun <@t> harthosp.org Wed Mar 24 13:11:26 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] DDC IHC Message-ID: Is anyone doing immunohistochemistry for DCC (deleted in colorectal cancer)? If so, where do you get your antibody? Thanks! Richard Cartun From cormier <@t> MIT.EDU Wed Mar 24 13:35:43 2004 From: cormier <@t> MIT.EDU (Kathleen Cormier) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Goodpasture's carbol fuchsin stain Message-ID: <5.2.1.1.2.20040324142851.00ad58e8@hesiod> Greetings! I have a researcher who is requesting a Goodpasture's carbol fuchsin stain on rabbit kidneys to stain protoza. (encephalitozoon cuniculi or the old old name renal nosematosis). My research indicates giemsa and gram would be acceptable as well, but, I can find no reference for the Goodpasture's carbol fuchsin. I have exhausted histonet archives and my histo books here. Can anyone point me in the direction of a procedure for the Goodpasture's? Thanks!!! Kathy DCM MIT From jqb7 <@t> cdc.gov Wed Mar 24 13:59:10 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Goodpasture's carbol fuchsin stain Message-ID: Do you think they meant the MacCallum-Goodpasture stain? Jeanine Bartlett, HT(ASCP) Centers for Disease Control Infectious Disease Pathology Activity 1600 Clifton Road, MS/G-32 Atlanta, GA 30333 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Cormier Sent: Wednesday, March 24, 2004 2:36 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Goodpasture's carbol fuchsin stain Greetings! I have a researcher who is requesting a Goodpasture's carbol fuchsin stain on rabbit kidneys to stain protoza. (encephalitozoon cuniculi or the old old name renal nosematosis). My research indicates giemsa and gram would be acceptable as well, but, I can find no reference for the Goodpasture's carbol fuchsin. I have exhausted histonet archives and my histo books here. Can anyone point me in the direction of a procedure for the Goodpasture's? Thanks!!! Kathy DCM MIT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Dorothy.L.Webb <@t> HealthPartners.Com Wed Mar 24 14:22:48 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] RE: Histonet Digest, Vol 4, Issue 23 Message-ID: You can postfix smears with formalin fumes by using a coplin jar with formalin soaked gauze in the cover for 20 minutes. Or, you can fix the smears in equal parts of hydrogen peroxide and methyl alcohol for 15 minutes. These both help to have the cells adhere to the slide. Hope this helps! Dorothy Webb, Regions Hospital, St. Paul, MN. -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Friday, March 19, 2004 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 4, Issue 23 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. NCSHT Spring Meeting (Delorise Williams) 2. need help with IHC detection of human cells in mouse tissue (Garlits, John) 3. (no subject) (Yang Wang) 4. Extra tissues in LCM capture (Yang Wang) 5. RE: Iron stain on frozen sections (Tony Henwood) 6. RE: Placental section ihc... (Rena Fail) 7. Using H & E slides on the Benchmark XT (Joe Nocito) 8. DAB-Mn histochemistry for superoxide: help to remove background (Subratab) 9. "Animal" person seeking clinical folks help! (Angela McNabola) 10. Re: "Animal" person seeking clinical folks help! (NICK KIRK) ---------------------------------------------------------------------- Message: 1 Date: Thu, 18 Mar 2004 18:46:08 -0500 From: "Delorise Williams" Subject: [Histonet] NCSHT Spring Meeting To: Cc: Delorise Williams Message-ID: <12816CB59E68184F9F5F28063E68B047285DC0@xsrvr.ciit.org> Content-Type: text/plain; charset="iso-8859-1" The North Carolina Society OF Histopathology Technologists presents the 2004 Spring Meeting on April 23 &24 at the Hilton Wilmington Riverside in Wilmington NC. Below is a summary of the program: Friday: April 23, 2004 Using Histological Techniques for Protein and mRNA Expression Analysis in Pharmaceutical Research and 8:15-9:15AM Drug Discovery Stephanie Broka, BS GlaxoSmithKline, Research Triangle Park, NC 9:15 - 10:15 AM Basic Immunology and Immunohistochemistry Godfrey Guerzon , MS,MBA, HT, DLM (ASCP) New Hanover Regional Medical Center,Wilmington, NC 11:00 - 12:00 Organize for Success Beth Moore,MT (ASCP) DLM, MBA New Hanover Regional Medical Center, Wilmington, NC WORKSHOP # 1 (1:30-4:30) HT (ASCP) Examination Readiness-The Practical Exam-Robert Lott, HTL(ASCP) Baptist Health System,Birmingham, Al WORKSHOP # 2 (1:30-4:30) Preparation of Tissue for Evaluation of Sexually Dimorphic Nuclei in Rat Brain-Melanie Struve, BS and Renee Thacker,BS, HT(ASCP) CIIT Centers for Health Research, Research Triangle Park, NC Saturday: April 24,2004 WORKSHOP # 3 (8:15-12:00) HT (ASCP) Examination Readiness-The Written Exam-Robert Lott, HTL (ASCP) Baptist Health Systems, Birmingham, Al WORKSHOP #4 (8:15-12:00) Catalyzed Signal Amplification system-Leslie Sells, BS, HTL, Technical Support Representative, DakoCytomation WORKSHOP # 5 (1:00-4:30) Dyes and Tissue: Chemistry Mechanism of Staining-Bert Dotson,MBA, HTL (ASCP) Duke University Medical Center, Durham, NC WORKSHOP # 6 (1:00-4:30) Microwave: Routine Histological Application-Theory to Practice Jim Milios, International Applications Specialist, Milestone Srl., Italy Delorise Williams CIIT Centers for Health Research PO Box 12137 Research Triangle Park, NC 27709 (919) 558-1200 Voice Mail-(919) 558-1252 Fax-(919) 558-1300 ------------------------------ Message: 2 Date: Thu, 18 Mar 2004 17:51:40 -0600 From: "Garlits, John" Subject: [Histonet] need help with IHC detection of human cells in mouse tissue To: Message-ID: <1E0CC447E59C974CA5C7160D2A2854EC238ABF@SJMEMXMB04.stjude.sjcrh.local> Content-Type: text/plain; charset="iso-8859-1" Hi, I was wondering if anybody could help me out. We want to look for human cells engrafted in mice, particularly in formalin-fixed, acid decalcified, paraffin-embedded tissue. I have so far tried two anti-human beta-2 microglobulin antibodies. The first I tried has been used in human/sheep transplant tissue, and I tried it, but unfortunately it is an IgM kappa, so it failed with the Biogenex mouse-on-mouse IHC kit, which it turns out is not made for IgM antibodies. If anybody is aware of a good way to use a mouse IgM on both human and mouse tissue, I'd love to know. I have looked for a good secondary antibody for this purpose, but have had no luck so far. The second antibody I tried is from Novocastra and distributed by Vector Labs. It is a rabbit polyclonal to human beta-2-microglobulin. The trouble I had was nonspecific binding in control mouse tissue. I was thinking to use some of the Biogenex mouse-on-mouse kit reagents to help block mouse antigens, but I do not know if this would actually work. The second problem with this antibody was that I did not always see positive marking in human tissue (especially osteocytes), which I thought should be pretty well all positive since it is human tissue. Is the expression of beta-2-microglobulin so variable? Has anyone tried any other general human cell marker in other animals? I did a quick search for beta actin, but it seems most of those cross-react with mouse. Thank you! John Garlits, M.S. Senior Research Technician Hematology Oncology Division Experimental Hematology Department St. Jude Children's Research Hospital 332 N Lauderdale Memphis, TN 38108 ------------------------------ Message: 3 Date: Thu, 18 Mar 2004 18:53:28 -0500 From: Yang Wang Subject: [Histonet] (no subject) To: "histonet@lists.utsouthwestern.edu" Message-ID: <1079654008.405a367856f5c@webmail.tufts.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi, dear histonet friends: We need help! We have been working on the conditions for LCM for about 7 months, which is still very frustrating:-(.We tried to capture single cells from intestinal tissues. However, the major problem we are facing now is that we always get extra tissues from the capture. (We could pull off a whole villi when we capture a single cellL) .We used the standard protocols provided by "ARCTURUS" for the tissue section and dehydration. Here is what we have done: 1.We cut 5um frozen sections and do either "pre" or "post" fixation in 4% paraformaldehyde. a. "Pre": fix 2h on ice before frozen, since we are working on GFP expressing tissue, which required pre-fixation. b. " Post": fix 15-30 min after frozen 2.The slides will then be dehydrated by: 75% ethyl 30s, 95% ethy 30s, 100% ethyl 1min and xylene 5min, then air dry 30min. We always pull off extra tissues, which adjacent to the cell interested. We tried treat slides and cap with "ARCTURUS prep strip". But it didn't help. We are wondering whether we need specific treatment with slides (we use fisher superfrost plus)? Or our procedures are not proper? Thanks a lot for help! Yang Wang New England Medical Center Yang.wang@tufts.edu ------------------------------ Message: 4 Date: Thu, 18 Mar 2004 18:56:48 -0500 From: Yang Wang Subject: [Histonet] Extra tissues in LCM capture To: "histonet@lists.utsouthwestern.edu" Cc: Yang.Wang@tufts.edu Message-ID: <1079654208.405a3740c18b2@webmail.tufts.edu> Content-Type: text/plain; charset=ISO-8859-1 Hi, dear histonet friends: We need help! We have been working on the conditions for LCM for about 7 months, which is still very frustrating:-(.We tried to capture single cells from intestinal tissues. However, the major problem we are facing now is that we always get extra tissues from the capture. (We could pull off a whole villi when we capture a single cellL) .We used the standard protocols provided by "ARCTURUS" for the tissue section and dehydration. Here is what we have done: 1.We cut 5um frozen sections and do either "pre" or "post" fixation in 4% paraformaldehyde. a. "Pre": fix 2h on ice before frozen, since we are working on GFP expressing tissue, which required pre-fixation. b. " Post": fix 15-30 min after frozen 2.The slides will then be dehydrated by: 75% ethyl 30s, 95% ethy 30s, 100% ethyl 1min and xylene 5min, then air dry 30min. We always pull off extra tissues, which adjacent to the cell interested. We tried to treat slides and cap with "ARCTURUS prep strip". But it didn't help. We are wondering whether we need specific treatment with slides (we use fisher superfrost plus)? Or our procedures are not proper? Thanks a lot for help! Yang Wang New England Medical Center Yang.wang@tufts.edu ------------------------------ Message: 5 Date: Fri, 19 Mar 2004 11:38:27 +1100 From: Tony Henwood Subject: RE: [Histonet] Iron stain on frozen sections To: "'Connolly, Brett M'" , "'HISTONET' (histonet@lists.utsouthwestern.edu)" Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E167@simba.kids> Content-Type: text/plain; charset="iso-8859-1" Brett, Following is an excerpt from a paper I published a year or so ago: Henwood, A.F., (2002) "Microwave Perl's Stain for urgent frozen sections" Aust J Med Sc 23(2):68-69). Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html MICROWAVE PERLS' STAIN FOR URGENT FROZEN SECTIONS Anthony F. Henwood Laboratory Manager, Histopathology, The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA Abstract A rapid Perls' Stain for hemosiderin using a microwave oven is described. It takes one minute to perform and is particularly suitable for urgent frozen section diagnosis. Introduction Urgent frozen section diagnosis is usually restricted to the interpretation of haematoxylin and eosin stained sections (1). With the increased utilisation of microwave modifications of routine special stains, it is possible to perform a range of such stains on frozen sections (1). To be applicable to urgent frozen section diagnosis, a special stain needs to be rapid, preferably taking 1-2 minutes to perform, easy to prepare, using stock solutions with few preparation steps and produce results equal to those obtained on paraffin sections. The following microwave Perls' technique for hemosiderin takes about one minute to perform. Method Frozen sections can be fixed in Shoobridge's fixative (10ml concentrated formalin in 30ml absolute alcohol) (2) or 95% alcohol. Kennedy and Forbes (1) have also successfully used Wolman's fixative (5% acetic acid in absolute ethanol). 1. Rinse fixed frozen sections in water and then place in a coplin jar containing 20m1 each of 2% Potassium Ferrocyanide and 2% Hydrochloric acid. Place in the microwave oven and microwave for 20 seconds at 650 watts. Slides can be left in this solution for up to 3 minutes. Remove slides if the solution begins to turn cloudy. 2. Rinse slides in water. 3. Counterstain using agitation, with eosin (as routinely used for the frozen section HE) for 5-7 seconds. Other counterstains such as Nuclear Fast Red (3) or 0.25% basic fuchsin (4) can also be used, though eosin is readily available at the frozen section site and works quite well. Since the frozen block is usually fixed and processed to paraffin, sections of this block should also be stained with the routine Perls' as a quality control procedure. Discussion Iron in the body is stored in the forms of hemosiderin (ferric hydroxide polymer) or ferritin (a ferrous iron-protein complex) (5). Iron in tissues occurs mainly in the ferric state (6,7). Microscopically, hemosiderin appears similar to other yellow to brown pigments, such as melanin and fine carbon dust. Macrophages can contain any of these pigments. Heavily pigmented macrophages, often masking the cell nucleus, can be confused with malignant melanoma. The identification of the pigment as being hemosiderin, especially at the time of frozen section, is diagnostically useful. The rapid Perls' stain described above uses the same solution as used by Schaffner (4). This solution is easier to prepare than others reported in the literature (3). The stain takes approximately one minute to perform and gives results equivalent to those obtained in paraffin sections. It is especially applicable to frozen section diagnosis. References: 1. Kennedy, A., Foulis, A.K. (1989) "Use of microwave oven improves morphological and staining of cryostat sections", J.Clin.Pathol. 42:101-105. 2. Shoobridge, M.P.K., (1978) "Improving frozen sections by wet fixation", (Abstract) Pathology 10:195. 3. Brian, N.T., (1983) "Rapid Metallic Histological staining using the microwave oven", J.Histotechnol. 6(3): 125-129. 4. Schaffner, R., (1986) "The Perls' Iron staining procedure for use in the microwave oven using a temperature probe", J.Histotechnol. 9(2): 107-108. 5. Barka, T., Anderson, P.J., (1963) "Histochemistry: Theory, practice and bibliography" Harper & Row Publishers Inc, New York, p172-174. 6. Davenport, H.A., (1961) "Histological and Histochemical Technics" W.B. Saunders Co., Philadelphia, 280-284. 7. Gabe, M., (1976) "Histological Techniques" Masson, Paris, p311-317. -----Original Message----- From: Connolly, Brett M [mailto:brett_connolly@merck.com] Sent: Friday, 19 March 2004 6:03 AM To: 'HISTONET' (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Iron stain on frozen sections Along with the bone marrow thread..... Has anyone had experience with iron stains on frozen sections? What fixative? Procedure? Thx, Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP26A-3000 PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-652-2075 e-mail. brett_connolly@merck.com ---------------------------------------------------------------------------- -- Notice: This e-mail message, together with any attachments, contains information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station, New Jersey, USA 08889), and/or its affiliates (which may be known outside the United States as Merck Frosst, Merck Sharp & Dohme or MSD and in Japan, as Banyu) that may be confidential, proprietary copyrighted and/or legally privileged. It is intended solely for the use of the individual or entity named on this message. If you are not the intended recipient, and have received this message in error, please notify us immediately by reply e-mail and then delete it from your system. ---------------------------------------------------------------------------- -- _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 6 Date: Thu, 18 Mar 2004 22:09:52 -0500 From: Rena Fail Subject: RE: [Histonet] Placental section ihc... To: nyilmaz@mersin.edu.tr, histonet@lists.utsouthwestern.edu Message-ID: <010501c40d5f$cde15f50$dc10a6a5@rena> Content-Type: text/plain; charset=us-ascii We are on occasion asked to stain placenta tissue for IHC. We use non-fat milk as a protein block. No problems with background Rena Fail -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of nyilmaz@mersin.edu.tr Sent: Thursday, March 18, 2004 3:29 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Placental section ihc... We are using placental tissue for immunohistochemistry, and have some problems about background staining especially with serum remnants in the vessels. Does anybody have ideas about this problem. If you inform us we'd be greatly appreciate... Nejat Yilmaz PhD Mersin University Medical School _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Fri, 19 Mar 2004 06:51:05 -0600 From: "Joe Nocito" Subject: [Histonet] Using H & E slides on the Benchmark XT To: "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Morning Histoland, We have acquired the Ventana Benchmark XT and I really need some help. When we were performing the slides manually, we were able to take an H&E slide, take off the coverslip and run an immuno on it. The doctors fell in love with this since we processes a lot of cervical biopsies and sometimes the lesion is not there on recuts. We have had a problem with H&E slides not picking up the immuno staining. I set up different programs using the wet slide protocol, but I'm having problems still. Would anyone be willing to share their protocol with me? Thanks Joe Nocito, BS, HT (ASCP) QIHC Histology Manager Pathology Reference Lab San Antonio, TX 78205 ***************************************Notice******************************* ************* This e-mail, including attachments, contains information that is confidential and may be legally privileged. This e-mail, including atachments, constitutes non-public information inteneded to be conveyed only to the designated recipient(s). If you are not an intended recipient,please delete this e-mail, including attachments, and notify me. The unauthorized use, dissemination, distribution or reproduction of this e-mail, including attachments, is prohibited and may be unlawful. **************************************************************************** ************* ------------------------------ Message: 8 Date: Fri, 19 Mar 2004 20:20:32 +0600 From: Subratab Subject: [Histonet] DAB-Mn histochemistry for superoxide: help to remove background To: Message-ID: <200403191425.i2JEPrGC023459@mailout.proshikanet.com> Content-Type: text/plain; charset="iso-8859-1"; Dear histo-experts I am trying to stain superoxide producing cells on frozen tissue sections of rat kidney. I am using Brigg's (and Karnovsky) cytochemical method in unfixed tissue. The reaction medium contains 1 mg/ml DAB, 1 mM sodium azide and 0.5 mM MnCl2 in 100mM Tris-HCl (pH 7.2). Incubation time 30 minutes at 37 degree C. (The principle of the reaction is that intracellular superoxide will oxidize Mn++ to Mn+++; then Mn+++ will in turn cause DAB oxidation and precipitaion). MY PROBLEM: I am getting profuse DAB staining throughout the tubulo-interstitial area (glomerulus is clean). Most probably its from endogenous peroxidase activity. So I tried to block endo perox by H2O2 (and also by sodium azide, and H2O2+azide) before incubation with reaction medium. This way I have removed tubulo-interstitial DAB staining. But I am not getting any positive stain. I suspect that my blocking step is somehow damaging (blocking) superoxide producing enzymes as well. At this stage I need some expert opinion. How can I specifically block endogenous peroxidase keeping other enzymes intact? Hope to get some effective ways from histo-experts. Thanks in advance Subrata Biswas MD Lab de Fisiopatologia Renal Uni of Campinas, SP, Brazil. ------------------------------ Message: 9 Date: Fri, 19 Mar 2004 09:31:13 -0500 From: Angela McNabola Subject: [Histonet] "Animal" person seeking clinical folks help! To: histonet@lists.utsouthwestern.edu Cc: Maura Broggi Message-ID: Content-Type: text/plain; charset=US-ASCII Hi all, I soliciting protocols that any of you may be willing to share on how to make slides suitable for IHC using FNA's. I am looking for how to best make slide smears (I guess!). Clinical sites will be collecting samples from patients and sending the slides to my lab for staining/evaluation. We are fairly new to this, even new to processing human samples, so any help you can provide would be greatly appreciated. Also, keep in mind that we potentially may be receiving samples from all over the world, so the easier the better since we will be asking seeral sites to do it all the same way thanks in advance! Angela McNabola, MS, HT(ACSP)QIHC, SLS, RLATG Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 angela.mcnabola.b@bayer.com ------------------------------ Message: 10 Date: Fri, 19 Mar 2004 15:25:09 +0000 (GMT) From: NICK KIRK Subject: Re: [Histonet] "Animal" person seeking clinical folks help! To: Angela McNabola , Histonet Message-ID: <20040319152509.87494.qmail@web86308.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Angela Here's a useful recipe/technique for cytological material. The immuno results are very good, plus you have the bonus of being able to keep any residual material for a longer period. RECIPE for PEG/IMS fixative 3% Polyethylene glycol in 50% Industrial methylated spirit (IMS) {=99% ethanol}. --------------------------------- Method 1. Spin cells down 2. Perform blood lysis (if required) with Ammonium chloride solution 3. Re-suspend in PEG/IMS fixative 4. Leave to fix for 2 hours 5. Make cytospins 6. Air dry completely 7. Place in 95% I.M.S. for at least 10 minutes to remove PEG 8. Immunostain Any left over material can be stored in the PEG/IMS fixative for prolonged periods of time. Hope this is of some help Nick Kirk Histopathology Hinchingbrooke Hospital Huntingdon England Angela McNabola wrote: Hi all, I soliciting protocols that any of you may be willing to share on how to make slides suitable for IHC using FNA's. I am looking for how to best make slide smears (I guess!). Clinical sites will be collecting samples from patients and sending the slides to my lab for staining/evaluation. We are fairly new to this, even new to processing human samples, so any help you can provide would be greatly appreciated. Also, keep in mind that we potentially may be receiving samples from all over the world, so the easier the better since we will be asking seeral sites to do it all the same way thanks in advance! Angela McNabola, MS, HT(ACSP)QIHC, SLS, RLATG Bayer Healthcare 400 Morgan Lane West Haven, CT 06516 angela.mcnabola.b@bayer.