From gu.lang <@t> gmx.at Sat Apr 1 02:18:23 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Apr 1 02:18:20 2006 Subject: [Histonet] Decal. of bone marrow core biopsies Message-ID: <000301c65564$d8396b10$eeeea8c0@SERVER01> Hello histonetters, I would be interested in the ratio how many labs do "acid" and how many do "neutral" decalcification on bone marrow core biopsies. Here in Austria most labs use the neutral way with Tris buffered EDTA, as we did until a year ago. Now we use 10% formic acid. Perhaps otherwhere in Histoland it looks different? Can anyone give me an estimation? Thanks Gudrun Lang Akh Linz, Austria From wangche <@t> auburn.edu Sat Apr 1 23:25:00 2006 From: wangche <@t> auburn.edu (Chengming Wang) Date: Sat Apr 1 23:25:15 2006 Subject: [Histonet] My frustrating experience with double fluorescence staining! Message-ID: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> Dear All, I am new with histology, and posted yesterday a message of: Help needed!!! Immunofluorescence double staining with mouse lung. Thanks a lot for the several helpful responses. As asked, I am here putting on my concise protocols for your check: Sampling: Cut a small cut along the mouse trachea, injected neg 50 (similar to OCT). Then quick freeze with liquid nitrogen, and the stored @ -80; After section with 6 um, slides were fixed with icy acetone for 5 minutes. Quick dry, and then stored @ -20 till use. Staining: 1. Washing 3 X 5 mins; Blocking Buffer (10% Rabbit serum; 5% BSA; 0.1% Tween 20) for 1 hour at room temperature; 2. Tapping away the blocking buffer, and applied first antibody for 1 hour; then wash 3 x 5 min' 3. 2nd antibody 1 hour; washing 3 x 5min; 4. Mounting media, read slides. Count stained with DAPI-mounting media. 5. For double staining, I did the same thing, except using mixed first antibodies and mixed second conjugated antibodies. Designing: Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit conjugated with Alexa Fluor 488; Stain 2: Rat anti-mouse antibody, and donkey anti-rat conjugated Alexa fluor 594; My problems: The individual staining signals are good. But there is some match color between two channels. For example, I could see weak and similar green signal when I applied Alexa 594. Basically, I could see the weak but true alveoli structure of mouse lung. Then I performed double staining, I could see lots of signal match between red and green channels, which should not be true. In this way, I could see basic alveoli structure of mouse lung in both green and red channels. By the way, I tried to red the slides before staining and after blocking, the autofluorescence is very very weak. I am really frustrating with this situations, and please feel free to tell me how I could figure out this problem. Any of your suggestions and comments are very much appreciated. Thanks, ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Chengming Wang DVM, M.S., Ph.D. Department of Pathobiology College of Veterinary Medicine Auburn University 264 Greene Hall Auburn, AL 36849-5519 Voice: (334) 844-2624 Email: wangche@auburn.edu From louise.renton <@t> gmail.com Mon Apr 3 06:17:00 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Mon Apr 3 06:17:11 2006 Subject: [Histonet] humidity chambers Message-ID: Hi all what are folks using as humidity chambers for immuno slides (manual method). Are they predominanty made in-house or are there ready made commercial products out there? best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. From jtrynak <@t> hotmail.com Mon Apr 3 06:32:26 2006 From: jtrynak <@t> hotmail.com (John Rynak) Date: Mon Apr 3 06:32:33 2006 Subject: [Histonet] Networking for Field Support Specialist - Histopathology TEXAS & Tennessee Regio Message-ID: SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for several Field Support Specialist - Histopathology, in the following regions : Southwest Region (TX Houston or Dallas ) and TN This position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: · Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; · Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; · Customer support Help-line for remote problem solving; · Designing and performing experiments to investigate and solve tough technical applications problems; and · Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: · Excellent problem solving and analytical skills · The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. · A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) · Practical experience in bench-level histopathology, including clinical immunohistochemistry · Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 From pmcardle <@t> ebsciences.com Mon Apr 3 10:16:53 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Mon Apr 3 10:17:02 2006 Subject: [Histonet] humidity chambers In-Reply-To: References: Message-ID: <44313C65.4060104@ebsciences.com> Hello Louise: I'm a vendor; check out http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H2801 Best regards, Phil McArdle louise renton wrote: > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson From Heather.A.Harper <@t> pcola.med.navy.mil Mon Apr 3 10:20:15 2006 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Mon Apr 3 10:22:57 2006 Subject: [Histonet] Thanks for the Input Message-ID: I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather From mauger <@t> email.chop.edu Mon Apr 3 10:42:36 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Mon Apr 3 10:43:25 2006 Subject: [Histonet] humidity chambers Message-ID: Louise, If you are staining only a few slides(about 10),a plastic slide box(100 count) works well. Place slides flat across the top. You can close the lid, and keep them moist! Jo >>> Phil McArdle 04/03/06 11:16 AM >>> Hello Louise: I'm a vendor; check out http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H2801 Best regards, Phil McArdle louise renton wrote: > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Mon Apr 3 10:49:29 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Mon Apr 3 10:49:58 2006 Subject: [Histonet] Thanks for the Input Message-ID: I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vazquezr <@t> ohsu.edu Mon Apr 3 11:29:17 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Mon Apr 3 11:29:37 2006 Subject: [Histonet] Thanks for the Input Message-ID: Sounds like you both work hard...I have been on both sides of the fence...keep up the good work...she needs all the support you can give her civilian or military! Just my two cents... Robyn OHSU >>> "Jackie M O'Connor" 4/3/2006 8:49 AM >>> I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@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 jtaylor <@t> meriter.com Mon Apr 3 12:44:16 2006 From: jtaylor <@t> meriter.com (Taylor, Jean) Date: Mon Apr 3 12:44:20 2006 Subject: [Histonet] Helicobacter pylori Message-ID: <328CBAE62F31C642B422970E879DFADC093DC4@pcwex01> Has anyone who's using the immuno for helicobacter encountered known positive cases by H&E to stain negative for immuno? I'm just wondering how sensitive the antibody is. The cases I've tested with many helico stain up nicely, but about 10 cases that were diagnosed as rare/occasional/scattered helicobacter are negative. I'm using the Dako antibody and have done testing with enzyme digestion as well as antigen retrieval, LSAB+ and EnVision+. Any suggestions would be great. Thanks, Jean Taylor, HT(ASCP)QIHC Immunochemistry GML - Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, April 03, 2006 12:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 3 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. humidity chambers (louise renton) 2. Networking for Field Support Specialist - Histopathology TEXAS & Tennessee Regio (John Rynak) 3. Re: humidity chambers (Phil McArdle) 4. Thanks for the Input (Heather.A.Harper@pcola.med.navy.mil) 5. Re: humidity chambers (Joanne Mauger) 6. Re: Thanks for the Input (Jackie M O'Connor) 7. Re: Thanks for the Input (Robyn Vazquez) ---------------------------------------------------------------------- Message: 1 Date: Mon, 3 Apr 2006 13:17:00 +0200 From: "louise renton" Subject: [Histonet] humidity chambers To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi all what are folks using as humidity chambers for immuno slides (manual method). Are they predominanty made in-house or are there ready made commercial products out there? best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. ------------------------------ Message: 2 Date: Mon, 03 Apr 2006 07:32:26 -0400 From: "John Rynak" Subject: [Histonet] Networking for Field Support Specialist - Histopathology TEXAS & Tennessee Regio To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for several Field Support Specialist - Histopathology, in the following regions : Southwest Region (TX Houston or Dallas ) and TN This position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: * Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; * Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; * Customer support Help-line for remote problem solving; * Designing and performing experiments to investigate and solve tough technical applications problems; and * Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: * Excellent problem solving and analytical skills * The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. * A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) * Practical experience in bench-level histopathology, including clinical immunohistochemistry * Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 ------------------------------ Message: 3 Date: Mon, 03 Apr 2006 11:16:53 -0400 From: Phil McArdle Subject: Re: [Histonet] humidity chambers To: louise renton , "Histonet (E-mail)" Message-ID: <44313C65.4060104@ebsciences.com> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello Louise: I'm a vendor; check out http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H28 01 Best regards, Phil McArdle louise renton wrote: > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson ------------------------------ Message: 4 Date: Mon, 3 Apr 2006 10:20:15 -0500 From: Heather.A.Harper@pcola.med.navy.mil Subject: [Histonet] Thanks for the Input To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather ------------------------------ Message: 5 Date: Mon, 03 Apr 2006 11:42:36 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] humidity chambers To: ,, Message-ID: Content-Type: text/plain; charset=US-ASCII Louise, If you are staining only a few slides(about 10),a plastic slide box(100 count) works well. Place slides flat across the top. You can close the lid, and keep them moist! Jo >>> Phil McArdle 04/03/06 11:16 AM >>> Hello Louise: I'm a vendor; check out http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H28 01 Best regards, Phil McArdle louise renton wrote: > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 3 Apr 2006 10:49:29 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Thanks for the Input To: Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 03 Apr 2006 09:29:17 -0700 From: "Robyn Vazquez" Subject: Re: [Histonet] Thanks for the Input To: Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Sounds like you both work hard...I have been on both sides of the fence...keep up the good work...she needs all the support you can give her civilian or military! Just my two cents... Robyn OHSU >>> "Jackie M O'Connor" 4/3/2006 8:49 AM >>> I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 3 *************************************** From Kristopher.Kalleberg <@t> unilever.com Mon Apr 3 12:46:03 2006 From: Kristopher.Kalleberg <@t> unilever.com (Kalleberg, Kristopher) Date: Mon Apr 3 12:46:09 2006 Subject: [Histonet] re: humididty chambers Message-ID: <0E6BC087F70F9C47ACFF2C203D6E329C7F12BE@NTRSEVS30002.s3.ms.unilever.com> A company called Market Lab offers a nice one for around $150. It is called a staining station. It has rubber runners that the slides sit on above the water filled base and a removable cover. It holds up to 20 slides. There is a also a screw in the base used for drainage. After time, water may drip through this screw and leak out onto the work area. All that is needed to remedy this small problem is to wrap the screw with teflon tape 2 or 3 times, screw the plug back in, and the problem is forever solved. From mauger <@t> email.chop.edu Mon Apr 3 13:15:32 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Mon Apr 3 13:16:17 2006 Subject: [Histonet] re: humididty chambers Message-ID: A much better one, though more expensive(450.00) is the slidemaster, now sold by ScyTek Laboratories(www.scytek.com). It has 2 slide racks holding 10 slides each, and can be tipped for rinsing. I use these every day, and they are great. Jo >>> "Kalleberg, Kristopher" 04/03/06 1:46 PM >>> A company called Market Lab offers a nice one for around $150. It is called a staining station. It has rubber runners that the slides sit on above the water filled base and a removable cover. It holds up to 20 slides. There is a also a screw in the base used for drainage. After time, water may drip through this screw and leak out onto the work area. All that is needed to remedy this small problem is to wrap the screw with teflon tape 2 or 3 times, screw the plug back in, and the problem is forever solved. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Luis.Chiriboga <@t> med.nyu.edu Mon Apr 3 13:10:41 2006 From: Luis.Chiriboga <@t> med.nyu.edu (Luis Chiriboga) Date: Mon Apr 3 13:17:05 2006 Subject: [Histonet] Time and Effort Message-ID: Hi everyone Does anyone know of a reference or resource that has calculated or derived the time and effort for performing different tasks in histology? It can be based on per-sample or per-slide basis. anyone have any ideas? TIA Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 From tkngflght <@t> yahoo.com Mon Apr 3 13:21:54 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Mon Apr 3 13:21:57 2006 Subject: [Histonet] Time and Effort In-Reply-To: Message-ID: <20060403182154.93683.qmail@web50901.mail.yahoo.com> Luis-- There used to be a standard for time established by The College of American Pathologist. We used it years ago to work out productivity and called it CAP Accounting. It allowed so many minutes for an H&E, so many for a blank, each special had an allottment, etc. I don't have a copy but maybe one of the other members remembers and can dig up an archive? It was accurate to a degree and a helpful baseline. Cheryl Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. From bob-meyer <@t> northwestern.edu Mon Apr 3 13:46:44 2006 From: bob-meyer <@t> northwestern.edu (bob-meyer@northwestern.edu) Date: Mon Apr 3 13:46:46 2006 Subject: [Histonet] Tissue graft coming off of slide Message-ID: <20060403184644.2FECE35C65@casbah.it.northwestern.edu> Just wondering if anyone has any ideas to keep ePTFE (I will ask researcher what this stands for) graft tissue from porcine from coming off of slides during antigen retrieval. Actually, it is starting to come off even after deparaffinization. I told the researcher this is our last hope that someone on the Histonet might have an idea. Here is a summary of what I have tried. I used silanized slides (from Dako) with an overnight bake at 57C for tissue adherance plus an additional 10 minute 10% NBF soak to fix the tissue firmly on the slide after deparaffinization and before antigen retrieval. I have used low temperature antigen retrieval (LTAR) in the past for delicate tissue, but if the tissue comes off after deparaffinization what is the point. Is there something else out there that I can try? Thanks for any and all help. Bob Meyer Sr. Research Technologist Northwestern University Chicago, IL 60445 312-908-5546 From llewllew <@t> shaw.ca Mon Apr 3 14:13:33 2006 From: llewllew <@t> shaw.ca (Bryan Llewellyn) Date: Mon Apr 3 14:14:34 2006 Subject: [Histonet] Time and Effort References: <20060403182154.93683.qmail@web50901.mail.yahoo.com> Message-ID: <000801c65752$b3780580$690a4246@yourlk4rlmsu> That document was based on the Canadian Workload Data Recording system, a program of the Canadian Federal Ministry of Health. It is used in every publicly funded hospital laboratory in Canada for monthly workload reporting and is kept up to date. Contact a Canadian hospital laboratory and I'm sure you could get a copy of the latest incarnation. Bryan Llewellyn ----- Original Message ----- From: "Cheryl" To: "Luis Chiriboga" ; "HISTONET" Sent: Monday, April 03, 2006 11:21 AM Subject: Re: [Histonet] Time and Effort > Luis-- > > There used to be a standard for time established by The College of > American Pathologist. We used it years ago to work out productivity and > called it CAP Accounting. It allowed so many minutes for an H&E, so many > for a blank, each special had an allottment, etc. I don't have a copy but > maybe one of the other members remembers and can dig up an archive? > > It was accurate to a degree and a helpful baseline. > > Cheryl > > > > > Cheryl Kerry, HT(ASCP) > Full Staff Inc. > Staffing the AP Lab by helping one Tech at a time. > 281.883.7704 c > 281.852.9457 o > admin@fullstaff.org > > Sign up for the FREE monthly newsletter AP News--updates, tricks of the > trade and current issues for Anatomic Pathology Clinical Labs. Send a > 'subscribe' request to APNews@fullstaff.org. Please include your name and > specialty in the body of the email. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From PMonfils <@t> Lifespan.org Mon Apr 3 14:17:56 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Apr 3 14:18:11 2006 Subject: [Histonet] humidity chambers Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176B7@lsexch.lsmaster.lifespan.org> For many years I have used the same home-made device consisting of two 14x18" fiberglass cafeteria trays, one of which has eight 10 ml serological pipettes epoxied to the inside surface, forming four double rows, each of which can support 9 slides. So the whole apparatus comfortably holds 36 slides (40 if I crowd them a bit). The second tray, without pipettes, serves as the cover. The thing has worked fine for me for 20 years, so I have never had reason to purchase anything else. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > louise renton > Sent: Monday, April 3, 2006 4:17 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] humidity chambers > > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Rcartun <@t> harthosp.org Mon Apr 3 14:18:01 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Mon Apr 3 14:18:41 2006 Subject: [Histonet] Helicobacter pylori Message-ID: In my experience, I am not aware of any H&E positive cases that were negative by IHC. Are your pathologists absolutely certain that these are bugs? Did you repeat these cases? How many sections were evaluated? Could you still see the organisms on your negative control? The problem with the DAKO antibody is specificity, not sensitivity (it labels non-H. pylori organisms). I guess you could send sections from these cases to another laboratory and see what results they get. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Taylor, Jean" 04/03/06 01:44PM >>> Has anyone who's using the immuno for helicobacter encountered known positive cases by H&E to stain negative for immuno? I'm just wondering how sensitive the antibody is. The cases I've tested with many helico stain up nicely, but about 10 cases that were diagnosed as rare/occasional/scattered helicobacter are negative. I'm using the Dako antibody and have done testing with enzyme digestion as well as antigen retrieval, LSAB+ and EnVision+. Any suggestions would be great. Thanks, Jean Taylor, HT(ASCP)QIHC Immunochemistry GML - Madison, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Monday, April 03, 2006 12:09 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 3 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. humidity chambers (louise renton) 2. Networking for Field Support Specialist - Histopathology TEXAS & Tennessee Regio (John Rynak) 3. Re: humidity chambers (Phil McArdle) 4. Thanks for the Input (Heather.A.Harper@pcola.med.navy.mil) 5. Re: humidity chambers (Joanne Mauger) 6. Re: Thanks for the Input (Jackie M O'Connor) 7. Re: Thanks for the Input (Robyn Vazquez) ---------------------------------------------------------------------- Message: 1 Date: Mon, 3 Apr 2006 13:17:00 +0200 From: "louise renton" Subject: [Histonet] humidity chambers To: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=ISO-8859-1 Hi all what are folks using as humidity chambers for immuno slides (manual method). Are they predominanty made in-house or are there ready made commercial products out there? best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. ------------------------------ Message: 2 Date: Mon, 03 Apr 2006 07:32:26 -0400 From: "John Rynak" Subject: [Histonet] Networking for Field Support Specialist - Histopathology TEXAS & Tennessee Regio To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; format="flowed" SciGenium's Client is experiencing tremendous growth within their North American operations. This growth is due to the recent launch of their FDA approved automated histology instrumentation system, and the growing antibody and reagent consumable product line for the histology market. Due to the new launch of these products they are currently looking for several Field Support Specialist - Histopathology, in the following regions : Southwest Region (TX Houston or Dallas ) and TN This position provides technical applications support for our next generation range of automated histopathology products. This is a very hands on role where your key tasks will include: * Building an in-depth understanding of our new product technologies and their applications in a diagnostic histopathology environment; * Customer liaison by providing scientific and laboratory expertise for in-field installations, training and trouble-shooting; * Customer support Help-line for remote problem solving; * Designing and performing experiments to investigate and solve tough technical applications problems; and * Preparing problem resolution reports and imparting your findings and knowledge to internal and field based Product Engineers and Technical Managers. To be successful in this role you will possess: * Excellent problem solving and analytical skills * The ability to communicate with front-end users, technical design engineers and pragmatic sales force members. * A tertiary qualification in a relevant science stream (eg. Medical Laboratory Science) * Practical experience in bench-level histopathology, including clinical immunohistochemistry * Demonstrated use of automated scientific instrumentation BS/ MS in Life Sciences and ASCP Certification Interest in using PC based software and maintenance of automated scientific instrumentation will be highly advantageous. In addition, you are extremely detailed and analytically minded with a strong sense of customer focus. Interested parties should forward a resume to resume@scigenium.com Send to : SciGenium PO 380916 Cambridge, MA 02238 ------------------------------ Message: 3 Date: Mon, 03 Apr 2006 11:16:53 -0400 From: Phil McArdle Subject: Re: [Histonet] humidity chambers To: louise renton , "Histonet (E-mail)" Message-ID: <44313C65.4060104@ebsciences.com> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello Louise: I'm a vendor; check out http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H28 01 Best regards, Phil McArdle louise renton wrote: > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson ------------------------------ Message: 4 Date: Mon, 3 Apr 2006 10:20:15 -0500 From: Heather.A.Harper@pcola.med.navy.mil Subject: [Histonet] Thanks for the Input To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather ------------------------------ Message: 5 Date: Mon, 03 Apr 2006 11:42:36 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] humidity chambers To: ,, Message-ID: Content-Type: text/plain; charset=US-ASCII Louise, If you are staining only a few slides(about 10),a plastic slide box(100 count) works well. Place slides flat across the top. You can close the lid, and keep them moist! Jo >>> Phil McArdle 04/03/06 11:16 AM >>> Hello Louise: I'm a vendor; check out http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H28 01 Best regards, Phil McArdle louise renton wrote: > Hi all > > what are folks using as humidity chambers for immuno slides (manual > method). Are they predominanty made in-house or are there ready made > commercial products out there? > > best regards > -- > Louise Renton > Bone Research Unit > University of the Witwatersrand > Johannesburg > South Africa > "Does a conceited cowboy, only ride on pompous grass?. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Mon, 3 Apr 2006 10:49:29 -0500 From: "Jackie M O'Connor" Subject: Re: [Histonet] Thanks for the Input To: Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="us-ascii" I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 7 Date: Mon, 03 Apr 2006 09:29:17 -0700 From: "Robyn Vazquez" Subject: Re: [Histonet] Thanks for the Input To: Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=us-ascii Sounds like you both work hard...I have been on both sides of the fence...keep up the good work...she needs all the support you can give her civilian or military! Just my two cents... Robyn OHSU >>> "Jackie M O'Connor" 4/3/2006 8:49 AM >>> I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 3 *************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Apr 3 14:32:46 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Apr 3 14:32:57 2006 Subject: [Histonet] Tissue graft coming off of slide Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176B8@lsexch.lsmaster.lifespan.org> For sections of synthetics I usually use a chrome alum gelatin product in the water bath. I used to make my own but in recent years have relied upon "Sta-On", manufactured by Surgipath. You have two major problems facing you. First, the need for antigen retrieval. Most of my work involving synthetic materials has not required antigen retrieval. The Sta-On has worked very well for me in most cases but I'm not sure how well it will stand up to boiling. Secondly, the particular synthetic you are working with, PTFE (polytetrafluoroethylene), which is marketed under the familiar trade name "Teflon", is specifically designed not to stick to anything! Still, I have had pretty good success with PTFE implants by using a fairly high concentration of Sta-On in the water bath, followed by drying on an 80 degree slide warmer before overnight drying at 60 degrees - but again, without antigen retrieval. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > bob-meyer@northwestern.edu > Reply To: bob-meyer@northwestern.edu > Sent: Monday, April 3, 2006 11:46 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue graft coming off of slide > > > > > > > Just wondering if anyone has any ideas to keep ePTFE (I will ask > researcher what this stands for) > graft tissue from porcine from coming off of slides during antigen > retrieval. Actually, it is > starting to come off even after deparaffinization. I told the researcher > this is our last hope > that someone on the Histonet might have an idea. Here is a summary of > what I have tried. I used > silanized slides (from Dako) with an overnight bake at 57C for tissue > adherance plus an additional > 10 minute 10% NBF soak to fix the tissue firmly on the slide after > deparaffinization and before > antigen retrieval. I have used low temperature antigen retrieval (LTAR) > in the past for delicate > tissue, but if the tissue comes off after deparaffinization what is the > point. Is there something > else out there that I can try? Thanks for any and all help. > > Bob Meyer > Sr. Research Technologist > Northwestern University > Chicago, IL 60445 > 312-908-5546 > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From PMonfils <@t> Lifespan.org Mon Apr 3 14:41:48 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Apr 3 14:42:00 2006 Subject: [Histonet] Tissue graft coming off of slide - P.S. Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176B9@lsexch.lsmaster.lifespan.org> P.S. I just recalled another method that gave me good results with synthetics, which I still use occasionally, namely PVA (polyvinyl acetate) in the water bath, followed by drying as described in my previous post. PVA makes the water bath milky white, which makes it more difficult to see the sections floating on it - which is why I prefer the chrome alum gelatin - but it is an excellent adhesive for difficult materials. It can be purchased in the form of "Elmer's Glue-All" or "Sobo Glue" or various other brands. Not all "white glues" are PVA however, so read the product label. Some people also use the tan-colored "carpenter's wood glue" which is available in hardware stores. I haven't used this myself, and am not sure of the chemical makeup of the product. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > bob-meyer@northwestern.edu > Reply To: bob-meyer@northwestern.edu > Sent: Monday, April 3, 2006 11:46 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue graft coming off of slide > > > Just wondering if anyone has any ideas to keep ePTFE (I will ask > researcher what this stands for) > graft tissue from porcine from coming off of slides during antigen > retrieval. Actually, it is > starting to come off even after deparaffinization. I told the researcher > this is our last hope > that someone on the Histonet might have an idea. Here is a summary of > what I have tried. I used > silanized slides (from Dako) with an overnight bake at 57C for tissue > adherance plus an additional > 10 minute 10% NBF soak to fix the tissue firmly on the slide after > deparaffinization and before > antigen retrieval. I have used low temperature antigen retrieval (LTAR) > in the past for delicate > tissue, but if the tissue comes off after deparaffinization what is the > point. Is there something > else out there that I can try? Thanks for any and all help. > > Bob Meyer > Sr. Research Technologist > Northwestern University > Chicago, IL 60445 > 312-908-5546 > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From Janet.Bonner <@t> FLHOSP.ORG Mon Apr 3 15:33:58 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Mon Apr 3 15:36:07 2006 Subject: [Histonet] Tissue graft coming off of slide References: <09C945920A6B654199F7A58A1D7D1FDE017176B8@lsexch.lsmaster.lifespan.org> Message-ID: "Gold" slides are also excellent - they are the ultimate in adherence! Once a section is on the slide, you can't even get it off to reposition it!! I believe we get them through Allegiance or Fisher Scientific. Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Monfils, Paul Sent: Mon 4/3/2006 3:32 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Tissue graft coming off of slide For sections of synthetics I usually use a chrome alum gelatin product in the water bath. I used to make my own but in recent years have relied upon "Sta-On", manufactured by Surgipath. You have two major problems facing you. First, the need for antigen retrieval. Most of my work involving synthetic materials has not required antigen retrieval. The Sta-On has worked very well for me in most cases but I'm not sure how well it will stand up to boiling. Secondly, the particular synthetic you are working with, PTFE (polytetrafluoroethylene), which is marketed under the familiar trade name "Teflon", is specifically designed not to stick to anything! Still, I have had pretty good success with PTFE implants by using a fairly high concentration of Sta-On in the water bath, followed by drying on an 80 degree slide warmer before overnight drying at 60 degrees - but again, without antigen retrieval. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > bob-meyer@northwestern.edu > Reply To: bob-meyer@northwestern.edu > Sent: Monday, April 3, 2006 11:46 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue graft coming off of slide > > > > > > > Just wondering if anyone has any ideas to keep ePTFE (I will ask > researcher what this stands for) > graft tissue from porcine from coming off of slides during antigen > retrieval. Actually, it is > starting to come off even after deparaffinization. I told the researcher > this is our last hope > that someone on the Histonet might have an idea. Here is a summary of > what I have tried. I used > silanized slides (from Dako) with an overnight bake at 57C for tissue > adherance plus an additional > 10 minute 10% NBF soak to fix the tissue firmly on the slide after > deparaffinization and before > antigen retrieval. I have used low temperature antigen retrieval (LTAR) > in the past for delicate > tissue, but if the tissue comes off after deparaffinization what is the > point. Is there something > else out there that I can try? Thanks for any and all help. > > Bob Meyer > Sr. Research Technologist > Northwestern University > Chicago, IL 60445 > 312-908-5546 > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 Janet.Bonner <@t> FLHOSP.ORG Mon Apr 3 15:36:14 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Mon Apr 3 15:36:56 2006 Subject: [Histonet] Tissue graft coming off of slide - P.S. References: <09C945920A6B654199F7A58A1D7D1FDE017176B9@lsexch.lsmaster.lifespan.org> Message-ID: Be careful of Elmer's glue, it's been known to be positive for a fungus! ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Monfils, Paul Sent: Mon 4/3/2006 3:41 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] Tissue graft coming off of slide - P.S. P.S. I just recalled another method that gave me good results with synthetics, which I still use occasionally, namely PVA (polyvinyl acetate) in the water bath, followed by drying as described in my previous post. PVA makes the water bath milky white, which makes it more difficult to see the sections floating on it - which is why I prefer the chrome alum gelatin - but it is an excellent adhesive for difficult materials. It can be purchased in the form of "Elmer's Glue-All" or "Sobo Glue" or various other brands. Not all "white glues" are PVA however, so read the product label. Some people also use the tan-colored "carpenter's wood glue" which is available in hardware stores. I haven't used this myself, and am not sure of the chemical makeup of the product. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > bob-meyer@northwestern.edu > Reply To: bob-meyer@northwestern.edu > Sent: Monday, April 3, 2006 11:46 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue graft coming off of slide > > > Just wondering if anyone has any ideas to keep ePTFE (I will ask > researcher what this stands for) > graft tissue from porcine from coming off of slides during antigen > retrieval. Actually, it is > starting to come off even after deparaffinization. I told the researcher > this is our last hope > that someone on the Histonet might have an idea. Here is a summary of > what I have tried. I used > silanized slides (from Dako) with an overnight bake at 57C for tissue > adherance plus an additional > 10 minute 10% NBF soak to fix the tissue firmly on the slide after > deparaffinization and before > antigen retrieval. I have used low temperature antigen retrieval (LTAR) > in the past for delicate > tissue, but if the tissue comes off after deparaffinization what is the > point. Is there something > else out there that I can try? Thanks for any and all help. > > Bob Meyer > Sr. Research Technologist > Northwestern University > Chicago, IL 60445 > 312-908-5546 > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@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 azdudley <@t> hotmail.com Mon Apr 3 15:41:24 2006 From: azdudley <@t> hotmail.com (anita dudley) Date: Mon Apr 3 15:41:34 2006 Subject: [Histonet] CAP Immuno Survey Slides In-Reply-To: <8E7AD740937B954F947F0DB4467EFEE056E973@mail.srhs-pa.org> Message-ID: I had trouble with the labels also, hope they leave them off next time, not a good idea. anita dudley, mobile alabama >From: "Jones, Laura" >To: "Histonet (E-mail)" >Subject: [Histonet] CAP Immuno Survey Slides >Date: Thu, 30 Mar 2006 12:04:30 -0500 > >Hello Histoland. We were wondering if anyone else who may have >participated >in the recent CAP survey had experienced any problems with the labels on >the >slides. We performed some of the survey by hand (our machine is down!) >using the Shandon Sequenza coverplates, and it seemed like the label >adhesive was blocking the reagents from passing onto the slide. We then >repeated some of the slides on a demo machine, after verifying our >antibodies, and even then it seemed like that adhesive was interfering with >the reagents. We also lost any info we had written on the slide labels, >even with the impervious markers, during automated processing. > >We didn't remember our survey slides ever having labels before, and are >just >curious if anyone else had encountered this. Thanks in advance! > >The Histochicks at Sharon Regional, Sharon, PA > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Apr 3 15:44:41 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Apr 3 15:44:44 2006 Subject: [Histonet] Time and Effort In-Reply-To: Message-ID: <20060403204441.81181.qmail@web61212.mail.yahoo.com> Hi Luis: I am attaching directly to you the text of a Workshop for the Florida Siciety of Histotechnology that I have just finished dealing with several issues, including this one. Ren? J. Luis Chiriboga wrote: Hi everyone Does anyone know of a reference or resource that has calculated or derived the time and effort for performing different tasks in histology? It can be based on per-sample or per-slide basis. anyone have any ideas? TIA Luis ____________________________________ Luis Chiriboga Ph.D. NYU Cancer Institute and Bellevue Hospital Center New York University School Of Medicine Department Of Pathology 4W27 462 First Avenue New York, N.Y. 10016 W(212) 562-4667. F(212) 263-2041 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From PMonfils <@t> Lifespan.org Mon Apr 3 17:28:31 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Apr 3 17:28:43 2006 Subject: [Histonet] Tissue graft coming off of slide - P.S. Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176BA@lsexch.lsmaster.lifespan.org> Hi Gayle, Yes, you are right. "PVA" is commonly used in reference to polyvinyl alcohol, and to polyvinyl acetate as well, so we should be sure which compound we are talking about. Interesting that polyvinyl alcohol can be used as a releasing agent while polyvinyl acetate is an adhesive. In print I believe I have seen the designation PVAc in reference to polyvinyl acetate, but in speech, in my experience, people commonly refer to both compounds as PVA. > ---------- > From: Gayle Callis > Sent: Monday, April 3, 2006 12:57 PM > To: Monfils, Paul > Subject: RE: [Histonet] Tissue graft coming off of slide - P.S. > > I always thought PVA stood for poly vinyl alcohol? AT least it is on the > > OCT cryoembedding media bottle. > > At 01:41 PM 4/3/2006, you wrote: > >P.S. I just recalled another method that gave me good results with > >synthetics, which I still use occasionally, namely PVA (polyvinyl > acetate) > >in the water bath, followed by drying as described in my previous post. > PVA > >makes the water bath milky white, which makes it more difficult to see > the > >sections floating on it - which is why I prefer the chrome alum gelatin - > >but it is an excellent adhesive for difficult materials. It can be > >purchased in the form of "Elmer's Glue-All" or "Sobo Glue" or various > other > >brands. Not all "white glues" are PVA however, so read the product > label. > >Some people also use the tan-colored "carpenter's wood glue" which is > >available in hardware stores. I haven't used this myself, and am not > sure > >of the chemical makeup of the product. > > > > > ---------- > > > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > > > bob-meyer@northwestern.edu > > > Reply To: bob-meyer@northwestern.edu > > > Sent: Monday, April 3, 2006 11:46 AM > > > To: Histonet@lists.utsouthwestern.edu > > > Subject: [Histonet] Tissue graft coming off of slide > > > > > > > > > Just wondering if anyone has any ideas to keep ePTFE (I will ask > > > researcher what this stands for) > > > graft tissue from porcine from coming off of slides during antigen > > > retrieval. Actually, it is > > > starting to come off even after deparaffinization. I told the > researcher > > > this is our last hope > > > that someone on the Histonet might have an idea. Here is a summary of > > > what I have tried. I used > > > silanized slides (from Dako) with an overnight bake at 57C for tissue > > > adherance plus an additional > > > 10 minute 10% NBF soak to fix the tissue firmly on the slide after > > > deparaffinization and before > > > antigen retrieval. I have used low temperature antigen retrieval > (LTAR) > > > in the past for delicate > > > tissue, but if the tissue comes off after deparaffinization what is > the > > > point. Is there something > > > else out there that I can try? Thanks for any and all help. > > > > > > Bob Meyer > > > Sr. Research Technologist > > > Northwestern University > > > Chicago, IL 60445 > > > 312-908-5546 > > > > > > > > > > > > > > > > > > > > > > > > _______________________________________________ > > > Histonet mailing list > > > Histonet@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 > 406 994-4303 (FAX) > > > From kaleid11 <@t> yahoo.com Mon Apr 3 20:54:05 2006 From: kaleid11 <@t> yahoo.com (Adam Perry) Date: Mon Apr 3 20:54:07 2006 Subject: [Histonet] My frustrating experience with double fluorescence staining! In-Reply-To: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> Message-ID: <20060404015405.12079.qmail@web30414.mail.mud.yahoo.com> Shouldn't your blocking buffer use donkey serum, since both of your secondaries are generated in donkey? I would think that the rabbit serum block with your first secondary antibody (donkey anti-rabbit) would produce high background and non-specific staining... Adam Department of Physiology and Biophysics University of Illinois at Chicago Chicago, IL 60612 (312)413-0737 Blocking Buffer (10% Rabbit serum; 5% BSA; 0.1% Tween 20) for 1 hour at room temperature; Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit conjugated with Alexa Fluor 488; Stain 2: Rat anti-mouse antibody, and donkey anti-rat conjugated Alexa fluor 594; --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. From drmoses111 <@t> comcast.net Mon Apr 3 21:36:00 2006 From: drmoses111 <@t> comcast.net (drmoses111@comcast.net) Date: Mon Apr 3 21:36:04 2006 Subject: [Histonet] Sukura Express Hepatocyte? Message-ID: <040420060236.18496.4431DB8F000F3259000048402200750330CECECE9C0A9C01039D0B@comcast.net> We recently got a Sukura Express microwave processor. So far, our immunos look pretty close to our routine slides, except for Dako's hepatocyte antibody. This antibody will not work on the liver cores processed on the Sukura. This is a hardy and easy to use antibody, it works well on autopsy or surgical tissue ,with HEIR or without. I have never had a problem till now. Does anybody have any ideas or similar experiences? From louise.renton <@t> gmail.com Tue Apr 4 01:48:51 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Tue Apr 4 01:48:59 2006 Subject: [Histonet] thanks Message-ID: Thanks to all those who helped me with my failing memory re: immuno. This is a great resource, one that I am daily grateful for. Thanks especially to gayle for her enormous immuno experience and her patient replies to my hesitant queries best regards -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. From modernarnis73 <@t> juno.com Tue Apr 4 05:56:58 2006 From: modernarnis73 <@t> juno.com (modernarnis73@juno.com) Date: Tue Apr 4 05:58:31 2006 Subject: [Histonet] Thank-you for Barrier slides suggestions Message-ID: <20060404.035723.19860.838842@webmail52.lax.untd.com> I just wanted to thank everyone who sent info. on different barrier slides. Thank-you, Enoch ________________________________________________________________________ Try Juno Platinum for Free! Then, only $9.95/month! Unlimited Internet Access with 1GB of Email Storage. Visit http://www.juno.com/value to sign up today! From andromeda_tm <@t> libero.it Tue Apr 4 06:33:59 2006 From: andromeda_tm <@t> libero.it (Massimo) Date: Tue Apr 4 06:34:06 2006 Subject: [Histonet] Cutting angle with flat razor blade Message-ID: <000501c657db$ab773b10$6aed1c97@SN300208440005> Hi all, I wonder if someone could give me some advice about the best cutting angle for specimens (mouse tissues) embedded into wax (Paraplast - melting point 56?C) at room temperature (about 18?C) with a microtome flat razor blade. Thank you in advance. Best Regards, Massimo From andromeda_tm <@t> libero.it Tue Apr 4 06:33:59 2006 From: andromeda_tm <@t> libero.it (Massimo) Date: Tue Apr 4 06:39:29 2006 Subject: [Histonet] Cutting angle with flat razor blade Message-ID: <000501c657db$ab773b10$6aed1c97@SN300208440005> Hi all, I wonder if someone could give me some advice about the best cutting angle for specimens (mouse tissues) embedded into wax (Paraplast - melting point 56?C) at room temperature (about 18?C) with a microtome flat razor blade. Thank you in advance. Best Regards, Massimo From DDDeltour <@t> mar.med.navy.mil Tue Apr 4 07:32:00 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Tue Apr 4 07:32:41 2006 Subject: [Histonet] Thanks for the Input Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE1E@marxchg03.mar.med.navy.mil> I work in this environment everyday. I see both sides (military and civilian) taking shots at each other instead of working as a team. I am soon leaving the service and you could not pay me enough money to supervise or work in a military facility. I understand Heather and her frustration of being left to do the job BUT the military requires its members to do things outside of the job to get promoted. That is a fact. I am living it. I see people in my own field get promoted because they did some BS bake sale but they can't even do a GMS. You can either find a way to work it out or find a new job. As for the military Pathologist....well they are in charge. They like that. Everything will change again when the next one transfers in. That is part of being in a military facility. I have seen it for years. The best thing to do is go with the flow or just go. I am going. Good luck. Douglas D. Deltour HT(ASCP) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Monday, April 03, 2006 12:29 PM To: Jackie.O'Connor@abbott.com; Harper, Heather A., CIV Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Thanks for the Input Sounds like you both work hard...I have been on both sides of the fence...keep up the good work...she needs all the support you can give her civilian or military! Just my two cents... Robyn OHSU >>> "Jackie M O'Connor" 4/3/2006 8:49 AM >>> I am thankful and grateful for anyone who ever signed up for military service - ever - regardless of their job description. Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 04/03/2006 10:20 AM To: histonet@lists.utsouthwestern.edu cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: [Histonet] Thanks for the Input I want to thank everybody who responded to my message titled..Need Input. I work for the DOD and I miss simply having a system in place. I agree one has to be flexible, but when you work with military, they are entitled to 1 hr lunch and 1 hour of PT. My tech works 7-4 and I work 6-2. I embed, she cuts and stains, I gross, accession, order supplies, admit bodies in and out of the morgue, set up for autopsies, frozens, and when my tech has to get pulled to do her other command duties, it leaves me holding the bag. It does get over whelming, and every 2-3 yrs, I get new pathologists rotating through. I have worked with 7 pathologists in 6 yrs, and reservists, and this one I just do not know what to think. Just keeping an open mind. Thanks again everybody. Be thankful you are in the civilian world. Heather _______________________________________________ Histonet mailing list Histonet@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 AFeatherstone <@t> KaleidaHealth.Org Tue Apr 4 08:26:26 2006 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Tue Apr 4 08:26:31 2006 Subject: [Histonet] breast processing Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, April 02, 2006 13:01 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 2 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. My frustrating experience with double fluorescence staining! (Chengming Wang) ---------------------------------------------------------------------- Message: 1 Date: Sat, 01 Apr 2006 23:25:00 -0600 From: "Chengming Wang" Subject: [Histonet] My frustrating experience with double fluorescence staining! To: Message-ID: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> Content-Type: text/plain; charset=US-ASCII Dear All, I am new with histology, and posted yesterday a message of: Help needed!!! Immunofluorescence double staining with mouse lung. Thanks a lot for the several helpful responses. As asked, I am here putting on my concise protocols for your check: Sampling: Cut a small cut along the mouse trachea, injected neg 50 (similar to OCT). Then quick freeze with liquid nitrogen, and the stored @ -80; After section with 6 um, slides were fixed with icy acetone for 5 minutes. Quick dry, and then stored @ -20 till use. Staining: 1. Washing 3 X 5 mins; Blocking Buffer (10% Rabbit serum; 5% BSA; 0.1% Tween 20) for 1 hour at room temperature; 2. Tapping away the blocking buffer, and applied first antibody for 1 hour; then wash 3 x 5 min' 3. 2nd antibody 1 hour; washing 3 x 5min; 4. Mounting media, read slides. Count stained with DAPI-mounting media. 5. For double staining, I did the same thing, except using mixed first antibodies and mixed second conjugated antibodies. Designing: Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit conjugated with Alexa Fluor 488; Stain 2: Rat anti-mouse antibody, and donkey anti-rat conjugated Alexa fluor 594; My problems: The individual staining signals are good. But there is some match color between two channels. For example, I could see weak and similar green signal when I applied Alexa 594. Basically, I could see the weak but true alveoli structure of mouse lung. Then I performed double staining, I could see lots of signal match between red and green channels, which should not be true. In this way, I could see basic alveoli structure of mouse lung in both green and red channels. By the way, I tried to red the slides before staining and after blocking, the autofluorescence is very very weak. I am really frustrating with this situations, and please feel free to tell me how I could figure out this problem. Any of your suggestions and comments are very much appreciated. Thanks, ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Chengming Wang DVM, M.S., Ph.D. Department of Pathobiology College of Veterinary Medicine Auburn University 264 Greene Hall Auburn, AL 36849-5519 Voice: (334) 844-2624 Email: wangche@auburn.edu ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 2 *************************************** 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 jqb7 <@t> cdc.gov Tue Apr 4 08:35:02 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Tue Apr 4 08:41:49 2006 Subject: [Histonet] breast processing Message-ID: When I worked in a hospital, we would have the pathologists gross the tissue as small as they could and then place it in PenFix overnight. (If it needed further grossing it could be done at this time since the PenFix helped to firm the tissue up a bi and it could be left in PenFix for a while longer.) Then it would go into formalin overnight. The amount of time in PenFix was directly related to how large the tissue sample for processing was. Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Featherstone, Annette Sent: Tuesday, April 04, 2006 9:26 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] breast processing Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, April 02, 2006 13:01 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 2 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. My frustrating experience with double fluorescence staining! (Chengming Wang) ---------------------------------------------------------------------- Message: 1 Date: Sat, 01 Apr 2006 23:25:00 -0600 From: "Chengming Wang" Subject: [Histonet] My frustrating experience with double fluorescence staining! To: Message-ID: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> Content-Type: text/plain; charset=US-ASCII Dear All, I am new with histology, and posted yesterday a message of: Help needed!!! Immunofluorescence double staining with mouse lung. Thanks a lot for the several helpful responses. As asked, I am here putting on my concise protocols for your check: Sampling: Cut a small cut along the mouse trachea, injected neg 50 (similar to OCT). Then quick freeze with liquid nitrogen, and the stored @ -80; After section with 6 um, slides were fixed with icy acetone for 5 minutes. Quick dry, and then stored @ -20 till use. Staining: 1. Washing 3 X 5 mins; Blocking Buffer (10% Rabbit serum; 5% BSA; 0.1% Tween 20) for 1 hour at room temperature; 2. Tapping away the blocking buffer, and applied first antibody for 1 hour; then wash 3 x 5 min' 3. 2nd antibody 1 hour; washing 3 x 5min; 4. Mounting media, read slides. Count stained with DAPI-mounting media. 5. For double staining, I did the same thing, except using mixed first antibodies and mixed second conjugated antibodies. Designing: Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit conjugated with Alexa Fluor 488; Stain 2: Rat anti-mouse antibody, and donkey anti-rat conjugated Alexa fluor 594; My problems: The individual staining signals are good. But there is some match color between two channels. For example, I could see weak and similar green signal when I applied Alexa 594. Basically, I could see the weak but true alveoli structure of mouse lung. Then I performed double staining, I could see lots of signal match between red and green channels, which should not be true. In this way, I could see basic alveoli structure of mouse lung in both green and red channels. By the way, I tried to red the slides before staining and after blocking, the autofluorescence is very very weak. I am really frustrating with this situations, and please feel free to tell me how I could figure out this problem. Any of your suggestions and comments are very much appreciated. Thanks, ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Chengming Wang DVM, M.S., Ph.D. Department of Pathobiology College of Veterinary Medicine Auburn University 264 Greene Hall Auburn, AL 36849-5519 Voice: (334) 844-2624 Email: wangche@auburn.edu ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 2 *************************************** 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 kemlo.rogerson <@t> waht.swest.nhs.uk Tue Apr 4 08:54:36 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Apr 4 08:53:55 2006 Subject: [Histonet] breast processing Message-ID: Prolonged fixation, thinner blocks, and extended processing. Unfixed fat doesn't process very well mainly in part I guess as you can't cut the blocks thin enough; there's some strange equation concerning time for fluids to penetrate, but basically the thicker the block then the time of penetration rises exponentially. You could also try going back into a dehydration fluid after 'clearing' in xylene as that is a fat solvent; so you remove the fat and expose more tissue to be dehydrated. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "Following the line of least resistance makes both men and rivers crooked" This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Featherstone, Annette [mailto:AFeatherstone@KaleidaHealth.Org] Sent: Tuesday, April 04, 2006 2:26 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] breast processing Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Sunday, April 02, 2006 13:01 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 2 Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. My frustrating experience with double fluorescence staining! (Chengming Wang) ---------------------------------------------------------------------- Message: 1 Date: Sat, 01 Apr 2006 23:25:00 -0600 From: "Chengming Wang" Subject: [Histonet] My frustrating experience with double fluorescence staining! To: Message-ID: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> Content-Type: text/plain; charset=US-ASCII Dear All, I am new with histology, and posted yesterday a message of: Help needed!!! Immunofluorescence double staining with mouse lung. Thanks a lot for the several helpful responses. As asked, I am here putting on my concise protocols for your check: Sampling: Cut a small cut along the mouse trachea, injected neg 50 (similar to OCT). Then quick freeze with liquid nitrogen, and the stored @ -80; After section with 6 um, slides were fixed with icy acetone for 5 minutes. Quick dry, and then stored @ -20 till use. Staining: 1. Washing 3 X 5 mins; Blocking Buffer (10% Rabbit serum; 5% BSA; 0.1% Tween 20) for 1 hour at room temperature; 2. Tapping away the blocking buffer, and applied first antibody for 1 hour; then wash 3 x 5 min' 3. 2nd antibody 1 hour; washing 3 x 5min; 4. Mounting media, read slides. Count stained with DAPI-mounting media. 5. For double staining, I did the same thing, except using mixed first antibodies and mixed second conjugated antibodies. Designing: Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit conjugated with Alexa Fluor 488; Stain 2: Rat anti-mouse antibody, and donkey anti-rat conjugated Alexa fluor 594; My problems: The individual staining signals are good. But there is some match color between two channels. For example, I could see weak and similar green signal when I applied Alexa 594. Basically, I could see the weak but true alveoli structure of mouse lung. Then I performed double staining, I could see lots of signal match between red and green channels, which should not be true. In this way, I could see basic alveoli structure of mouse lung in both green and red channels. By the way, I tried to red the slides before staining and after blocking, the autofluorescence is very very weak. I am really frustrating with this situations, and please feel free to tell me how I could figure out this problem. Any of your suggestions and comments are very much appreciated. Thanks, ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Chengming Wang DVM, M.S., Ph.D. Department of Pathobiology College of Veterinary Medicine Auburn University 264 Greene Hall Auburn, AL 36849-5519 Voice: (334) 844-2624 Email: wangche@auburn.edu ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 2 *************************************** 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 tkngflght <@t> yahoo.com Tue Apr 4 09:09:02 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Tue Apr 4 09:09:15 2006 Subject: [Histonet] breast processing In-Reply-To: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Message-ID: <20060404140902.17481.qmail@web50909.mail.yahoo.com> Annette-- Having run into this all over the place--there are several solutions (other than cutting thinner and processing longer) that come to mind....in order of simplicity: 1. Have you seen the TALL plastic cassettes? They come about twice as deep as traditional cassettes and give a lot more room for fat and yucky tissues. You'll have to police your grossers not to slack and cut thicker, and the sections will warp but if you hold them down while embedding this is a minor irritation, not a problem You can use the cassettes to embed or switch to the shorter ones that take less storage space. Most of the companies that make cassettes have these in at least one color and they are usefull for things you don't want to trim down for cross section (eyes), as well. 2. On really goopy breasts we used to double process. This makes NO sense but it worked...you'll want to try it on some reserve signed out breast /fatty colon before trying it on current open cases. Instead of embedding the cassettes when they come off the machines, we re-ran them just as they were through the end of the process. Redoing the last 100% alchohols, xylenes and again through the paraffins, WITHOUT taking off the original paraffins and then embedding them allowed for same day turnout (though late) but GOOD solid wax-impregnated sections. We scheduled a special process program and rotated the solutions after taking the breast and fatty colon stuff off the machines to prevent any contamination to the next runs. We've done this for large polyps and other medium sized tissues that didn't fix or process the first time into the middles as well--but please run a test first to make sure it will work for your variables. 3. Another old-fashioned method is to use, instead of cassettes, the old metal perforated baskets and group your tissues. They are like mini-petri dishes made out of metal and can hold a number of sections in one basket. find any tech who's been working for more than 25 years and they'll describe them to you better than this. (remember paper tags, guys!!) You'll need to include a paper tag with the case number in pencil for ID. These might be hard to find and will change your cassetting a bit but again--the tissues float more freely, have more solution in contact with the surface and are better processed for the effort. Run these baskets through the purge cycle to clean or run under hot soapy water before using again...I have no idea if they are still being made but most old labs have a drawer full somewhere. Hope this helps-- Cheryl "Featherstone, Annette" wrote: Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. From jstn192 <@t> yahoo.com Tue Apr 4 10:31:17 2006 From: jstn192 <@t> yahoo.com (Justin Thomas) Date: Tue Apr 4 10:31:21 2006 Subject: [Histonet] Microtome knife sharpening/ plastic microwave accessories Message-ID: <20060404153117.28033.qmail@web35709.mail.mud.yahoo.com> Just to let everyone know: There is finally a company out there that sharpens microtome knifes (both stainless steel & tungsten) and manufactures plastic microwave accessories at a low price. I send all our knifes to them and I am extremely satisfied with the customer service I receive, along with the pricing. Also, they have made a number of accessories for us and I have nothing but good words to say about them. The accessory's work well and with no problem. The best thing about them is the company offers a warranty with each accessory. If you wish, email me back with your contact information and I will pass it on to the sharpening and MW accessory company. Thanks, Dr. Thomas --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From PIXLEYSK <@t> UCMAIL.UC.EDU Tue Apr 4 11:06:31 2006 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Tue Apr 4 11:06:35 2006 Subject: [Histonet] Homemade humidity chamber Message-ID: Dear Louise: We make our own. We use a Tupperware container. Then we purchase plastic ceiling "tiles" from the hardware store. The ones we get look like a white plastic grid, with the holes approximately 1 cm by 1 cm. Comes in sheets about 4 feet by 3 feet wide, for very low cost. Use pliers to cut into pieces that fit into the container. We put down two pieces of this in the Tupperware container, and then you can fill with water up to about a cm deep. On top of the tile pieces, we put a metal slide tray (holds 20 slides, came in a set with a wooden box and many trays). If I want to do more than 20, I put down one tray, set single pieces of the ceiling tile on the corners and set a second tray on top. With two tiles in there, you can put up to a cm of water in, ensuring humidity. Sarah Pixley Univ. Cincinnati, Ohio From gcallis <@t> montana.edu Tue Apr 4 11:17:00 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Apr 4 11:17:13 2006 Subject: [Histonet] Microtome knife sharpening/ plastic microwave accessories In-Reply-To: <20060404153117.28033.qmail@web35709.mail.mud.yahoo.com> References: <20060404153117.28033.qmail@web35709.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20060404101545.01b4e140@gemini.msu.montana.edu> As a histonetter, and NOT a vendor, you can give the information out without getting flamed, although I like vendor commentary and answers to questions. At 09:31 AM 4/4/2006, you wrote: >Just to let everyone know: > > There is finally a company out there that sharpens microtome knifes > (both stainless steel & tungsten) and manufactures plastic microwave > accessories at a low price. I send all our knifes to them and I am > extremely satisfied with the customer service I receive, along with the > pricing. Also, they have made a number of accessories for us and I have > nothing but good words to say about them. The accessory's work well and > with no problem. The best thing about them is the company offers a > warranty with each accessory. If you wish, email me back with your > contact information and I will pass it on to the sharpening and MW > accessory company. > > Thanks, > Dr. Thomas > > >--------------------------------- >New Yahoo! Messenger with Voice. Call regular phones from your PC and save >big. >_______________________________________________ >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 406 994-4303 (FAX) From kcai <@t> prosci-inc.com Tue Apr 4 11:26:58 2006 From: kcai <@t> prosci-inc.com (karen Cai) Date: Tue Apr 4 11:23:55 2006 Subject: [Histonet] Cleaning Tissue embedding center Message-ID: <000601c65804$98c9e700$7d01a8c0@prosci.com> Hello, I have a Tissue Tek II tissue embedding center. Is there anybody know how to clean up the paraffin inside. I think maybe the paraffin inside get contaminated. Thanks in advance, Best, Karen -- No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.3.5/300 - Release Date: 4/3/2006 From Rcartun <@t> harthosp.org Tue Apr 4 12:13:52 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Tue Apr 4 12:14:30 2006 Subject: [Histonet] PTH Message-ID: Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From naje1972 <@t> yahoo.com Tue Apr 4 13:10:55 2006 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Tue Apr 4 13:11:01 2006 Subject: [Histonet] Humedity chamber Message-ID: <20060404181055.97672.qmail@web33006.mail.mud.yahoo.com> Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. From ROrr <@t> enh.org Tue Apr 4 13:23:54 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Tue Apr 4 13:24:00 2006 Subject: [Histonet] Fatty breast processing Message-ID: HI Annette, You have gotten quite a few responses to your question. I have just one more. When we get huge pieces of fatty breast that is unfixed, we place the block in the embedder, melt it and let it "soak" in the paraffin for 2 hours. I have left them in this paraffin bath as long as 4 hours but am unsure of any damage that could be done,( I haven't seen any) But 2 seems to be as long as we need. I would think that this process would cause damage, but since the tissue is unworkable in the first place, this can only help. This soaking in paraffin seems to do the trick, whether it's helping infiltration or simply dehydrating, I imagine it's doing a little bit of both. After this process, the blocks are much easier to cut. I don't see any problem with my IHC stains especially for ER/PR, Her2. These all seem to stain consistently. Ultimately, you would want to run your samples as others have recommended before it gets to the embedding stage. Hope this helps, Becky Becky Orr, CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 tm@libero.it> Message: 5 Date: Tue, 4 Apr 2006 09:26:26 -0400 From: "Featherstone, Annette" Subject: [Histonet] breast processing To: Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Content-Type: text/plain; charset="iso-8859-1" Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT From Dorothy.L.Webb <@t> HealthPartners.Com Tue Apr 4 13:25:14 2006 From: Dorothy.L.Webb <@t> HealthPartners.Com (Webb, Dorothy L) Date: Tue Apr 4 13:25:25 2006 Subject: [Histonet] neutrophils Message-ID: <0E394B648E5284478A6CCB78E5AFDA270171FD2A@hpes1.HealthPartners.int> Does anyone know of a stain for neutrophils? I believe I had read that staining with chloroacetate esterase is the way to go, but, I am not familiar with that stain. Does anyone know of a company that would have the stain in a kit?? This is for a research project and am rather stumped! Thanks, as always, fellow Histonetters for your help and support!!!! ________________________________________ 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 PMonfils <@t> Lifespan.org Tue Apr 4 13:37:46 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Apr 4 13:37:52 2006 Subject: [Histonet] neutrophils Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176BC@lsexch.lsmaster.lifespan.org> I use the Naphthol AS-D Chloroacetate Esterase Kit from Sigma (cat# 91C-1KT) on frozen sections. It's easy to use and very reliable. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Webb, > Dorothy L > Sent: Tuesday, April 4, 2006 11:25 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] neutrophils > > Does anyone know of a stain for neutrophils? I believe I had read that > staining with chloroacetate esterase is the way to go, but, I am not > familiar with that stain. Does anyone know of a company that would > have the stain in a kit?? This is for a research project and am rather > stumped! Thanks, as always, fellow Histonetters for your help and > support!!!! > ________________________________________ > This e-mail and any files transmitted with it are confidential and are > intended solely for the use of the individual or entity to whom they are > addressed. If you are not the intended recipient or the individual > responsible for delivering the e-mail to the intended recipient, please be > advised that you have received this e-mail in error and that any use, > dissemination, forwarding, printing, or copying of this e-mail is strictly > prohibited. > > If you have received this e-mail in error, please immediately notify the > HealthPartners Support Center by telephone at (952) 967-6600. You will be > reimbursed for reasonable costs incurred in notifying us. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From gpbnas <@t> yahoo.es Tue Apr 4 13:58:33 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Tue Apr 4 13:58:37 2006 Subject: [Histonet] Sirius red with low contrast between cells cytoplasm and stained fibers Message-ID: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From mauger <@t> email.chop.edu Tue Apr 4 14:49:36 2006 From: mauger <@t> email.chop.edu (Joanne Mauger) Date: Tue Apr 4 14:51:21 2006 Subject: [Histonet] Humedity chamber Message-ID: Cynthia, It's www.scytek.com Jo >>> cynthia haynes 04/04/06 2:10 PM >>> Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lowman034 <@t> yahoo.com Tue Apr 4 15:19:41 2006 From: lowman034 <@t> yahoo.com (Dave Low) Date: Tue Apr 4 15:19:44 2006 Subject: [Histonet] Thanks for the Input In-Reply-To: <3F500F8B416C554EBB21FF16642F72E959CE1E@marxchg03.mar.med.navy.mil> Message-ID: <20060404201941.10561.qmail@web32004.mail.mud.yahoo.com> Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From jstn192 <@t> yahoo.com Tue Apr 4 15:22:18 2006 From: jstn192 <@t> yahoo.com (Justin Thomas) Date: Tue Apr 4 15:22:21 2006 Subject: [Histonet] RE: Microtome knife sharpening service/ Microwave accessories Message-ID: <20060404202218.93567.qmail@web35701.mail.mud.yahoo.com> Just to let everyone know: There is finally a company out there that sharpens microtome knifes (both stainless steel & tungsten) and manufactures plastic microwave accessories at a low price. I send all our knifes to them and I am extremely satisfied with the customer service I receive, along with the pricing. Also, they have made a number of accessories for us and I have nothing but good words to say about them. The accessory's work well and with no problem. The best thing about them is the company offers a warranty with each accessory. If you wish, email me back with your contact information and I will pass it on to the sharpening and MW accessory company. Thanks, Dr. Thomas --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From dharclerode <@t> cytoritx.com Tue Apr 4 15:36:57 2006 From: dharclerode <@t> cytoritx.com (Donna Harclerode) Date: Tue Apr 4 15:35:51 2006 Subject: [Histonet] Humidity chamber Message-ID: <3DE0F644E093DF4BAE80C254176696A5090706@mp-mailserver.macropore.com> Thermo Electron makes a great system with a humidity chamber and a coverplate that I have used for many years. The slides are held with the plastic coverplate(the coverplates are suppose to be disposable, but I rinse them with only DI water and reuse them many times) I use 100ul of antibody (primary, secondary- etc, normally, but have managed to use as little as 90ul) to cover almost the whole slide. For buffer wash I fill up the top of the chamber with a squeeze bottle of PBS and allow to run through in about 5 minutes. I load the slides add primary, wash, secondary etc and only take them out for chromagen staining. The racks worked great when I had the positive control at the top of the slide and the test below. I do fluorescence secondaries in the coverplates, but I not do DAB in the coverplates, but flat on the counter. For HRP frozens I use the Dako endogenous peroxidase block in the Sequenza racks. I also use the Dako avidin biotin block in the coverplates when necessary. I usually incubate primary overnight in the fridge and the chambers stay humid for at least 48 hours. It takes a bit of practice to load the plates, but it is so worth it. Sequenza Racks - # 73310017 (Holds 10 coverplate assemblies) Coverplates - 25/pk = # 72110017 50/pk = # 7219950 250/case = # 72110013 http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_19578.pdf 4th page Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 5416 dharclerode@cytoritx.com From Eric <@t> ategra.com Tue Apr 4 16:13:06 2006 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Tue Apr 4 16:01:25 2006 Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW Message-ID: Fellow-Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) Here are some of my Hottest Histology Jobs: 1. Ohio (Southwestern Ohio) (Full-time, Perm, Histo Manager) 2. Northern New Jersey (Full-time, Perm and/or Temp, Bench Histo Tech, 1st shift) 3. Rhode Island - Providence (Full-time, Perm, Bench Histo Tech, 2nd Shift) 4. Northern New Jersey (Full-time, Perm, Bench Histo Tech) 5. Ohio (Southwestern Ohio) (Full-time, Perm, Bench Histo Tech) 6. New York (Long Island) (Full-time, Perm, SUPERVISOR Histo Tech) 7. Virginia(South - Coastal) (Full-time, Perm, Bench Histo Tech) 8. Virginia - Metro DC (Full-time, Perm, Lead Histo Tech) 9. South Florida (Full-time, Perm, Supervisor Histo Tech) 10. South Florida (Full-time, Perm, Bench Histo Tech) 11. Mass (Boston) (Part-time, Perm, Bench Histo Tech) 12. West Viriginia (Full-time, Perm, Bench Histo Tech) The clients are currently interviewing Histology candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- Note: this message is intended for: Fellow-Histonetters at histonet@lists.utsouthwestern.edu ---------------------------------------------------------------- ---------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------------- From gpbnas <@t> yahoo.es Tue Apr 4 16:22:43 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Tue Apr 4 16:22:48 2006 Subject: [Histonet] Hematoxylin staining of frozen samples, where are the nuclei? Message-ID: <20060404212243.32475.qmail@web26209.mail.ukl.yahoo.com> Dear histonetters, When I stain samples placed on VWR frosted slides with Gills hematoxylin, nuclei are nicely stained. However, when the same samples are counterstained after passing through all necessary steps involved in immunohistochemistry, hematoxylin staining of nuclei is simply horrible. Is it possible that nuclei are lost in the different washing steps? Please put forward any suggestions you might have. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From LRaff <@t> lab.uropartners.com Tue Apr 4 16:23:23 2006 From: LRaff <@t> lab.uropartners.com (Lester Raff) Date: Tue Apr 4 16:23:30 2006 Subject: [Histonet] Qualifications for Histology Supervisor Message-ID: <5DA1CA5D0B98A84985B545A24423B822A7AE@UPLAB01.uplab.local> Hi all, We are preparing for our first CAP inspection and want to make sure that my very qualified histology supervisor meets CLIA regs. Does anyone have a copy of the old federal regs 42CFR493.1427 that defined minimal qualifications for general supervisor? Thanks, Lester J. Raff, MD Medical Director UroPartners, LLC Laboratory ph: 708-486-0076 fax: 708-486-0080 From lrichey <@t> u.washington.edu Tue Apr 4 16:41:19 2006 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Tue Apr 4 16:41:35 2006 Subject: [Histonet] humidity chambers In-Reply-To: References: Message-ID: <4432E7FF.4090702@u.washington.edu> We buy Fisher brand absorbant paper, which is plastic coated on one side. We have it spread flat on the counter and wet with water. We put the slides flat on the paper, and cover them with a cafeteria tray. This works well for 100-200 slides a day. louise renton wrote: >Hi all > >what are folks using as humidity chambers for immuno slides (manual >method). Are they predominanty made in-house or are there ready made >commercial products out there? > >best regards >-- >Louise Renton >Bone Research Unit >University of the Witwatersrand >Johannesburg >South Africa >"Does a conceited cowboy, only ride on pompous grass?. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From lrichey <@t> u.washington.edu Tue Apr 4 16:49:48 2006 From: lrichey <@t> u.washington.edu (Lori Richey) Date: Tue Apr 4 16:49:52 2006 Subject: [Histonet] breast processing In-Reply-To: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> References: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Message-ID: <4432E9FC.2080100@u.washington.edu> Alcoholic Formalin or Penfix in the processor helps some, if the tissue is grossed in at a reasonable thickness.. Featherstone, Annette wrote: >Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. > >Annette Featherstone HT/MLT > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >histonet-request@lists.utsouthwestern.edu >Sent: Sunday, April 02, 2006 13:01 >To: histonet@lists.utsouthwestern.edu >Subject: Histonet Digest, Vol 29, Issue 2 > > >Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > >To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > >You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > >When replying, please edit your Subject line so it is more specific >than "Re: Contents of Histonet digest..." > > >Today's Topics: > > 1. My frustrating experience with double fluorescence staining! > (Chengming Wang) > > >---------------------------------------------------------------------- > >Message: 1 >Date: Sat, 01 Apr 2006 23:25:00 -0600 >From: "Chengming Wang" >Subject: [Histonet] My frustrating experience with double fluorescence > staining! >To: >Message-ID: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> >Content-Type: text/plain; charset=US-ASCII > >Dear All, > >I am new with histology, and posted yesterday a message of: Help >needed!!! Immunofluorescence double staining with mouse lung. Thanks a >lot for the several helpful responses. As asked, I am here putting on >my concise protocols for your check: > >Sampling: Cut a small cut along the mouse trachea, injected neg 50 >(similar to OCT). Then quick freeze with liquid nitrogen, and the stored >@ -80; After section with 6 um, slides were fixed with icy acetone for >5 minutes. Quick dry, and then stored @ -20 till use. > >Staining: 1. Washing 3 X 5 mins; Blocking Buffer (10% Rabbit serum; 5% >BSA; 0.1% Tween 20) for 1 hour at room temperature; > 2. Tapping away the blocking buffer, and applied first >antibody for 1 hour; then wash 3 x 5 min' > 3. 2nd antibody 1 hour; washing 3 x 5min; > 4. Mounting media, read slides. Count stained with >DAPI-mounting media. > 5. For double staining, I did the same thing, except >using mixed first antibodies and mixed second conjugated antibodies. > >Designing: Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit >conjugated with Alexa Fluor 488; > Stain 2: Rat anti-mouse antibody, and donkey anti-rat >conjugated Alexa fluor 594; > >My problems: The individual staining signals are good. But there is >some match color between two channels. For example, I could see weak and >similar green signal when I applied Alexa 594. Basically, I could see >the weak but true alveoli structure of mouse > lung. > Then I performed double staining, I could see >lots of signal match between red and green channels, which should not be >true. In this way, I could see basic alveoli structure of mouse > lung in both green and red channels. By the way, >I tried to red the slides before staining and after blocking, the >autofluorescence is very very weak. > >I am really frustrating with this situations, and please feel free to >tell me how I could figure out this problem. Any of your suggestions and >comments are very much appreciated. > >Thanks, > > > >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Chengming Wang >DVM, M.S., Ph.D. >Department of Pathobiology >College of Veterinary Medicine >Auburn University >264 Greene Hall >Auburn, AL 36849-5519 > >Voice: (334) 844-2624 >Email: wangche@auburn.edu > > > > >------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest, Vol 29, Issue 2 >*************************************** > > >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 banaw <@t> HSC.EDU.KW Tue Apr 4 16:50:47 2006 From: banaw <@t> HSC.EDU.KW (Anwar Al-Banaw) Date: Tue Apr 4 16:50:38 2006 Subject: [Histonet] Teaching Microtomes Message-ID: <59A3C2F593DF494294C4E0E8FFDC9028143D9F@MAIL.HSC.EDU.KW> Hi, I am planning to get few new manual rotary microtomes for teaching purposes. I heard about "accu-cut" from Tissue-Tek Sakura, have a simple microtome but I want the review of ppl used this machine. The other choices that I have are from Leica and Shandon. Regards A AlBanaw Medical Lab Sci. Kuwait University Kuwait From jkiernan <@t> uwo.ca Tue Apr 4 16:58:33 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Apr 4 16:57:34 2006 Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers In-Reply-To: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> References: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> Message-ID: <4432EC09.3010504@uwo.ca> Instead of your steps 3 and 4, rinse in slightly acidified water. (Your 0.01% HCl should be OK; I use 0.2-0.5% acetic; about 30 s with agitation). You can do 2 rinses if there's too much colour in the water. This step does not extract any bound dye. Shake off as much acidified water as you can before the next step. For your step 5 go directly into the first of 3 changes of 100% alcohol. Agitate slides for about 30 sec in each, then clear (xylene X2) and apply a coverslip. You lose some of the picric acid in the dehydration, but there's enough left in the section to give a yellow colour to everything except collagen. Nuclei are also yellow, unless you stain them black before the picri-sirius red. The long stay in the staining solution extracts even Weigert's iron-haematoxylin, so a prior nuclear stain needs to be decidedly overdone. Junqueira at al (1979, Histochem. J. 11:447-455) thoroughly studied this method; the paper is well worth reading. John Kiernan Anatomy, UWO London, Canada ------------------- Guillermo Palao wrote: > > Hello all: > > I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. > > My protocol is as follows: > > 1. Deparafinize and hydrate samples > 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid > 3. Wash 2 min 0.01 N HCl > 4. Rinse in water > 5. Dehydrate and cover slides > > I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. > > Guillermo > > > --------------------------------- > > LLama Gratis a cualquier PC del Mundo. > Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. > http://es.voice.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jenbug812 <@t> aol.com Tue Apr 4 17:14:10 2006 From: jenbug812 <@t> aol.com (jenbug812@aol.com) Date: Tue Apr 4 17:14:18 2006 Subject: [Histonet] Feedback on Milestone Medical's RHS Series Microwave Tissue Processor Message-ID: <8C8264C3B5C9264-1088-88C3@mblk-d50.sysops.aol.com> Just wondering if anyone is currently using this tissue processor and if so, can you provide any feedback, good or bad? My company is conducting demos with this product and I would like to be prepared for the sales rep. Thanks From AnthonyH <@t> chw.edu.au Tue Apr 4 17:27:17 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Tue Apr 4 17:27:24 2006 Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Message-ID: Drop step 3 and 4 (picric acid is soluble in water) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Guillermo Palao Sent: Wednesday, 5 April 2006 4:59 AM To: Histonet Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From katri <@t> cogeco.ca Tue Apr 4 18:05:39 2006 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Tue Apr 4 18:05:38 2006 Subject: [Histonet] Hematoxylin staining of frozen samples, where are the nuclei? References: <20060404212243.32475.qmail@web26209.mail.ukl.yahoo.com> Message-ID: <004b01c6583c$4a92e220$6a9a9618@Katri> Guillermo, Tissues go through a long procedure with immuno and the most detrimental for all tissue components is the heat retrieval and to a lesser degree enzyme digestion. The less well fixed and processed tissue suffers most: connective tissue is distorted and nuclei stain very poorly. Hope this explains your problem, Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Guillermo Palao" To: "Histonet" Sent: Tuesday, April 04, 2006 5:22 PM Subject: [Histonet] Hematoxylin staining of frozen samples,where are the nuclei? > Dear histonetters, > > When I stain samples placed on VWR frosted slides with Gills hematoxylin, > nuclei are nicely stained. However, when the same samples are > counterstained after passing through all necessary steps involved in > immunohistochemistry, hematoxylin staining of nuclei is simply horrible. > Is it possible that nuclei are lost in the different washing steps? Please > put forward any suggestions you might have. > > Guillermo > > > --------------------------------- > > LLama Gratis a cualquier PC del Mundo. > Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. > http://es.voice.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JMacDonald <@t> mtsac.edu Wed Apr 5 00:31:56 2006 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Wed Apr 5 00:32:21 2006 Subject: [Histonet] neutrophils Message-ID: Napththol AS-D Chloroacetate Esterase is used to identify granulo cytes. This procedure can be done on FFPE tissue,or blood smears.&nbs usually done on < -----histonet-bounces To: Histonet@lists.utsouthwestern.edu Fr Sent by: Date: 04/04/2006 11:25AM S Does anyone know of a stain for neutro read that staining with chloroacetate ester not familiar with that stain. Does would have the stain in a kit?? &nb and am rather stumped! Thanks, a and support!!!! ___ ______________________ 5F__ This e-mail and any files transmitt and are intended solely for the use of the indi to whom they are addressed. If you are not the intended re or the individual responsible for delivering the e-mail to the inte e-mail in err printing, or copying of thi If you have received this e-mail in notify the HealthPartners Support Center by tele 967-6600. You will be reimbursed for reasonable costs incurr notifying us. _________________ ______________________ 5F__ Histonet mailing list Histonet@lists.utsouthwestern.e [1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet" From DDDeltour <@t> mar.med.navy.mil Wed Apr 5 04:15:57 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Wed Apr 5 04:16:25 2006 Subject: [Histonet] Thanks for the Input Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE25@marxchg03.mar.med.navy.mil> It seems lately some people take bits and pieces of posts and twist them around for argument sake. I am sure you are not quoting me?? I can speak on the part of the Navy only. Maybe I should have stated that on my last post. As for the Navy ...While the others are out selling cookies others must stay behind and pick up the slack. These cookie sellers place the bake sale on the evaluation which earns them the bullet for a higher mark. The higher mark means a better chance for promotion. A more experienced and certified tech may not have the time to sell these cookies for whatever reason or because they care more about the REAL job than playing Martha Stewart. Green food coloring is not a counterstain. -----Original Message----- From: Dave Low [mailto:lowman034@yahoo.com] Sent: Tuesday, April 04, 2006 4:20 PM To: Deltour, Douglas D. (HM2); vazquezr@ohsu.edu; Jackie.O'Connor@abbott.com; Harper, Heather A., CIV Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks for the Input Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Apr 5 04:33:20 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Apr 5 04:32:51 2006 Subject: [Histonet] Thanks for the Input Message-ID: Geez I'm losing this!!!! Does the army make cakes? I thought they shot rifles and things. You get promoted if you can make a good cake but not if you shoot straight? Lost it totally. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "Following the line of least resistance makes both men and rivers crooked" This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: DDDeltour@mar.med.navy.mil [mailto:DDDeltour@mar.med.navy.mil] Sent: Wednesday, April 05, 2006 10:16 AM To: lowman034@yahoo.com; vazquezr@ohsu.edu; Jackie.O'Connor@abbott.com; Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks for the Input It seems lately some people take bits and pieces of posts and twist them around for argument sake. I am sure you are not quoting me?? I can speak on the part of the Navy only. Maybe I should have stated that on my last post. As for the Navy ...While the others are out selling cookies others must stay behind and pick up the slack. These cookie sellers place the bake sale on the evaluation which earns them the bullet for a higher mark. The higher mark means a better chance for promotion. A more experienced and certified tech may not have the time to sell these cookies for whatever reason or because they care more about the REAL job than playing Martha Stewart. Green food coloring is not a counterstain. -----Original Message----- From: Dave Low [mailto:lowman034@yahoo.com] Sent: Tuesday, April 04, 2006 4:20 PM To: Deltour, Douglas D. (HM2); vazquezr@ohsu.edu; Jackie.O'Connor@abbott.com; Harper, Heather A., CIV Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks for the Input Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AFeatherstone <@t> KaleidaHealth.Org Wed Apr 5 06:05:58 2006 From: AFeatherstone <@t> KaleidaHealth.Org (Featherstone, Annette) Date: Wed Apr 5 06:06:11 2006 Subject: [Histonet] validation for CAP Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA5515B@KALEXMB02.KaleidaHealth.org> We have just purchased 2 new Dako immuno stainers and I was wondering what CAP requirements are for validation. The company says that both instruments are validated against each other and therefore you can validate the antibodies on either stainer. I hope this is fact. Annette Featherstone HT/MLT MT(HEW) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, April 05, 2006 05:34 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 6 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. PTH (Richard Cartun) 2. Humedity chamber (cynthia haynes) 3. Fatty breast processing (Orr, Rebecca) 4. neutrophils (Webb, Dorothy L) 5. RE: neutrophils (Monfils, Paul) 6. Sirius red with low contrast between cells cytoplasm and stained fibers (Guillermo Palao) 7. Re: Humedity chamber (Joanne Mauger) 8. RE: Thanks for the Input (Dave Low) 9. RE: Microtome knife sharpening service/ Microwave accessories (Justin Thomas) 10. Humidity chamber (Donna Harclerode) 11. Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW (Eric Dye (ext 223)) 12. Hematoxylin staining of frozen samples, where are the nuclei? (Guillermo Palao) 13. Qualifications for Histology Supervisor (Lester Raff) 14. Re: humidity chambers (Lori Richey) 15. Re: breast processing (Lori Richey) 16. Teaching Microtomes (Anwar Al-Banaw) 17. Re: Sirius red with low contrast between cells cytoplasm andstained fibers (John Kiernan) 18. Feedback on Milestone Medical's RHS Series Microwave Tissue Processor (jenbug812@aol.com) 19. RE: Sirius red with low contrast between cells cytoplasm andstained fibers (Tony Henwood) 20. Re: Hematoxylin staining of frozen samples, where are the nuclei? (Katri Tuomala) 21. Re: neutrophils (Jennifer MacDonald) 22. RE: Thanks for the Input (DDDeltour@mar.med.navy.mil) 23. RE: Thanks for the Input (Kemlo Rogerson) ---------------------------------------------------------------------- Message: 1 Date: Tue, 04 Apr 2006 13:13:52 -0400 From: "Richard Cartun" Subject: [Histonet] PTH To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ------------------------------ Message: 2 Date: Tue, 4 Apr 2006 11:10:55 -0700 (PDT) From: cynthia haynes Subject: [Histonet] Humedity chamber To: Histonet@lists.utsouthwestern.edu Message-ID: <20060404181055.97672.qmail@web33006.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. ------------------------------ Message: 3 Date: Tue, 4 Apr 2006 13:23:54 -0500 From: "Orr, Rebecca" Subject: [Histonet] Fatty breast processing To: Message-ID: Content-Type: text/plain; charset="US-ASCII" HI Annette, You have gotten quite a few responses to your question. I have just one more. When we get huge pieces of fatty breast that is unfixed, we place the block in the embedder, melt it and let it "soak" in the paraffin for 2 hours. I have left them in this paraffin bath as long as 4 hours but am unsure of any damage that could be done,( I haven't seen any) But 2 seems to be as long as we need. I would think that this process would cause damage, but since the tissue is unworkable in the first place, this can only help. This soaking in paraffin seems to do the trick, whether it's helping infiltration or simply dehydrating, I imagine it's doing a little bit of both. After this process, the blocks are much easier to cut. I don't see any problem with my IHC stains especially for ER/PR, Her2. These all seem to stain consistently. Ultimately, you would want to run your samples as others have recommended before it gets to the embedding stage. Hope this helps, Becky Becky Orr, CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 tm@libero.it> Message: 5 Date: Tue, 4 Apr 2006 09:26:26 -0400 From: "Featherstone, Annette" Subject: [Histonet] breast processing To: Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Content-Type: text/plain; charset="iso-8859-1" Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT ------------------------------ Message: 4 Date: Tue, 04 Apr 2006 13:25:14 -0500 From: "Webb, Dorothy L" Subject: [Histonet] neutrophils To: Histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA270171FD2A@hpes1.HealthPartners.int> Content-Type: text/plain; charset="US-ASCII" Does anyone know of a stain for neutrophils? I believe I had read that staining with chloroacetate esterase is the way to go, but, I am not familiar with that stain. Does anyone know of a company that would have the stain in a kit?? This is for a research project and am rather stumped! Thanks, as always, fellow Histonetters for your help and support!!!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 5 Date: Tue, 4 Apr 2006 14:37:46 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] neutrophils To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176BC@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I use the Naphthol AS-D Chloroacetate Esterase Kit from Sigma (cat# 91C-1KT) on frozen sections. It's easy to use and very reliable. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Webb, > Dorothy L > Sent: Tuesday, April 4, 2006 11:25 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] neutrophils > > Does anyone know of a stain for neutrophils? I believe I had read that > staining with chloroacetate esterase is the way to go, but, I am not > familiar with that stain. Does anyone know of a company that would > have the stain in a kit?? This is for a research project and am rather > stumped! Thanks, as always, fellow Histonetters for your help and > support!!!! > ________________________________________ > This e-mail and any files transmitted with it are confidential and are > intended solely for the use of the individual or entity to whom they are > addressed. If you are not the intended recipient or the individual > responsible for delivering the e-mail to the intended recipient, please be > advised that you have received this e-mail in error and that any use, > dissemination, forwarding, printing, or copying of this e-mail is strictly > prohibited. > > If you have received this e-mail in error, please immediately notify the > HealthPartners Support Center by telephone at (952) 967-6600. You will be > reimbursed for reasonable costs incurred in notifying us. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 6 Date: Tue, 4 Apr 2006 20:58:33 +0200 (CEST) From: Guillermo Palao Subject: [Histonet] Sirius red with low contrast between cells cytoplasm and stained fibers To: Histonet Message-ID: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com ------------------------------ Message: 7 Date: Tue, 04 Apr 2006 15:49:36 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] Humedity chamber To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Cynthia, It's www.scytek.com Jo >>> cynthia haynes 04/04/06 2:10 PM >>> Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 4 Apr 2006 13:19:41 -0700 (PDT) From: Dave Low Subject: RE: [Histonet] Thanks for the Input To: DDDeltour@mar.med.navy.mil, vazquezr@ohsu.edu, Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060404201941.10561.qmail@web32004.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 9 Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) From: Justin Thomas Subject: [Histonet] RE: Microtome knife sharpening service/ Microwave accessories To: histonet@lists.utsouthwestern.edu Message-ID: <20060404202218.93567.qmail@web35701.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Just to let everyone know: There is finally a company out there that sharpens microtome knifes (both stainless steel & tungsten) and manufactures plastic microwave accessories at a low price. I send all our knifes to them and I am extremely satisfied with the customer service I receive, along with the pricing. Also, they have made a number of accessories for us and I have nothing but good words to say about them. The accessory's work well and with no problem. The best thing about them is the company offers a warranty with each accessory. If you wish, email me back with your contact information and I will pass it on to the sharpening and MW accessory company. Thanks, Dr. Thomas --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. ------------------------------ Message: 10 Date: Tue, 4 Apr 2006 13:36:57 -0700 From: "Donna Harclerode" Subject: [Histonet] Humidity chamber To: Message-ID: <3DE0F644E093DF4BAE80C254176696A5090706@mp-mailserver.macropore.com> Content-Type: text/plain; charset="us-ascii" Thermo Electron makes a great system with a humidity chamber and a coverplate that I have used for many years. The slides are held with the plastic coverplate(the coverplates are suppose to be disposable, but I rinse them with only DI water and reuse them many times) I use 100ul of antibody (primary, secondary- etc, normally, but have managed to use as little as 90ul) to cover almost the whole slide. For buffer wash I fill up the top of the chamber with a squeeze bottle of PBS and allow to run through in about 5 minutes. I load the slides add primary, wash, secondary etc and only take them out for chromagen staining. The racks worked great when I had the positive control at the top of the slide and the test below. I do fluorescence secondaries in the coverplates, but I not do DAB in the coverplates, but flat on the counter. For HRP frozens I use the Dako endogenous peroxidase block in the Sequenza racks. I also use the Dako avidin biotin block in the coverplates when necessary. I usually incubate primary overnight in the fridge and the chambers stay humid for at least 48 hours. It takes a bit of practice to load the plates, but it is so worth it. Sequenza Racks - # 73310017 (Holds 10 coverplate assemblies) Coverplates - 25/pk = # 72110017 50/pk = # 7219950 250/case = # 72110013 http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_19578.pdf 4th page Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 5416 dharclerode@cytoritx.com ------------------------------ Message: 11 Date: Tue, 4 Apr 2006 17:13:06 -0400 From: Eric Dye (ext 223) Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW To: Histonetters Message-ID: Content-Type: text/plain Fellow-Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) Here are some of my Hottest Histology Jobs: 1. Ohio (Southwestern Ohio) (Full-time, Perm, Histo Manager) 2. Northern New Jersey (Full-time, Perm and/or Temp, Bench Histo Tech, 1st shift) 3. Rhode Island - Providence (Full-time, Perm, Bench Histo Tech, 2nd Shift) 4. Northern New Jersey (Full-time, Perm, Bench Histo Tech) 5. Ohio (Southwestern Ohio) (Full-time, Perm, Bench Histo Tech) 6. New York (Long Island) (Full-time, Perm, SUPERVISOR Histo Tech) 7. Virginia(South - Coastal) (Full-time, Perm, Bench Histo Tech) 8. Virginia - Metro DC (Full-time, Perm, Lead Histo Tech) 9. South Florida (Full-time, Perm, Supervisor Histo Tech) 10. South Florida (Full-time, Perm, Bench Histo Tech) 11. Mass (Boston) (Part-time, Perm, Bench Histo Tech) 12. West Viriginia (Full-time, Perm, Bench Histo Tech) The clients are currently interviewing Histology candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- Note: this message is intended for: Fellow-Histonetters at histonet@lists.utsouthwestern.edu ---------------------------------------------------------------- ---------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------------- ------------------------------ Message: 12 Date: Tue, 4 Apr 2006 23:22:43 +0200 (CEST) From: Guillermo Palao Subject: [Histonet] Hematoxylin staining of frozen samples, where are the nuclei? To: Histonet Message-ID: <20060404212243.32475.qmail@web26209.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Dear histonetters, When I stain samples placed on VWR frosted slides with Gills hematoxylin, nuclei are nicely stained. However, when the same samples are counterstained after passing through all necessary steps involved in immunohistochemistry, hematoxylin staining of nuclei is simply horrible. Is it possible that nuclei are lost in the different washing steps? Please put forward any suggestions you might have. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com ------------------------------ Message: 13 Date: Tue, 4 Apr 2006 16:23:23 -0500 From: "Lester Raff" Subject: [Histonet] Qualifications for Histology Supervisor To: Message-ID: <5DA1CA5D0B98A84985B545A24423B822A7AE@UPLAB01.uplab.local> Content-Type: text/plain; charset="us-ascii" Hi all, We are preparing for our first CAP inspection and want to make sure that my very qualified histology supervisor meets CLIA regs. Does anyone have a copy of the old federal regs 42CFR493.1427 that defined minimal qualifications for general supervisor? Thanks, Lester J. Raff, MD Medical Director UroPartners, LLC Laboratory ph: 708-486-0076 fax: 708-486-0080 ------------------------------ Message: 14 Date: Tue, 04 Apr 2006 14:41:19 -0700 From: Lori Richey Subject: Re: [Histonet] humidity chambers To: louise renton Cc: Histonet@lists.utsouthwestern.edu Message-ID: <4432E7FF.4090702@u.washington.