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 4, Issue 23 *************************************** ________________________________________ 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 bryand <@t> netbistro.com Wed Mar 24 14:39:50 2004 From: bryand <@t> netbistro.com (Bryan Llewellyn) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Goodpasture's carbol fuchsin stain References: <5.2.1.1.2.20040324142851.00ad58e8@hesiod> Message-ID: <002901c411e0$279ec6c0$99a5b5d0@yourlk4rlmsu41> It is in Lillie's book as the Goodpasture-Perrin method for influenza organisms, encephalitozoa and toxoplasmata (Arch. Path. 36:568, 1943) Carbol aniline fuchsin: Basic fuchsin 0.59 g 30% alcohol 100 mL Analine 1 mL Phenol 1 g Fix in Zenker or Orth, or formalin fix and post chrome with 2.5% potassium dichromate for 2 days. Process in paraffin and section as usual. 1. Bring sections to water with xylene and ethanol. 2. If Zenker fixed, remove mercury pigment. 3. Place in carbol ailine fuchsin for 5 minutes at 70C, or steaming on a hot plate. 4. Rinse quickly with tap water. 5. Decolourise with strong formalin (40%) a few drops at a time until no more colour is removed (15-20 minutes) 6. Rinse with tap water. 7. Counterstain with saturated aqueous picric acid for 1 minute. 8. Dehydrate with ethanol, clear with xylene and mount with Clarite (or something similar). Results: Encephalitozoa - blue black Toxoplasma chromatin - brown red Influenza bacilli - blue Nuclei - light red Cytoplasm - pink yellow Erythrocytes - bright yellow Bryan Llewellyn ----- Original Message ----- From: "Kathleen Cormier" To: Sent: Wednesday, March 24, 2004 11:35 AM Subject: [Histonet] Goodpasture's carbol fuchsin stain > Greetings! > > I have a researcher who is requesting a Goodpasture's carbol fuchsin stain > on rabbit kidneys to stain protoza. (encephalitozoon cuniculi or the old > old name renal nosematosis). My research indicates giemsa and gram would be > acceptable as well, but, I can find no reference for the Goodpasture's > carbol fuchsin. I have exhausted histonet archives and my histo books here. > Can anyone point me in the direction of a procedure for the Goodpasture's? > Thanks!!! > > > Kathy > > DCM MIT > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Dorothy.L.Webb <@t> HealthPartners.Com Wed Mar 24 14:37:10 2004 From: Dorothy.L.Webb <@t> HealthPartners.Com (Dorothy.L.Webb@HealthPartners.Com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] smears for IHC Message-ID: Whenever we do IHC on smears, we fix the smears in equal parts of hydrogen peroxide and methyl alcohol for 15 minutes. Another method that can be used is to place smears in a coplin jar and place 37%formaldehyde soaked gauze in the lid. Cover the smears for 20 minutes. Hope this helps! Dorothy Webb at Regions Hospital, St. Paul, Minn, -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, March 23, 2004 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 4, Issue 27 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. Atoska Gentry/ B Plus (Joyce Cline) 2. Ventana staining Avidin Biotin Block (Featherstone, Annette) 3. Re: Ventana staining Avidin Biotin Block (Lilia Muolo) 4. RE: Ventana staining Avidin Biotin Block (GUTIERREZ, JUAN) 5. Wendy England/ iron smears (Joyce Cline) 6. IHC position in San Francisco bay area. (Morken, Tim - Labvision) 7. Small Animal atlas (Volumes 1 and 2) are back in print! (Gayle Callis) 8. RE: Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's (Tony Henwood) 9. please remove me from your mailing list (Anna Bozzano) 10. RE: Web site for Cytology (Kemlo Rogerson) 11. subscribe (Anne Van Binsbergen) 12. Mouse Lung (Starkus, Laurie) 13. RE: help with elastic staining (Featherstone, Annette) 14. Problems with c-fos immediate early gene labelling (komo@bu.edu) 15. RE: CD98 and PLGF (Featherstone, Annette) 16. Re: Problems with c-fos immediate early gene labelling (Geoff McAuliffe) 17. FW: Job opening - Wildlife Conservation Society (McAloose, Dee) 18. IHC on blood and cytology smears (ICC) (Jan Shivers) 19. NYSHS 2004 Annual Meeting (Luis Chiriboga) ---------------------------------------------------------------------- Message: 1 Date: Mon, 22 Mar 2004 13:15:40 -0500 From: "Joyce Cline" Subject: [Histonet] Atoska Gentry/ B Plus To: "Histonet" Message-ID: <006901c41039$af661660$1d2a14ac@wchsys.org> Content-Type: text/plain; charset="iso-8859-1" Hi Atoska I get my B Plus from Market Lab, order # ML0407, Phone # 800-237-3604. I do not know it's components, other than formaldehyde. The company that makes it is Advanced Biomedical Reagents & Technologies. ------------------------------ Message: 2 Date: Mon, 22 Mar 2004 13:47:13 -0500 From: "Featherstone, Annette" Subject: [Histonet] Ventana staining Avidin Biotin Block To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. ------------------------------ Message: 3 Date: Mon, 22 Mar 2004 14:21:12 -0500 From: Lilia Muolo Subject: Re: [Histonet] Ventana staining Avidin Biotin Block To: AFeatherstone@KaleidaHealth.Org, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hi, our lab only uses the A /B blocking on the Ventana if necessary but usually we don't need to use it. We use the Discovery here but I do remember using the A/B blocking more often when we had the Nexes. I would imagine that the Benchmark would be like the Discovery. Lil Muolo Cancer Institute of New Jersey >>> "Featherstone, Annette" 3/22/04 1:47:13 PM >>> hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 22 Mar 2004 13:21:59 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Ventana staining Avidin Biotin Block To: "Featherstone, Annette" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="utf-8" We use it on everything. It's a lot easier than having a different protocol for aech type of tissue. Juan -----Original Message----- From: Featherstone, Annette [mailto:AFeatherstone@KaleidaHealth.Org] Sent: Mon 3/22/2004 12:47 PM To: Histonet (E-mail) Cc: Subject: [Histonet] Ventana staining Avidin Biotin Block hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 22 Mar 2004 15:56:30 -0500 From: "Joyce Cline" Subject: [Histonet] Wendy England/ iron smears To: "Histonet" Message-ID: <001f01c41050$277dcb00$1d2a14ac@wchsys.org> Content-Type: text/plain; charset="iso-8859-1" I use an old fashioned way of fixing the bone marrow smears. We had trouble with no reaction on our smears. Now we use a coplin jar with a small piece of paper towel in the bottom, drop 10% buffered formalin to saturate the paper towel but not to reach the blood on the slides. Leave the slides in for 10 minutes and stain normally. We have not had any trouble with an iron reaction since using this method. But do not leave the slides in more than 15 minutes max. ------------------------------ Message: 6 Date: Mon, 22 Mar 2004 14:30:56 -0800 From: "Morken, Tim - Labvision" Subject: [Histonet] IHC position in San Francisco bay area. To: "Histology Net List Server (E-Mail)" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA217741F@usca0082k08.labvision.apogent.com> Content-Type: text/plain Lab Vision, a fast-growing biotech company located in Fremont, California, just southeast of San Francisco, has an opening for an Immunohistochemistry Technologist in the Quality Control laboratory. The position entails testing of manufactured lots of antibody and detection reagents as well as prospective reagents. The applicant must have extensive experience in immunohistochemistry, especially in development of new antibodies and other IHC reagents. Advancement is limited only by personal initiative. Certification by ASCP as HT or HTL and/or QIHC is preferred. Salary is negotiable. Lab Vision (www.labvision.com) is a world-leading manufacturer of IHC-automation instruments and immunohistochemistry reagents for the pathology laboratory and biological research. Lab Vision is an Apogent company (www.apogent.com) If interested please send resume by email to: Tim Morken Product Development Lab Vision / Neomarkers 47790 Westinghouse Dr. Fremont, CA 94539 USA PH: 510-991-2840 FAX: 510-991-2826 email: tpmorken@labvision.com www.labvision.com ------------------------------ Message: 7 Date: Mon, 22 Mar 2004 16:12:28 -0700 From: Gayle Callis Subject: [Histonet] Small Animal atlas (Volumes 1 and 2) are back in print! To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040322161228.00bcc478@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Dear All, For those of you who work with or dissect laboratory animals and are in need of an atlas to guide the way. After many years of frustration and sadness when these atlases went out of print, they are back on the market. I saw the new atlas today, and couldn't wait to order it. A huge silent hooray! There are two color atlases (volume 1 is for rabbit and guinea pig and Volume 2 is rat, mouse, and Golden hamster. Pricing for Vol 1 and Vol 2 are $139 per atlas, and that is a deal. ISBN:0702027030 Title:Colour Atlas of Anatomy of Small Laboratory Animals, Volume 2, mouse, rat, hamster Price:$139.00 Go to Elsevier website to order, it is very user friendly, excellent. I just gave/ordered myself an early birthday gift! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 8 Date: Tue, 23 Mar 2004 11:22:18 +1100 From: Tony Henwood Subject: RE: [Histonet] Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's To: "'Boswell'" , Histonet@lists.utsouthwestern.edu Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E16A@simba.kids> Content-Type: text/plain; charset="iso-8859-1" Dear Boswell, The following Cherukian's modification works very well. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html Document Procedure: Phosphotungstic Acid Haematoxylin Principle: Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Use brain sections and section of muscle. A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined. Reagents: 1. Eosin: - Warning: Flammable - see MSDS Eosin Y, water soluble (CI 145380) 0.5g Distilled Water 10ml 80% Ethanol 190ml Working: 10ml stock and Before use add 50ul glacial acetic acid. 2. 1% Periodic Acid 3. PTAH solution Haematoxylin (CI 75290) 0.5g Phosphotungstic Acid 10g Distilled water 500ml Dissolve solid ingredients in separate portions of the water. Use gentle heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium permanganate to ripen. The stain is ready to use. Procedure: 1. Dewax and hydrate sections to 80% alcohol. 2. Place slides in eosin for 30 seconds. 3. Wash slides in distilled water for a few seconds. 4. Place slides in 1% periodic acid for 20 minutes. 5. Wash slides in water for 3 minutes. 6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30 minutes on. 7. Dehydrate, clear and mount. Results: Dendrites, nuclei, fibrin, platelets and muscle - blue Red blood cells and collagen - red. Notes: Reference: 1. Cherukian, C.J., Histologic. 8(4); 105, (1977). 2. Luna, L., Histologic. 5(2); 66, (1975). -----Original Message----- From: Boswell [mailto:kcboswell@grandecom.net] Sent: Saturday, 20 March 2004 7:30 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using Zenker's Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain without using Zenker's. We have looked everywhere and are unable to find it. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 9 Date: Tue, 23 Mar 2004 10:14:40 +0100 From: Anna Bozzano Subject: [Histonet] please remove me from your mailing list To: histonet@lists.utsouthwestern.edu Message-ID: <3.0.1.32.20040323101440.0120fd88@cucafera.icm.csic.es> Content-Type: text/plain; charset="us-ascii" please remove me from your mailing list ------------------------------ Message: 10 Date: Tue, 23 Mar 2004 09:21:37 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Web site for Cytology To: "'Kathleen Boozer'" , Message-ID: <000001c410b8$3f376b80$48362850@KEMLOS> Content-Type: text/plain; charset="us-ascii" Cytopathnet.org I think. It sort of comes and goes depending on the variety of virus it becomes infected with. I'm always amused that it becomes regularly infected by viruses; sort of gives you a feeling of its authenticity. I think it is negative for the oncogenic variety but hope it has regular tests including a silicone biopsy. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Boozer Sent: 17 March 2004 17:13 To: histonet@pathology.swmed.edu Subject: [Histonet] Web site for Cytology Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 23 Mar 2004 12:59:07 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] subscribe To: Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E01F18A95@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="iso-8859-1" ------------------------------ Message: 12 Date: Tue, 23 Mar 2004 07:56:38 -0500 From: "Starkus, Laurie" Subject: [Histonet] Mouse Lung To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="ISO-8859-1" I've recently started working with mouse lung and was given a protocol for processing. The protocol calls for 0.85% NaCl for 60 minutes at 4 degrees celsius. Then a 1:1 0.85% NaCl to Absolute ethanol for 30 minutes at room temp. Then the processing looks rather normal. Since my experience is with human tissue, I've never seen a sodium chloride step in processing. What does it do? And, is it really necessary? This is mouse lung infected with TB or cryptococcus. Thanks in advance. ------------------------------ Message: 13 Date: Tue, 23 Mar 2004 08:34:14 -0500 From: "Featherstone, Annette" Subject: RE: [Histonet] help with elastic staining To: 'Patsy Ruegg' , Deb Stults , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" When doing the pentachrome stain do not over differentiate the elastic too much in the ferric chloride, keep it kind of dark. The acid solutions that follow continue to lighten the elastic fibers. Annette FeatherstoneHT/MLT Kaleida Health -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 14:19 To: Deb Stults; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] help with elastic staining I have this problem when I do a Pentachrome stain for elastic fibers, the fibers stain fine when I first do the hematoxylin but as I continue with the rest of the stain the fiber stain is lost, I have remedied this by repeating the hematoxylin again at the end of the staining process to get back the fiber staining. Try it. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Deb Stults Sent: Friday, March 19, 2004 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help with elastic staining We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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. ------------------------------ Message: 14 Date: Tue, 23 Mar 2004 09:48:31 -0500 From: komo@bu.edu Subject: [Histonet] Problems with c-fos immediate early gene labelling To: histonet@lists.utsouthwestern.edu Message-ID: <1080053311.40604e3f585a5@www.bu.edu> Content-Type: text/plain I'm hoping I could get some helpful advice on what might cause my c-fos IHC labelling to work well in one lab, but not my own. I am running the protocol on rat brains using a hydrogen peroxide/methanol step for endogenous peroxidase blocking, a normal goat serum blocking step, a c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated secondary, a streptavidin horeradish peroxidase, and a nickle enhanced DAB reaction. Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. Does anyone has any suggestions as to why this might be? One person at the other lab suggested that possibly the wells that we run the tissue in were not being cleaned properly (we soak them in bleach while the other lab uses just pex). Could this have any effect? Thanks for any advice you can provide!! Rob Komorowski ------------------------------ Message: 15 Date: Tue, 23 Mar 2004 09:50:21 -0500 From: "Featherstone, Annette" Subject: RE: [Histonet] CD98 and PLGF To: 'Patsy Ruegg' , "Histonet@Pathology. Swmed. Edu" Cc: "Ihcrg@Yahoogroups. Com" Message-ID: Content-Type: text/plain; charset="iso-8859-1" YES!!I am! Or was... I used a pepsin pretreatment @37* for 15 minutes, Block with a protein block, primary antibody 1:100 for 1hr, Goat secondary 30 min rt, Alk phos 30 minutes rt, and whatever chromgen you use for alk phos, rt 40 min. My Plgf is from Santa Cruz. Annette Featherstone HT/MLT -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 13:58 To: Patsy Ruegg; Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: RE: [Histonet] CD98 and PLGF I guess no one is using cd98 and/or Placental Growth Factor on ffpe tissue???? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, March 17, 2004 10:03 AM To: Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: [Histonet] CD98 and PLGF I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160 and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies work in ffpe tissue? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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. ------------------------------ Message: 16 Date: Tue, 23 Mar 2004 10:24:38 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] Problems with c-fos immediate early gene labelling To: komo@bu.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <406080E6.2080605@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Ron: komo@bu.edu wrote: >I'm hoping I could get some helpful advice on what might cause my c-fos >IHC labelling to work well in one lab, but not my own. I am running the >protocol on rat brains using a hydrogen peroxide/methanol step for >endogenous peroxidase blocking, a normal goat serum blocking step, a >c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated >secondary, a streptavidin horeradish peroxidase, and a nickle enhanced >DAB reaction. > >Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. > When you say you are using the "exact same antibodies, chemicals, and ..... " do you mean the same bottle of reagent? You are carrying the bottle or vial of reagent from one lab to another? If not, look at the batch of antibody, your peroxide, your DAB, etc. If you are storing antibody in a freezer at home it is being subjected to freeze-thaw cycles (unless you have a very old freezer that does not defrost itself on a timer). If your bottle of peroxide is more than a few months old it may be in the process of turning into water, peroxide does that. I have had DAB work one day and fail the next. Since you have found that the brain slices stain well in the "other lab", the problem must lie with your home lab. Since bleach reacts with DAB I would avoid using staining dishes treated with bleach. Geoff >Does anyone has any suggestions as to why this might be? One person >at the other lab suggested that possibly the wells that we run the tissue in >were not being cleaned properly (we soak them in bleach while the other >lab uses just pex). Could this have any effect? > >Thanks for any advice you can provide!! > >Rob Komorowski > > > >_______________________________________________ >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 ********************************************** ------------------------------ Message: 17 Date: Tue, 23 Mar 2004 10:46:32 -0500 From: "McAloose, Dee" Subject: [Histonet] FW: Job opening - Wildlife Conservation Society To: Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD6D3@WOLF.wcs.org> Content-Type: text/plain; charset="iso-8859-1" > Hello, > > I'd like to announce an immediate opening for a full-time histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity and in becoming part of our team! > > Thanks for listening and we look forward to hearing from you. > > D McAloose, VMD, Dipl ACVP > Head, Department of Pathology > Wildlife Conservation Society > Bronx, NY 10464 > (phone) 718-220-7105 > (fax) 718-220-7126 > dmcaloose@wcs.org > > ------------------------------ Message: 18 Date: Tue, 23 Mar 2004 10:21:54 -0600 From: "Jan Shivers" Subject: [Histonet] IHC on blood and cytology smears (ICC) To: "histonet" Message-ID: <006301c410f2$f45d1ed0$78065486@vdl220FAC> Content-Type: text/plain; charset="iso-8859-1" Could someone please send me a protocol for performing IHC/ICC on smears (I normally only do FFPE slides)? I'd like info mostly on if the slides have to be coated/charged , smears air-dried or fixed (and in what) for how long, and if you do any modification of the immuno protocol, such as blocking endogenous peroxidase later in the procedure (and with what concentration of H2O2), and/or changing incubation times/dilutions. I've done smears on very rare occasions, but have had trouble getting the cells to remain on the uncoated slides that the smears are being made on. Thank you very much in advance. Jan Shivers Univ of Minnesota Vet Diag Lab shive003@tc.umn.edu ------------------------------ Message: 19 Date: Tue, 23 Mar 2004 12:02:46 -0500 From: Luis Chiriboga Subject: [Histonet] NYSHS 2004 Annual Meeting To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" The 2004 New York State Histotechnology Society annual meeting will be held at the Holiday Inn in beautiful Saratoga Springs, New York from Friday April 30th through Saturday May 1st. This years meeting theme is "Histology is no mystery: Let New York State solve your problems". An e-mail mini program is available upon request by replying to this message. For a program and registration packet, please contact Judy LaDuc at 518-897-2247 or jaladuc@capital.net We hope to see you there. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 4, Issue 27 *************************************** ________________________________________ 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. 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From jqb7 <@t> cdc.gov Wed Mar 24 14:41:14 2004 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Goodpasture's carbol fuchsin stain Message-ID: Also in Sheehan's. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bryan Llewellyn Sent: Wednesday, March 24, 2004 3:40 PM To: Histonet; Kathleen Cormier Subject: Re: [Histonet] Goodpasture's carbol fuchsin stain It is in Lillie's book as the Goodpasture-Perrin method for influenza organisms, encephalitozoa and toxoplasmata (Arch. Path. 36:568, 1943) Carbol aniline fuchsin: Basic fuchsin 0.59 g 30% alcohol 100 mL Analine 1 mL Phenol 1 g Fix in Zenker or Orth, or formalin fix and post chrome with 2.5% potassium dichromate for 2 days. Process in paraffin and section as usual. 1. Bring sections to water with xylene and ethanol. 2. If Zenker fixed, remove mercury pigment. 3. Place in carbol ailine fuchsin for 5 minutes at 70C, or steaming on a hot plate. 4. Rinse quickly with tap water. 5. Decolourise with strong formalin (40%) a few drops at a time until no more colour is removed (15-20 minutes) 6. Rinse with tap water. 7. Counterstain with saturated aqueous picric acid for 1 minute. 8. Dehydrate with ethanol, clear with xylene and mount with Clarite (or something similar). Results: Encephalitozoa - blue black Toxoplasma chromatin - brown red Influenza bacilli - blue Nuclei - light red Cytoplasm - pink yellow Erythrocytes - bright yellow Bryan Llewellyn ----- Original Message ----- From: "Kathleen Cormier" To: Sent: Wednesday, March 24, 2004 11:35 AM Subject: [Histonet] Goodpasture's carbol fuchsin stain > Greetings! > > I have a researcher who is requesting a Goodpasture's carbol fuchsin > stain on rabbit kidneys to stain protoza. (encephalitozoon cuniculi or > the old old name renal nosematosis). My research indicates giemsa and > gram would be > acceptable as well, but, I can find no reference for the Goodpasture's > carbol fuchsin. I have exhausted histonet archives and my histo books here. > Can anyone point me in the direction of a procedure for the > Goodpasture's? Thanks!!! > > > Kathy > > DCM MIT > > > _______________________________________________ > Histonet mailing list > Histonet@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 gentras <@t> vetmed.auburn.edu Wed Mar 24 14:56:51 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] 20% PFA/ 0.1M PB Message-ID: <6.0.1.1.0.20040324145419.0259a490@mailhost.vetmed.auburn.edu> Hello All, will someone please share with me the shelf life of in house 20% PFA/ 0.1M Phosphate Buffer stored at 4C? Thanks, Atoska Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From jmitchell <@t> neurology.wisc.edu Wed Mar 24 14:53:52 2004 From: jmitchell <@t> neurology.wisc.edu (Mitchell (Jean A.)) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Tina Anderson or Sherri Coffman Message-ID: <061D5F3CF2DFDF4EADF5611CC1218C4512A700@nrl-lorenz.neurology.wisc.edu> Sherri or Tina: You contacted me concerning the one-step gomori stain on frozen muscle and I wrote down your phone number incorrectly - could you please contact me again.(hopefully one of you subscribes to histonet) Or if anyone knows a contact phone number for either one of these histotechs could you please forward it on to me. Thanks, Jean Mitchell, BS, HT (ASCP) University of Wisconsin Hospital & Clinics Department of Neurology Madison, WI From histo007 <@t> hotmail.com Wed Mar 24 15:33:49 2004 From: histo007 <@t> hotmail.com (Jim Ball) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] coated slides Message-ID: I recently read an article in NSH( Maybe 3 to 4 issues back ) about a new type of coating a company developed for slides. I was wondering if any one out there has tried this new wonder coating. The article did not give a supplier,but they did furnish a number where you could get more information, but I do not have access to this publication at the present time, but do plan on checking out the NSH site to see if I can find the article, but in the mean time if any one has tried them please post your observations after using these slides. I have just recently got back into having to cut nails again and every coating I can think of. Nothing I have done has prevented the nails from being washed off of the slides during the staing procedure. The tricks I have tried are listed. Coated my own slides Air dried sections over night at RT and 37 degrees Used Gum Mastic Miicro waved the slides Elmer's Glue Used thinner sections to reduce the surface area the reagents could work on Slung a dead cat over my head while chanting a Tibetan folk song When I say all coatings I mean Salinized, Positive coated, and one other I canot remember the name of at the present time. Jim Ball histo007@hotmail.com _________________________________________________________________ Find a broadband plan that fits. Great local deals on high-speed Internet access. https://broadband.msn.com/?pgmarket=en-us/go/onm00200360ave/direct/01/ From gcallis <@t> montana.edu Wed Mar 24 15:54:12 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Shelf life of 20% PFA/ 0.1M PB In-Reply-To: <6.0.1.1.0.20040324145419.0259a490@mailhost.vetmed.auburn.e du> Message-ID: <3.0.6.32.20040324145412.00bc39b8@gemini.msu.montana.edu> I have never made a 20% solution, rather a working/stock 4% (4 gm PFA in 100 ml Dulbeccos PBS) In general, this is a common concentration for most PFA fixations and we tend to dilute the 4% for lesser concentration. Some people like their PFA fresh, then use immediately. We store 4% PFA for several months, but if there is any cloudiness/turbidity in solution, it is tossed. Months is a variable (not to mention people handling the reagent - the piggy factor!) - we have had 4% PFA go bad within a month and good as long as 6 months. A lot depends on how often people dip into the reagent, if they are careless about sticking pipettes into stock or leave at RT for hours before returning to refrig. Personal rule: store in a clean, clear glass bottle and check for turbid, cloudy solution before each use. Toss if that cloudy flocculant ppt shows up. This was discussed a couple of years ago on Histonet, you might want to go into archives. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Barry.R.Rittman <@t> uth.tmc.edu Wed Mar 24 16:22:22 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] smears for IHC Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FD3E@UTHEVS3.mail.uthouston.edu> A point worth mentioning here is that fixation of smears can be accomplished using this vapor method with a variety of fixatives, formaldehyde, glutaraldehyde, acetone, ethanol, osmium tetroxide etc. The major advantage of vapor fixation is that the tissue is not exposed to the vehicle containing the fixative. Because of this thin layers such as smears tend to stay on the slides better than if placing these in a solution of the fixative. It also eliminates the need to worry about osmolarity, concentration and pH. It is however, only useful for thin layers, cryostat sections etc. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dorothy.L.Webb@HealthPartners.Com Sent: Wednesday, March 24, 2004 2:37 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] smears for IHC Whenever we do IHC on smears, we fix the smears in equal parts of hydrogen peroxide and methyl alcohol for 15 minutes. Another method that can be used is to place smears in a coplin jar and place 37%formaldehyde soaked gauze in the lid. Cover the smears for 20 minutes. Hope this helps! Dorothy Webb at Regions Hospital, St. Paul, Minn, -----Original Message----- From: histonet-request@lists.utsouthwestern.edu [mailto:histonet-request@lists.utsouthwestern.edu] Sent: Tuesday, March 23, 2004 12:00 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 4, Issue 27 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. Atoska Gentry/ B Plus (Joyce Cline) 2. Ventana staining Avidin Biotin Block (Featherstone, Annette) 3. Re: Ventana staining Avidin Biotin Block (Lilia Muolo) 4. RE: Ventana staining Avidin Biotin Block (GUTIERREZ, JUAN) 5. Wendy England/ iron smears (Joyce Cline) 6. IHC position in San Francisco bay area. (Morken, Tim - Labvision) 7. Small Animal atlas (Volumes 1 and 2) are back in print! (Gayle Callis) 8. RE: Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's (Tony Henwood) 9. please remove me from your mailing list (Anna Bozzano) 10. RE: Web site for Cytology (Kemlo Rogerson) 11. subscribe (Anne Van Binsbergen) 12. Mouse Lung (Starkus, Laurie) 13. RE: help with elastic staining (Featherstone, Annette) 14. Problems with c-fos immediate early gene labelling (komo@bu.edu) 15. RE: CD98 and PLGF (Featherstone, Annette) 16. Re: Problems with c-fos immediate early gene labelling (Geoff McAuliffe) 17. FW: Job opening - Wildlife Conservation Society (McAloose, Dee) 18. IHC on blood and cytology smears (ICC) (Jan Shivers) 19. NYSHS 2004 Annual Meeting (Luis Chiriboga) ---------------------------------------------------------------------- Message: 1 Date: Mon, 22 Mar 2004 13:15:40 -0500 From: "Joyce Cline" Subject: [Histonet] Atoska Gentry/ B Plus To: "Histonet" Message-ID: <006901c41039$af661660$1d2a14ac@wchsys.org> Content-Type: text/plain; charset="iso-8859-1" Hi Atoska I get my B Plus from Market Lab, order # ML0407, Phone # 800-237-3604. I do not know it's components, other than formaldehyde. The company that makes it is Advanced Biomedical Reagents & Technologies. ------------------------------ Message: 2 Date: Mon, 22 Mar 2004 13:47:13 -0500 From: "Featherstone, Annette" Subject: [Histonet] Ventana staining Avidin Biotin Block To: "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="iso-8859-1" hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. ------------------------------ Message: 3 Date: Mon, 22 Mar 2004 14:21:12 -0500 From: Lilia Muolo Subject: Re: [Histonet] Ventana staining Avidin Biotin Block To: AFeatherstone@KaleidaHealth.Org, histonet@pathology.swmed.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Hi, our lab only uses the A /B blocking on the Ventana if necessary but usually we don't need to use it. We use the Discovery here but I do remember using the A/B blocking more often when we had the Nexes. I would imagine that the Benchmark would be like the Discovery. Lil Muolo Cancer Institute of New Jersey >>> "Featherstone, Annette" 3/22/04 1:47:13 PM >>> hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 4 Date: Mon, 22 Mar 2004 13:21:59 -0600 From: "GUTIERREZ, JUAN" Subject: RE: [Histonet] Ventana staining Avidin Biotin Block To: "Featherstone, Annette" , "Histonet (E-mail)" Message-ID: Content-Type: text/plain; charset="utf-8" We use it on everything. It's a lot easier than having a different protocol for aech type of tissue. Juan -----Original Message----- From: Featherstone, Annette [mailto:AFeatherstone@KaleidaHealth.Org] Sent: Mon 3/22/2004 12:47 PM To: Histonet (E-mail) Cc: Subject: [Histonet] Ventana staining Avidin Biotin Block hi Just wondering if anyone out there using the Ventana can tell me if they are using Avidin Biotin Block (BlockerA and BlockerB) on all their ABC system antibodies. Our institution is only using them on a select few and I think we should use it on all of them.Thanks for your response Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Mon, 22 Mar 2004 15:56:30 -0500 From: "Joyce Cline" Subject: [Histonet] Wendy England/ iron smears To: "Histonet" Message-ID: <001f01c41050$277dcb00$1d2a14ac@wchsys.org> Content-Type: text/plain; charset="iso-8859-1" I use an old fashioned way of fixing the bone marrow smears. We had trouble with no reaction on our smears. Now we use a coplin jar with a small piece of paper towel in the bottom, drop 10% buffered formalin to saturate the paper towel but not to reach the blood on the slides. Leave the slides in for 10 minutes and stain normally. We have not had any trouble with an iron reaction since using this method. But do not leave the slides in more than 15 minutes max. ------------------------------ Message: 6 Date: Mon, 22 Mar 2004 14:30:56 -0800 From: "Morken, Tim - Labvision" Subject: [Histonet] IHC position in San Francisco bay area. To: "Histology Net List Server (E-Mail)" Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA217741F@usca0082k08.labvision.apogent.co m> Content-Type: text/plain Lab Vision, a fast-growing biotech company located in Fremont, California, just southeast of San Francisco, has an opening for an Immunohistochemistry Technologist in the Quality Control laboratory. The position entails testing of manufactured lots of antibody and detection reagents as well as prospective reagents. The applicant must have extensive experience in immunohistochemistry, especially in development of new antibodies and other IHC reagents. Advancement is limited only by personal initiative. Certification by ASCP as HT or HTL and/or QIHC is preferred. Salary is negotiable. Lab Vision (www.labvision.com) is a world-leading manufacturer of IHC-automation instruments and immunohistochemistry reagents for the pathology laboratory and biological research. Lab Vision is an Apogent company (www.apogent.com) If interested please send resume by email to: Tim Morken Product Development Lab Vision / Neomarkers 47790 Westinghouse Dr. Fremont, CA 94539 USA PH: 510-991-2840 FAX: 510-991-2826 email: tpmorken@labvision.com www.labvision.com ------------------------------ Message: 7 Date: Mon, 22 Mar 2004 16:12:28 -0700 From: Gayle Callis Subject: [Histonet] Small Animal atlas (Volumes 1 and 2) are back in print! To: Histonet@lists.utsouthwestern.edu Message-ID: <3.0.6.32.20040322161228.00bcc478@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii" Dear All, For those of you who work with or dissect laboratory animals and are in need of an atlas to guide the way. After many years of frustration and sadness when these atlases went out of print, they are back on the market. I saw the new atlas today, and couldn't wait to order it. A huge silent hooray! There are two color atlases (volume 1 is for rabbit and guinea pig and Volume 2 is rat, mouse, and Golden hamster. Pricing for Vol 1 and Vol 2 are $139 per atlas, and that is a deal. ISBN:0702027030 Title:Colour Atlas of Anatomy of Small Laboratory Animals, Volume 2, mouse, rat, hamster Price:$139.00 Go to Elsevier website to order, it is very user friendly, excellent. I just gave/ordered myself an early birthday gift! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) ------------------------------ Message: 8 Date: Tue, 23 Mar 2004 11:22:18 +1100 From: Tony Henwood Subject: RE: [Histonet] Phosphotungstic Acid Hematoxylin stain without usi ng Zenker's To: "'Boswell'" , Histonet@lists.utsouthwestern.edu Message-ID: <1CF2E2E5BB36D5119E7A0008C791F3740800E16A@simba.kids> Content-Type: text/plain; charset="iso-8859-1" Dear Boswell, The following Cherukian's modification works very well. Tony Henwood JP, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 http://www.histosearch.com/homepages/TonyHenwood/default.html http://us.geocities.com/tonyhenwoodau/index.html Document Procedure: Phosphotungstic Acid Haematoxylin Principle: Cherukian's modification employs an eosin solution that stains the erythrocytes red and differentiates them from the blue fibrin. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Controls: Use brain sections and section of muscle. A good stain will demonstrate the dendrites as blue where as in a bad stain they appear light grey to salmon in colour. Nuclei, fibrin, platelets and muscle will be blue, red cells and collagen appear red. Muscle striations should be well defined. Reagents: 1. Eosin: - Warning: Flammable - see MSDS Eosin Y, water soluble (CI 145380) 0.5g Distilled Water 10ml 80% Ethanol 190ml Working: 10ml stock and Before use add 50ul glacial acetic acid. 2. 1% Periodic Acid 3. PTAH solution Haematoxylin (CI 75290) 0.5g Phosphotungstic Acid 10g Distilled water 500ml Dissolve solid ingredients in separate portions of the water. Use gentle heat for Haematoxylin. Combine solutions when cool. Add 0.088g potassium permanganate to ripen. The stain is ready to use. Procedure: 1. Dewax and hydrate sections to 80% alcohol. 2. Place slides in eosin for 30 seconds. 3. Wash slides in distilled water for a few seconds. 4. Place slides in 1% periodic acid for 20 minutes. 5. Wash slides in water for 3 minutes. 6. Place slides in PTAH for 30-90 minutes in 60oC oven. Check from 30 minutes on. 7. Dehydrate, clear and mount. Results: Dendrites, nuclei, fibrin, platelets and muscle - blue Red blood cells and collagen - red. Notes: Reference: 1. Cherukian, C.J., Histologic. 8(4); 105, (1977). 2. Luna, L., Histologic. 5(2); 66, (1975). -----Original Message----- From: Boswell [mailto:kcboswell@grandecom.net] Sent: Saturday, 20 March 2004 7:30 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Phosphotungstic Acid Hematoxylin stain without using Zenker's Does anyone have a procedure for Phosphotungstic Acid Hematoxylin stain without using Zenker's. We have looked everywhere and are unable to find it. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This footnote also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 9 Date: Tue, 23 Mar 2004 10:14:40 +0100 From: Anna Bozzano Subject: [Histonet] please remove me from your mailing list To: histonet@lists.utsouthwestern.edu Message-ID: <3.0.1.32.20040323101440.0120fd88@cucafera.icm.csic.es> Content-Type: text/plain; charset="us-ascii" please remove me from your mailing list ------------------------------ Message: 10 Date: Tue, 23 Mar 2004 09:21:37 -0000 From: "Kemlo Rogerson" Subject: RE: [Histonet] Web site for Cytology To: "'Kathleen Boozer'" , Message-ID: <000001c410b8$3f376b80$48362850@KEMLOS> Content-Type: text/plain; charset="us-ascii" Cytopathnet.org I think. It sort of comes and goes depending on the variety of virus it becomes infected with. I'm always amused that it becomes regularly infected by viruses; sort of gives you a feeling of its authenticity. I think it is negative for the oncogenic variety but hope it has regular tests including a silicone biopsy. Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS Tel: 0208 970 8414 Mob: 07830 196072 Mobile E-Mail kemlorogerson@3mail.com FAX & Answer Phone 0871 242 8094 E-mail Accounts: kemlo@tiscali.co.uk or kemlo1@btinternet.com Disclaimer: The information contained in this message and/or any attachments(s) may be of a private and confidential nature, and is intended solely for the attention of the addressee. If you have received this message in error or feel you should not have been the intended recipient, please return it and any attachments to the sender immediately. All messages relating to this communication should then be deleted from your system. Unauthorised usage, copying, disclosure or alteration of this message and/or attachment(s) is strictly prohibited. Barking, Havering and Redbridge Hospitals NHS Trust will not be held responsible for any direct or indirect damages which may arise from alteration of this message or any attachment(s), by a third party or resulting from the transmission of a virus. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen Boozer Sent: 17 March 2004 17:13 To: histonet@pathology.swmed.edu Subject: [Histonet] Web site for Cytology Is there something like the Histonet available for Cytology? I have a co-worker looking for perameters in quality control for her department as she documents screening errors. Thanks! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 11 Date: Tue, 23 Mar 2004 12:59:07 +0400 From: "Anne Van Binsbergen" Subject: [Histonet] subscribe To: Message-ID: <0C44F1AAEE47D54DA4210A60AB206F5E01F18A95@SKMCEMAIL.skmc.gov.ae> Content-Type: text/plain; charset="iso-8859-1" ------------------------------ Message: 12 Date: Tue, 23 Mar 2004 07:56:38 -0500 From: "Starkus, Laurie" Subject: [Histonet] Mouse Lung To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="ISO-8859-1" I've recently started working with mouse lung and was given a protocol for processing. The protocol calls for 0.85% NaCl for 60 minutes at 4 degrees celsius. Then a 1:1 0.85% NaCl to Absolute ethanol for 30 minutes at room temp. Then the processing looks rather normal. Since my experience is with human tissue, I've never seen a sodium chloride step in processing. What does it do? And, is it really necessary? This is mouse lung infected with TB or cryptococcus. Thanks in advance. ------------------------------ Message: 13 Date: Tue, 23 Mar 2004 08:34:14 -0500 From: "Featherstone, Annette" Subject: RE: [Histonet] help with elastic staining To: 'Patsy Ruegg' , Deb Stults , histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="iso-8859-1" When doing the pentachrome stain do not over differentiate the elastic too much in the ferric chloride, keep it kind of dark. The acid solutions that follow continue to lighten the elastic fibers. Annette FeatherstoneHT/MLT Kaleida Health -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 14:19 To: Deb Stults; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] help with elastic staining I have this problem when I do a Pentachrome stain for elastic fibers, the fibers stain fine when I first do the hematoxylin but as I continue with the rest of the stain the fiber stain is lost, I have remedied this by repeating the hematoxylin again at the end of the staining process to get back the fiber staining. Try it. Patsy -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Deb Stults Sent: Friday, March 19, 2004 11:48 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] help with elastic staining We ran into a problem today with an elastic stain on an artery biopsy. We're pretty sure it is due to a reagent problem, but not sure which one. The elastic fibers did not stain at all. We use Weigert's iron hematoxylin solutions A and B, which are new bottles and worked yesterday in another stain (we make up fresh before each use), the Resorcin Fuchsin working solution expired last week, and our new bottle (previously ordered) has not yet arrived, and the Van Gieson's solution is brand new just opened today. Since the fibers are not black, that makes me think that its the Weigert's, but since the Resorcin Fuchsin is expired, we weren't sure if it could be the problem. If anyone has any suggestions, please let me know asap. The pathologist doesn't want to wait for new reagent to come in. Thanks, Deb and Karen _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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. ------------------------------ Message: 14 Date: Tue, 23 Mar 2004 09:48:31 -0500 From: komo@bu.edu Subject: [Histonet] Problems with c-fos immediate early gene labelling To: histonet@lists.utsouthwestern.edu Message-ID: <1080053311.40604e3f585a5@www.bu.edu> Content-Type: text/plain I'm hoping I could get some helpful advice on what might cause my c-fos IHC labelling to work well in one lab, but not my own. I am running the protocol on rat brains using a hydrogen peroxide/methanol step for endogenous peroxidase blocking, a normal goat serum blocking step, a c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated secondary, a streptavidin horeradish peroxidase, and a nickle enhanced DAB reaction. Unfortunately, although I have all of the exact same antibodies, chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. Does anyone has any suggestions as to why this might be? One person at the other lab suggested that possibly the wells that we run the tissue in were not being cleaned properly (we soak them in bleach while the other lab uses just pex). Could this have any effect? Thanks for any advice you can provide!! Rob Komorowski ------------------------------ Message: 15 Date: Tue, 23 Mar 2004 09:50:21 -0500 From: "Featherstone, Annette" Subject: RE: [Histonet] CD98 and PLGF To: 'Patsy Ruegg' , "Histonet@Pathology. Swmed. Edu" Cc: "Ihcrg@Yahoogroups. Com" Message-ID: Content-Type: text/plain; charset="iso-8859-1" YES!!I am! Or was... I used a pepsin pretreatment @37* for 15 minutes, Block with a protein block, primary antibody 1:100 for 1hr, Goat secondary 30 min rt, Alk phos 30 minutes rt, and whatever chromgen you use for alk phos, rt 40 min. My Plgf is from Santa Cruz. Annette Featherstone HT/MLT -----Original Message----- From: Patsy Ruegg [mailto:pruegg@colobio.com] Sent: Friday, March 19, 2004 13:58 To: Patsy Ruegg; Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: RE: [Histonet] CD98 and PLGF I guess no one is using cd98 and/or Placental Growth Factor on ffpe tissue???? -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Patsy Ruegg Sent: Wednesday, March 17, 2004 10:03 AM To: Histonet@Pathology. Swmed. Edu Cc: Ihcrg@Yahoogroups. Com Subject: [Histonet] CD98 and PLGF I am seeking those who may have experience with cd98 (h-300) Santa Cruz-9160 and/or Abcam anti-PLGF ab9542 IHC on ffpe tissue??? Do these antibodies work in ffpe tissue? Patsy _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This email 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. ------------------------------ Message: 16 Date: Tue, 23 Mar 2004 10:24:38 -0800 From: Geoff McAuliffe Subject: Re: [Histonet] Problems with c-fos immediate early gene labelling To: komo@bu.edu Cc: histonet@lists.utsouthwestern.edu Message-ID: <406080E6.2080605@umdnj.edu> Content-Type: text/plain; format=flowed; charset=ISO-8859-1 Hi Ron: komo@bu.edu wrote: >I'm hoping I could get some helpful advice on what might cause my c-fos >IHC labelling to work well in one lab, but not my own. I am running the >protocol on rat brains using a hydrogen peroxide/methanol step for >endogenous peroxidase blocking, a normal goat serum blocking step, a >c-fos Ab-5 primary from oncogene, a goat-anti rabbit biotinylated >secondary, a streptavidin horeradish peroxidase, and a nickle enhanced >DAB reaction. > >Unfortunately, although I have all of the exact same antibodies, >chemicals, and concentrations as the lab from which the protocol was developed, the label is weak and not very specific when I run it at my home lab. In fact, I even took brain slices from the same rats that I was running at my lab and ran them at the lab where the protocol was developed, and found that the c-fos label was beautiful with a low background. > When you say you are using the "exact same antibodies, chemicals, and ..... " do you mean the same bottle of reagent? You are carrying the bottle or vial of reagent from one lab to another? If not, look at the batch of antibody, your peroxide, your DAB, etc. If you are storing antibody in a freezer at home it is being subjected to freeze-thaw cycles (unless you have a very old freezer that does not defrost itself on a timer). If your bottle of peroxide is more than a few months old it may be in the process of turning into water, peroxide does that. I have had DAB work one day and fail the next. Since you have found that the brain slices stain well in the "other lab", the problem must lie with your home lab. Since bleach reacts with DAB I would avoid using staining dishes treated with bleach. Geoff >Does anyone has any suggestions as to why this might be? One person >at the other lab suggested that possibly the wells that we run the tissue in >were not being cleaned properly (we soak them in bleach while the other >lab uses just pex). Could this have any effect? > >Thanks for any advice you can provide!! > >Rob Komorowski > > > >_______________________________________________ >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 ********************************************** ------------------------------ Message: 17 Date: Tue, 23 Mar 2004 10:46:32 -0500 From: "McAloose, Dee" Subject: [Histonet] FW: Job opening - Wildlife Conservation Society To: Message-ID: <9C26E1A5C9EE5F4B84216095CBE2E24A059DD6D3@WOLF.wcs.org> Content-Type: text/plain; charset="iso-8859-1" > Hello, > > I'd like to announce an immediate opening for a full-time > histotechnician in the pathology department at the Wildlife Conservation Society located at the Bronx Zoo. We provide the diagnostic pathology services to one of the largest zoological collections in the country. Applications are being accepted until May 15, 2004 or until the position is filled. Please contact Ms. Tawanda Williams (Human Resources; email: twilliams@wcs.org) if you're interested in this exciting opportunity and in becoming part of our team! > > Thanks for listening and we look forward to hearing from you. > > D McAloose, VMD, Dipl ACVP > Head, Department of Pathology > Wildlife Conservation Society > Bronx, NY 10464 > (phone) 718-220-7105 > (fax) 718-220-7126 > dmcaloose@wcs.org > > ------------------------------ Message: 18 Date: Tue, 23 Mar 2004 10:21:54 -0600 From: "Jan Shivers" Subject: [Histonet] IHC on blood and cytology smears (ICC) To: "histonet" Message-ID: <006301c410f2$f45d1ed0$78065486@vdl220FAC> Content-Type: text/plain; charset="iso-8859-1" Could someone please send me a protocol for performing IHC/ICC on smears (I normally only do FFPE slides)? I'd like info mostly on if the slides have to be coated/charged , smears air-dried or fixed (and in what) for how long, and if you do any modification of the immuno protocol, such as blocking endogenous peroxidase later in the procedure (and with what concentration of H2O2), and/or changing incubation times/dilutions. I've done smears on very rare occasions, but have had trouble getting the cells to remain on the uncoated slides that the smears are being made on. Thank you very much in advance. Jan Shivers Univ of Minnesota Vet Diag Lab shive003@tc.umn.edu ------------------------------ Message: 19 Date: Tue, 23 Mar 2004 12:02:46 -0500 From: Luis Chiriboga Subject: [Histonet] NYSHS 2004 Annual Meeting To: Histonet Message-ID: Content-Type: text/plain; charset="iso-8859-1" The 2004 New York State Histotechnology Society annual meeting will be held at the Holiday Inn in beautiful Saratoga Springs, New York from Friday April 30th through Saturday May 1st. This years meeting theme is "Histology is no mystery: Let New York State solve your problems". An e-mail mini program is available upon request by replying to this message. For a program and registration packet, please contact Judy LaDuc at 518-897-2247 or jaladuc@capital.net We hope to see you there. ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 4, Issue 27 *************************************** ________________________________________ 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. 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You will be reimbursed for reasonable costs incurred in notifying us. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Wed Mar 24 17:10:51 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] cryo preserving and sectioning of bone In-Reply-To: <20040323212051.81956.qmail@web13306.mail.yahoo.com> Message-ID: <3.0.6.32.20040324161051.00bc39b8@gemini.msu.montana.edu> One young fellow has fixed murine nasal turbinates in PLP overnight at 4C, changed fixative and fixed overnight again. It is advisable to perfuse the animal with PLP at euthanasia via heart before immersion into larger vol of PLP. After PLP fixation, he decalcifies in tetrasodium EDTA (1.25%) dissolved in 20% sucrose in Dulbecco's PBS. He adjusts the pH to 7 or so. Decalcification is done at 4C after drawing a vacuum to exclude air bubbles in nasal passages and allow EDTA better, close contact with turbinates. It can take up to 1 week or longer to decalcify. Snap freeze using a petri dish floating in layer of liquid nitrogen. Embed bone in OCT in a cryomold (Peel a way, plastic cryomold) and sit this inside petri dish until block is frozen. DO NOT LET LIQUID nitrogen get into petri dish, just let dish canoe in this with platform support, don't want mold to tip over into Liq N2. which can crack block from extreme cold. The EDTA decalcified bone cryosections without problems with excellent immunostaining. Advantage, decalcification while cryoprotecting with sucrose solution. We have one project (hamster turbinates, PLP fixed overnight, with EDTA upped to 5% concentration in 20% sucrose in DPBS, adjusted to pH 7.6 with glacial acetic acid. We chose to raise the pH a bit to speed up decalcification since EDTA decalcifies as a function of pH, with lower pH giving slower decalcification rate but staying within a working pH similar to buffers used during immunostaining - pH 7 to 7.6. Acid decalcification is avoided although should work IF one rinses acid out thoroughly and your antigens survive acid exposure. 20% to 30% sucrose cryoprotection will help rinse out any residual acid, 2 changes might even be better. One could also do tedious acid neutralization. We don't like the idea of residual acid in section trimmings sitting around inside the cryostat for potential corrosion of metal parts. We do a simple decalcification endpoint check, weight loss, weight gain method that puts us in the ball park for total calcium removal, it works with EDTA quite nicely and acid decalcification methods. Hopefully you have a FAXITRON for sensitive endpoint determinations. EDTA chemical determination is a pain to do, although a good idea with acid methods. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Wed Mar 24 17:16:56 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Mouse Histological Atlas, another book Message-ID: <3.0.6.32.20040324161656.00bc39b8@gemini.msu.montana.edu> Andrea asked for a histology atlas, not just the anatomy atlas. Here it is: Histological Atlas of the Laboratory Mouse, WD Gude, GE Cosgrove and GP Hirsch Plenum Press, NY and London, 1982 ISBN#0-306-40686-1 No price. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From dingj <@t> nicemice.cn Wed Mar 24 20:09:05 2004 From: dingj <@t> nicemice.cn (dingjun) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] How to remove my email address from the list Message-ID: <00c601c4120e$2648bdc0$ef64a8c0@djxmg> SGkgZGVhciBmZWxsb3dzDQpEb2VzIGFueW9uZSBrbm93IGhvdyB0byBhdm9pZCByZWNlaXZpbmcg YWxsIHRoZSBlbWFpbCBwb3N0ZWQgaGVyZT8gTXkgbWFpbCBib3ggY2FuJ3QgYmVhci4gV2hvIGNh biBoZWxwIG1lPyANClRoYW5rcy4NCkNoZWVycw0KDQpKdW4gDQpNb2RlbCBBbmltYWwgUmVzZWFy Y2ggQ2VudGVyKE1BUkMpDQpOYW5qaW5nIFVuaXZlcnNpdHkNCk5hbkppbmcgUC5SLkNoaW5hDQpQ b3N0YWwgQ29kZTogMjEwMDkzDQpQaG9uZTogODYtMjUtODY0MTUxMQ0KRmF4OiA4Ni0yNS04NjQx NTAwDQpodHRwOi8vd3d3Lm5pY2VtaWNlLmNuL2VuX2luZGV4Lmh0bQ0KDQogDQo= From Juan.Solon <@t> lshtm.ac.uk Thu Mar 25 05:26:05 2004 From: Juan.Solon <@t> lshtm.ac.uk (Juan Solon) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] CD4/CD25/TGFB Antibodies; Filter choices forLeica Message-ID: Sorry, I neglected to mention that I will be working with human samples. J Dr. Juan Antonio Solon MRC Keneba Field Station MRC Laboratories Gambia Atlantic Road, Fajara PO Box 273, Banjul The Gambia Email : juan.solon@lshtm.ac.uk Tel No: (++220) 541021 Fax No: (++220) 541022 / (++220) 496513 >>> Gayle Callis 03/24/04 4:41 PM >>> What species are you working with? Mouse? Human? At 09:36 AM 3/24/2004 +0000, you wrote: >Dear All, > >As part of a study on mucosal immunity in The Gambia, W. Africa, I plan to double stain antral and duodenal specimens for CD4 and CD25 & CD8 and CD25. Also, we would attempt to triple stain for TGF beta on these subsets. We have access to a Leica DMRXE although the filter cubes are all currently long pass suppression filters . I have both formalin-fixed and frozen sections of the antrum and the duodenum. > >I would like to ask for advise on several points: > >1. Can anyone share their experience on antibody combinations known to work for CD4-25, CD8-25 and triple staining for CD4-CD25-TGFB either with vectastain elite (peroxidase) or immunofluorescence. I have compiled a list of CD4, CD8, CD25, TGFB antibodies from Dako, Ancell, Novocastra, BD, and Serotec, but before purchasing any of these, I thought it would be wise to ask advise from the Histonet community. > >2. I would probably need to acquire different filter cubes to do simultaneous imaging of fluorescence. There are several suppliers I have encountered on the net (Omega Filters and Chroma Technology) * does anyone have experience on using filters from these companies on Leica microscopes (specifically the DMRXE). Any problems encountered with using third-party filters with Leica microscopes? > >Thanks. >Juan > > >Dr. Juan Antonio Solon >MRC Keneba Field Station >MRC Laboratories Gambia >Atlantic Road, Fajara >PO Box 273, Banjul >The Gambia >Email : juan.solon@lshtm.ac.uk >Tel No: (++220) 541021 >Fax No: (++220) 541022 / (++220) 496513 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From cormier <@t> MIT.EDU Thu Mar 25 04:33:58 2004 From: cormier <@t> MIT.EDU (Kathleen Cormier) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Goodpasture's carbol fuchsin stain In-Reply-To: Message-ID: <5.2.1.1.2.20040325053301.00ad5be8@hesiod> Great idea, I thought so too, until I remembered that the reference specifically stated "carbol fuchsin... At 02:59 PM 3/24/2004 -0500, you wrote: >Do you think they meant the MacCallum-Goodpasture stain? > >Jeanine Bartlett, HT(ASCP) >Centers for Disease Control >Infectious Disease Pathology Activity >1600 Clifton Road, MS/G-32 >Atlanta, GA 30333 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen >Cormier >Sent: Wednesday, March 24, 2004 2:36 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] Goodpasture's carbol fuchsin stain > > >Greetings! > >I have a researcher who is requesting a Goodpasture's carbol fuchsin >stain >on rabbit kidneys to stain protoza. (encephalitozoon cuniculi or the old > >old name renal nosematosis). My research indicates giemsa and gram would >be >acceptable as well, but, I can find no reference for the Goodpasture's >carbol fuchsin. I have exhausted histonet archives and my histo books >here. >Can anyone point me in the direction of a procedure for the >Goodpasture's? >Thanks!!! > > >Kathy > >DCM MIT > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marjorie.Lehman <@t> unilever.com Thu Mar 25 06:44:24 2004 From: Marjorie.Lehman <@t> unilever.com (marjorie lehman) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Goodpasture's carbol fuchsin stain Message-ID: Good Grief! Tell your researcher to come join us in the twenty-first century. I got the following from Kolmer & Boerner's Approved Laboratory Technic, published in 1941 Goodpasture's Stain Place prepared sections in the following stain for 10 to 30 minutes Alcohol.............100 c.c. Basic fuchsin.....0.59 gr. Aniline oil..........1.00 c.c. Phenol..............1.00 gm. Wash in water Place in 40% formalin for a few seconds (bright red color fades to clear rose) Wash in water Counterstain in saturated water solution of picric acid for 3-5 minutes (until section assumes a purplish-yellow color) Wash in water Differentiate in 95% alcohol (red appears and some red and some picric acid is washed out) Wash in water Stain in Sterling's gentian violet for 5 or more minutes gentian violet ......................5 gm. Alcohol (95%)....................10 c.c. Grind in a mortar and add Aniline oil ...........................2 c.c. Water (distilled) .................88 c.c. Let stand 1 or 2 days and filter Wash in water Place in Gram's iodine solution for 5 minutes Blot dry Place in aniline oil/xylene (equal parts) until no more color comes away Place in xylene and mount Gram-negative organisms stain red; Gram positive organisms stain blue; tissues stain in shades of red to purple. That ought to give your Safety Officer a nervous breakdown!! Marge -----Original Message----- From: Kathleen Cormier [SMTP:cormier@mit.edu] Sent: Thursday, March 25, 2004 5:34 AM To: Bartlett, Jeanine; histonet@pathology.swmed.edu Subject: RE: [Histonet] Goodpasture's carbol fuchsin stain Great idea, I thought so too, until I remembered that the reference specifically stated "carbol fuchsin... At 02:59 PM 3/24/2004 -0500, you wrote: >Do you think they meant the MacCallum-Goodpasture stain? > >Jeanine Bartlett, HT(ASCP) >Centers for Disease Control >Infectious Disease Pathology Activity >1600 Clifton Road, MS/G-32 >Atlanta, GA 30333 > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kathleen >Cormier >Sent: Wednesday, March 24, 2004 2:36 PM >To: histonet@pathology.swmed.edu >Subject: [Histonet] Goodpasture's carbol fuchsin stain > > >Greetings! > >I have a researcher who is requesting a Goodpasture's carbol fuchsin >stain >on rabbit kidneys to stain protoza. (encephalitozoon cuniculi or the old > >old name renal nosematosis). My research indicates giemsa and gram would >be >acceptable as well, but, I can find no reference for the Goodpasture's >carbol fuchsin. I have exhausted histonet archives and my histo books >here. >Can anyone point me in the direction of a procedure for the >Goodpasture's? >Thanks!!! > > >Kathy > >DCM MIT > > >_______________________________________________ >Histonet mailing list >Histonet@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 carmen_loiselle <@t> hotmail.com Thu Mar 25 10:41:25 2004 From: carmen_loiselle <@t> hotmail.com (carmen loiselle) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] C4d antibody Message-ID: Hello everyone, I just receive a call from a pathologist who wants the C4d antibody order from the company QUIDEL. Does anyone knows about this company and how to contact them. She also mentionned about the secondary antibody; ALEXA Fluor goat anti-mouse. It's to be used, of course, on frozen section . I am currently using the antibody C4d (BI-RC4D) from Biomedica , on paraffin section, and very happy with the result. But , she still wants to have the antibody mentionned above, to try. Can anybody help me, please ??? Thanks in advance _________________________________________________________________ MSN Messenger : discutez en direct avec vos amis ! http://messenger.fr.msn.ca/ From MAUGER <@t> email.chop.edu Thu Mar 25 10:53:22 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] C4d antibody Message-ID: Dear Histonet, I am preparing to post a notice for a job and would like to target histotechs from the Delaware valley area- PA, NJ, DE. I was wondering if the Histology Societies from those states have a mailing list available. If anyone knows how I can get these lists, or who to email directly with the request, please reply. Thanks, Jo Mauger >>> "carmen loiselle" 03/25/04 11:41AM >>> Hello everyone, I just receive a call from a pathologist who wants the C4d antibody order from the company QUIDEL. Does anyone knows about this company and how to contact them. She also mentionned about the secondary antibody; ALEXA Fluor goat anti-mouse. It's to be used, of course, on frozen section . I am currently using the antibody C4d (BI-RC4D) from Biomedica , on paraffin section, and very happy with the result. But , she still wants to have the antibody mentionned above, to try. Can anybody help me, please ??? Thanks in advance _________________________________________________________________ MSN Messenger : discutez en direct avec vos amis ! http://messenger.fr.msn.ca/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Thu Mar 25 11:20:43 2004 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Mouse Histological Atlas, another book Message-ID: <494304423C63E246A5CF87A3AEEB577011EE18@bumail01.barrynet.barry.edu> Gude's Histological Atlas of the Laboratory Mouse has been out of print for many years, and used copies are unavailable. Allen A. Smith, Ph.D. Professor of Anatomy School of Graduate Medical Sciences Barry University Miami Shores, FL -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gayle Callis Sent: Wednesday, March 24, 2004 6:17 PM To: andreah@imclone.com; Histonet@lists.utsouthwestern.edu Subject: [Histonet] Mouse Histological Atlas, another book Andrea asked for a histology atlas, not just the anatomy atlas. Here it is: Histological Atlas of the Laboratory Mouse, WD Gude, GE Cosgrove and GP Hirsch Plenum Press, NY and London, 1982 ISBN#0-306-40686-1 No price. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 gentras <@t> vetmed.auburn.edu Thu Mar 25 11:28:49 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Fwd: Shelf life of 20% PFA/ 0.1M PB Message-ID: <6.0.1.1.0.20040325112322.0259d2b0@mailhost.vetmed.auburn.edu> Special thanks to all. We use 4% PFA / 0.1M PB as our standard fixative for certain protocols. And I'm aware that it changes form after 2 weeks. But, I make my 4% from this 20% stock. And I apologize for an error in my original inquiry the 20% is made in distilled H20 and not 0.1M PB. Thanks again. Atoska >X-Sender: gcallis@gemini.msu.montana.edu >X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) >Date: Wed, 24 Mar 2004 14:54:12 -0700 >To: "Atoska S. Gentry" , > Histonet@lists.utsouthwestern.edu >From: Gayle Callis >Subject: Shelf life of 20% PFA/ 0.1M PB > >I have never made a 20% solution, rather a working/stock 4% (4 gm PFA in >100 ml Dulbeccos PBS) In general, this is a common concentration for most >PFA fixations and we tend to dilute the 4% for lesser concentration. > >Some people like their PFA fresh, then use immediately. We store 4% PFA >for several months, but if there is any cloudiness/turbidity in solution, >it is tossed. Months is a variable (not to mention people handling the >reagent - the piggy factor!) - we have had 4% PFA go bad within a month and >good as long as 6 months. A lot depends on how often people dip into the >reagent, if they are careless about sticking pipettes into stock or leave >at RT for hours before returning to refrig. > >Personal rule: store in a clean, clear glass bottle and check for turbid, >cloudy solution before each use. Toss if that cloudy flocculant ppt shows >up. This was discussed a couple of years ago on Histonet, you might want to >go into archives. > > >Gayle Callis >MT,HT,HTL(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University - Bozeman >PO Box 173610 >Bozeman MT 59717-3610 >406 994-6367 (lab with voice mail) >406 994-4303 (FAX) Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From gcallis <@t> montana.edu Thu Mar 25 11:38:07 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] C4d antibody In-Reply-To: Message-ID: <3.0.6.32.20040325103807.00bc9270@gemini.msu.montana.edu> Buy this secondary from Molecular Probes, that is the only source as far as I know unless you want to conjugate the Alexa dye to your own secondary antibody. Molecular Probes has kits to do conjugations. You did NOT say which Alexa fluor you want, their are many different Alexa fluorophore conjugates in different colors or if you want a FITC (fluorocein equivalent i.e FITC) then it would be goat antiMouse-Alexa 488. Molecular Probes has an extensive and wonderful website. If they don't have goat antiMouse Alexa 488, they will have other host secondary conjugated to their Alexa dyes. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From LElby <@t> hsh.org Thu Mar 25 13:32:53 2004 From: LElby <@t> hsh.org (Elby, Lynette) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Please unsubscribe Message-ID: <86EF8BDFC3E8D411BE6500508B2E176C0726C305@hshmail.hsh.org> Confidentiality Disclaimer: The information contained in this communication may be confidential, is intended for the use of the recipient named above, and may be legally privileged. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, or copying of this communication, or any of its contents, is strictly prohibited. If you received this communication in error, please resend this communication to the sender and delete the original message and any copy of it from your computer system. Thank You From juan.gutierrez <@t> christushealth.org Thu Mar 25 13:58:01 2004 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] C4d antibody Message-ID: Quidel Corporation 265 North Whisman Road Mountain View, CA 94043 (800)524-6318 Cat.