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed We buy Fisher brand absorbant paper, which is plastic coated on one side. We have it spread flat on the counter and wet with water. We put the slides flat on the paper, and cover them with a cafeteria tray. This works well for 100-200 slides a day. louise renton wrote: >Hi all > >what are folks using as humidity chambers for immuno slides (manual >method). Are they predominanty made in-house or are there ready made >commercial products out there? > >best regards >-- >Louise Renton >Bone Research Unit >University of the Witwatersrand >Johannesburg >South Africa >"Does a conceited cowboy, only ride on pompous grass?. > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 15 Date: Tue, 04 Apr 2006 14:49:48 -0700 From: Lori Richey Subject: Re: [Histonet] breast processing To: "Featherstone, Annette" Cc: histonet@lists.utsouthwestern.edu Message-ID: <4432E9FC.2080100@u.washington.edu> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Alcoholic Formalin or Penfix in the processor helps some, if the tissue is grossed in at a reasonable thickness.. Featherstone, Annette wrote: >Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. > >Annette Featherstone HT/MLT > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of >histonet-request@lists.utsouthwestern.edu >Sent: Sunday, April 02, 2006 13:01 >To: histonet@lists.utsouthwestern.edu >Subject: Histonet Digest, Vol 29, Issue 2 > > >Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > >To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > >You can reach the person managing the list at > histonet-owner@lists.utsouthwestern.edu > >When replying, please edit your Subject line so it is more specific >than "Re: Contents of Histonet digest..." > > >Today's Topics: > > 1. My frustrating experience with double fluorescence staining! > (Chengming Wang) > > >---------------------------------------------------------------------- > >Message: 1 >Date: Sat, 01 Apr 2006 23:25:00 -0600 >From: "Chengming Wang" >Subject: [Histonet] My frustrating experience with double fluorescence > staining! >To: >Message-ID: <442F0BCC020000AF0001AB19@groupwise1.duc.auburn.edu> >Content-Type: text/plain; charset=US-ASCII > >Dear All, > >I am new with histology, and posted yesterday a message of: Help >needed!!! Immunofluorescence double staining with mouse lung. Thanks a >lot for the several helpful responses. As asked, I am here putting on >my concise protocols for your check: > >Sampling: Cut a small cut along the mouse trachea, injected neg 50 >(similar to OCT). Then quick freeze with liquid nitrogen, and the stored >@ -80; After section with 6 um, slides were fixed with icy acetone for >5 minutes. Quick dry, and then stored @ -20 till use. > >Staining: 1. Washing 3 X 5 mins; Blocking Buffer (10% Rabbit serum; 5% >BSA; 0.1% Tween 20) for 1 hour at room temperature; > 2. Tapping away the blocking buffer, and applied first >antibody for 1 hour; then wash 3 x 5 min' > 3. 2nd antibody 1 hour; washing 3 x 5min; > 4. Mounting media, read slides. Count stained with >DAPI-mounting media. > 5. For double staining, I did the same thing, except >using mixed first antibodies and mixed second conjugated antibodies. > >Designing: Stain 1: Rabbit anti-mouse antibody, and donkey anti-rabbit >conjugated with Alexa Fluor 488; > Stain 2: Rat anti-mouse antibody, and donkey anti-rat >conjugated Alexa fluor 594; > >My problems: The individual staining signals are good. But there is >some match color between two channels. For example, I could see weak and >similar green signal when I applied Alexa 594. Basically, I could see >the weak but true alveoli structure of mouse > lung. > Then I performed double staining, I could see >lots of signal match between red and green channels, which should not be >true. In this way, I could see basic alveoli structure of mouse > lung in both green and red channels. By the way, >I tried to red the slides before staining and after blocking, the >autofluorescence is very very weak. > >I am really frustrating with this situations, and please feel free to >tell me how I could figure out this problem. Any of your suggestions and >comments are very much appreciated. > >Thanks, > > > >~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ >Chengming Wang >DVM, M.S., Ph.D. >Department of Pathobiology >College of Veterinary Medicine >Auburn University >264 Greene Hall >Auburn, AL 36849-5519 > >Voice: (334) 844-2624 >Email: wangche@auburn.edu > > > > >------------------------------ > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest, Vol 29, Issue 2 >*************************************** > > >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: 16 Date: Wed, 5 Apr 2006 00:50:47 +0300 From: "Anwar Al-Banaw" Subject: [Histonet] Teaching Microtomes To: Message-ID: <59A3C2F593DF494294C4E0E8FFDC9028143D9F@MAIL.HSC.EDU.KW> Content-Type: text/plain; charset="iso-8859-1" Hi, I am planning to get few new manual rotary microtomes for teaching purposes. I heard about "accu-cut" from Tissue-Tek Sakura, have a simple microtome but I want the review of ppl used this machine. The other choices that I have are from Leica and Shandon. Regards A AlBanaw Medical Lab Sci. Kuwait University Kuwait ------------------------------ Message: 17 Date: Tue, 04 Apr 2006 17:58:33 -0400 From: John Kiernan Subject: Re: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers To: Guillermo Palao Cc: Histonet Message-ID: <4432EC09.3010504@uwo.ca> Content-Type: text/plain; charset=iso-8859-1; format=flowed Instead of your steps 3 and 4, rinse in slightly acidified water. (Your 0.01% HCl should be OK; I use 0.2-0.5% acetic; about 30 s with agitation). You can do 2 rinses if there's too much colour in the water. This step does not extract any bound dye. Shake off as much acidified water as you can before the next step. For your step 5 go directly into the first of 3 changes of 100% alcohol. Agitate slides for about 30 sec in each, then clear (xylene X2) and apply a coverslip. You lose some of the picric acid in the dehydration, but there's enough left in the section to give a yellow colour to everything except collagen. Nuclei are also yellow, unless you stain them black before the picri-sirius red. The long stay in the staining solution extracts even Weigert's iron-haematoxylin, so a prior nuclear stain needs to be decidedly overdone. Junqueira at al (1979, Histochem. J. 11:447-455) thoroughly studied this method; the paper is well worth reading. John Kiernan Anatomy, UWO London, Canada ------------------- Guillermo Palao wrote: > > Hello all: > > I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. > > My protocol is as follows: > > 1. Deparafinize and hydrate samples > 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid > 3. Wash 2 min 0.01 N HCl > 4. Rinse in water > 5. Dehydrate and cover slides > > I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. > > Guillermo > > > --------------------------------- > > LLama Gratis a cualquier PC del Mundo. > Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. > http://es.voice.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 18 Date: Tue, 04 Apr 2006 18:14:10 -0400 From: jenbug812@aol.com Subject: [Histonet] Feedback on Milestone Medical's RHS Series Microwave Tissue Processor To: histonet@lists.utsouthwestern.edu Message-ID: <8C8264C3B5C9264-1088-88C3@mblk-d50.sysops.aol.com> Content-Type: text/plain; charset="us-ascii" Just wondering if anyone is currently using this tissue processor and if so, can you provide any feedback, good or bad? My company is conducting demos with this product and I would like to be prepared for the sales rep. Thanks ------------------------------ Message: 19 Date: Wed, 5 Apr 2006 08:27:17 +1000 From: "Tony Henwood" Subject: RE: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers To: "Guillermo Palao" , "Histonet" Message-ID: Content-Type: text/plain; charset="iso-8859-1" Drop step 3 and 4 (picric acid is soluble in water) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Guillermo Palao Sent: Wednesday, 5 April 2006 4:59 AM To: Histonet Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ------------------------------ Message: 20 Date: Tue, 4 Apr 2006 19:05:39 -0400 From: "Katri Tuomala" Subject: Re: [Histonet] Hematoxylin staining of frozen samples, where are the nuclei? To: "Guillermo Palao" , "Histonet" Message-ID: <004b01c6583c$4a92e220$6a9a9618@Katri> Content-Type: text/plain; format=flowed; charset="iso-8859-1"; reply-type=original Guillermo, Tissues go through a long procedure with immuno and the most detrimental for all tissue components is the heat retrieval and to a lesser degree enzyme digestion. The less well fixed and processed tissue suffers most: connective tissue is distorted and nuclei stain very poorly. Hope this explains your problem, Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Guillermo Palao" To: "Histonet" Sent: Tuesday, April 04, 2006 5:22 PM Subject: [Histonet] Hematoxylin staining of frozen samples,where are the nuclei? > Dear histonetters, > > When I stain samples placed on VWR frosted slides with Gills hematoxylin, > nuclei are nicely stained. However, when the same samples are > counterstained after passing through all necessary steps involved in > immunohistochemistry, hematoxylin staining of nuclei is simply horrible. > Is it possible that nuclei are lost in the different washing steps? Please > put forward any suggestions you might have. > > Guillermo > > > --------------------------------- > > LLama Gratis a cualquier PC del Mundo. > Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. > http://es.voice.yahoo.com > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 21 Date: Tue, 4 Apr 2006 22:31:56 -0700 From: Jennifer MacDonald Subject: Re: [Histonet] neutrophils To: "Webb, Dorothy L" Cc: Histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset="ISO-8859-1" Napththol AS-D Chloroacetate Esterase is used to identify granulo cytes. This procedure can be done on FFPE tissue,or blood smears.&nbs usually done on < -----histonet-bounces To: Histonet@lists.utsouthwestern.edu Fr Sent by: Date: 04/04/2006 11:25AM S Does anyone know of a stain for neutro read that staining with chloroacetate ester not familiar with that stain. Does would have the stain in a kit?? &nb and am rather stumped! Thanks, a and support!!!! ___ ______________________ 5F__ This e-mail and any files transmitt and are intended solely for the use of the indi to whom they are addressed. If you are not the intended re or the individual responsible for delivering the e-mail to the inte e-mail in err printing, or copying of thi If you have received this e-mail in notify the HealthPartners Support Center by tele 967-6600. You will be reimbursed for reasonable costs incurr notifying us. _________________ ______________________ 5F__ Histonet mailing list Histonet@lists.utsouthwestern.e [1]http://lists.utsouthwestern.edu/mailman/listinfo/histonet References 1. 3D"http://lists.utsouthwestern.edu/mailman/listinfo/histonet" ------------------------------ Message: 22 Date: Wed, 5 Apr 2006 05:15:57 -0400 From: DDDeltour@mar.med.navy.mil Subject: RE: [Histonet] Thanks for the Input To: lowman034@yahoo.com, vazquezr@ohsu.edu, Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE25@marxchg03.mar.med.navy.mil> Content-Type: text/plain It seems lately some people take bits and pieces of posts and twist them around for argument sake. I am sure you are not quoting me?? I can speak on the part of the Navy only. Maybe I should have stated that on my last post. As for the Navy ...While the others are out selling cookies others must stay behind and pick up the slack. These cookie sellers place the bake sale on the evaluation which earns them the bullet for a higher mark. The higher mark means a better chance for promotion. A more experienced and certified tech may not have the time to sell these cookies for whatever reason or because they care more about the REAL job than playing Martha Stewart. Green food coloring is not a counterstain. -----Original Message----- From: Dave Low [mailto:lowman034@yahoo.com] Sent: Tuesday, April 04, 2006 4:20 PM To: Deltour, Douglas D. (HM2); vazquezr@ohsu.edu; Jackie.O'Connor@abbott.com; Harper, Heather A., CIV Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks for the Input Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 23 Date: Wed, 5 Apr 2006 10:33:20 +0100 From: Kemlo Rogerson Subject: RE: [Histonet] Thanks for the Input To: "'DDDeltour@mar.med.navy.mil'" , lowman034@yahoo.com, vazquezr@ohsu.edu, Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain Geez I'm losing this!!!! Does the army make cakes? I thought they shot rifles and things. You get promoted if you can make a good cake but not if you shoot straight? Lost it totally. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "Following the line of least resistance makes both men and rivers crooked" This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: DDDeltour@mar.med.navy.mil [mailto:DDDeltour@mar.med.navy.mil] Sent: Wednesday, April 05, 2006 10:16 AM To: lowman034@yahoo.com; vazquezr@ohsu.edu; Jackie.O'Connor@abbott.com; Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks for the Input It seems lately some people take bits and pieces of posts and twist them around for argument sake. I am sure you are not quoting me?? I can speak on the part of the Navy only. Maybe I should have stated that on my last post. As for the Navy ...While the others are out selling cookies others must stay behind and pick up the slack. These cookie sellers place the bake sale on the evaluation which earns them the bullet for a higher mark. The higher mark means a better chance for promotion. A more experienced and certified tech may not have the time to sell these cookies for whatever reason or because they care more about the REAL job than playing Martha Stewart. Green food coloring is not a counterstain. -----Original Message----- From: Dave Low [mailto:lowman034@yahoo.com] Sent: Tuesday, April 04, 2006 4:20 PM To: Deltour, Douglas D. (HM2); vazquezr@ohsu.edu; Jackie.O'Connor@abbott.com; Harper, Heather A., CIV Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Thanks for the Input Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 6 *************************************** 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 Karen.Heckford <@t> CHW.edu Wed Apr 5 07:03:31 2006 From: Karen.Heckford <@t> CHW.edu (Heckford, Karen - SMMC-SF) Date: Wed Apr 5 07:03:46 2006 Subject: [Histonet] Thanks for the Input Message-ID: While I was in the Navy from 1985-1989 we made rank by passing tests and time in service not by doing community service for example bake sales. I never heard of such a thing. It was strictly on your ability to know your job. The Armed Services are now understaffed and underpaid. It is not easy being away from friends and family do what these people do. If it was not for them volunteering to go in to the Armed Services we would not be able to have a discussion like this on the internet. A proud Navy Veteran Karen Heckford HT (ASCP) From douglasgondo <@t> yahoo.com Wed Apr 5 07:12:42 2006 From: douglasgondo <@t> yahoo.com (Douglas Gondo) Date: Wed Apr 5 07:12:50 2006 Subject: [Histonet] Re: Histonet Digest, Vol 29, Issue 6 Message-ID: <20060405121242.57817.qmail@web31814.mail.mud.yahoo.com> please can anyone help I am histologist with a Bsc Uni. of Zimbabwe currently working for Namibia Institute of Pathology. my problem is I have seen that I am finding stained section that are not stained uniformly. Some areas are faint some dark as if there are holes in the sections. What could be the cause of this anormally? histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. PTH (Richard Cartun) 2. Humedity chamber (cynthia haynes) 3. Fatty breast processing (Orr, Rebecca) 4. neutrophils (Webb, Dorothy L) 5. RE: neutrophils (Monfils, Paul) 6. Sirius red with low contrast between cells cytoplasm and stained fibers (Guillermo Palao) 7. Re: Humedity chamber (Joanne Mauger) 8. RE: Thanks for the Input (Dave Low) 9. RE: Microtome knife sharpening service/ Microwave accessories (Justin Thomas) 10. Humidity chamber (Donna Harclerode) 11. Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW (Eric Dye (ext 223)) 12. Hematoxylin staining of frozen samples, where are the nuclei? (Guillermo Palao) 13. Qualifications for Histology Supervisor (Lester Raff) 14. Re: humidity chambers (Lori Richey) 15. Re: breast processing (Lori Richey) 16. Teaching Microtomes (Anwar Al-Banaw) 17. Re: Sirius red with low contrast between cells cytoplasm andstained fibers (John Kiernan) 18. Feedback on Milestone Medical's RHS Series Microwave Tissue Processor (jenbug812@aol.com) 19. RE: Sirius red with low contrast between cells cytoplasm andstained fibers (Tony Henwood) 20. Re: Hematoxylin staining of frozen samples, where are the nuclei? (Katri Tuomala) 21. Re: neutrophils (Jennifer MacDonald) 22. RE: Thanks for the Input (DDDeltour@mar.med.navy.mil) 23. RE: Thanks for the Input (Kemlo Rogerson) ---------------------------------------------------------------------- Message: 1 Date: Tue, 04 Apr 2006 13:13:52 -0400 From: "Richard Cartun" Subject: [Histonet] PTH To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ------------------------------ Message: 2 Date: Tue, 4 Apr 2006 11:10:55 -0700 (PDT) From: cynthia haynes Subject: [Histonet] Humedity chamber To: Histonet@lists.utsouthwestern.edu Message-ID: <20060404181055.97672.qmail@web33006.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. ------------------------------ Message: 3 Date: Tue, 4 Apr 2006 13:23:54 -0500 From: "Orr, Rebecca" Subject: [Histonet] Fatty breast processing To: Message-ID: Content-Type: text/plain; charset="US-ASCII" HI Annette, You have gotten quite a few responses to your question. I have just one more. When we get huge pieces of fatty breast that is unfixed, we place the block in the embedder, melt it and let it "soak" in the paraffin for 2 hours. I have left them in this paraffin bath as long as 4 hours but am unsure of any damage that could be done,( I haven't seen any) But 2 seems to be as long as we need. I would think that this process would cause damage, but since the tissue is unworkable in the first place, this can only help. This soaking in paraffin seems to do the trick, whether it's helping infiltration or simply dehydrating, I imagine it's doing a little bit of both. After this process, the blocks are much easier to cut. I don't see any problem with my IHC stains especially for ER/PR, Her2. These all seem to stain consistently. Ultimately, you would want to run your samples as others have recommended before it gets to the embedding stage. Hope this helps, Becky Becky Orr, CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 tm@libero.it> Message: 5 Date: Tue, 4 Apr 2006 09:26:26 -0400 From: "Featherstone, Annette" Subject: [Histonet] breast processing To: Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Content-Type: text/plain; charset="iso-8859-1" Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT ------------------------------ Message: 4 Date: Tue, 04 Apr 2006 13:25:14 -0500 From: "Webb, Dorothy L" Subject: [Histonet] neutrophils To: Histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA270171FD2A@hpes1.HealthPartners.int> Content-Type: text/plain; charset="US-ASCII" Does anyone know of a stain for neutrophils? I believe I had read that staining with chloroacetate esterase is the way to go, but, I am not familiar with that stain. Does anyone know of a company that would have the stain in a kit?? This is for a research project and am rather stumped! Thanks, as always, fellow Histonetters for your help and support!!!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 5 Date: Tue, 4 Apr 2006 14:37:46 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] neutrophils To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176BC@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I use the Naphthol AS-D Chloroacetate Esterase Kit from Sigma (cat# 91C-1KT) on frozen sections. It's easy to use and very reliable. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Webb, > Dorothy L > Sent: Tuesday, April 4, 2006 11:25 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] neutrophils > > Does anyone know of a stain for neutrophils? I believe I had read that > staining with chloroacetate esterase is the way to go, but, I am not > familiar with that stain. Does anyone know of a company that would > have the stain in a kit?? This is for a research project and am rather > stumped! Thanks, as always, fellow Histonetters for your help and > support!!!! > ________________________________________ > This e-mail and any files transmitted with it are confidential and are > intended solely for the use of the individual or entity to whom they are > addressed. If you are not the intended recipient or the individual > responsible for delivering the e-mail to the intended recipient, please be > advised that you have received this e-mail in error and that any use, > dissemination, forwarding, printing, or copying of this e-mail is strictly > prohibited. > > If you have received this e-mail in error, please immediately notify the > HealthPartners Support Center by telephone at (952) 967-6600. You will be > reimbursed for reasonable costs incurred in notifying us. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 6 Date: Tue, 4 Apr 2006 20:58:33 +0200 (CEST) From: Guillermo Palao Subject: [Histonet] Sirius red with low contrast between cells cytoplasm and stained fibers To: Histonet Message-ID: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com ------------------------------ Message: 7 Date: Tue, 04 Apr 2006 15:49:36 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] Humedity chamber To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Cynthia, It's www.scytek.com Jo >>> cynthia haynes 04/04/06 2:10 PM >>> Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 4 Apr 2006 13:19:41 -0700 (PDT) From: Dave Low Subject: RE: [Histonet] Thanks for the Input To: DDDeltour@mar.med.navy.mil, vazquezr@ohsu.edu, Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060404201941.10561.qmail@web32004.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 9 Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) From: Justin Thomas Subject: [Histonet] RE: Microtome knife sharpening service/ Microwave accessories To: histonet@lists.utsouthwestern.edu Message-ID: <20060404202218.93567.qmail@web35701.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Just to let everyone know: There is finally a company out there that sharpens microtome knifes (both stainless steel & tungsten) and manufactures plastic microwave accessories at a low price. I send all our knifes to them and I am extremely satisfied with the customer service I receive, along with the pricing. Also, they have made a number of accessories for us and I have nothing but good words to say about them. The accessory's work well and with no problem. The best thing about them is the company offers a warranty with each accessory. If you wish, email me back with your contact information and I will pass it on to the sharpening and MW accessory company. Thanks, Dr. Thomas --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. ------------------------------ Message: 10 Date: Tue, 4 Apr 2006 13:36:57 -0700 From: "Donna Harclerode" Subject: [Histonet] Humidity chamber To: Message-ID: <3DE0F644E093DF4BAE80C254176696A5090706@mp-mailserver.macropore.com> Content-Type: text/plain; charset="us-ascii" Thermo Electron makes a great system with a humidity chamber and a coverplate that I have used for many years. The slides are held with the plastic coverplate(the coverplates are suppose to be disposable, but I rinse them with only DI water and reuse them many times) I use 100ul of antibody (primary, secondary- etc, normally, but have managed to use as little as 90ul) to cover almost the whole slide. For buffer wash I fill up the top of the chamber with a squeeze bottle of PBS and allow to run through in about 5 minutes. I load the slides add primary, wash, secondary etc and only take them out for chromagen staining. The racks worked great when I had the positive control at the top of the slide and the test below. I do fluorescence secondaries in the coverplates, but I not do DAB in the coverplates, but flat on the counter. For HRP frozens I use the Dako endogenous peroxidase block in the Sequenza racks. I also use the Dako avidin biotin block in the coverplates when necessary. I usually incubate primary overnight in the fridge and the chambers stay humid for at least 48 hours. It takes a bit of practice to load the plates, but it is so worth it. Sequenza Racks - # 73310017 (Holds 10 coverplate assemblies) Coverplates - 25/pk = # 72110017 50/pk = # 7219950 250/case = # 72110013 http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_19578.pdf 4th page Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 5416 dharclerode@cytoritx.com ------------------------------ Message: 11 Date: Tue, 4 Apr 2006 17:13:06 -0400 From: Eric Dye (ext 223) Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW To: Histonetters Message-ID: Content-Type: text/plain Fellow-Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) Here are some of my Hottest Histology Jobs: 1. Ohio (Southwestern Ohio) (Full-time, Perm, Histo Manager) 2. Northern New Jersey (Full-time, Perm and/or Temp, Bench Histo Tech, 1st shift) 3. Rhode Island - Providence (Full-time, Perm, Bench Histo Tech, 2nd Shift) 4. Northern New Jersey (Full-time, Perm, Bench Histo Tech) 5. Ohio (Southwestern Ohio) (Full-time, Perm, Bench Histo Tech) 6. New York (Long Island) (Full-time, Perm, SUPERVISOR Histo Tech) 7. Virginia(South - Coastal) (Full-time, Perm, Bench Histo Tech) === message truncated === --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. From DDDeltour <@t> mar.med.navy.mil Wed Apr 5 08:47:00 2006 From: DDDeltour <@t> mar.med.navy.mil (DDDeltour@mar.med.navy.mil) Date: Wed Apr 5 08:47:28 2006 Subject: [Histonet] Thanks for the Input Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE27@marxchg03.mar.med.navy.mil> Karen you are correct that you take a test (E1-E6) but you are given more points toward that test if you have a better evaluation. They did away with time in rate long ago. The test I would be taking is a general medical test and shipboard stuff. You are not tested on your job, believe me. When you go up for E7 you are up on a board for selection. Anyway I have been doing this for 14 years and it is time to move on. Cookies anyone! :) I think that we are going way off topic for Histonet..... Next. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heckford, Karen - SMMC-SF Sent: Wednesday, April 05, 2006 8:04 AM To: Histonet (E-mail) Subject: [Histonet] Thanks for the Input While I was in the Navy from 1985-1989 we made rank by passing tests and time in service not by doing community service for example bake sales. I never heard of such a thing. It was strictly on your ability to know your job. The Armed Services are now understaffed and underpaid. It is not easy being away from friends and family do what these people do. If it was not for them volunteering to go in to the Armed Services we would not be able to have a discussion like this on the internet. A proud Navy Veteran Karen Heckford HT (ASCP) From fmoraes <@t> igc.gulbenkian.pt Wed Apr 5 09:03:55 2006 From: fmoraes <@t> igc.gulbenkian.pt (fmoraes@igc.gulbenkian.pt) Date: Wed Apr 5 09:04:00 2006 Subject: [Histonet] Immunohistochemistry with whole mouse embryos In-Reply-To: References: Message-ID: <1144245835.4433ce4bee61e@webmail.igc.gulbenkian.pt> Hi I am having problems to perform some whole mount stainings protocols in mouse embryos E10.5: 1) The whole mount staining protocol for PECAM (anti-mouse PECAM monoclonal antibody MEC13.3-Pharmigen) with mouse embryos E10.5 (10dpc). The color reaction (using DAB takes 10 min and the embryos are all black) however, after pos-developing washes I don't have any signal. I think is a question of penetrance of the Antibody. Do you have some tips to permeabilize the embryos? 2) If I want to do an immunofluorescence protocol with whole mount mouse embryos E10.5 (using a Cy3 conjugated monoclonal Antibody - red spectrum, Absorbance:552nm Emission:570nm) and I want to analyse the whole specimen under a confocal microscope. What should be the mounting medium more appropriated? Thank You very much Filipa Moraes Graduate Student Gulbenkian Institute of Science Portugal +315 214464525 From asmith <@t> mail.barry.edu Wed Apr 5 09:11:34 2006 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Wed Apr 5 09:11:41 2006 Subject: [Histonet] Hematoxylin staining of frozen samples, where are the nuclei? Message-ID: <5D2189E74151CC42BEC02906BA8996322B91BB@exchsrv01.barrynet.barry.edu> The fading after most retrieval procedures should not be very bad. Heat retrieval in citrate buffer causes very little nuclear fading in my tissues (and no fading of lymphocyte nuclei). Your staining and mounting procedure may be the problem. I find Vector's Nova Red compatible with (Harris) hematoxylin. Since Nova Red is insoluble in xylene, I can clear my tissue and mount in resin (Permount) so that the nuclear staining is more easily observed. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Katri Tuomala Sent: Tuesday, April 04, 2006 7:06 PM To: Guillermo Palao; Histonet Subject: Re: [Histonet] Hematoxylin staining of frozen samples,where are the nuclei? Guillermo, Tissues go through a long procedure with immuno and the most detrimental for all tissue components is the heat retrieval and to a lesser degree enzyme digestion. The less well fixed and processed tissue suffers most: connective tissue is distorted and nuclei stain very poorly. Hope this explains your problem, Katri Katri Tuomala Hamilton, Ontario, Canada ----- Original Message ----- From: "Guillermo Palao" To: "Histonet" Sent: Tuesday, April 04, 2006 5:22 PM Subject: [Histonet] Hematoxylin staining of frozen samples,where are the nuclei? > Dear histonetters, > > When I stain samples placed on VWR frosted slides with Gills hematoxylin, > nuclei are nicely stained. However, when the same samples are > counterstained after passing through all necessary steps involved in > immunohistochemistry, hematoxylin staining of nuclei is simply horrible. > Is it possible that nuclei are lost in the different washing steps? Please > put forward any suggestions you might have. > > Guillermo > > > --------------------------------- > > LLama Gratis a cualquier PC del Mundo. > Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. > http://es.voice.yahoo.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 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 vazquezr <@t> ohsu.edu Wed Apr 5 09:11:14 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Apr 5 09:11:48 2006 Subject: [Histonet] Re: Histonet Digest, Vol 29, Issue 6 Message-ID: it sounds like the paraffin is not melted in all the tissue sections. Robyn OHSU >>> "Douglas Gondo" 4/5/2006 5:12 AM >>> please can anyone help I am histologist with a Bsc Uni. of Zimbabwe currently working for Namibia Institute of Pathology. my problem is I have seen that I am finding stained section that are not stained uniformly. Some areas are faint some dark as if there are holes in the sections. What could be the cause of this anormally? histonet-request@lists.utsouthwestern.edu wrote: Send Histonet mailing list submissions to histonet@lists.utsouthwestern.edu To subscribe or unsubscribe via the World Wide Web, visit http://lists.utsouthwestern.edu/mailman/listinfo/histonet or, via email, send a message with subject or body 'help' to histonet-request@lists.utsouthwestern.edu You can reach the person managing the list at histonet-owner@lists.utsouthwestern.edu When replying, please edit your Subject line so it is more specific than "Re: Contents of Histonet digest..." Today's Topics: 1. PTH (Richard Cartun) 2. Humedity chamber (cynthia haynes) 3. Fatty breast processing (Orr, Rebecca) 4. neutrophils (Webb, Dorothy L) 5. RE: neutrophils (Monfils, Paul) 6. Sirius red with low contrast between cells cytoplasm and stained fibers (Guillermo Palao) 7. Re: Humedity chamber (Joanne Mauger) 8. RE: Thanks for the Input (Dave Low) 9. RE: Microtome knife sharpening service/ Microwave accessories (Justin Thomas) 10. Humidity chamber (Donna Harclerode) 11. Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW (Eric Dye (ext 223)) 12. Hematoxylin staining of frozen samples, where are the nuclei? (Guillermo Palao) 13. Qualifications for Histology Supervisor (Lester Raff) 14. Re: humidity chambers (Lori Richey) 15. Re: breast processing (Lori Richey) 16. Teaching Microtomes (Anwar Al-Banaw) 17. Re: Sirius red with low contrast between cells cytoplasm andstained fibers (John Kiernan) 18. Feedback on Milestone Medical's RHS Series Microwave Tissue Processor (jenbug812@aol.com) 19. RE: Sirius red with low contrast between cells cytoplasm andstained fibers (Tony Henwood) 20. Re: Hematoxylin staining of frozen samples, where are the nuclei? (Katri Tuomala) 21. Re: neutrophils (Jennifer MacDonald) 22. RE: Thanks for the Input (DDDeltour@mar.med.navy.mil) 23. RE: Thanks for the Input (Kemlo Rogerson) ---------------------------------------------------------------------- Message: 1 Date: Tue, 04 Apr 2006 13:13:52 -0400 From: "Richard Cartun" Subject: [Histonet] PTH To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ------------------------------ Message: 2 Date: Tue, 4 Apr 2006 11:10:55 -0700 (PDT) From: cynthia haynes Subject: [Histonet] Humedity chamber To: Histonet@lists.utsouthwestern.edu Message-ID: <20060404181055.97672.qmail@web33006.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. ------------------------------ Message: 3 Date: Tue, 4 Apr 2006 13:23:54 -0500 From: "Orr, Rebecca" Subject: [Histonet] Fatty breast processing To: Message-ID: Content-Type: text/plain; charset="US-ASCII" HI Annette, You have gotten quite a few responses to your question. I have just one more. When we get huge pieces of fatty breast that is unfixed, we place the block in the embedder, melt it and let it "soak" in the paraffin for 2 hours. I have left them in this paraffin bath as long as 4 hours but am unsure of any damage that could be done,( I haven't seen any) But 2 seems to be as long as we need. I would think that this process would cause damage, but since the tissue is unworkable in the first place, this can only help. This soaking in paraffin seems to do the trick, whether it's helping infiltration or simply dehydrating, I imagine it's doing a little bit of both. After this process, the blocks are much easier to cut. I don't see any problem with my IHC stains especially for ER/PR, Her2. These all seem to stain consistently. Ultimately, you would want to run your samples as others have recommended before it gets to the embedding stage. Hope this helps, Becky Becky Orr, CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 tm@libero.it> Message: 5 Date: Tue, 4 Apr 2006 09:26:26 -0400 From: "Featherstone, Annette" Subject: [Histonet] breast processing To: Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Content-Type: text/plain; charset="iso-8859-1" Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT ------------------------------ Message: 4 Date: Tue, 04 Apr 2006 13:25:14 -0500 From: "Webb, Dorothy L" Subject: [Histonet] neutrophils To: Histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA270171FD2A@hpes1.HealthPartners.int> Content-Type: text/plain; charset="US-ASCII" Does anyone know of a stain for neutrophils? I believe I had read that staining with chloroacetate esterase is the way to go, but, I am not familiar with that stain. Does anyone know of a company that would have the stain in a kit?? This is for a research project and am rather stumped! Thanks, as always, fellow Histonetters for your help and support!!!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 5 Date: Tue, 4 Apr 2006 14:37:46 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] neutrophils To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176BC@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I use the Naphthol AS-D Chloroacetate Esterase Kit from Sigma (cat# 91C-1KT) on frozen sections. It's easy to use and very reliable. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Webb, > Dorothy L > Sent: Tuesday, April 4, 2006 11:25 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] neutrophils > > Does anyone know of a stain for neutrophils? I believe I had read that > staining with chloroacetate esterase is the way to go, but, I am not > familiar with that stain. Does anyone know of a company that would > have the stain in a kit?? This is for a research project and am rather > stumped! Thanks, as always, fellow Histonetters for your help and > support!!!! > ________________________________________ > This e-mail and any files transmitted with it are confidential and are > intended solely for the use of the individual or entity to whom they are > addressed. If you are not the intended recipient or the individual > responsible for delivering the e-mail to the intended recipient, please be > advised that you have received this e-mail in error and that any use, > dissemination, forwarding, printing, or copying of this e-mail is strictly > prohibited. > > If you have received this e-mail in error, please immediately notify the > HealthPartners Support Center by telephone at (952) 967-6600. You will be > reimbursed for reasonable costs incurred in notifying us. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 6 Date: Tue, 4 Apr 2006 20:58:33 +0200 (CEST) From: Guillermo Palao Subject: [Histonet] Sirius red with low contrast between cells cytoplasm and stained fibers To: Histonet Message-ID: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com ------------------------------ Message: 7 Date: Tue, 04 Apr 2006 15:49:36 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] Humedity chamber To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Cynthia, It's www.scytek.com Jo >>> cynthia haynes 04/04/06 2:10 PM >>> Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 4 Apr 2006 13:19:41 -0700 (PDT) From: Dave Low Subject: RE: [Histonet] Thanks for the Input To: DDDeltour@mar.med.navy.mil, vazquezr@ohsu.edu, Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060404201941.10561.qmail@web32004.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 9 Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) From: Justin Thomas Subject: [Histonet] RE: Microtome knife sharpening service/ Microwave accessories To: histonet@lists.utsouthwestern.edu Message-ID: <20060404202218.93567.qmail@web35701.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Just to let everyone know: There is finally a company out there that sharpens microtome knifes (both stainless steel & tungsten) and manufactures plastic microwave accessories at a low price. I send all our knifes to them and I am extremely satisfied with the customer service I receive, along with the pricing. Also, they have made a number of accessories for us and I have nothing but good words to say about them. The accessory's work well and with no problem. The best thing about them is the company offers a warranty with each accessory. If you wish, email me back with your contact information and I will pass it on to the sharpening and MW accessory company. Thanks, Dr. Thomas --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. ------------------------------ Message: 10 Date: Tue, 4 Apr 2006 13:36:57 -0700 From: "Donna Harclerode" Subject: [Histonet] Humidity chamber To: Message-ID: <3DE0F644E093DF4BAE80C254176696A5090706@mp-mailserver.macropore.com> Content-Type: text/plain; charset="us-ascii" Thermo Electron makes a great system with a humidity chamber and a coverplate that I have used for many years. The slides are held with the plastic coverplate(the coverplates are suppose to be disposable, but I rinse them with only DI water and reuse them many times) I use 100ul of antibody (primary, secondary- etc, normally, but have managed to use as little as 90ul) to cover almost the whole slide. For buffer wash I fill up the top of the chamber with a squeeze bottle of PBS and allow to run through in about 5 minutes. I load the slides add primary, wash, secondary etc and only take them out for chromagen staining. The racks worked great when I had the positive control at the top of the slide and the test below. I do fluorescence secondaries in the coverplates, but I not do DAB in the coverplates, but flat on the counter. For HRP frozens I use the Dako endogenous peroxidase block in the Sequenza racks. I also use the Dako avidin biotin block in the coverplates when necessary. I usually incubate primary overnight in the fridge and the chambers stay humid for at least 48 hours. It takes a bit of practice to load the plates, but it is so worth it. Sequenza Racks - # 73310017 (Holds 10 coverplate assemblies) Coverplates - 25/pk = # 72110017 50/pk = # 7219950 250/case = # 72110013 http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_19578.pdf 4th page Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 5416 dharclerode@cytoritx.com ------------------------------ Message: 11 Date: Tue, 4 Apr 2006 17:13:06 -0400 From: Eric Dye (ext 223) Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW To: Histonetters Message-ID: Content-Type: text/plain Fellow-Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) Here are some of my Hottest Histology Jobs: 1. Ohio (Southwestern Ohio) (Full-time, Perm, Histo Manager) 2. Northern New Jersey (Full-time, Perm and/or Temp, Bench Histo Tech, 1st shift) 3. Rhode Island - Providence (Full-time, Perm, Bench Histo Tech, 2nd Shift) 4. Northern New Jersey (Full-time, Perm, Bench Histo Tech) 5. Ohio (Southwestern Ohio) (Full-time, Perm, Bench Histo Tech) 6. New York (Long Island) (Full-time, Perm, SUPERVISOR Histo Tech) 7. Virginia(South - Coastal) (Full-time, Perm, Bench Histo Tech) === message truncated === --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From maramydave <@t> aol.com Wed Apr 5 09:27:03 2006 From: maramydave <@t> aol.com (maramydave@aol.com) Date: Wed Apr 5 09:27:17 2006 Subject: [Histonet] PTH In-Reply-To: <200604051012.6324433d04f137@rly-xn02.mx.aol.com> References: <200604051012.6324433d04f137@rly-xn02.mx.aol.com> Message-ID: <8C826D4245C3947-1698-B98@FWM-D10.sysops.aol.com> Rich We had the same problem, Vector's antibody is good with PK. Mary Helie Message: 1 Date: Tue, 04 Apr 2006 13:13:52 -0400 From: "Richard Cartun" Subject: [Histonet] PTH To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From gcallis <@t> montana.edu Wed Apr 5 09:28:11 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Apr 5 09:28:24 2006 Subject: [Histonet] Re: Immunohistochemistry with whole mouse embryos In-Reply-To: <1144245835.4433ce4bee61e@webmail.igc.gulbenkian.pt> References: <1144245835.4433ce4bee61e@webmail.igc.gulbenkian.pt> Message-ID: <6.0.0.22.1.20060405081746.01b25720@gemini.msu.montana.edu> Prolong gold antifade ready to use, but is this going to flatten the embryo under a coverslip? If the CLSM is fitted with an inverted microscope, there are special petri dishes that have coverslip bottoms. You can float a stained embryos in PBS without having to use mounting media over the top of the embryos to prevent drying. It does not have to be antifade mounting media - but you can try both for scanning and z stack imaging. Personally, I prefer Alexa dyes for CLSM - they are brighter and do NOT photobleach as readily. Check out Molecular Probes Alexa Fluor selections on line. Some are going to far red fluorophores to avoid autofluorescence issues,- I suggest you look into that also. Permeabilizing can be done with a detergent, Tween 20 or Triton X 100, and with the thicker embryos, you may need to stain overnight at 4C to have adequate penetration of everything. I have seen this in the literature (methods and materials for both brain and embryo work). I am not sure of the concentrations. At 08:03 AM 4/5/2006, you wrote: >Hi > >I am having problems to perform some whole mount stainings protocols in >mouse embryos E10.5: > >1) The whole mount staining protocol for PECAM (anti-mouse PECAM >monoclonal antibody MEC13.3-Pharmigen) with mouse embryos E10.5 (10dpc). >The color reaction (using DAB takes 10 min and the embryos are all black) >however, after >pos-developing washes I don't have any signal. I think is a question of >penetrance of the Antibody. Do you have some tips to permeabilize the embryos? > >2) If I want to do an immunofluorescence protocol with whole mount mouse >embryos E10.5 (using a Cy3 conjugated monoclonal Antibody - red spectrum, >Absorbance:552nm Emission:570nm) and I want to analyse the whole specimen under >a confocal microscope. What should be the mounting medium more appropriated? 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 406 994-4303 (FAX) From cpomajzl <@t> cpllabs.com Wed Apr 5 09:52:12 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Wed Apr 5 09:49:24 2006 Subject: [Histonet] It's no wonder... Message-ID: <009b01c658c0$8f845f60$26fca8c0@CSP> ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. From dsantana <@t> pmaonline.com Wed Apr 5 09:27:39 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Wed Apr 5 09:53:51 2006 Subject: [Histonet] postal Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB07113959@MAILPMA> Does anyone remember several months ago about a new logo for NSH meeting.... something to do with histologist going postal. I have read a lot of emails lately that are making me a little concerned about the mental health of my conrads in the histo field. Take it easy people I think you are all great! Diane Santana PMA Haverhill, Mass. ( A simple histo tech with nothing special, just loves her job) From cking <@t> rallansci.com Wed Apr 5 09:54:09 2006 From: cking <@t> rallansci.com (King, Curtis - RAS) Date: Wed Apr 5 09:54:17 2006 Subject: [Histonet] It's no wonder... Message-ID: <490C1EC04BA10F4891494D0ED756AC9303C94D66@usmi0012k03.rallansci.apogent.com> Well said Chris. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Chris Pomajzl Sent: Wednesday, April 05, 2006 10:52 AM To: HISTONET Subject: [Histonet] It's no wonder... ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From michael.owen <@t> fda.hhs.gov Wed Apr 5 10:02:20 2006 From: michael.owen <@t> fda.hhs.gov (Owen, Michael P) Date: Wed Apr 5 10:02:47 2006 Subject: [Histonet] postal Message-ID: Perhaps I am off-base with this comment, but I prefer the term "postal" not being used when referencing work rage and work violence. One of my best friends was an employee of the U.S. Postal Service. She was shot and killed by a customer of her branch in 1998. If my memory serves me correctly, work violence is no longer primarily found in U.S. Postal Service facilities. Diana, my post is not meant as a flaming message to you. Thanks for your informative comments. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov From tkngflght <@t> yahoo.com Wed Apr 5 10:05:44 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Wed Apr 5 10:05:49 2006 Subject: [Histonet] Corporate culture--lab culture... In-Reply-To: <009b01c658c0$8f845f60$26fca8c0@CSP> Message-ID: <20060405150544.545.qmail@web50902.mail.yahoo.com> Hi Chris and all the Histofolk-- There is a huge article in Inc. Magazine this month regarding corporate culture and how it forms in direct response to the managers and directors of a facility....Chris's response below is an EXACT reflection on the culture found in his lab, and the company around him. I have a new-to traveling temp tech helping in his facility while he hires to accomodate growth. She calls me every THREE DAYS to tell me how GREAT it is to be there and how happy, helpful and FUN the people are even with the large work volume. Like Chris, I imagine every single piece of tissue to be from my Grandpa or my child--the patient is likely one of those things to someone. We have amazing skill, we help make a difference for every single patient, we are paid to do complex tasks and yes, wash dishes!! Wouldn't we all like to walk IN to work with a smile? The most germaine point? It IS a choice. Make it daily. Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. From rjbuesa <@t> yahoo.com Wed Apr 5 10:13:21 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Apr 5 10:13:32 2006 Subject: [Histonet] validation for CAP In-Reply-To: <9B4A77DF11463E4FB723D484214AE9BCA5515B@KALEXMB02.KaleidaHealth.org> Message-ID: <20060405151321.25216.qmail@web61211.mail.yahoo.com> Annette: Although I used Dako autostainers for years to my entire satisfaction, I find the argument from the company as an example of "circular (invalid) reasoning". That one instrument is "validated" against the other does not preclude the fact that BOTH could be wrong in their "intervalidation". I don't buy the argument and neither should you. Each one is a different unit that could malfunction while the other is OK. I would validate each, and not one against the other (regardless of CAP). Just a thought! Ren? J. "Featherstone, Annette" wrote: We have just purchased 2 new Dako immuno stainers and I was wondering what CAP requirements are for validation. The company says that both instruments are validated against each other and therefore you can validate the antibodies on either stainer. I hope this is fact. Annette Featherstone HT/MLT MT(HEW) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, April 05, 2006 05:34 To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 6 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. PTH (Richard Cartun) 2. Humedity chamber (cynthia haynes) 3. Fatty breast processing (Orr, Rebecca) 4. neutrophils (Webb, Dorothy L) 5. RE: neutrophils (Monfils, Paul) 6. Sirius red with low contrast between cells cytoplasm and stained fibers (Guillermo Palao) 7. Re: Humedity chamber (Joanne Mauger) 8. RE: Thanks for the Input (Dave Low) 9. RE: Microtome knife sharpening service/ Microwave accessories (Justin Thomas) 10. Humidity chamber (Donna Harclerode) 11. Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW (Eric Dye (ext 223)) 12. Hematoxylin staining of frozen samples, where are the nuclei? (Guillermo Palao) 13. Qualifications for Histology Supervisor (Lester Raff) 14. Re: humidity chambers (Lori Richey) 15. Re: breast processing (Lori Richey) 16. Teaching Microtomes (Anwar Al-Banaw) 17. Re: Sirius red with low contrast between cells cytoplasm andstained fibers (John Kiernan) 18. Feedback on Milestone Medical's RHS Series Microwave Tissue Processor (jenbug812@aol.com) 19. RE: Sirius red with low contrast between cells cytoplasm andstained fibers (Tony Henwood) 20. Re: Hematoxylin staining of frozen samples, where are the nuclei? (Katri Tuomala) 21. Re: neutrophils (Jennifer MacDonald) 22. RE: Thanks for the Input (DDDeltour@mar.med.navy.mil) 23. RE: Thanks for the Input (Kemlo Rogerson) ---------------------------------------------------------------------- Message: 1 Date: Tue, 04 Apr 2006 13:13:52 -0400 From: "Richard Cartun" Subject: [Histonet] PTH To: Message-ID: Content-Type: text/plain; charset=US-ASCII Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax ------------------------------ Message: 2 Date: Tue, 4 Apr 2006 11:10:55 -0700 (PDT) From: cynthia haynes Subject: [Histonet] Humedity chamber To: Histonet@lists.utsouthwestern.edu Message-ID: <20060404181055.97672.qmail@web33006.