# A213 Murine monoclonal anti-human C4d Juan -----Original Message----- From: carmen loiselle [mailto:carmen_loiselle@hotmail.com] Sent: Thu 3/25/2004 10:41 AM To: histonet@pathology.swmed.edu Cc: Subject: [Histonet] C4d antibody Hello everyone, I just receive a call from a pathologist who wants the C4d antibody order from the company QUIDEL. Does anyone knows about this company and how to contact them. She also mentionned about the secondary antibody; ALEXA Fluor goat anti-mouse. It's to be used, of course, on frozen section . I am currently using the antibody C4d (BI-RC4D) from Biomedica , on paraffin section, and very happy with the result. But , she still wants to have the antibody mentionned above, to try. Can anybody help me, please ??? Thanks in advance _________________________________________________________________ MSN Messenger : discutez en direct avec vos amis ! http://messenger.fr.msn.ca/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From MAUGER <@t> email.chop.edu Thu Mar 25 14:23:04 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Job Opening Message-ID: The Childrens' Hospital of Philadelphia is seeking candidates for a position in our Core Histology Lab.This full time position offers competitive salary and excellent benefits package. HT(ASCP) certification or equivalent by experience is required Duties will include basic histology on research specimens,as well as special stains and immunohistochemistry. Excellent cutting skills, good troubleshooting ability, and computer literacy are a must. A highly motivated individual who can communicate and interact well with different research groups is urged to reply. Please forward your resume to: pawelb@email.chop.edu From gentras <@t> vetmed.auburn.edu Thu Mar 25 15:51:19 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:43 2005 Subject: Fwd: Re: [Histonet] Fwd: Shelf life of 20% Message-ID: <6.0.1.1.0.20040325154233.02596d60@mailhost.vetmed.auburn.edu> Yes, you're correct but, I'm not sure if I will be able to determine stability by visualization alone; because my 4% doesn't get cloudy after 2 weeks I've just been advised that it converts to formalin after that period of time. It is a pain to go into solution but thankfully the protocol I use recommends heating it until it reaches 65C then adding 3.6 ml of 1N NaOH to help dissolve the PFA. After cooling I Q.S. and dilute to 4% with Sorensen's Buffer. Will most definitely let you know if I get it figured out. >X-Sender: gcallis@gemini.msu.montana.edu >X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) >Date: Thu, 25 Mar 2004 14:28:37 -0700 >To: "Atoska S. Gentry" >From: Gayle Callis >Subject: Re: [Histonet] Fwd: Shelf life of 20% PFA/ 0.1M PB > >Sounds like an experiment about to happen! Can you get powder into >solution at 20% concentration, a solubility consideration? > >I make 4% weeks (usually 500 mls) in advance per than answer I gave, doing >it a day ahead is sometimes a bother - but it is a gamble to see if it >stays viable. My EM tech made up 4% PFA in a Sorenson phosphate buffer and >he taught me the cloudy solution trick. He kept his for weeks also, and >you know how picky EM is -----. Interesting that I can make it up, split >the batch I made, send one batch to a lab and keep other in my lab. Theirs >goes bad in a couple of weeks (lots of whining and long faces!) and mine >stays good for months. One gal using it is sloppy and probably the reason >theirs always goes bad! Consequently, I don't volunteer to make it up for >them too often. > >If you find out shelf life on 20%, let me know. Have fun - > >Gayle Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From Linresearch <@t> aol.com Fri Mar 26 05:15:01 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] (no subject) Message-ID: <129.3d966017.2d956ab5@aol.com> Hello, Can anyone recommend antibodies to differentiate necrotic and apoptotic cells? I have used the tunnel assay and gotten good results in labeling both type of cells. If someone with experience with Caspase 3, could share their supplier and protocol for this Ab, it would be greatly appreciated. Also, is there a specific marker for necrotic cells? Thanks, Lin From c.m.vanderloos <@t> amc.uva.nl Fri Mar 26 05:55:44 2004 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] RE: CD4/CD25/TGFB Antibodies; Filter choices for Leica Message-ID: <3de9093dcc81.3dcc813de909@amc.uva.nl> Dear Juan, Persumed your investigation is on human tissues I have the following suggestions for you. For double staining of CD4/CD25 and CD8/CD25 I would strongly recommend you to focus on cryostat sections for two reasons. In the first place I have tested a couple of CD25 antibodies that suppose to work on FFPE sections but so far I am not very impressed. CD25 staining on cryo's seems much more reliable to me (DAKO, Becton&Dickinson, etc.). In the second place, the performance of fluorescence staining techniques on FFPE sections is not as ideal because of autofluorescence of formaldehyde-fixed tissues. Again, acetone-fixed cryostat sections do not share this type of problem. For a protocol you may test this: 1. CD4 or CD8 2. GAM/fluorochrome 1 3. Normal Mouse Serum 4. CD25/fluorochrome 2 On the otherhand the IHC staining of TGFB on cryo's, should be considered as tricky and perhaps even impossible (despite many papers that have been published...). The point is that TGFB is a small protein that tends to leak away from your cryo's (see: JHC 49:699-709, 2001 "IHC detection of IFNgamma: fake or fact?"). Although I don't have personal experience with TGFB, I have heard about problems about the IHC detection completeley similar to our IFNgamma IHC visualization failure. TGFB is perhaps detected much more reliable using optimally fixed FFPE sections. Given of course that the antigen is surviving the embedding procedure. Lots of success, Chris van der Loos, PhD Dept. of Pathology Academical Medical Center Amsterdam - The Netherlands ----- Original Message ----- >From Juan Solon Date Wed, 24 Mar 2004 09:36:30 +0000 To histonet@lists.utsouthwestern.edu Subject [Histonet] CD4/CD25/TGFB Antibodies; Filter choices for Leica Dear All, As part of a study on mucosal immunity in The Gambia, W. Africa, I plan to double stain antral and duodenal specimens for CD4 and CD25 & CD8 and CD25. Also, we would attempt to triple stain for TGF beta on these subsets. We have access to a Leica DMRXE although the filter cubes are all currently long pass suppression filters . I have both formalin-fixed and frozen sections of the antrum and the duodenum. I would like to ask for advise on several points: 1. Can anyone share their experience on antibody combinations known to work for CD4-25, CD8-25 and triple staining for CD4-CD25-TGFB either with vectastain elite (peroxidase) or immunofluorescence. I have compiled a list of CD4, CD8, CD25, TGFB antibodies from Dako, Ancell, Novocastra, BD, and Serotec, but before purchasing any of these, I thought it would be wise to ask advise from the Histonet community. 2. I would probably need to acquire different filter cubes to do simultaneous imaging of fluorescence. There are several suppliers I have encountered on the net (Omega Filters and Chroma Technology) * does anyone have experience on using filters from these companies on Leica microscopes (specifically the DMRXE). Any problems encountered with using third-party filters with Leica microscopes? Thanks. Juan Dr. Juan Antonio Solon MRC Keneba Field Station MRC Laboratories Gambia Atlantic Road, Fajara PO Box 273, Banjul The Gambia Email : juan.solon@lshtm.ac.uk Tel No: (++220) 541021 Fax No: (++220) 541022 / (++220) 496513 From yichaowu <@t> hotmail.com Fri Mar 26 08:15:51 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] C4d antibody Message-ID: Hi,loiselle, Yes, A213 from Quidel in USA. 20 minutes of 88% formic acid pretreatment would let the A213 applicable on paraffin-embedded sections too. Yichao WU Research Institute of Nephrology Jinling Hospital Nanjing China _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail From Sue.Kapoor <@t> uhsi.org Fri Mar 26 09:39:14 2004 From: Sue.Kapoor <@t> uhsi.org (Kapoor, Sue) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] correlating HPV Message-ID: <61E9F2400F53D5119CFC00508B44E33B019F5577@khmcexch.uhsi.org> Hello Histoland, I'm going to start HPV ISH on Ventana's BenchMark and I'm wondering how others have correlated their results. Did you use known (bought) controls or did you send out to have testing repeated at a reference lab? BTW, how many out in histoland run both high risk and low risk on every case? I have two pathologist that only want high risk done while one wants both on every case. Thanks in advance, Sue Kapoor, HT (ASCP) Histology Coordinator Kenosha Medical Center Kenosha, WI 262-653-5570 From cormier <@t> MIT.EDU Fri Mar 26 12:34:22 2004 From: cormier <@t> MIT.EDU (Kathleen Cormier) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] h pylori and h hepaticus Message-ID: <5.2.1.1.2.20040326132939.00ad0030@hesiod> Hey Gang, I just thought that I would throw this question out there, what antibody source/ antibody is everyone currently using for H pylori? Any one doing IHC for h hepaticus? We work w/ FFPE mouse parts, and were wondering what was being currently being used out there in the wild wild world of histology.... Thanks! Kathy Cormier Div Comp Med MIT From gcallis <@t> montana.edu Fri Mar 26 13:15:15 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] A wonderful article on bone samples and a SAFE bone saw! Message-ID: <3.0.6.32.20040326121515.00bd7a98@gemini.msu.montana.edu> CAP Today, March 2004 had a delightful, informative article on handling bone samples/calcified tissues. The key info was a bone band saw, relatively inexpensive at approx $810, Mar-Med Bone Band Saw, with diamond blade, water cooled to eliminate bone dust (no aerosol!) and blade will not amputate users fingers. Author pressed his gloved finger against the saw blade without breaking skin! It is the size of a microscope, quiet, cuts very thin slices, easy to clean, change blade, and a selection of saw blades (probably fine to coarse) - a dream saw for clinical/veterinary labs in need of a useful, safe instrument to obtain a sample suitable for fixation, decalcification,etc. Mar-Med Inc, PO Box 361201, Cleveland OH 44136, 440-572-5175. Sorry, no website listed. The article was filled with clever inexpensive ways/tools to hold bone samples during slicing plus other hints on handling difficult bone samples. Dr. Haber, author, can be reached at slhaber@stanford.edu - he might forward a copy of this publication IF you do not get CAP Today. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From ryaskovich <@t> dir.nidcr.nih.gov Fri Mar 26 14:01:39 2004 From: ryaskovich <@t> dir.nidcr.nih.gov (Yaskovich, Ruth A (NIH/NIDCR)) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] A wonderful article on bone samples and a SAFE bon e saw! Message-ID: <8F3AB322628548428A992EFB0E80F5D3A2EF85@nihexchange8.nih.gov> Gayle, I'll be speaking about this saw at the N.S.H. meeting in Toronto. The workshop is entitled Cutting it the Hard Way! I love this saw! Ruth -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Friday, March 26, 2004 2:15 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] A wonderful article on bone samples and a SAFE bone saw! CAP Today, March 2004 had a delightful, informative article on handling bone samples/calcified tissues. The key info was a bone band saw, relatively inexpensive at approx $810, Mar-Med Bone Band Saw, with diamond blade, water cooled to eliminate bone dust (no aerosol!) and blade will not amputate users fingers. Author pressed his gloved finger against the saw blade without breaking skin! It is the size of a microscope, quiet, cuts very thin slices, easy to clean, change blade, and a selection of saw blades (probably fine to coarse) - a dream saw for clinical/veterinary labs in need of a useful, safe instrument to obtain a sample suitable for fixation, decalcification,etc. Mar-Med Inc, PO Box 361201, Cleveland OH 44136, 440-572-5175. Sorry, no website listed. The article was filled with clever inexpensive ways/tools to hold bone samples during slicing plus other hints on handling difficult bone samples. Dr. Haber, author, can be reached at slhaber@stanford.edu - he might forward a copy of this publication IF you do not get CAP Today. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From padunnje <@t> iupui.edu Fri Mar 26 14:18:23 2004 From: padunnje <@t> iupui.edu (Dunn-Jena, Patsy A) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] A wonderful article on bone samples and a SAFE bone saw! Message-ID: I have been using this saw for a couple of years now and the ease of use for someone needing to do simple one cuts is excellent. It cuts bone, teeth, orthodontic resin, methyl methacrylate, plastic slides. You just change the blades to fit your needs. Patsy Dunn-Jena, RVT, LAT, HT(ASCP) Indiana University School of Dentistry Mineralized Tissue and Histology Research Laboratory Indianapolis, IN padunnje@iupui.edu -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gayle Callis Sent: Friday, March 26, 2004 2:15 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] A wonderful article on bone samples and a SAFE bone saw! CAP Today, March 2004 had a delightful, informative article on handling bone samples/calcified tissues. The key info was a bone band saw, relatively inexpensive at approx $810, Mar-Med Bone Band Saw, with diamond blade, water cooled to eliminate bone dust (no aerosol!) and blade will not amputate users fingers. Author pressed his gloved finger against the saw blade without breaking skin! It is the size of a microscope, quiet, cuts very thin slices, easy to clean, change blade, and a selection of saw blades (probably fine to coarse) - a dream saw for clinical/veterinary labs in need of a useful, safe instrument to obtain a sample suitable for fixation, decalcification,etc. Mar-Med Inc, PO Box 361201, Cleveland OH 44136, 440-572-5175. Sorry, no website listed. The article was filled with clever inexpensive ways/tools to hold bone samples during slicing plus other hints on handling difficult bone samples. Dr. Haber, author, can be reached at slhaber@stanford.edu - he might forward a copy of this publication IF you do not get CAP Today. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gentras <@t> vetmed.auburn.edu Fri Mar 26 15:35:38 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:43 2005 Subject: Fwd: RE: Re: [Histonet] Fwd: Shelf life of 20% Message-ID: <6.0.1.1.0.20040326153214.025a78b0@mailhost.vetmed.auburn.edu> Thanks George, currently I am making my paraformaldehyde fresh I was just curious about other possibilities in the event that I am face with a situation whereby it would be beneficial. In this life I'm learning that exploring all options pays off more often than not. >From: "George Cole" >To: "'Atoska S. Gentry'" >Subject: RE: Re: [Histonet] Fwd: Shelf life of 20% >Date: Fri, 26 Mar 2004 08:50:54 -0800 >X-Mailer: Microsoft Outlook, Build 10.0.2627 >Importance: Normal > >Atoska; >I'm not being prudish----the again, maybe I am--- when I >mention:----with any reagent that MIGHT go bad between uses---I always >mixed it fresh just before use. There seems to be a lot of time spent >questioning back and forth on the Histonet about when mixtures go >bad----this old retired histotech noticed about 35 years ago, that I >was worrying and fretting about freshness and usefulness of mixtures >that took less time to make than it did to worry about. Those reagents >that were bonified safe pre mixed, I used. They were very few. Most of >my routines began with mixing stains fresh in the quantity that would >give good results with the number of sides I had to stain. Once this >became a habit, it was far less complicated than crossing my fingers >about the potency of my reagents and, as a result, quality raised its >droopy head. The habit of dragging one's feet comes upon us kind of >sneaky-like--- from behind. Being a little bit bright about routine >tasks saves a lot of scrapes caused by dragging your self around. >georgecole@ev1.net > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Atoska >S. Gentry >Sent: Thursday, March 25, 2004 1:51 PM >To: Histonet >Subject: Fwd: Re: [Histonet] Fwd: Shelf life of 20% > > >Yes, you're correct but, I'm not sure if I will be able to determine >stability by visualization alone; because my 4% doesn't get cloudy after >2 >weeks I've just been advised that it converts to formalin after that >period >of time. It is a pain to go into solution but thankfully the protocol I >use >recommends heating it until it reaches 65C then adding 3.6 ml of 1N NaOH >to >help dissolve the PFA. After cooling I Q.S. and dilute to 4% with >Sorensen's Buffer. Will most definitely let you know if I get it figured >out. > > >X-Sender: gcallis@gemini.msu.montana.edu > >X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) > >Date: Thu, 25 Mar 2004 14:28:37 -0700 > >To: "Atoska S. Gentry" > >From: Gayle Callis > >Subject: Re: [Histonet] Fwd: Shelf life of 20% PFA/ 0.1M PB > > > >Sounds like an experiment about to happen! Can you get powder into > >solution at 20% concentration, a solubility consideration? > > > >I make 4% weeks (usually 500 mls) in advance per than answer I gave, >doing > >it a day ahead is sometimes a bother - but it is a gamble to see if it > >stays viable. My EM tech made up 4% PFA in a Sorenson phosphate buffer >and > >he taught me the cloudy solution trick. He kept his for weeks also, >and > >you know how picky EM is -----. Interesting that I can make it up, >split > >the batch I made, send one batch to a lab and keep other in my lab. >Theirs > >goes bad in a couple of weeks (lots of whining and long faces!) and >mine > >stays good for months. One gal using it is sloppy and probably the >reason > >theirs always goes bad! Consequently, I don't volunteer to make it up >for > >them too often. > > > >If you find out shelf life on 20%, let me know. Have fun - > > > >Gayle > >Atoska S. Gentry B.S., HT(ASCP) >Research Assistant III >Scott-Ritchey Research Center >College of Veterinary Medicine >Auburn University, AL 36849 >Phone# (334)844-5579 Fax# (334)844-5850 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From jizv66 <@t> hotmail.com Fri Mar 26 18:08:31 2004 From: jizv66 <@t> hotmail.com (Jorge Ivan Zapata Valencia) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] About H2O2 and CD Markers Message-ID: Hello everybody. I have a simple question: do someone know the effect, if any, of the different concentrations, and times, of H2O2 on the CD markers (Paraffin and/or frozen sections)? I have read that some people use 0.01%, others 0.1% and others 3% for different times, usually 10 or 30 minutes. What could happen if I use a longer time? Thank You very much. JORGE IVAN ZAPATA _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail From Linresearch <@t> aol.com Sat Mar 27 16:13:36 2004 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Abs for Apoptotic Cells Message-ID: <1da.1d8aeece.2d975690@aol.com> Hello, Can anyone recommend antibodies to? differentiate necrotic and apoptotic cells? I have used the tunnel assay and gotten good results in labeling both type of cells. If someone with experience with Caspase 3, could share their supplier and protocol for this Ab,? it would be greatly appreciated. Also, is there a specific marker for necrotic cells? Thanks, Lin From amosbrooks <@t> earthlink.net Sat Mar 27 19:01:39 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Giant Microbes In-Reply-To: References: Message-ID: <6.0.0.22.0.20040327190011.01b71220@pop.earthlink.net> Hi, This is just too cute! http://www.giantmicrobes.com Take a look, Amos From amosbrooks <@t> earthlink.net Sat Mar 27 19:07:14 2004 From: amosbrooks <@t> earthlink.net (Amos Brooks) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] H2O2 on CD markers In-Reply-To: References: Message-ID: <6.0.0.22.0.20040327190223.01b9ede8@pop.earthlink.net> Jorge, I don't have any rule of thumb to go by, merely evidence that there is an effect. Our CD4 works best if you do the H2O2 block prior to Hi pH antigen retrieval (pH 10.0 vs 7.6). In fact if you forget and block after retrieval it may not label at all. It's worth doing some experimentation on. Amos Brooks At 12:00 PM 3/27/2004, you wrote: >Hello everybody. >I have a simple question: do someone know the effect, if any, of the >different concentrations, and times, of H2O2 on the CD markers (Paraffin >and/or frozen sections)? I have read that some people use 0.01%, others 0.1% >and others 3% for different times, usually 10 or 30 minutes. What could >happen if I use a longer time? >Thank You very much. > >JORGE IVAN ZAPATA From is135475 <@t> bcm.tmc.edu Sat Mar 27 19:41:48 2004 From: is135475 <@t> bcm.tmc.edu (Isaiah G. Schauer) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] CD31/PECAM-1 Message-ID: <142C8790-8059-11D8-B01A-003065D60DC4@bcm.tmc.edu> Histonetters, Our lab has used Pharmingen's CD31 antibody (cat# 01951A/D) with great results for a couple years for paraffin-embedded mouse tissue IHC. Recently, they decided to "sublet" the production of this antibody to a company called Cymbus, which sells the antibody at 1/10th the original protein concentration. We've called Pharmingen regarding this, and they staunchly maintain that there is no difference between their antibody and the Cymbus rendition. We now have to use a 1:5 dilution, on mouse xenograft tumors fixed in 4% paraformaldehyde 16 hrs at 4C. The cost of repeatedly using this Ab has now become prohibitive for the lab. Can anyone recommend a better CD31/PECAM-1 antibody for mouse xenograft IHC on 4% PF-fixed, paraffin-embedded tissue? Histonet archive searches have turned up Santa Cruz (cat# sc-1506), Dako (cat# M0823, clone JC-70A) and Serotec (cat# MCA1334G) as possible suppliers, but I wanted to hear from someone with paraformaldehyde-fixed, mouse tissue experience. We don't currently use enzymatic digestion prior to staining, but I've read on Histonet of people using proteinase K, pronase or trypsin at varying temperatures for varying times for CD31 staining. Might anyone recommend a starting time/temp for my particular tissue type/fixation that could allow me to increase the 1:5 dilution of the Pharmingen antibody? Thanks for your help and time. Sincerely, Isaiah Schauer 3rd year graduate student Lab of David R. Rowley, Ph.D. Baylor College of Medicine Department of Molecular and Cellular Biology One Baylor Plaza, room 326D Houston, TX 77030 lab: 713-798-6221 fax: 713-790-1275 From p_bourne_14526 <@t> yahoo.com Sat Mar 27 20:11:44 2004 From: p_bourne_14526 <@t> yahoo.com (Patricia Bourne) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] H2O2 on CD markers In-Reply-To: <6.0.0.22.0.20040327190223.01b9ede8@pop.earthlink.net> Message-ID: <20040328021144.55873.qmail@web10004.mail.yahoo.com> Very interesting, we just did a test blocking with 3% H2O2 after primary and the results were much better and sharper. We use Dako TRS at ph6.1 for 40 minutes in the steamer. Put the slides on the Dako stainer do the primary incubations, wash , and then do the quenching. Try it you just might like it. We are now testing other CD markers to see if they too are improved. Pat --------------------------------- Do you Yahoo!? Yahoo! Finance Tax Center - File online. File on time. From georgecole <@t> ev1.net Sun Mar 28 03:05:01 2004 From: georgecole <@t> ev1.net (George Cole) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] Sigma 221 alkaline buffer Message-ID: <000001c414a3$c1573840$0b4dbad0@hppav> Histotechs; Mary North, the great gal who replaced me when my Grandfather Clock read---?Retire----?, tells me that the great Alkaline Buffer once listed as 221 in the Sigma catalog is now listed as Alkaline Buffer A 9226. This is the buffer that can replace the old Veronal Buffer usually listed as a component of ATP-ase and Acid Phosphatase. This old buffer contains Nembutal, a sodium salt of Pentobarbital, and requires a controlled substance license to order. 221 gives brilliant results in those two procedures, rendering the Old Dope Buffer pass?. georgecole@ev1.net From Rcartun <@t> harthosp.org Sun Mar 28 10:38:46 2004 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] h pylori and h hepaticus Message-ID: I use DakoCytomation's polyclonal antibody (B0471) for H. pylori identification in formalin-fixed human tissue. It works very well. It does cross-react with H. heilmannii; I don't have experience with other types of Helicobacter. I have also noticed weak cross-reactions with non-Helicobacter organisms from time to time. I have also used Novocastra's monoclonal Campylobacter jejuni antibody to label H. pylori. It works very well and does not cross-react with non-Campylobacter/Helicobacter organisms. Richard Cartun >>> Kathleen Cormier 03/26/04 01:34PM >>> Hey Gang, I just thought that I would throw this question out there, what antibody source/ antibody is everyone currently using for H pylori? Any one doing IHC for h hepaticus? We work w/ FFPE mouse parts, and were wondering what was being currently being used out there in the wild wild world of histology.... Thanks! Kathy Cormier Div Comp Med MIT _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From siksik03 <@t> comcast.net Sun Mar 28 14:48:00 2004 From: siksik03 <@t> comcast.net (Steven E. Slap) Date: Fri Sep 16 15:22:43 2005 Subject: [Histonet] A wonderful article on bone samples and a SAFE bone saw! In-Reply-To: <3.0.6.32.20040326121515.00bd7a98@gemini.msu.montana.edu> References: <3.0.6.32.20040326121515.00bd7a98@gemini.msu.montana.edu> Message-ID: Dear Gayle & HistoNetters I have worked extensively with the Mar-Med bone saw and it is every bit as good as the article claims. The website is http://www.mar-med.com. best regards, Steven Slap At 12:15 PM -0700 3/26/04, Gayle Callis wrote: >CAP Today, March 2004 had a delightful, informative article on handling >bone samples/calcified tissues. The key info was a bone band saw, >relatively inexpensive at approx $810, Mar-Med Bone Band Saw, with diamond >blade, water cooled to eliminate bone dust (no aerosol!) and blade will not >amputate users fingers. Author pressed his gloved finger against the saw >blade without breaking skin! > >It is the size of a microscope, quiet, cuts very thin slices, easy to >clean, change blade, and a selection of saw blades (probably fine to >coarse) - a dream saw for clinical/veterinary labs in need of a useful, >safe instrument to obtain a sample suitable for fixation, >decalcification,etc. > >Mar-Med Inc, PO Box 361201, Cleveland OH 44136, 440-572-5175. Sorry, no >website listed. > >The article was filled with clever inexpensive ways/tools to hold bone >samples during slicing plus other hints on handling difficult bone samples. > Dr. Haber, author, can be reached at slhaber@stanford.edu - he might >forward a copy of this publication IF you do not get CAP Today. From John.ROPHAEL <@t> svhm.org.au Mon Mar 29 00:15:37 2004 From: John.ROPHAEL <@t> svhm.org.au (ROPHAEL John (SVHM)) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Query Message-ID: Dear Sir / Madam I was wondering if anyone had protocols for the following 1 ? IHC of paraffin embedded (PFA fixed) mouse tissue using NG2 Ab to demonstrate pericytes 2 IHC of paraffin embedded (PFA fixed) mouse tissue using Pref-1 Ab to demonstrate preadipocytes Many thanks John Rophael MBBS Hons -------------- next part -------------- Disclaimer : The contents of this e-mail including any attachments are intended only for the person or entity to which this e-mail is addressed and may contain confidential, privileged and/or commercially sensitive material. If you are not, or believe you may not be, the intended recipient, please advise the sender immediately by return e-mail, delete this e-mail and destroy any copies. From MAUGER <@t> email.chop.edu Mon Mar 29 06:44:53 2004 From: MAUGER <@t> email.chop.edu (Joanne Mauger) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] CD31/PECAM-1 Message-ID: To Isaiah' I used CD31 from Pharmingen also, about 6 years ago. I used a 1:50 dilution after retreieval with pepsin for 30-40 min. at 37C(depending on length of fixation. If the tumors are human cells in mice, the Dako anti human CD31 will probably work at about the same dilution, with pepsin as well. Jo >>> "Isaiah G. Schauer" 03/27/04 08:41PM >>> Histonetters, Our lab has used Pharmingen's CD31 antibody (cat# 01951A/D) with great results for a couple years for paraffin-embedded mouse tissue IHC. Recently, they decided to "sublet" the production of this antibody to a company called Cymbus, which sells the antibody at 1/10th the original protein concentration. We've called Pharmingen regarding this, and they staunchly maintain that there is no difference between their antibody and the Cymbus rendition. We now have to use a 1:5 dilution, on mouse xenograft tumors fixed in 4% paraformaldehyde 16 hrs at 4C. The cost of repeatedly using this Ab has now become prohibitive for the lab. Can anyone recommend a better CD31/PECAM-1 antibody for mouse xenograft IHC on 4% PF-fixed, paraffin-embedded tissue? Histonet archive searches have turned up Santa Cruz (cat# sc-1506), Dako (cat# M0823, clone JC-70A) and Serotec (cat# MCA1334G) as possible suppliers, but I wanted to hear from someone with paraformaldehyde-fixed, mouse tissue experience. We don't currently use enzymatic digestion prior to staining, but I've read on Histonet of people using proteinase K, pronase or trypsin at varying temperatures for varying times for CD31 staining. Might anyone recommend a starting time/temp for my particular tissue type/fixation that could allow me to increase the 1:5 dilution of the Pharmingen antibody? Thanks for your help and time. Sincerely, Isaiah Schauer 3rd year graduate student Lab of David R. Rowley, Ph.D. Baylor College of Medicine Department of Molecular and Cellular Biology One Baylor Plaza, room 326D Houston, TX 77030 lab: 713-798-6221 fax: 713-790-1275 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mprice26 <@t> juno.com Mon Mar 29 07:17:31 2004 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] TSH Brochure Message-ID: <20040329.051820.11628.73138@webmail24.nyc.untd.com> Histonetters, Can someone send me a brochure for the TSH (April 15-19). I need the hotel info etc. to turn in for one of my employees to be able to go to. Is it available online? Thank you. Marsha Price ________________________________________________________________ The best thing to hit the Internet in years - Juno SpeedBand! Surf the Web up to FIVE TIMES FASTER! Only $14.95/ month - visit www.juno.com to sign up today! From rjr6 <@t> psu.edu Mon Mar 29 08:54:44 2004 From: rjr6 <@t> psu.edu (Roberta Horner) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Multi-chambered cassettes Message-ID: <007b01c4159d$c5b1d030$8861ba92@padlspsu.psu.edu> I am looking for multi-chambered cassettes. I have one now that I don't like and would like to try something different. What all companies sell these? Thank you, Roberta Horner HT/HTL Animal Diagnostic Lab Penn State University From gcallis <@t> montana.edu Mon Mar 29 09:31:34 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] About H2O2 and CD Markers In-Reply-To: Message-ID: <3.0.6.32.20040329083134.00bfb910@gemini.msu.montana.edu> We prefer to use the least concentration of hydrogen peroxide, and always in a PBS rather than methanol solvent when blocking endogenous peroxidase in FROZEN SECTIONS. You will find that the higher the concentration of H2O2, the more likely frozen sections (which is minimally fixed with acetone or even acetone/absolute alcohol) will be chewed off the slide. The section was totally worthless with badly damaged morphology. This happened when we first set up immunostaining. We sawa bubble appear on top of the section when more concentrated H2O2 blockers are applied. That led us to the two blocking methods below. We use either the DAKO S2001 blocker that contains buffer, 0.03% hydrogen peroxide and a bit of sodium azide and is used for 10 minutes, although the package insert indicates less, we have done 10 to 15 min without problems. It is very gentle and effective. The best blocking is done by a glucose oxidase method, which preserves morphology in frozen sections AND quenches peroxidase and pseudoperoxidases completely. This is by far our favorite endogenous peroxidase blocker. You should try higher concentrations and observe IF your frozen sections come off the slides or damaged by the peroxide. When this happens, then you should consider a gentler method. Methanol is NEVER used with CD marker staining, it is known to cause diminshed staining of these markers, and was discussed by people staining paraffin sections and experienced this problem. They switched to buffer with hydrogen peroxide. Methanolic bridges or cross links occur - this is also found in the literature (Elias books). Paraffin sections are much more robust than frozen sections, and paraffin withstand stronger peroxide concetrations although methanol is still avoided for murine CD markers. You can avoid peroxidase blocking entirely IF you use alkaline phosphatase enzyme methods. At 12:08 AM 3/27/2004 +0000, you wrote: >Hello everybody. >I have a simple question: do someone know the effect, if any, of the >different concentrations, and times, of H2O2 on the CD markers (Paraffin >and/or frozen sections)? I have read that some people use 0.01%, others 0.1% >and others 3% for different times, usually 10 or 30 minutes. What could >happen if I use a longer time? >Thank You very much. > >JORGE IVAN ZAPATA > >_________________________________________________________________ >Help STOP SPAM with the new MSN 8 and get 2 months FREE* >http://join.msn.com/?page=features/junkmail > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Mar 29 09:40:49 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] H2O2 on CD markers Message-ID: <3.0.6.32.20040329084049.00c09160@gemini.msu.montana.edu> >Date: Mon, 29 Mar 2004 08:40:21 -0700 >To: Amos Brooks >From: Gayle Callis >Subject: Re: [Histonet] H2O2 on CD markers >In-Reply-To: <6.0.0.22.0.20040327190223.01b9ede8@pop.earthlink.net> >References: > >Amos, > >Are you working with human or mouse tissues? Murine CD4 and CD8 is a total failure with formalin fixed paraffin tissues, and no retrieval or digestion recovers these antigens. They do work, however with zinc TRIS buffer fixation where no retrieval is needed, but endogenous peroxidase cannot be successfully/totally blocked with this fixation. Alkaline phosphatase methods work well with ZnTRIS to avoid peroxidase blocking. This is why we stay with frozen sections for murine CD4 and CD8. > > >At 07:07 PM 3/27/2004 -0600, you wrote: >>Jorge, >> I don't have any rule of thumb to go by, merely evidence that >>there is an effect. Our CD4 works best if you do the H2O2 block prior to Hi >>pH antigen retrieval (pH 10.0 vs 7.6). In fact if you forget and block >>after retrieval it may not label at all. It's worth doing some >>experimentation on. >>Amos Brooks >> >>At 12:00 PM 3/27/2004, you wrote: >>>Hello everybody. >>>I have a simple question: do someone know the effect, if any, of the >>>different concentrations, and times, of H2O2 on the CD markers (Paraffin >>>and/or frozen sections)? I have read that some people use 0.01%, others 0.1% >>>and others 3% for different times, usually 10 or 30 minutes. What could >>>happen if I use a longer time? >>>Thank You very much. >>> >>>JORGE IVAN ZAPATA >> >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> From lleach <@t> uic.edu Mon Mar 29 09:50:35 2004 From: lleach <@t> uic.edu (Lu Leach) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Versican Message-ID: <6.0.1.1.2.20040329094426.02357948@tigger.uic.edu> Dear Histonetters, Does anyone have a protocol for the Versican antibody otherwise known as Chondroitin Sulfate Proteoglycan 2? The specification sheet is not very helpful and the technical representative didn't have much infomation to offer. I was also curious about what folks use for a control. Any help would be greatly appreciated. Thank You, Lu Leach University of Illinois at Chicago lleach@uic.edu 312-996-3869 From gcallis <@t> montana.edu Mon Mar 29 09:58:23 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] CD31/PECAM-1 In-Reply-To: <142C8790-8059-11D8-B01A-003065D60DC4@bcm.tmc.edu> Message-ID: <3.0.6.32.20040329085823.00bfb910@gemini.msu.montana.edu> Isaiah, Another possibility is make the antibody from cell line clone (ATCC). The supernate can be used for staining, undiluted, and can contain around 10 ug/ml of antibody. We do this frequently with antibodies we use in large quantities. At 07:41 PM 3/27/2004 -0600, you wrote: >Histonetters, > >Our lab has used Pharmingen's CD31 antibody (cat# 01951A/D) with great >results for a couple years for paraffin-embedded mouse tissue IHC. >Recently, they decided to "sublet" the production of this antibody to a >company called Cymbus, which sells the antibody at 1/10th the original >protein concentration. We've called Pharmingen regarding this, and they >staunchly maintain that there is no difference between their antibody >and the Cymbus rendition. Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Luis.Chiriboga <@t> med.nyu.edu Mon Mar 29 10:10:34 2004 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] About H2O2 and CD Markers In-Reply-To: Message-ID: I have used up to 10% H2O2(5 minutes Max) for extremely bloody cell blocks (FFPE) but wasn't looking at CD markers. Routinely use ~3% for B&T cell markers on FFPE and haven't noticed any deleterious effects. I have also used for (<3%diluted in PBS, no methanol) for frozen sections CD staining. Just need to be careful, bubbling doesn't lift off section. You will need to determine time and concentration to best suit your tissue and needs. Hope this helps Luis -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jorge Ivan Zapata Valencia Sent: Friday, March 26, 2004 7:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] About H2O2 and CD Markers Hello everybody. I have a simple question: do someone know the effect, if any, of the different concentrations, and times, of H2O2 on the CD markers (Paraffin and/or frozen sections)? I have read that some people use 0.01%, others 0.1% and others 3% for different times, usually 10 or 30 minutes. What could happen if I use a longer time? Thank You very much. JORGE IVAN ZAPATA _________________________________________________________________ Help STOP SPAM with the new MSN 8 and get 2 months FREE* http://join.msn.com/?page=features/junkmail _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Mar 29 10:08:35 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Multi-chambered cassettes In-Reply-To: <007b01c4159d$c5b1d030$8861ba92@padlspsu.psu.edu> Message-ID: <3.0.6.32.20040329090835.00bfb910@gemini.msu.montana.edu> Two sources we have used Fisher and Evergreen Scientific (Evergreen should have samples available if you call them). Evergreen has a better selection for different numbers of chambers. They come 2, 4, 6, 8 chambers. Question: what is it about multichamber cassettes you don't like? We have to insure they do NOT float, that they fill and sink to bottom of fixative container. This was true of brands tried from both sources. At 09:54 AM 3/29/2004 -0500, you wrote: >I am looking for multi-chambered cassettes. I have one now that I don't >like and would like to try something different. What all companies sell >these? > >Thank you, > >Roberta Horner HT/HTL > >Animal Diagnostic Lab > >Penn State University > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From arunams <@t> interchange.ubc.ca Mon Mar 29 10:33:52 2004 From: arunams <@t> interchange.ubc.ca (Aruna Mahendra Somasiri) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Immunos for CD4 and CD8 In-Reply-To: <3.0.6.32.20040329083134.00bfb910@gemini.msu.montana.edu> Message-ID: HI every one, I have a problem with my CD4 and CD8 staining on frozen sections. Even after 3% H2O2, and IgG blocks I get very hight levels of background in my IgG control slides. I amd using frozen mouse spleen sections. Any help woud be appreciated. Thanks Aruna SOmasiri From yichaowu <@t> hotmail.com Mon Mar 29 10:48:54 2004 From: yichaowu <@t> hotmail.com (yichao wu) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] C4d antibody Message-ID: Hi, Doug, I used the concentration of 1:250 dilution in PBS. Yichao >From: "Doug Geddes" >To: >Subject: RE: [Histonet] C4d antibody >Date: Mon, 29 Mar 2004 10:54:35 -0500 > >Hi Yichao, > >What concentration do you use your antibody at. > >Doug Geddes BSc, MLT >London Health Sciences Centre >London, Ontario, Canada > > > >>> "yichao wu" 03/26/04 09:15AM >>> >Hi,loiselle, > >Yes, A213 from Quidel in USA. > >20 minutes of 88% formic acid pretreatment would let the A213 applicable on >paraffin-embedded sections too. > >Yichao WU > >Research Institute of Nephrology >Jinling Hospital >Nanjing China > >_________________________________________________________________ >Help STOP SPAM with the new MSN 8 and get 2 months FREE* >http://join.msn.com/?page=features/junkmail > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _________________________________________________________________ Add photos to your e-mail with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From ampippen <@t> duke.edu Mon Mar 29 11:32:46 2004 From: ampippen <@t> duke.edu (Anne Pippen) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] human Y-chromosome antibody Message-ID: <047b01c415b3$d9d65c50$c8391098@CARDIOANNE429> Hi there, Are any of you aware of a human Y-chromosome antibody? If so, what company offers this product? Anne From ramona.tolliver <@t> yale.edu Mon Mar 29 12:37:13 2004 From: ramona.tolliver <@t> yale.edu (Ramona Tolliver) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Yale University - Opening Message-ID: <6.0.1.1.2.20040329133608.01db4df0@email.med.yale.edu> Good afternoon, Yale University has a Histology Manager (1st shift) position available from 7:00 a.m. - 3:00 p.m. If you know of anyone who may be interested in an outstanding growth opportunity in a department committed to excellence in patient care, teaching, and discovery, please forward this information on to them. Salary is commensurate with experience. Relocation assistance is available. Inquiries should be sent to ramona.tolliver@yale.edu. Thank you for your time! Ramona Tolliver Ramona E. Tolliver Human Resource Manager Yale University School of Medicine Department of Pathology 310 Cedar Street New Haven, CT 06520-8023 Telephone: (203) 785-6689 Fax: (203) 785-7303 From ASelf <@t> gmhsc.com Mon Mar 29 13:58:36 2004 From: ASelf <@t> gmhsc.com (Amy Self) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] DFA-TP Message-ID: Histonetters, I am looking for a lab that performs DFA-TP (direct flourescent antibody to TP) on smears prepared from syphillis lesions. If anyone out there in histoworld does this test or knows where I can send these slides to, to get this test done please let me know. Thanks in advance for your help....... Amy Self Georgetown Memorial Hospital 843-527-7179 Note: 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 paw555 <@t> yahoo.com Mon Mar 29 15:01:57 2004 From: paw555 <@t> yahoo.com (pam plumlee) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] acrylic block files Message-ID: <20040329210157.98900.qmail@web11605.mail.yahoo.com> Hi all: Can anyone refer me to a company that makes/sells the acrylic bench top block files? I've used them for years and never had the need to order new ones. Thanks, Pam __________________________________ Do you Yahoo!? Yahoo! Finance Tax Center - File online. File on time. http://taxes.yahoo.com/filing.html From mrl0627 <@t> mail.ecu.edu Mon Mar 29 15:48:18 2004 From: mrl0627 <@t> mail.ecu.edu (mrl0627@mail.ecu.edu) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Versican Message-ID: <4327112.1080596898546.JavaMail.peete@onestop9> Hello Lu; I have had great results with 2 anti-mouse antiversicans raised in rabbit by Chemicon. (GAG A and GAG B) in paraffin embedded tissue fixed either in Carnoy's, Dent's or 4% para. No HIER required, just Chondroitinase ABC digestion. Will be glad to share my protocol if you want. Maureen Loomer Master's candidate East Carolina University Dept of Biology -----Original Message----- From: "Lu Leach" <lleach@uic.edu> To: <histonet@lists.utsouthwestern.edu> Sent: Mon, 29 Mar 2004 10:50:35 -0500 Subject: [Histonet] Versican From plxja <@t> nottingham.ac.uk Mon Mar 29 15:51:30 2004 From: plxja <@t> nottingham.ac.uk (Joelle Alcock) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Apoptosis detection Message-ID: Hi all I am looking for an IHC technique to detect apoptosis cells on testis sections. I have previously used the Tunnel assay but would like an alternative as often meiotic cells are also labelled and I want to avoid this. I have seen the Histone labelling assay by UPSTATE but I think that is mainly for ICC not IHC. Thanks for any help Joelle Alcock Institute of Genetics Queens Medical Centre Nottingham, UK From nakagawa <@t> umn.edu Mon Mar 29 16:20:41 2004 From: nakagawa <@t> umn.edu (Yasushi Nakagawa) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] AO 925 Knife sharpener Message-ID: Hi We recently bought a used AO925 microtome knife sharpener. We found that the sharpening kit that we used to use for other knife sharpener (PATHCO Microtome knife sharpening kit, PA-2000) is not deep enough to accommodate the back-and-forth movement of the base plate of AO925; the knife gets out of the abrasive sheet during the sharpening process, which is not acceptable. Does anybody know which sharpening kit (or abrasive sheets) fit this microtome? And where can we buy them? We also did not get the manual for the microtome. If somebody has it and can send it by email or by fax, that would be great. Thank you. Yasushi Nakagawa -- Yasushi Nakagawa, M.D., Ph.D. Assistant Professor, Department of Neuroscience Faculty member, Stem Cell Institute University of Minnesota Medical School Department of Neuroscience 6-145 Jackson Hall, 321 Church Street SE. Minneapolis, MN55455 Phone 1-612-625-4497 (office), 1-612-626-0674 (lab) Fax: 1-612-626-2553 (lab) From bhewlett <@t> cogeco.ca Mon Mar 29 18:01:03 2004 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Apoptosis detection References: Message-ID: <000601c415ea$18493d00$6500a8c0@mainbox> Joelle, Apoptosis is apoptosis and may well be part of the meiotic process. You may wish to consider APAF1 or the one of the caspase antibodies from Novacastra as an alternative to TUNEL. You could also check with Neil Hand in the pathology IHC lab at Queens for his recommendation. Regards, Bryan ----- Original Message ----- From: "Joelle Alcock" To: Sent: Monday, March 29, 2004 4:51 PM Subject: [Histonet] Apoptosis detection Hi all I am looking for an IHC technique to detect apoptosis cells on testis sections. I have previously used the Tunnel assay but would like an alternative as often meiotic cells are also labelled and I want to avoid this. I have seen the Histone labelling assay by UPSTATE but I think that is mainly for ICC not IHC. Thanks for any help Joelle Alcock Institute of Genetics Queens Medical Centre Nottingham, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Myri37 <@t> aol.com Tue Mar 30 01:15:30 2004 From: Myri37 <@t> aol.com (Myri37@aol.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] cells makers of periodontal ligament Message-ID: <6A464983.328468B1.0005167B@aol.com> hello everyone am looking for specific markers to identify cells in cell population extracted from teeth (cementoblasts, PDL fibroblasts, gingival fibroblasts, Malassez cells, epithelial jonction cells...) i would like testing markers cells by FACS, or immunostaining Coul you help me, do you konw any specific marker ? thanking you very much for anyhelp Myriam baali Natural implant From Jackie.O'Connor <@t> abbott.com Tue Mar 30 07:06:11 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Apoptosis detection Message-ID: The Caspase 3 antibody demonstrates only apoptotic cells. Jackie O'Connor Abbott Laboratories Abbott Park, IL "Joelle Alcock" Sent by: histonet-bounces@lists.utsouthwestern.edu 03/29/2004 03:51 PM To: cc: Subject: [Histonet] Apoptosis detection Hi all I am looking for an IHC technique to detect apoptosis cells on testis sections. I have previously used the Tunnel assay but would like an alternative as often meiotic cells are also labelled and I want to avoid this. I have seen the Histone labelling assay by UPSTATE but I think that is mainly for ICC not IHC. Thanks for any help Joelle Alcock Institute of Genetics Queens Medical Centre Nottingham, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Tue Mar 30 08:07:58 2004 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] AO 925 Knife sharpener Message-ID: Hi Dr. Nakagawa, The AO925 came from Leica's old American Optical group. The 925, 903, and 930 all used a selenoid to push the knife against the glass plate and tended to move around the films. I am mailing a copy of the old instruction manual. Wet abrasives are available to replace films. 14041819700 Coarse Abrasive, 14041819701 Fine Abrasive and 14041819702 Hone Glass compound are still available for the sharpeners. If you have further questions, please feel free to phone me. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 Yasushi Nakagawa To: Sent by: cc: histonet-bounces@lists.utsouth Subject: [Histonet] AO 925 Knife sharpener western.edu 03/29/2004 04:20 PM Hi We recently bought a used AO925 microtome knife sharpener. We found that the sharpening kit that we used to use for other knife sharpener (PATHCO Microtome knife sharpening kit, PA-2000) is not deep enough to accommodate the back-and-forth movement of the base plate of AO925; the knife gets out of the abrasive sheet during the sharpening process, which is not acceptable. Does anybody know which sharpening kit (or abrasive sheets) fit this microtome? And where can we buy them? We also did not get the manual for the microtome. If somebody has it and can send it by email or by fax, that would be great. Thank you. Yasushi Nakagawa -- Yasushi Nakagawa, M.D., Ph.D. Assistant Professor, Department of Neuroscience Faculty member, Stem Cell Institute University of Minnesota Medical School Department of Neuroscience 6-145 Jackson Hall, 321 Church Street SE. Minneapolis, MN55455 Phone 1-612-625-4497 (office), 1-612-626-0674 (lab) Fax: 1-612-626-2553 (lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________ From AFeatherstone <@t> KaleidaHealth.Org Tue Mar 30 07:57:01 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] missing specimens Message-ID: I would like to know what procedures are being done for specimens that are missing. (say no specimen in container). How is everyone handling the situation and what exact steps are taken at the time of discovery up until writing the report. Thanks much. Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 AFeatherstone <@t> KaleidaHealth.Org Tue Mar 30 07:56:01 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Missing specimens Message-ID: I would like to know what procedures are being done for specimens that are missing. (say no specimen in container). How is everyone handling the situation and what exact steps are taken at the time of discovery up until writing the report. Thanks much. Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 MDiCarlo <@t> KaleidaHealth.Org Tue Mar 30 09:10:42 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] AO 925 Knife sharpener Message-ID: I faxed a copy of the manual. -----Original Message----- From: Yasushi Nakagawa [mailto:nakagawa@umn.edu] Sent: Monday, March 29, 2004 17:21 To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] AO 925 Knife sharpener Hi We recently bought a used AO925 microtome knife sharpener. We found that the sharpening kit that we used to use for other knife sharpener (PATHCO Microtome knife sharpening kit, PA-2000) is not deep enough to accommodate the back-and-forth movement of the base plate of AO925; the knife gets out of the abrasive sheet during the sharpening process, which is not acceptable. Does anybody know which sharpening kit (or abrasive sheets) fit this microtome? And where can we buy them? We also did not get the manual for the microtome. If somebody has it and can send it by email or by fax, that would be great. Thank you. Yasushi Nakagawa -- Yasushi Nakagawa, M.D., Ph.D. Assistant Professor, Department of Neuroscience Faculty member, Stem Cell Institute University of Minnesota Medical School Department of Neuroscience 6-145 Jackson Hall, 321 Church Street SE. Minneapolis, MN55455 Phone 1-612-625-4497 (office), 1-612-626-0674 (lab) Fax: 1-612-626-2553 (lab) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 veronese13 <@t> hotmail.com Tue Mar 30 11:02:23 2004 From: veronese13 <@t> hotmail.com (Denisa S. Melichian) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Eosinophils & Fibroblasts Message-ID: Hello Everyone, Does anyone have a successful protocol for staining eosionphils and fibroblasts in skin tissue? If so could you possibly share? Thank you, Denisa Melichian Research, Dept. of Medicine UIHMC _________________________________________________________________ [1]MSN Toolbar provides one-click access to Hotmail from any Web page FREE download! References 1. http://g.msn.com/8HMBENUS/2740??PS= From bill501 <@t> mindspring.com Tue Mar 30 11:08:28 2004 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Missing specimens In-Reply-To: References: Message-ID: We do the following: 1. Have 3 observers confirm the specimen is missing and a statement of this made in the gross description. The fluid may be filtered and submitted. 2. Call the hospital and surgeon and discuss the specimen. Could it have ended up in another patient's jar? etc 3. Put out a report stating the results of these efforts. Dx= No specimen received... 4. Mark the case to be discussed in our QA/QC and surgery meeting. Bill At 8:56 AM -0500 3/30/04, Featherstone, Annette wrote: >I would like to know what procedures are being done for specimens that are >missing. (say no specimen in container). How is everyone handling the >situation and what exact steps are taken at the time of discovery up until >writing the report. Thanks much. -- ______________ Bill Blank, MD Heartland Lab, Inc From Dixon.Leslie <@t> mayo.edu Tue Mar 30 12:11:00 2004 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] adhesive for paraffin sections Message-ID: Good morning all, I have received some old paraffin sections from another country. They do not appear to have been treated in anyway. I plan on doing some immunos that involve HIER on these sections. Is there anything other than celloidin that I can use to insure these specimens remain on the slides during staining? Thanks in advance, Leslie Mayo Clinic Research Histology From MDiCarlo <@t> KaleidaHealth.Org Tue Mar 30 12:36:37 2004 From: MDiCarlo <@t> KaleidaHealth.Org (DiCarlo, Margaret) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] adhesive for paraffin sections Message-ID: Fred Underwood told me to about using 5% titebond.(made from Titebond II wood glue that you buy in a hardwood store)- I use it for all my bone slides and the sections stay on 98% of the time during H&E staining. Using acid cleaned slides, I use a disposable pipette and drop a few drops on my slide and spread it with a kimwipe and air dry a few seconds before picking up section from the waterbath. Why not just use superfrost plus charged slides? Peggy DiCarlo HT(ASCP) -----Original Message----- From: Dixon, Leslie E. [mailto:Dixon.Leslie@mayo.edu] Sent: Tuesday, March 30, 2004 13:11 To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] adhesive for paraffin sections Good morning all, I have received some old paraffin sections from another country. They do not appear to have been treated in anyway. I plan on doing some immunos that involve HIER on these sections. Is there anything other than celloidin that I can use to insure these specimens remain on the slides during staining? Thanks in advance, Leslie Mayo Clinic Research Histology _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 dennijc <@t> vetmed.auburn.edu Tue Mar 30 12:04:01 2004 From: dennijc <@t> vetmed.auburn.edu (John C. Dennis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Fixation for lectin work Message-ID: <5.1.1.5.1.20040330120205.017c83e8@mailhost.vetmed.auburn.edu> Dear Histonetters: Does anyone know why 10% buffered formaldehyde may not be suitable for lectin labeling? John C. Dennis From psanquin <@t> lugo.usc.es Tue Mar 30 13:09:32 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Fixation for lectin work In-Reply-To: <5.1.1.5.1.20040330120205.017c83e8@mailhost.vetmed.auburn.e du> Message-ID: <3.0.6.32.20040330210932.007aab80@pop.lugo.usc.es> I do not know the reason, but I remember that few years ago that fact caused me chronic headache. Just after switching to Acetic-Alcohol fixed material I got nice labelling. Bouin fluid is also an excellent fixative for lectin studies. Sometimes aldehydes work in some tissues, so before working with lectins it is advisable to compare different fixatives. Cheers Pablo Sanchez At 12:04 p.m. 30/03/04 -0600, you wrote: >Dear Histonetters: > >Does anyone know why 10% buffered formaldehyde may not be suitable for >lectin labeling? > >John C. Dennis > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From rschoon <@t> email.unc.edu Tue Mar 30 13:19:55 2004 From: rschoon <@t> email.unc.edu (Robert Schoonhoven) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Fixation for lectin work References: <3.0.6.32.20040330210932.007aab80@pop.lugo.usc.es> Message-ID: <4069C85B.7020105@email.unc.edu> Must be more to it then that. Bouin's is made up with formaldehyde. Robert Schoonhoven Pablo S?nchez Quinteiro wrote: >I do not know the reason, but I remember that few years ago that fact >caused me chronic headache. Just after switching to Acetic-Alcohol fixed >material I got nice labelling. Bouin fluid is also an excellent fixative >for lectin studies. > >Sometimes aldehydes work in some tissues, so before working with lectins it >is advisable to compare different fixatives. > >Cheers > >Pablo Sanchez > >At 12:04 p.m. 30/03/04 -0600, you wrote: > > >>Dear Histonetters: >> >>Does anyone know why 10% buffered formaldehyde may not be suitable for >>lectin labeling? >> >>John C. Dennis >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@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 Tue Mar 30 13:56:30 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Fixation for lectin work Message-ID: <566FB0B522443D43AF02D2ADBE35A6F06359DB@UTHEVS3.mail.uthouston.edu> Pablo Bouin's fixative can be a good fixative for some IHC applications but I cannot agree that it is good for lectin studies. I did some studies on lectin binding severl year ago and found that the best fixative to be alcohol and acetone. Alcohol acetic probably is OK too. An important point here however is that there is considerable variation depending on the tissue and also on the lectin that is used. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Robert Schoonhoven Sent: Tue 3/30/2004 1:19 PM To: Pablo S?nchez Quinteiro Cc: Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Fixation for lectin work Must be more to it then that. Bouin's is made up with formaldehyde. Robert Schoonhoven Pablo S?nchez Quinteiro wrote: >I do not know the reason, but I remember that few years ago that fact >caused me chronic headache. Just after switching to Acetic-Alcohol fixed >material I got nice labelling. Bouin fluid is also an excellent fixative >for lectin studies. > >Sometimes aldehydes work in some tissues, so before working with lectins it >is advisable to compare different fixatives. > >Cheers > >Pablo Sanchez > >At 12:04 p.m. 30/03/04 -0600, you wrote: > > >>Dear Histonetters: >> >>Does anyone know why 10% buffered formaldehyde may not be suitable for >>lectin labeling? >> >>John C. Dennis >> >> >>_______________________________________________ >>Histonet mailing list >>Histonet@lists.utsouthwestern.edu >>http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Mar 30 13:57:38 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Fixation for lectin work In-Reply-To: <5.1.1.5.1.20040330120205.017c83e8@mailhost.vetmed.auburn.e du> Message-ID: <3.0.6.32.20040330125738.00bdfd18@gemini.msu.montana.edu> John, Formalin works. Have done frozen sections using acetone/alcohol, acetone fixation and these work also. If you do formalin or paraformaldehyde fixation, use trypsin digestion. The only problem I can see with lectins and formalin is immunofluorescent staining and the autofluorescence created by aldehyde fixation. I also have a wonderful review of autofluorescence (although they wrote it for GFP), it still tells sources of this problem along with fixation considerations and still useful for other fluorescent technics. If you want, I will be glad to attach to you privately. There is a wonderful book on Lectin Immunohistochemistry from Springer-Verlag, very cheap that deals with all lectins and tells what they stain (more than one thing is not uncommon). There are buffer considerations - avoiding PBS with certain lectins may be necessary and you will have to use the Lectin Buffer TRIS with Ca and Mg added. Also, E-Y Laboratories puts out a freebie Lectin book, with list of all lectins and their inhibiting sugars, as does Vector. Vector catalog supplies the concentration on inhibition sugar with lectin to create your negative control. > Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Dixon.Leslie <@t> mayo.edu Tue Mar 30 14:20:16 2004 From: Dixon.Leslie <@t> mayo.edu (Dixon, Leslie E.) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Adhesive for paraffin sections Message-ID: Thank you to all who have already replied. To clarify, I have received the slides already cut on plain slides. Thanks, Leslie From plucas <@t> biopath.org Tue Mar 30 13:58:36 2004 From: plucas <@t> biopath.org (Paula Lucas) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Cytology Staining QC Message-ID: <002e01c41691$631a1820$6c01a8c0@new001> Our lab is starting up the Cytology section of our laboratory and I have a question about Quality Control. In histology, we run a control (tissue section) on our H&E auto-stainer before the first run of the day. How are the quality control standards for cytology staining performed at your lab? Thank you. Paula Lucas Bio-Path Medical Group From psanquin <@t> lugo.usc.es Tue Mar 30 15:35:03 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Fixation for lectin work Message-ID: <3.0.6.32.20040330233503.0080e950@pop.lugo.usc.es> John, I am glad to give you my input. It is constrained to my experience in nervous tissue, testis and nasal cavity of mammals as well as in the brain of fishes. I have got good results with Paraformaldehyde just in fishes. Bouin worked superbly in any case. As Barry points there is huge variations depending on the tissue, lectin and even on the species. For that reason I counsel always to try the widest possible number of fixatives. Barry, on what tissue did you do your study? Regards Pablo At 01:16 p.m. 30/03/04 -0600, you wrote: >Dear Pablo Sanchez > >Thank you for responding. Could you expand a bit on your statement that >"aldehydes work in some tissues"? > >I appreciate you time and help in this. > >Regards, > >John Carroll Dennis >Anatomy, Physiology, and Pharmacology >109 Greene Hall >Auburn University, AL 36849 > From kstrati <@t> wisc.edu Tue Mar 30 17:33:25 2004 From: kstrati <@t> wisc.edu (KATERINA STRATI) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Decalcifying mouse heads Message-ID: <56a68a567f7b.567f7b56a68a@wiscmail.wisc.edu> We are interested in decalcifying whole heads from adult mice in order to examine histological sections throughout the oral cavity and esophagus. Are you aware what would be appropriate for such a large piece of tissue, or how long it would take to decalcify? In the future we would like to have the option of performing immunohistochemistry on our decalcified samples. Do you have any experience with IHC on such large decalcified samples? From badesuyi <@t> hotmail.com Tue Mar 30 18:34:23 2004 From: badesuyi <@t> hotmail.com (Banjo Adesuyi) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] NEED INFO ON HER-2, ER & PR Message-ID: Hi, Please we are planning to start running the HER-2, ER and PR in our Pathology Dept, at Val Verde Regional Medical Center, Del Rio, TX, and we were wondering if we could get info as per what is going to cost us in order to start(i.e interms of the equipment and the reagents). Please you can direct your response(s) to my boss,the Laboratory Director, MR RODNEY PAGAN MT(ASCP). Tel# 830-775-8566 Ext.3641. E-Mail: RPAGAN@VVRMC.ORG. Thanking you all for the usual co-operation. Histologically Yours, Banjo Adesuyi, B.Sc, HT(ASCP) Pathology Dept, Val Verde Regional Medical Center, Del Rio, TX 78840. _________________________________________________________________ Add photos to your messages with MSN 8. Get 2 months FREE*. http://join.msn.com/?page=features/featuredemail From caroline.stott <@t> anatomy.otago.ac.nz Tue Mar 30 20:13:32 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Decalcifying mouse heads Message-ID: <5.2.1.1.0.20040331140917.025ae1e0@anatomy.otago.ac.nz> Hi there, We used formic acid to decalcify our mouse heads. They didn't take too long (1 week) although I cant remember how long precisely. And yes they were used for immuno studies. We had a few studies, one for temporal bones and another for the whole head. They worked fine. Caroline We are interested in decalcifying whole heads from adult mice in order to examine histological sections throughout the oral cavity and esophagus. Are you aware what would be appropriate for such a large piece of tissue, or how long it would take to decalcify? In the future we would like to have the option of performing immunohistochemistry on our decalcified samples. Do you have any experience with IHC on such large decalcified samples? Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From ESerrano <@t> imim.es Wed Mar 31 02:09:38 2004 From: ESerrano <@t> imim.es (Serrano del Pozo, Erika) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Apoptosis detection -Caspase 3- Message-ID: <10708933FD0CD511BCFE000102BE5F72025FAD04@hpserv.imim.es> Hi Jackie, As Joelle, I am looking for an ICC technique to detect apoptosis cells, instead mine are muscular cells (C2C12). I am really interested in the Caspase 3 antibody. Could you tell me which commercial antibody do you use and the protocol to perform the ICC ( I?m used to perform IHC not ICC, and I?m not really sure if there is any "triky" step). Thanks in advance. > Erika Serrano > > Centre de Regulaci? Gen?mica > Differenciation and Cancer Programme > Barcelona -SPAIN- > > > ---------- > From: Jackie.O'Connor@abbott.com > Sent: Tuesday, March 30, 2004 15:06 PM > To: Joelle Alcock > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Apoptosis detection > > The Caspase 3 antibody demonstrates only apoptotic cells. > > Jackie O'Connor > Abbott Laboratories > Abbott Park, IL > > > > > > > > > "Joelle Alcock" > Sent by: histonet-bounces@lists.utsouthwestern.edu > 03/29/2004 03:51 PM > > > To: > cc: > Subject: [Histonet] Apoptosis detection > > > Hi all > > I am looking for an IHC technique to detect apoptosis cells on testis > sections. I have previously used the Tunnel assay but would like an > alternative as often meiotic cells are also labelled and I want to avoid > this. > > I have seen the Histone labelling assay by UPSTATE but I think that is > mainly for ICC not IHC. > > Thanks for any help > > Joelle Alcock > > Institute of Genetics > Queens Medical Centre > Nottingham, UK > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From ree3 <@t> leicester.ac.uk Wed Mar 31 03:58:45 2004 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] of mice and men Message-ID: "Antibodies that label paraffin-embedded mouse tissues:A Collaborative Endeavour".....Mikaelian et al, TOXICOLOGIC PATHOLOGY, 32:181-191, 2004. .........a good read for those spending their lives as I do!! Richard Edwards MRC TOX UNIT....U.K.. From psanquin <@t> lugo.usc.es Wed Mar 31 04:51:31 2004 From: psanquin <@t> lugo.usc.es (Pablo =?iso-8859-1?Q?S=E1nchez?= Quinteiro) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Decalcifying mouse heads In-Reply-To: <56a68a567f7b.567f7b56a68a@wiscmail.wisc.edu> Message-ID: <3.0.6.32.20040331125131.007c1100@pop.lugo.usc.es> Currently we are employing in adult mice whole heads "TBD-1 Rapid-decalcifier" from Shandon. At room temperature and with gentle shaking it takes about five hours. Better to check periodically with a blade. We have not yet used this samples for immunohistochemistry, just for lectin histochemical labelling and it works. Bye Pablo At 05:33 p.m. 30/03/04 -0600, you wrote: > > >We are interested in decalcifying whole heads from adult mice in order to examine histological sections throughout the oral cavity and esophagus. Are you aware what would be appropriate for such a large piece of tissue, or how long it would take to decalcify? In the future we would like to have the option of performing immunohistochemistry on our decalcified samples. Do you have any experience with IHC on such large decalcified samples? > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From AFeatherstone <@t> KaleidaHealth.Org Wed Mar 31 05:03:18 2004 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Microwave for antigen retrieval Message-ID: Are people still using the microwave for anitgen retrieval? I thought steamers and pressure cookers pretty much replaced that method. Annette Featherstone HT/MLT Kaleida Health Buffalo General Hospital 100 High St Buffalo NY 14203 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 garygill <@t> dcla.com Wed Mar 31 07:43:09 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Cytology Staining QC Message-ID: Paula: There is no generally accepted QA, not QC, method for confirming stain performance in cytology. However, I will email you individually a 6-page MS Word document, which I authored and was published in SCAN (semi-annual publication of National Association of Cytologists in England) that describes using buccal smears for this purpose and related applications. I'd attach the file to this email, but I believe this Listserv rejects attachments. If anyone else would like a copy, email me individually and I'll email you a copy in my reply. Gary Gill -----Original Message----- From: Paula Lucas [mailto:plucas@biopath.org] Sent: Tuesday, March 30, 2004 2:59 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cytology Staining QC Our lab is starting up the Cytology section of our laboratory and I have a question about Quality Control. In histology, we run a control (tissue section) on our H&E auto-stainer before the first run of the day. How are the quality control standards for cytology staining performed at your lab? Thank you. Paula Lucas Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From garygill <@t> dcla.com Wed Mar 31 07:43:09 2004 From: garygill <@t> dcla.com (Gary Gill) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Cytology Staining QC Message-ID: Paula: There is no generally accepted QA, not QC, method for confirming stain performance in cytology. However, I will email you individually a 6-page MS Word document, which I authored and was published in SCAN (semi-annual publication of National Association of Cytologists in England) that describes using buccal smears for this purpose and related applications. I'd attach the file to this email, but I believe this Listserv rejects attachments. If anyone else would like a copy, email me individually and I'll email you a copy in my reply. Gary Gill -----Original Message----- From: Paula Lucas [mailto:plucas@biopath.org] Sent: Tuesday, March 30, 2004 2:59 PM To: histonet@pathology.swmed.edu Cc: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cytology Staining QC Our lab is starting up the Cytology section of our laboratory and I have a question about Quality Control. In histology, we run a control (tissue section) on our H&E auto-stainer before the first run of the day. How are the quality control standards for cytology staining performed at your lab? Thank you. Paula Lucas Bio-Path Medical Group _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nyilmaz <@t> mersin.edu.tr Wed Mar 31 07:22:53 2004 From: nyilmaz <@t> mersin.edu.tr (=?iso-8859-9?Q?nejat_y=FDlmaz?=) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Urgent Free Image Analyse Software Needed Message-ID: <007b01c41723$5b2b11e0$4c01a8c0@Hislab2> Dear Histonetters... We'd like to measure color density of ihc applied sections within a short time. Is there anybody knows any source or link that we can download a free software for this purpose. We tried to measure it with scionimage but we had problems with density calibration of the programme. Any other suggestions will be appreciated... Dr. Necat YILMAZ From biovetlab <@t> biovet.se Wed Mar 31 08:22:26 2004 From: biovetlab <@t> biovet.se (Lab BioVet AB) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Toluidine Blue Message-ID: <004601c4172b$97de5720$9b64a8c0@YLVA> We have used the same stain protocols for years for mast cells (metachromatic staining) and now we have some problems. We have received a new batch of salt from Polysciences (Have used the salt from Polyscinces Toluidine Blue O C.I 52040 purified for years) Stain protocols we have done is 1% Toluidine blue solved in 20% ethanol 20 minutes. Rinse in 95% ethanol Dehydrate quickly through 95% Abs. alcohols Clear in Xylene. The metachromatic is not purple the are pale blue or even blue-green. Hope for some input. BioVet AB Britt-Marie Smedberg biovetlab@biovet.se From gcallis <@t> montana.edu Wed Mar 31 09:55:18 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Decalcifying mouse heads In-Reply-To: <56a68a567f7b.567f7b56a68a@wiscmail.wisc.edu> Message-ID: <3.0.6.32.20040331085518.00bce8d0@gemini.msu.montana.edu> You did not say which antigens you are trying to stain for. For immunohistochemistry, we totally fix nasal turbinate portion of heads. It is best to perfuse animal with a fixative via heart. Animal is anesthetized deeply so heart beats, and PBS is introduced via left ventricle (hope I got that side correctly) to rinse out blood, then fixative is injected via the same route. This is really not difficult to do. We enjoy success with PLP fixative, suitable for immunohistochemistry. After perfusion, the head (skin, muscles, brain, eyes, lower jaw with tongue must be removed), then turbinates are immersed in PLP overnight with one change of PLP (freshly made) for a second overnight immersion. Use at least 20 times volume of fixative to size of sample, don't skimp on this. We apply vacuum to suck air bubbles out turbinates to insure fixative good contact with nasal mucosa. To decalcify, turbinates are immersed in 1.25% to 5% TETRASODIUM EDTA dissolved in Dulbeccos PBS with pH adjusted to 7.6 with glacial acetic acid. Please note WHICH EDTA we use. The head takes anywhere from 1 week to 2 weeks in this decalcifier. We cut off incisors and do a weight loss/weight gain decalcification endpoint check UNLESS you have a FAXITRON for xray endpoint check. These heads are destined a prion and other prion IHC, embedded in paraffin. EDTA should be rinsed out with buffer - several changes. For frozen sections of turbinates, everything remains the same except tetrasodium EDTA is 1.25% in DPBS containing 10% sucrose (I gave that figure as 20% before, but 10% is the norm) until the head is decalcified. Head is rinsed in 10% sucrose in DPBS, embedded in OCT, and snap frozen using petri dish floating on layer of liquid nitrogen. It cryosections beautifully. Decalcification in this EDTA solution is carried out at 4C, with daily checks (very easy). The sample is intially suspended in decalcifier and vacuum is applied once again to pull any air out of turbinates. Decalcification time is long, but success of IHC is alive and well. You can either use a huge volume of decalcifier OR change it daily. If we only need routine H&E and no IHC on paraffin embedded turbinates, then we decalcify in 10 - 15% formic acid, usually on NBF fixed bone (we let heads fix longer, several days), do the same endpoint test or you can use chemical test. Head decalcifies in a couple of days, but is controlled to avoid overexposure to acid. For IHC, you may want to use buffered formic acid (approx 4.5% formic with either sodium citrate or sodium formate, easily made up in lab or commercial source) which is gentler and touted for IHC. EDTA is more protective of antigens, but some are robust enough to withstand formic acid, and on occasion, HCl. I suggest you determine the effects of your chosen decalcifier on your IHC staining BEFORE you need option of IHC rather than get caught with decalcified bone and no results at all. For paraffin processing of turbinates, we do extended schedule processing using Tissue Prep 2, a harder paraffin and only 1 change of xylene, and 1 change of Clearite 3 or Propar to avoid hardening bone excessively. We have done NBF fixed whole rat and mouse heads with lower jaws intact using 10 - 15% formic acid with success, just rinse these in running water for a hour and do extended processing. For some murine antibodies, fussy CD markers in particular, we frequently do CryoJane tape transfer on undecalcified bone to avoid all aldehyde fixatives. Whew, what a long lecture! Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From gentras <@t> vetmed.auburn.edu Wed Mar 31 10:00:50 2004 From: gentras <@t> vetmed.auburn.edu (Atoska S. Gentry) Date: Fri Sep 16 15:22:44 2005 Subject: Fwd: Re: [Histonet] Decalcifying mouse heads Message-ID: <6.0.1.1.0.20040331100022.02599070@mailhost.vetmed.auburn.edu> please what percent formic acid? >X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 >Date: Wed, 31 Mar 2004 14:13:32 +1200 >To: Histonet@lists.utsouthwestern.edu >From: Caroline Stott >X-scanner: scanned by Inflex 1.0.12.7 >X-Scan-Signature: 05628b3a735271e1629bc35bb8abda07 >Cc: >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.3 >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: 976d3788468f6168a965852c1e2d4d6f >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >Subject: Re: [Histonet] Decalcifying mouse heads >X-Spam-Checker-Version: SpamAssassin 2.63 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: >X-Spam-Status: No, hits=0.0 required=6.5 tests=none autolearn=no version=2.63 >X-SA-Exim-Version: 3.1 (built Tue Oct 14 16:21:02 CDT 2003) >X-SA-Exim-Scanned: Yes > >Hi there, >We used formic acid to decalcify our mouse heads. They didn't take too >long (1 week) although I cant remember how long precisely. And yes they >were used for immuno studies. We had a few studies, one for temporal >bones and another for the whole head. They worked fine. >Caroline > > >We are interested in decalcifying whole heads from adult mice in order to >examine histological sections throughout the oral cavity and esophagus. >Are you aware what would be appropriate for such a large piece of tissue, >or how long it would take to decalcify? In the future we would like to >have the option of performing immunohistochemistry on our decalcified >samples. Do you have any experience with IHC on such large decalcified samples? > > > >Caroline Stott > >Histology Service Unit >Medical School >University of Otago >Dunedin >(03) 479 7152 > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Atoska S. Gentry B.S., HT(ASCP) Research Assistant III Scott-Ritchey Research Center College of Veterinary Medicine Auburn University, AL 36849 Phone# (334)844-5579 Fax# (334)844-5850 From mward <@t> wfubmc.edu Wed Mar 31 10:37:46 2004 From: mward <@t> wfubmc.edu (Martha Ward) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] CISH for Her2 Message-ID: <61135F0455D33347B5AAE209B903A304076A4E04@EXCHVS2.medctr.ad.wfubmc.edu> One of my doctors would like to look into doing CISH for the Her2 oncogene. Has anyone had any experience with this procedure? He gave me information for a probe and detection system from Zymed. Any information would be greatly appreciated. Martha Ward Wake Forest University Baptist Medical Center From DRG <@t> Stowers-Institute.org Wed Mar 31 13:51:11 2004 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Mouse pancreas Message-ID: Hi Histonetters, I am looking for a good tissue processing schedule for mouse pancreas. I am currently using the following schedule and having trouble with sectioning the pancreas, the H&E's look cracked and parched as if my water bath is too hot. The temperature on my water bath is 42-43 degrees Celsius, I have a thermometer also that I check the temp with. I use Tissue Prep paraffin for infiltration and embedding. Fixed by the researcher in 4% PFA 24 hours rinsed in distilled water 5' x 3. 70% Alcohol- 30 min. Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour 30 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From Jackie.O'Connor <@t> abbott.com Wed Mar 31 14:13:07 2004 From: Jackie.O'Connor <@t> abbott.com (Jackie.O'Connor@abbott.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Mouse pancreas Message-ID: The processing schedule you are using would make fibroid uterus brittle. What is prosoft? Anyway - 30 minutes in two changes each of (ethanol) 70%, 95%, Absolute, Xylene, and 2 changes of paraffin at 55 minutes each should do you just fine - for just pancreas - since it is so delicate. I use the above schedule (45 minutes each station) for routine mice tissue and xenografts with beautiful results. Jacqueline M. O'Connor HT(ASCP) Abbott Laboratories Global Pharmaceutical Research and Development Discovery Chemotherapeutics "Grant, Debra" Sent by: histonet-bounces@lists.utsouthwestern.edu 03/31/2004 01:51 PM To: "Histonet" cc: Subject: [Histonet] Mouse pancreas Hi Histonetters, I am looking for a good tissue processing schedule for mouse pancreas. I am currently using the following schedule and having trouble with sectioning the pancreas, the H&E's look cracked and parched as if my water bath is too hot. The temperature on my water bath is 42-43 degrees Celsius, I have a thermometer also that I check the temp with. I use Tissue Prep paraffin for infiltration and embedding. Fixed by the researcher in 4% PFA 24 hours rinsed in distilled water 5' x 3. 70% Alcohol- 30 min. Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour Prosoft- 1 hour 30 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Clearite 3- 1 hour 15 min. Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Paraffin- 1 hour Thanks in advance! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Michael.Owen <@t> fda.gov Wed Mar 31 14:16:58 2004 From: Michael.Owen <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Loeffler's Alkaline Methylene Blue Message-ID: Dear Histonet Users, I am interested in receiving all of the formulations for Loeffler's Alkaline Methylene Blue available. Its uses include visualization of Corynebacterium diphtheria and Mycobacterium leprae. I have already visited Dr. John Kiernan's Histochemistry FAQ, the FDA CFSAN BAM Online Reagents Manual, the Hardy Diagnostics Web site, and the Histonet Archives. If you know of any formulations different from the sources listed above, please send them to me at your earliest convenience. Thank you in advance for your assistance. Sincerely, The FDA Lab Rat Michael P. Owen, Regulatory Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From Michael.Owen <@t> fda.gov Wed Mar 31 14:34:15 2004 From: Michael.Owen <@t> fda.gov (Owen, Michael P) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Histonet: Loeffler's Alkaline Methylene Blue Message-ID: Dear Histonet Users, I am interested in receiving all of the formulations for Loeffler's Alkaline Methylene Blue available. Its uses include visualization of Corynebacterium diphtheria and Mycobacterium leprae. I have already visited Dr. John Kiernan's Histochemistry FAQ, the FDA CFSAN BAM Online Reagents Manual, the Hardy Diagnostics Web site, and the Histonet Archives. If you know of any formulations different from the sources listed above, please send them to me at your earliest convenience. Thank you in advance for your assistance. Sincerely, Michael P. Owen The FDA Lab Rat Michael P. Owen, Regulatory Microbiologist FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021 Phone: 425-483-4865 E-Mail: michael.owen@fda.gov From DRG <@t> Stowers-Institute.org Wed Mar 31 14:54:55 2004 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Mouse pancreas processing schedule Message-ID: Hi all, For those that wanted to know Prosoft is an alcohol substitute from Anatech and is made with: -ether and ester derivatives of propylene glycol, and propanol. Hope this helps! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From jcline <@t> wchsys.org Wed Mar 31 14:56:33 2004 From: jcline <@t> wchsys.org (Joyce Cline) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] er/pr charging Message-ID: <003d01c41762$a7d36760$1d2a14ac@wchsys.org> Does everyone use the CPT code 88342 for charging the er/pr immuno's. Or is there another code that is used for the technical component? ***** 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 Jenny-Oblander <@t> omrf.ouhsc.edu Wed Mar 31 15:00:46 2004 From: Jenny-Oblander <@t> omrf.ouhsc.edu (Jenny Oblander) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Procedure help with dermal papilla Message-ID: Hi Guys, I need your help in tracing down a procedure, antibodies etc for a research project one of my investigators is doing. They would like to differentiate keratinocytes from dermal papilla fibroblasts. Does anyone have experience with TGF-beta-RII or IL-1-RI? Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 From la.sebree <@t> hosp.wisc.edu Wed Mar 31 15:14:37 2004 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] er/pr charging Message-ID: We used to use 88342 but now use 88361. 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: Joyce Cline [mailto:jcline@wchsys.org] Sent: Wednesday, March 31, 2004 2:57 PM To: Histonet Subject: [Histonet] er/pr charging Does everyone use the CPT code 88342 for charging the er/pr immuno's. Or is there another code that is used for the technical component? ***** 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caroline.stott <@t> anatomy.otago.ac.nz Wed Mar 31 15:52:23 2004 From: caroline.stott <@t> anatomy.otago.ac.nz (Caroline Stott) Date: Fri Sep 16 15:22:44 2005 Subject: Fwd: Re: [Histonet] Decalcifying mouse heads Message-ID: <5.2.1.1.0.20040401094556.020deec0@anatomy.otago.ac.nz> We use 10% formic acid. We use the chemical test to check if decalcification is complete. That is 5ml of ammonium hydroxide added to 5ml of the bone solution and add 5ml saturated ammonium oxalate. If the solution is milky white, there is still calcium present. So keep changing the solution every second day until it is finished. Caroline Caroline Stott Histology Service Unit Medical School University of Otago Dunedin (03) 479 7152 From ihc <@t> unipathllc.com Wed Mar 31 15:49:55 2004 From: ihc <@t> unipathllc.com (UniPath IHC) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] LTC4 SYNTHASE, EG2 Message-ID: <000001c4176a$1ab3a6d0$4500a8c0@unipath02> Does anyone know where to find antibodies to LTC4 synthase or EG2? Thanks, Brianna Jackson, BS, QIHC UniPath, LLC bjackson@unipathllc.com 303-512-2220 From gcallis <@t> montana.edu Wed Mar 31 16:48:28 2004 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Mouse pancreas Message-ID: <3.0.6.32.20040331154828.00be4610@gemini.msu.montana.edu> >Is the Prosoft a gradient? Nothing wrong with your waterbath, it is the processing schedule. Your little mouse tissue is processed in a schedule for much bigger tissues, and is probably overexposed to dehydrants. Included is a schedule next to yours with explanation of equivalent to our schedule, cut back times in dehydrants, clearant and paraffins and if still friable, dry and cracked, cut back even more. Be sure you trim block and soak in water, you can try warm then cold, or warm then ice water if needed. What you are doing is removing too much of the bound water attached to tissue proteins along with free water in tissue spaces. >> > 70% Alcohol- 30 min. equivalant is 70% 30 min >30 min >Prosoft- 1 hour " " 80% 30 min >15 min >Prosoft- 1 hour " " 95% 15 min >15 min >Prosoft- 1 hour " " 95% 15 min >30 min >Prosoft- 1 hour " " 95% 30 min >30 min >Prosoft- 1 hour " " 100% 30 min >30 min >Prosoft- 1 hour 30 min. " " 100% 30 min >45 min or 1 hour >Clearite 3- 1 hour 15 min. >45 min or 1 hour >Clearite 3- 1 hour 15 min. >elminate >Clearite 3- 1 hour 15 min. or adjust Clearite changes to be 20, 20, and 45 min >30 min >Paraffin- 1 hour >30 min >Paraffin- 1 hour >30 min >Paraffin- 1 hour >30 min >Paraffin- 1 hour >for a total of 2 hours in paraffin at no more than 60C. >> Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 (lab with voice mail) 406 994-4303 (FAX) From Barry.R.Rittman <@t> uth.tmc.edu Wed Mar 31 16:50:28 2004 From: Barry.R.Rittman <@t> uth.tmc.edu (Barry R Rittman) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Procedure help with dermal papilla Message-ID: <566FB0B522443D43AF02D2ADBE35A6F077FD5B@UTHEVS3.mail.uthouston.edu> Jenny Why not use pan cytokeratin? Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jenny Oblander Sent: Wednesday, March 31, 2004 3:01 PM To: Histonet (E-mail) Subject: [Histonet] Procedure help with dermal papilla Hi Guys, I need your help in tracing down a procedure, antibodies etc for a research project one of my investigators is doing. They would like to differentiate keratinocytes from dermal papilla fibroblasts. Does anyone have experience with TGF-beta-RII or IL-1-RI? Thanks Jenny J.Oblander, HT (A.S.C.P.) Comparative Medicine Oklahoma Medical Research Foundation MS#32 825 NE 13th St. Okc,Ok 73104 jenny-oblander@omrf.ouhsc.edu 405-271-7083 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DRG <@t> Stowers-Institute.org Wed Mar 31 16:56:41 2004 From: DRG <@t> Stowers-Institute.org (Grant, Debra) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Mouse pancreas processing Message-ID: Thanks to all for the pancreas protocols, it seems I need to shorten my times in all stations of the processor. I will try this. Thanks again! Debby Grant Research Technician II Histology Core Facility Stowers Institute for Medical Research 1000 E. 50th Street Kansas City, MO 64110 drg@stowers-institute.org From ASamra <@t> mrl.ubc.ca Wed Mar 31 17:32:20 2004 From: ASamra <@t> mrl.ubc.ca (Amrit Samra) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Lectin IHC Message-ID: I am doing lectin staining for frozen guinea pig tissues. I have tried different fixitives; bouin, 10% formalin, acetone, ethanol, B5 and different buffers; TBS, PBS and lectin buffer. I am still not able to get a positive staining for my known positive. The last trial I did was fixing in bouin and washing with TBS. any help would be greatly appreciated. Jenny Amrit Samra Histology Lab,the iCAPTURE Centre St.Paul's Hospital 1081 Burrard Street,Vancouver,BC Canada V6Z 1Y6 Phone: 604-682-2344 extension 62703 From kstrati <@t> wisc.edu Wed Mar 31 18:09:46 2004 From: kstrati <@t> wisc.edu (KATERINA STRATI) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Decalcifying mouse heads Message-ID: <658073650c1f.650c1f658073@wiscmail.wisc.edu> Thank you to everyone for useful advice Katerina Strati Lambert Lab 222 McArdle Lab 1400 University Ave. Madison, WI 53706 (608) 262-6407 kstrati@wisc.edu ----- Original Message ----- From: KATERINA STRATI Date: Tuesday, March 30, 2004 5:33 pm Subject: [Histonet] Decalcifying mouse heads > > > We are interested in decalcifying whole heads from adult mice in > order to examine histological sections throughout the oral cavity > and esophagus. Are you aware what would be appropriate for such a > large piece of tissue, or how long it would take to decalcify? In > the future we would like to have the option of performing > immunohistochemistry on our decalcified samples. Do you have any > experience with IHC on such large decalcified samples? > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From DMBCMP <@t> aol.com Wed Mar 31 19:13:18 2004 From: DMBCMP <@t> aol.com (DMBCMP@aol.com) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] Validation-Cyto Non-Gyn to Thin Prep Message-ID: <67.2570733f.2d9cc6ae@aol.com> Hello All: The director of my lab (pathologist) and my supervisor (Cytotech) have asked me to post this question on HistoNet. Has anyone done validation for converting non-gyn cytology specimens to Thin Prep? Did you run parallel samples .....how many?,etc. Any input will be graciously welcomed. Thanks very much. Dannie Blake, HT Fresno Community Hospital Fresno, Ca. From ernestinemiddleton <@t> yahoo.ca Wed Mar 31 20:19:15 2004 From: ernestinemiddleton <@t> yahoo.ca (Ernestine Middleton) Date: Fri Sep 16 15:22:44 2005 Subject: [Histonet] ARISTAN STAINER Message-ID: <20040401021915.65884.qmail@web41607.mail.yahoo.com> Hi; For those who are using the Aristan stainer; will you please send me you procedure for gram and wathin starry. Thank you, Ernestine Middleton, Manager, HT/HTL Montefiore Med. Ct. Bronx, NY 718-920-4157 718-547-1920 fax --------------------------------- Post your free ad now! Yahoo! Canada Personals