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. ------------------------------ Message: 3 Date: Tue, 4 Apr 2006 13:23:54 -0500 From: "Orr, Rebecca" Subject: [Histonet] Fatty breast processing To: Message-ID: Content-Type: text/plain; charset="US-ASCII" HI Annette, You have gotten quite a few responses to your question. I have just one more. When we get huge pieces of fatty breast that is unfixed, we place the block in the embedder, melt it and let it "soak" in the paraffin for 2 hours. I have left them in this paraffin bath as long as 4 hours but am unsure of any damage that could be done,( I haven't seen any) But 2 seems to be as long as we need. I would think that this process would cause damage, but since the tissue is unworkable in the first place, this can only help. This soaking in paraffin seems to do the trick, whether it's helping infiltration or simply dehydrating, I imagine it's doing a little bit of both. After this process, the blocks are much easier to cut. I don't see any problem with my IHC stains especially for ER/PR, Her2. These all seem to stain consistently. Ultimately, you would want to run your samples as others have recommended before it gets to the embedding stage. Hope this helps, Becky Becky Orr, CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 tm@libero.it> Message: 5 Date: Tue, 4 Apr 2006 09:26:26 -0400 From: "Featherstone, Annette" Subject: [Histonet] breast processing To: Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA55154@KALEXMB02.KaleidaHealth.org> Content-Type: text/plain; charset="iso-8859-1" Does anyone have any suggestions for better tissue processing for fatty tissue such as breast? We are experiencing insufficent dehydration, clearing and infiltration. Annette Featherstone HT/MLT ------------------------------ Message: 4 Date: Tue, 04 Apr 2006 13:25:14 -0500 From: "Webb, Dorothy L" Subject: [Histonet] neutrophils To: Histonet@lists.utsouthwestern.edu Message-ID: <0E394B648E5284478A6CCB78E5AFDA270171FD2A@hpes1.HealthPartners.int> Content-Type: text/plain; charset="US-ASCII" Does anyone know of a stain for neutrophils? I believe I had read that staining with chloroacetate esterase is the way to go, but, I am not familiar with that stain. Does anyone know of a company that would have the stain in a kit?? This is for a research project and am rather stumped! Thanks, as always, fellow Histonetters for your help and support!!!! ________________________________________ This e-mail and any files transmitted with it are confidential and are intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient or the individual responsible for delivering the e-mail to the intended recipient, please be advised that you have received this e-mail in error and that any use, dissemination, forwarding, printing, or copying of this e-mail is strictly prohibited. If you have received this e-mail in error, please immediately notify the HealthPartners Support Center by telephone at (952) 967-6600. You will be reimbursed for reasonable costs incurred in notifying us. ------------------------------ Message: 5 Date: Tue, 4 Apr 2006 14:37:46 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] neutrophils To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176BC@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I use the Naphthol AS-D Chloroacetate Esterase Kit from Sigma (cat# 91C-1KT) on frozen sections. It's easy to use and very reliable. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Webb, > Dorothy L > Sent: Tuesday, April 4, 2006 11:25 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] neutrophils > > Does anyone know of a stain for neutrophils? I believe I had read that > staining with chloroacetate esterase is the way to go, but, I am not > familiar with that stain. Does anyone know of a company that would > have the stain in a kit?? This is for a research project and am rather > stumped! Thanks, as always, fellow Histonetters for your help and > support!!!! > ________________________________________ > This e-mail and any files transmitted with it are confidential and are > intended solely for the use of the individual or entity to whom they are > addressed. If you are not the intended recipient or the individual > responsible for delivering the e-mail to the intended recipient, please be > advised that you have received this e-mail in error and that any use, > dissemination, forwarding, printing, or copying of this e-mail is strictly > prohibited. > > If you have received this e-mail in error, please immediately notify the > HealthPartners Support Center by telephone at (952) 967-6600. You will be > reimbursed for reasonable costs incurred in notifying us. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 6 Date: Tue, 4 Apr 2006 20:58:33 +0200 (CEST) From: Guillermo Palao Subject: [Histonet] Sirius red with low contrast between cells cytoplasm and stained fibers To: Histonet Message-ID: <20060404185833.93523.qmail@web26202.mail.ukl.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com ------------------------------ Message: 7 Date: Tue, 04 Apr 2006 15:49:36 -0400 From: "Joanne Mauger" Subject: Re: [Histonet] Humedity chamber To: , Message-ID: Content-Type: text/plain; charset=US-ASCII Cynthia, It's www.scytek.com Jo >>> cynthia haynes 04/04/06 2:10 PM >>> Hello all! Some one put a website for a humedity chamber for Immunohistochemistry staining. I think it was www.scyetex.com. or something like that, would please put the site address on the histonet again. Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 4 Apr 2006 13:19:41 -0700 (PDT) From: Dave Low Subject: RE: [Histonet] Thanks for the Input To: DDDeltour@mar.med.navy.mil, vazquezr@ohsu.edu, Jackie.O'Connor@abbott.com, Heather.A.Harper@pcola.med.navy.mil Cc: histonet@lists.utsouthwestern.edu Message-ID: <20060404201941.10561.qmail@web32004.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Douglas, Military personnel DO NOT make rank by working at some bake sales! Are you insinuating Master Chief Petty Officers, Sergeant Majors, and Chief Master Sargeants made rank selling cookies? Rubbish! I am an Air Force Master Sargeant, worked in histology for over 17 years out of my 20+ years service. I worked at the Armed Forces Institute of Pathology in Washington DC from 1997-2000 and had the pleasure to work for the Army, Navy, and civilians histology technicians and managers. The promotions I did see were from hard work not only from the job but to the military and civilian communities. It's called the whole person concept! In the future please screen your e-mail for appropriateness before you broadcast to a large audience like histonet. Dave Low, MSgt,USAF HT(ASCP)QIHC --- DDDeltour@mar.med.navy.mil wrote: > I work in this environment everyday. I see both > sides (military and > civilian) taking shots at each other instead of > working as a team. I am soon > leaving the service and you could not pay me enough > money to supervise or > work in a military facility. I understand Heather > and her frustration of > being left to do the job BUT the military requires > its members to do things > outside of the job to get promoted. That is a fact. > I am living it. I see > people in my own field get promoted because they did > some BS bake sale but > they can't even do a GMS. You can either find a way > to work it out or find a > new job. As for the military Pathologist....well > they are in charge. They > like that. Everything will change again when the > next one transfers in. That > is part of being in a military facility. I have seen > it for years. The best > thing to do is go with the flow or just go. I am > going. Good luck. > > Douglas D. Deltour HT(ASCP) > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] > On Behalf Of Robyn > Vazquez > Sent: Monday, April 03, 2006 12:29 PM > To: Jackie.O'Connor@abbott.com; Harper, Heather A., > CIV > Cc: histonet@lists.utsouthwestern.edu > Subject: Re: [Histonet] Thanks for the Input > > Sounds like you both work hard...I have been on both > sides of the > fence...keep up the good work...she needs all the > support you can give her > civilian or military! > Just my two cents... > > Robyn > OHSU > > >>> "Jackie M O'Connor" > 4/3/2006 8:49 AM >>> > I am thankful and grateful for anyone who ever > signed up for military > service - ever - regardless of their job > description. > > > > > > Heather.A.Harper@pcola.med.navy.mil > Sent by: histonet-bounces@lists.utsouthwestern.edu > 04/03/2006 10:20 AM > > > To: histonet@lists.utsouthwestern.edu > cc: (bcc: Jackie M > O'Connor/LAKE/GPRD/ABBOTT) > Subject: [Histonet] Thanks for the > Input > > > I want to thank everybody who responded to my > message titled..Need Input. > I > work for the DOD and I miss simply having a system > in place. I agree one > has > to be flexible, but when you work with military, > they are entitled to 1 hr > lunch and 1 hour of PT. My tech works 7-4 and I work > 6-2. I embed, she > cuts > and stains, I gross, accession, order supplies, > admit bodies in and out of > the morgue, set up for autopsies, frozens, and when > my tech has to get > pulled to do her other command duties, it leaves me > holding the bag. It > does get over whelming, and every 2-3 yrs, I get new > pathologists rotating > through. I have worked with 7 pathologists in 6 yrs, > and reservists, and > this one I just do not know what to think. Just > keeping an open mind. > Thanks > again everybody. Be thankful you are in the civilian > world. > > > > Heather > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 9 Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) From: Justin Thomas Subject: [Histonet] RE: Microtome knife sharpening service/ Microwave accessories To: histonet@lists.utsouthwestern.edu Message-ID: <20060404202218.93567.qmail@web35701.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Just to let everyone know: There is finally a company out there that sharpens microtome knifes (both stainless steel & tungsten) and manufactures plastic microwave accessories at a low price. I send all our knifes to them and I am extremely satisfied with the customer service I receive, along with the pricing. Also, they have made a number of accessories for us and I have nothing but good words to say about them. The accessory's work well and with no problem. The best thing about them is the company offers a warranty with each accessory. If you wish, email me back with your contact information and I will pass it on to the sharpening and MW accessory company. Thanks, Dr. Thomas --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. ------------------------------ Message: 10 Date: Tue, 4 Apr 2006 13:36:57 -0700 From: "Donna Harclerode" Subject: [Histonet] Humidity chamber To: Message-ID: <3DE0F644E093DF4BAE80C254176696A5090706@mp-mailserver.macropore.com> Content-Type: text/plain; charset="us-ascii" Thermo Electron makes a great system with a humidity chamber and a coverplate that I have used for many years. The slides are held with the plastic coverplate(the coverplates are suppose to be disposable, but I rinse them with only DI water and reuse them many times) I use 100ul of antibody (primary, secondary- etc, normally, but have managed to use as little as 90ul) to cover almost the whole slide. For buffer wash I fill up the top of the chamber with a squeeze bottle of PBS and allow to run through in about 5 minutes. I load the slides add primary, wash, secondary etc and only take them out for chromagen staining. The racks worked great when I had the positive control at the top of the slide and the test below. I do fluorescence secondaries in the coverplates, but I not do DAB in the coverplates, but flat on the counter. For HRP frozens I use the Dako endogenous peroxidase block in the Sequenza racks. I also use the Dako avidin biotin block in the coverplates when necessary. I usually incubate primary overnight in the fridge and the chambers stay humid for at least 48 hours. It takes a bit of practice to load the plates, but it is so worth it. Sequenza Racks - # 73310017 (Holds 10 coverplate assemblies) Coverplates - 25/pk = # 72110017 50/pk = # 7219950 250/case = # 72110013 http://www.thermo.com/eThermo/CMA/PDFs/Product/productPDF_19578.pdf 4th page Donna Harclerode, HT, (ASCP), HTL, QIHC Scientist / Immunohistochemistry Cytori Therapeutics 3020 Callan Rd. San Diego, CA 92121 858-458-0900 ext 5416 dharclerode@cytoritx.com ------------------------------ Message: 11 Date: Tue, 4 Apr 2006 17:13:06 -0400 From: Eric Dye (ext 223) Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed- Intervirewing and Hiring NOW To: Histonetters Message-ID: Content-Type: text/plain Fellow-Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) === message truncated === --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. From ree3 <@t> leicester.ac.uk Wed Apr 5 10:16:52 2006 From: ree3 <@t> leicester.ac.uk (Edwards, R.E.) Date: Wed Apr 5 10:17:06 2006 Subject: [Histonet] It's no wonder... Message-ID: it seems to me that the writers of the extensive and rambling off topic eeees might have not enough work to do, unlike me, who only has time for 29 words!!!! Cheers Workaholicric. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Chris Pomajzl Sent: 05 April 2006 15:52 To: HISTONET Subject: [Histonet] It's no wonder... ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From TJJ <@t> Stowers-Institute.org Wed Apr 5 10:29:13 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Wed Apr 5 10:29:54 2006 Subject: [Histonet] Reminder - when replying to digest messages Message-ID: A gentle reminder, dear folks... If you receive the Histonet messages in digest form and click reply in your email program, the body of all the messages are included in your reply. As digest users, you well know what this means in trying to sift through the new messages, especially after someone has clicked "reply" to reply to yours (including the messages again). I've finally figured out how to find my place again when the messages are numbered 1, 2, 3, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 4, 5, 1, 2, 3, 4, 5, 6, 7, 8....etc., but it's getting to be a problem again and sometimes I just don't have the patience to sift through them. Please remember to delete the list of messages in your reply before typing in your message and sending, or set your software to not include them in your reply. Best wishes, Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From gpbnas <@t> yahoo.es Wed Apr 5 10:50:32 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Wed Apr 5 10:53:02 2006 Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers In-Reply-To: Message-ID: <20060405155032.40643.qmail@web26212.mail.ukl.yahoo.com> Thanks, you advice going straight from sirius red to 70% ethanol or maybe to 100% as suggested in other reply, but how do I get rid of the excess red stuff on the slide? Best regards, Guillermo Tony Henwood escribi?: Drop step 3 and 4 (picric acid is soluble in water) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Guillermo Palao Sent: Wednesday, 5 April 2006 4:59 AM To: Histonet Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From mlm11 <@t> cornell.edu Wed Apr 5 11:00:35 2006 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Wed Apr 5 11:00:41 2006 Subject: [Histonet] tendon processing Message-ID: <5.2.1.1.2.20060405115331.00c5d140@postoffice9.mail.cornell.edu> Hello Histonet, Please, will someone send a tried and true protocol for paraffin processing of tendon? These people won't trim the pieces thin enough and the cassettes sit up off the molds. The first few I used xylene and have since used Propar and they are that much better. I still could use any other helpful hints. Thanks muchly. Do you charge more tendon? I find I'm going through knives. Thanks for all your help. Mary Lou Norman From tpmorken <@t> labvision.com Wed Apr 5 11:16:09 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Wed Apr 5 11:16:17 2006 Subject: [Histonet] California Society meeting, May 18-21 Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D80B@usca0082k08.labvision.apogent.com> Registration information for the 30th Annual California Society for Histotechology Symposium/Convention in Costa Mesa can be downloaded from the CSH website at www.californiahistology.org Program outline: THURSDAY, MAY 18TH MORNING WORKSHOPS, 8 AM TO 11:30 AM #1 NEW VISION OF H&E STAIN WHEN DYES HAVE BEEN APPLIED VICE VERSA - PART 1 Aricadiy I. Brusilovskiy, MD, PhD, HT(ASCP) Dr. AIB's Private Histology Lab #2 LEAN THINKING IN PATHOLOGY Stephen Jones, MS QIHC US Labs Randy Stephens US Labs #3 COMMUNICATION SKILLS FOR THE MODERN PATHOLOGY LABORATORY Carlos Gentry, BS, HT (ASCP), QIHC Genzyme Corporation Lora-Ann Gentry, BA HT (ASCP) Torrance Memorial Medical Center THURSDAY, MAY 18TH AFTERNOON WORKSHOPS, 1:30PM- 5:00PM #4 NEW VISION OF H&E STAIN WHEN DYES HAVE BEEN APPLIED VICE VERSA- PART II Aricadiy I. Brusilovskiy, MD, PhD, HT (ASCP) Dr .AIB's Private Histology Lab #5 WHAT STAINS FOR WHAT TUMOR??? Alvin W. Martin, M.D. University of Louisville Sharen Lear, HT (ASCP), HTL, QIHC University of Louisville #6 TECHNIQUE FOR PAINFUL SENSORY NEUROPATHY EVALUATION WITH SKIN Jean Mitchell HT (ASCP) University of Wisconsin Hospital & Clinics FRIDAY, MAY 19TH MORNING WORKSHOPS, 8AM T0 11:30AM #7 MULTI-LEVEL TESTING: A WORKING ALGORITHM FOR IHC ASSAY DEVELOPMENT AND VALIDATION Carlos Gentry, BS, HT (ASCP), QIHC Genzyme Corporation Daisy Joseph, BA, HT/HTL (ASCP) Genzyme Corporation #8 "IF YOU HAVE THE MUSCLE: I HAVE THE NERVE" Jean Mitchell HT (ASCP) University of Wisconsin Hospital & Clinics #9 INTRODUCTION OF VARIOUS CYTOPREPARATORY TECHNIQUES AND PAPANICOLAOU STAINING METHOD IN PREPARING CYTOLOGY SPECIMENS Claro Y.Masangcay, BA, CT(ASCP) Loma Linda Medical Center Kelly Liu, BS CT (ASCP) Loma Linda Medical Center FRIDAY, MAY 19TH AFTERNOON WORKSHOPS, 1:30PM TO 5:00PM #10 "WORKING SMARTER NOT HARDER" Debra Cobb, MBA, HT(ASCP) KKB Consulting Services #11 HAVE YOU HEARD??? THE REGULATIONS FOR SHIPPING MEDICAL SPECIMENS HAVE CHANGED - AGAIN! Linda Durbin EXAKT Technologies, Inc., Oklahoma #12 IMMUNOHISTOCHEMICAL STAINING AND ITS USE IN HEMATOPATHOLOGY Alvin W. Martin, MD University of Louisville, Kentucky Sheron Lear, HT(ASCP), HTL, QIHC University of Louisville, Kentucky SATURDAY, MAY 20TH MORNING WORKSHOPS, 8AM TO 11:30PM #13 WHEN YOUR TRUSTY MICROTOME JUST WON'T CUT IT....... Linda Durbin EXAKT Technologies, Inc., Oklahoma Level:Basic Limit: Unlimited #14 HEREDITARY BASIS OF COLORECTAL CARCINOMA Holly Yamada, MD Hemediagnostics, Inc. Level: Intermediate and Advanced Limited: Unlimited #15 FROZEN SECTION TECHNIQUE AND NEW METHODS FOR CRYOEMBEDDING TISSUES Stephen Peters, MD Hackensack University Medical Center Level: Basic Limit: Unlimited SATURDAY, MAY 20TH EDUCATIONAL MEMORIAL WORKSHOP, 2:00PM TO 5:00PM #16 CSH WELCOMES DR. CLIVE TAYLOR TO THE OC MOLECULAR MORPHOLOGY; DIAGNOSTIC APPLICATION OF IMMUNOHISTOCHEMISTRY AND IN SITU HYBRIDIZATION IN 2006 SUNDAY, MAY 21ST MORNING WORKSHOP, 8AM TO 11:30AM #17 TSE's IN THE UNITED STATES: WHAT ARE YOUR RISKS? Greg Anderson, HTL (ASCP) QIHC Ventana Medical Systems, Inc. #18 APPLICATIONS OF HISTOTECHNOLOGY IN VETERINARY DIAGNOSTICS Francisco Uzal, DVM, MS, PhD University of California, Davis Meredith Rhea, BS University of California, Davis Jenee Odani, DVM University of California, Davis #19 THE ART AND SCIENCE OF IHC ON MOUSE AND RAT TISSUE David Tacha, HTL(ASCP), Ph.D BioCare Medical Tim Morken From liz <@t> premierlab.com Wed Apr 5 11:22:14 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Wed Apr 5 11:20:33 2006 Subject: [Histonet] tendon processing In-Reply-To: <5.2.1.1.2.20060405115331.00c5d140@postoffice9.mail.cornell.edu> Message-ID: <002801c658cd$193d5e30$0300a8c0@Chlipala> Mary Lou We process quite a bit of human tendon samples and it can be challenging. The pieces we receive are not that thick but we process between biopsy pads to help keep the tissue flat. We use a longer processing cycle (1 to 1.5 hours in each station). When sectioning you need to be very careful when rough trimming in so you don't tear the fibers, let soak on cold ice for about 30 minutes to an hour and then section. We use new blades. We place the cut sections flat on a warm hot plate (paraffin does not melt) overnight and then stain the following day. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mary Lou Norman Sent: Wednesday, April 05, 2006 10:01 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] tendon processing Hello Histonet, Please, will someone send a tried and true protocol for paraffin processing of tendon? These people won't trim the pieces thin enough and the cassettes sit up off the molds. The first few I used xylene and have since used Propar and they are that much better. I still could use any other helpful hints. Thanks muchly. Do you charge more tendon? I find I'm going through knives. Thanks for all your help. Mary Lou Norman _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________ NOD32 1.1473 (20060405) Information __________ This message was checked by NOD32 antivirus system. http://www.eset.com From gcallis <@t> montana.edu Wed Apr 5 11:20:34 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Apr 5 11:20:49 2006 Subject: [Histonet] tendon processing In-Reply-To: <5.2.1.1.2.20060405115331.00c5d140@postoffice9.mail.cornell .edu> References: <5.2.1.1.2.20060405115331.00c5d140@postoffice9.mail.cornell.edu> Message-ID: <6.0.0.22.1.20060405101222.01b5ad38@gemini.msu.montana.edu> Buy the deeper metal molds for embedding oversize tissues. You can get these from Fisher, VWR, Thermo, etc to fit your cassettes. If you use high profile disposable blades, you get less chatter, more stability during paraffin sections (tendons, decalcified bone, and very fibrous tissues) . High profiles are just as sharp as low profile disposable blades. Tendon may be a bit drier, so a gentle soak on warm then cold water may help, beware of overdehydration and never add heat to dehydration and clearing steps, that only dries out these tough tissues even more. Propar should work fine and less hardening to this tissue or any other tissue, just make sure you rotate this out more frequently as this xylene substitute is a bit more sensitive to water carryover, xylene tends to clear water a bit better, but keeping Propar fresh with more frequent changes helps - this is dependent on how many tissues you put through a processor in a week? What do you mean by "charge more tendon"? or is that change knives more often when sectioning tendon? You should be able to use disposable blades rather than permanent c profile knives (if that is what you are using?) At 10:00 AM 4/5/2006, you wrote: >Hello Histonet, > >Please, will someone send a tried and true protocol for paraffin >processing of tendon? These people won't trim the pieces thin enough and >the cassettes sit up off the molds. The first few I used xylene and have >since used Propar and they are that much better. I still could use any >other helpful hints. Thanks muchly. >Do you charge more tendon? I find I'm going through knives. > >Thanks for all your help. >Mary Lou Norman > > > >_______________________________________________ >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 406 994-4303 (FAX) From Jackie.O'Connor <@t> abbott.com Wed Apr 5 11:26:02 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Wed Apr 5 11:27:09 2006 Subject: [Histonet] postal Message-ID: Unfortunately, "postal" has fallen into the English Language to define just that - just like KleenexTM means any tissue, CrayolaTM is a catch-all phrase for any crayon, etc, etc. Just don't take it out on my good friend, Steve Postl. Jackie O' "Owen, Michael P" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/05/2006 10:02 AM To: "'Histonet'" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] postal Perhaps I am off-base with this comment, but I prefer the term "postal" not being used when referencing work rage and work violence. One of my best friends was an employee of the U.S. Postal Service. She was shot and killed by a customer of her branch in 1998. If my memory serves me correctly, work violence is no longer primarily found in U.S. Postal Service facilities. Diana, my post is not meant as a flaming message to you. Thanks for your informative comments. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Wed Apr 5 11:35:02 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Wed Apr 5 11:40:47 2006 Subject: [Histonet] postal Message-ID: It's even in the dictionary: go postal Slang To become extremely angry or deranged, especially in an outburst of violence -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Wednesday, April 05, 2006 12:26 PM To: Owen, Michael P Cc: 'Histonet'; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] postal Unfortunately, "postal" has fallen into the English Language to define just that - just like KleenexTM means any tissue, CrayolaTM is a catch-all phrase for any crayon, etc, etc. Just don't take it out on my good friend, Steve Postl. Jackie O' "Owen, Michael P" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/05/2006 10:02 AM To: "'Histonet'" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] postal Perhaps I am off-base with this comment, but I prefer the term "postal" not being used when referencing work rage and work violence. One of my best friends was an employee of the U.S. Postal Service. She was shot and killed by a customer of her branch in 1998. If my memory serves me correctly, work violence is no longer primarily found in U.S. Postal Service facilities. Diana, my post is not meant as a flaming message to you. Thanks for your informative comments. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From funderwood <@t> mcohio.org Wed Apr 5 11:47:09 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Wed Apr 5 11:47:36 2006 Subject: [Histonet] postal Message-ID: Isn't it time we stamp out postal behavior. >>> "Bartlett, Jeanine" 04/05/06 12:35PM >>> It's even in the dictionary: go postal Slang To become extremely angry or deranged, especially in an outburst of violence -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jackie M O'Connor Sent: Wednesday, April 05, 2006 12:26 PM To: Owen, Michael P Cc: 'Histonet'; histonet-bounces@lists.utsouthwestern.edu Subject: RE: [Histonet] postal Unfortunately, "postal" has fallen into the English Language to define just that - just like KleenexTM means any tissue, CrayolaTM is a catch-all phrase for any crayon, etc, etc. Just don't take it out on my good friend, Steve Postl. Jackie O' "Owen, Michael P" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/05/2006 10:02 AM To: "'Histonet'" cc: (bcc: Jackie M O'Connor/LAKE/GPRD/ABBOTT) Subject: RE: [Histonet] postal Perhaps I am off-base with this comment, but I prefer the term "postal" not being used when referencing work rage and work violence. One of my best friends was an employee of the U.S. Postal Service. She was shot and killed by a customer of her branch in 1998. If my memory serves me correctly, work violence is no longer primarily found in U.S. Postal Service facilities. Diana, my post is not meant as a flaming message to you. Thanks for your informative comments. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov _______________________________________________ Histonet mailing list Histonet@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 julien_lambreydesouza <@t> uqar.qc.ca Wed Apr 5 11:51:19 2006 From: julien_lambreydesouza <@t> uqar.qc.ca (Julien Lambrey de Souza) Date: Wed Apr 5 11:51:24 2006 Subject: [Histonet] Mayer's hematox Message-ID: <7bc655d477fe96232fcc967de2f2eea5@uqar.qc.ca> Hello, Does anyone have the protocol for Mayer's hematox from Drury and Wallington's book (Carleton's histological techniques)? I need to compare with other protocols I have. Thanks' Julien Lambrey de Souza Research assistant, Evolutionary biology University of Quebec at Rimouski Tel: (418) 723-1986 #1714 Fax: (418) 724-1849 From vazquezr <@t> ohsu.edu Wed Apr 5 11:55:14 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Apr 5 11:56:13 2006 Subject: [Histonet] It's no wonder... Message-ID: I second it... Robyn >>> "King, Curtis - RAS" 4/5/2006 7:54 AM >>> Well said Chris. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Chris Pomajzl Sent: Wednesday, April 05, 2006 10:52 AM To: HISTONET Subject: [Histonet] It's no wonder... ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Wed Apr 5 12:09:43 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Apr 5 12:09:44 2006 Subject: [Histonet] Mayer's hematox References: <7bc655d477fe96232fcc967de2f2eea5@uqar.qc.ca> Message-ID: <4433F9D7.527A6ECF@uwo.ca> >From the 4th edition (1967), p.127: "Stain for 5-10 minutes." -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Julien Lambrey de Souza wrote: > > Hello, > > Does anyone have the protocol for Mayer's hematox from Drury and > Wallington's book (Carleton's histological techniques)? I need to > compare with other protocols I have. > > Thanks' > > Julien Lambrey de Souza > Research assistant, Evolutionary biology > University of Quebec at Rimouski > > Tel: (418) 723-1986 #1714 > Fax: (418) 724-1849 > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Wed Apr 5 12:45:01 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Apr 5 12:45:07 2006 Subject: [Histonet] It's no wonder... Message-ID: It is not unusual for people to vent their feeling regarding their job. A study several years ago concluded that people that complain are at least interested in their job. Apathy on the other hand gets us nowhere (as seen with our political system and lack of votes). Venting is I believe a healthy thing and prevents internal pressure building up, providing that there is a limit to venting. I think that it is difficult not to be frustrated when we see the valuable contribution that the profession makes and yet often sees little reward. We are all expected to do more work in the same time and many times the service/hospital etc. may benefit financially but this does not seem to trickle down to the professionals that are actually doping the work. Please do not think that this is unique to this profession. Just look at the -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Wednesday, April 05, 2006 11:55 AM To: cpomajzl@cpllabs.com; histonet@lists.utsouthwestern.edu; cking@rallansci.com Subject: RE: [Histonet] It's no wonder... I second it... Robyn >>> "King, Curtis - RAS" 4/5/2006 7:54 AM >>> Well said Chris. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Chris Pomajzl Sent: Wednesday, April 05, 2006 10:52 AM To: HISTONET Subject: [Histonet] It's no wonder... ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dsantana <@t> pmaonline.com Wed Apr 5 12:51:02 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Wed Apr 5 12:52:52 2006 Subject: [Histonet] It's no wonder Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB0711395A@MAILPMA> That's why we have Happy Hour.....LOL From Barry.R.Rittman <@t> uth.tmc.edu Wed Apr 5 12:54:29 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Wed Apr 5 12:54:34 2006 Subject: [Histonet] It's no wonder... Message-ID: I apologize, my wrist caught one of the computer buttons before I finished.... It is not unusual for people to vent their feeling regarding their job. A study several years ago concluded that people that complain are at least interested in their job. Apathy on the other hand gets us nowhere (as seen with our political system and lack of votes). Venting is I believe a healthy thing and prevents internal pressure building up, providing that there is a limit to venting. I think that it is difficult not to be frustrated when we see the valuable contribution that the profession makes and yet often sees little reward. We are all expected to do more work in the same time and many times the service/hospital etc. may benefit financially but this does not seem to trickle down to the professionals that are actually doping the work. Please do not think that this is unique to this profession. Just look at higher education where the student faculty ratio has been increasing for several year. I believe that we should channel this love of the profession into something positive. If we want to have better recognition, better pay and conditions it is up to us. We need to inform the general public, legislators etc. about what we do and its value to the communities we serve. So the bottom line is get it off your chest - you will feel a lot better but also get off your butt and write your comments to the above groups. Above all be have constructive suggestions. No good complaining unless you have constructive comments on how to improve the situation. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Wednesday, April 05, 2006 11:55 AM To: cpomajzl@cpllabs.com; histonet@lists.utsouthwestern.edu; cking@rallansci.com Subject: RE: [Histonet] It's no wonder... I second it... Robyn >>> "King, Curtis - RAS" 4/5/2006 7:54 AM >>> Well said Chris. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Chris Pomajzl Sent: Wednesday, April 05, 2006 10:52 AM To: HISTONET Subject: [Histonet] It's no wonder... ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From specialstainsqueen <@t> hotmail.com Wed Apr 5 12:56:49 2006 From: specialstainsqueen <@t> hotmail.com (Sherri Anderson) Date: Wed Apr 5 12:56:55 2006 Subject: [Histonet] RE: Microtome knife sharpening service/ Microwaveaccessories Message-ID: Over the past few weeks, I've read several of your messages with similar content to your most recent email -- without naming the company, you endorse the company's services and offer to provide or pass along info if someone contacts you directly. Unless you are a vendor trying to disguise yourself....or unless you're receiving some sort of "kickback" from this company by referring potential customers (i.e. Histonetters who have contacted you), why do you just not name the company so that all subscribers can benefit? I don't mean to offend you in any way, but why not share the company name and info with everyone? It would benefit the company as well as Histonet subscribers. Sincerely, Sherri >From: Justin Thomas >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] RE: Microtome knife sharpening service/ >Microwaveaccessories >Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) > >Just to let everyone know: > > There is finally a company out there that sharpens microtome knifes >(both stainless steel & tungsten) and manufactures plastic microwave >accessories at a low price. I send all our knifes to them and I am >extremely satisfied with the customer service I receive, along with the >pricing. Also, they have made a number of accessories for us and I have >nothing but good words to say about them. The accessory's work well and >with no problem. The best thing about them is the company offers a >warranty with each accessory. If you wish, email me back with your contact >information and I will pass it on to the sharpening and MW accessory >company. > > Thanks, > Dr. Thomas > > >--------------------------------- >New Yahoo! Messenger with Voice. Call regular phones from your PC and save >big. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From histology <@t> gradymem.org Wed Apr 5 13:14:58 2006 From: histology <@t> gradymem.org (histology@gradymem.org) Date: Wed Apr 5 13:18:34 2006 Subject: [Histonet] RE: Microtome knife sharpening service/ Microwaveaccessories Message-ID: <25226fc2525e02.2525e0225226fc@onenet.net> I don't know if this is the company he is talking about, but Coleman Manfacturing at coleman_manufacturing@yahoo.com has inexpensive microwave accessories and will custom build them for you. Dr. Thomas is probabaly just a satisfied customer that doesn't want to use the histonet to advertise to the masses unless you are interested. Angie Barnett, HTL(ASCP) Grady Memorial Hospital Pathology Department 405/224-2258 histology@gradymem.org ----- Original Message ----- From: Sherri Anderson Date: Wednesday, April 5, 2006 12:56 pm Subject: RE: [Histonet] RE: Microtome knife sharpening service/ Microwaveaccessories > Over the past few weeks, I've read several of your messages with > similar > content to your most recent email -- without naming the company, > you endorse > the company's services and offer to provide or pass along info if > someone > contacts you directly. > > Unless you are a vendor trying to disguise yourself....or unless > you're > receiving some sort of "kickback" from this company by referring > potential > customers (i.e. Histonetters who have contacted you), why do you > just not > name the company so that all subscribers can benefit? > I don't mean to offend you in any way, but why not share the > company name > and info with everyone? It would benefit the company as well as > Histonet > subscribers. > > Sincerely, > Sherri > > > >From: Justin Thomas > >To: histonet@lists.utsouthwestern.edu > >Subject: [Histonet] RE: Microtome knife sharpening service/ > >Microwaveaccessories > >Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) > > > >Just to let everyone know: > > > > There is finally a company out there that sharpens microtome knifes > >(both stainless steel & tungsten) and manufactures plastic microwave > >accessories at a low price. I send all our knifes to them and I am > >extremely satisfied with the customer service I receive, along > with the > >pricing. Also, they have made a number of accessories for us and > I have > >nothing but good words to say about them. The accessory's work > well and > >with no problem. The best thing about them is the company offers a > >warranty with each accessory. If you wish, email me back with > your contact > >information and I will pass it on to the sharpening and MW accessory > >company. > > > > Thanks, > > Dr. Thomas > > > > > >--------------------------------- > >New Yahoo! Messenger with Voice. Call regular phones from your PC > and save > >big. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ > Don?t just search. Find. Check out the new MSN Search! > http://search.msn.click-url.com/go/onm00200636ave/direct/01/ > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From michael.owen <@t> fda.hhs.gov Wed Apr 5 13:42:45 2006 From: michael.owen <@t> fda.hhs.gov (Owen, Michael P) Date: Wed Apr 5 13:53:57 2006 Subject: [Histonet] Apologies for April 5th Message Message-ID: Dear List Members, First, I apologize for spelling Diane's name incorrectly. Second, I apologize if my comment was considered as an attack. It was not intended at all to attack Diane nor anyone else on this list. I am usually a positive person. I try to show this attitude by usually staying out of the heated discussions on this list. I did not know the aforementioned term is now in general use. I appreciate the earlier reference to the dictionary entry. I have apologized to Diane off-list. I received a slightly negative e-mail from a list user whom said I was off-base. Again, I state I did not mean to make the discussion more volatile. On a more positive note, I highly recommend the book below (available from Amazon.com for only $10.74 U.S.). After reading the recent messages on this forum, the laboratories that are placing too many demands on their workers should follow the concepts in this book instead of following status quo. (The book is also helpful to workers for improving their own conditions and their job searches when improving their current position is no longer an option -- the main focus of the text) Corporate Confidential: 50 Secrets Your Company Doesn't Want You to Know -- and What to Do About Them (Paperback) by Cynthia Shapiro Paperback: 224 pages Publisher: St. Martin's Griffin (September 1, 2005) Language: English ISBN: 0312337361 Product Dimensions: 8.6 x 6.3 x 0.6 inches Shipping Weight: 7.2 ounces. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov From michael.owen <@t> fda.hhs.gov Wed Apr 5 14:15:17 2006 From: michael.owen <@t> fda.hhs.gov (Owen, Michael P) Date: Wed Apr 5 14:30:08 2006 Subject: [Histonet] Staining Bacterial Spores Message-ID: Dear List Members, Are there any solutions used in your field that kill bacterial spores? Does Zenker's Solution kill spores with a high degree of success? Thanks in advance for your feedback. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov From brett_connolly <@t> merck.com Wed Apr 5 14:30:01 2006 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Wed Apr 5 14:30:24 2006 Subject: [Histonet] Job Opportunity - Pennsylvania Message-ID: <355C35514FEAC9458F75947F5270974D67CE02@usctmx1103.merck.com> Histonetters, We have a new position available in my lab. I encourage any interested parties to follow the link below to our Career site and enter the job # (PHA000612) in the search box foe the application details Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com PHA000612 - Research Biologist / Staff Biologist / Biologist (Imaging) Merck develops breakthrough medicines and treatments that offer a new lease on life. At Merck, improving patient health isn't just what we do. It's who we are, sharing a passion for life that brings out the best in a diverse workforce. That's why Merck is recognized as one of the world's leading research-based pharmaceutical companies. The Department of Imaging at the Merck Research Laboratories in West Point, PA is seeking an in vitro histologist to join the group. The successful candidate will be expected to work as part of a team using cutting edge imaging technologies to develop and evaluate non clinical models to support drug discovery projects. Experience with a variety of histological methodologies such as specimen fixation, embedding, sectioning, immunocytochemistry and immunofluorescence, and the ability to work effectively both independently and in a team environment is essential. Practical expertise with image analysis systems would be advantageous. Under general scientific supervision, the successful candidate will be primarily responsible for the generation and analysis of experimental data. Qualifications for this position include a BS or MS degree in Biological science and ASCP certification as a histotechnician (HT) or histotechnologist (HTL) is required. The qualified individual must be highly motivated, conscientious, meticulous with expertise in immunohistological techniques as evidenced by several years of laboratory experience. She/he also needs to be computer literate, have excellent organizational and communication skills and enjoy working in dynamic teams. Our commitment to our employees resonates in the benefits we offer including competitive compensation, tuition reimbursement, work-life balance initiatives, on-site child care at many of our locations and opportunities for personal and professional enrichment. Join us and become a part of our commitment and our legacy which continues to deliver novel medicines to the people that need them the most. To be considered for this position, please visit our career site at http://www.merck.com/careers/search_jobs.html to create a profile and submit your CV for job number PHA000612. Discovery of Great Drugs demands continual discovery of Great People... Merck is an equal opportunity employer--proudly embracing diversity in all of its manifestations. ------------------------------------------------------------------------------ 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 ploykasek <@t> phenopath.com Wed Apr 5 14:30:35 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Wed Apr 5 14:30:58 2006 Subject: [Histonet] Breast fixatives Message-ID: I understand the need for well-fixed & processed breast tissue. Also, there is always the competing turn around time pressure. However, I would like to toss in an observation about using alcoholic formalin. If you are planning on doing Her2neu testing by IHC, the use of alcoholic formalin will artificially increase the signal. Look carefully at the normal ducts in breast tissue fixed with this method & you will usually see signal in the normal ducts. Also, if you are running a FDA approved method, usually the fixation method is specified & this may not conform with it. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From specialstainsqueen <@t> hotmail.com Wed Apr 5 14:33:01 2006 From: specialstainsqueen <@t> hotmail.com (Sherri Anderson) Date: Wed Apr 5 14:33:09 2006 Subject: [Histonet] RE: Microtome knife sharpening service/ Microwaveaccessories In-Reply-To: <25226fc2525e02.2525e0225226fc@onenet.net> Message-ID: Dear Angie- Thanks for the reply. Dr. Thomas also replied to me. I wasn't trying to offend him or anyone else, I just didn't really understand why a customer wouldn't recommend a company by name (I realize that vendors are not to use this forum to advertise their own products...and rightfully so...but a lot of general subscribers either praise or pass along critical comments regarding specific brands and products -- and that truly benefits all customers). I apologize to both Dr. Thomas and the subscibers in general if my email was accusatory in nature. I just really didn't understand (with the exception of the two reasons I listed in my original email) why a non-vendor wouldn't praise a company by name. I didn't mean it in any other way. Thank you. Sincerely, Sherri >From: histology@gradymem.org >To: Sherri Anderson >CC: jstn192@yahoo.com, histonet@lists.utsouthwestern.edu >Subject: Re: RE: [Histonet] RE: Microtome knife sharpening service/ >Microwaveaccessories >Date: Wed, 05 Apr 2006 13:14:58 -0500 > >I don't know if this is the company he is talking about, but Coleman >Manfacturing at coleman_manufacturing@yahoo.com has inexpensive >microwave accessories and will custom build them for you. > >Dr. Thomas is probabaly just a satisfied customer that doesn't want to >use the histonet to advertise to the masses unless you are interested. > >Angie Barnett, HTL(ASCP) >Grady Memorial Hospital >Pathology Department >405/224-2258 >histology@gradymem.org > > >----- Original Message ----- >From: Sherri Anderson >Date: Wednesday, April 5, 2006 12:56 pm >Subject: RE: [Histonet] RE: Microtome knife sharpening service/ >Microwaveaccessories > > Over the past few weeks, I've read several of your messages with > > similar > > content to your most recent email -- without naming the company, > > you endorse > > the company's services and offer to provide or pass along info if > > someone > > contacts you directly. > > > > Unless you are a vendor trying to disguise yourself....or unless > > you're > > receiving some sort of "kickback" from this company by referring > > potential > > customers (i.e. Histonetters who have contacted you), why do you > > just not > > name the company so that all subscribers can benefit? > > I don't mean to offend you in any way, but why not share the > > company name > > and info with everyone? It would benefit the company as well as > > Histonet > > subscribers. > > > > Sincerely, > > Sherri > > > > > > >From: Justin Thomas > > >To: histonet@lists.utsouthwestern.edu > > >Subject: [Histonet] RE: Microtome knife sharpening service/ > > >Microwaveaccessories > > >Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) > > > > > >Just to let everyone know: > > > > > > There is finally a company out there that sharpens microtome >knifes > > >(both stainless steel & tungsten) and manufactures plastic >microwave > > >accessories at a low price. I send all our knifes to them and I am > > >extremely satisfied with the customer service I receive, along > > with the > > >pricing. Also, they have made a number of accessories for us and > > I have > > >nothing but good words to say about them. The accessory's work > > well and > > >with no problem. The best thing about them is the company offers a > > >warranty with each accessory. If you wish, email me back with > > your contact > > >information and I will pass it on to the sharpening and MW accessory > > >company. > > > > > > Thanks, > > > Dr. Thomas > > > > > > > > >--------------------------------- > > >New Yahoo! Messenger with Voice. Call regular phones from your PC > > and save > > >big. > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _________________________________________________________________ > > Don’t just search. Find. Check out the new MSN Search! > > http://search.msn.click-url.com/go/onm00200636ave/direct/01/ > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From rodger_65489 <@t> yahoo.com Wed Apr 5 09:20:12 2006 From: rodger_65489 <@t> yahoo.com (Roger Smithwell) Date: Wed Apr 5 14:36:12 2006 Subject: [Histonet] ATTN: Anyone who wants Milestone microwave accessories (almost free) Message-ID: <20060405142012.85992.qmail@web38307.mail.mud.yahoo.com> Fellow Histonetters: We have two new extra milestone accessory histomodule kits in our Lab. A 110 cassette and a 30 cassette; it has everything with it just as it came from Milestone. We have both that are used under vacuum and under non- vac. We have never used them and they are only taking up space. If you want them email me back with your contact info and the best time to get in touch with you. Best Regards, Dr. Rodger __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From qbta <@t> aol.com Wed Apr 5 15:01:33 2006 From: qbta <@t> aol.com (qbta@aol.com) Date: Wed Apr 5 15:01:48 2006 Subject: [Histonet] Slide identifiers Message-ID: <8C82702DF23DF9C-444-3514@mblk-d32.sysops.aol.com> Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. From froyer <@t> bitstream.net Wed Apr 5 15:04:26 2006 From: froyer <@t> bitstream.net (Ford Royer) Date: Wed Apr 5 15:04:40 2006 Subject: [Histonet] RE: Microtome knife sharpening service/Microwaveaccessories In-Reply-To: Message-ID: <002f01c658ec$242553b0$6f01a80a@fords> Yet isn't it curious that when someone is not satisfied with a certain vendor's product or service that they do not hesitate to name that vendor and flame them unmercifully on this List? But when someone is completely satisfied and pleased with the product or service from a vendor, it is felt that that vendor's name should be kept secret to avoid advertising/endorsing? Negative news (complaints) is not "advertising," yet positive news is? Strange... ~ Ford ...let the flaming begin. Ford M. Royer, MT(ASCP) Minnesota Medical Specialists, Inc. 7177 Madison Ave. W. Golden Valley, MN 55427-3601 email -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Sherri Anderson Sent: Wednesday, April 05, 2006 2:33 PM To: histology@gradymem.org Cc: histonet@lists.utsouthwestern.edu; jstn192@yahoo.com Subject: Re: RE: [Histonet] RE: Microtome knife sharpening service/Microwaveaccessories Dear Angie- Thanks for the reply. Dr. Thomas also replied to me. I wasn't trying to offend him or anyone else, I just didn't really understand why a customer wouldn't recommend a company by name (I realize that vendors are not to use this forum to advertise their own products...and rightfully so...but a lot of general subscribers either praise or pass along critical comments regarding specific brands and products -- and that truly benefits all customers). I apologize to both Dr. Thomas and the subscibers in general if my email was accusatory in nature. I just really didn't understand (with the exception of the two reasons I listed in my original email) why a non-vendor wouldn't praise a company by name. I didn't mean it in any other way. Thank you. Sincerely, Sherri >From: histology@gradymem.org >To: Sherri Anderson >CC: jstn192@yahoo.com, histonet@lists.utsouthwestern.edu >Subject: Re: RE: [Histonet] RE: Microtome knife sharpening service/ >Microwaveaccessories >Date: Wed, 05 Apr 2006 13:14:58 -0500 > >I don't know if this is the company he is talking about, but Coleman >Manfacturing at coleman_manufacturing@yahoo.com has inexpensive >microwave accessories and will custom build them for you. > >Dr. Thomas is probabaly just a satisfied customer that doesn't want to >use the histonet to advertise to the masses unless you are interested. > >Angie Barnett, HTL(ASCP) >Grady Memorial Hospital >Pathology Department >405/224-2258 >histology@gradymem.org > > >----- Original Message ----- >From: Sherri Anderson >Date: Wednesday, April 5, 2006 12:56 pm >Subject: RE: [Histonet] RE: Microtome knife sharpening service/ >Microwaveaccessories > > Over the past few weeks, I've read several of your messages with > > similar > > content to your most recent email -- without naming the company, > > you endorse > > the company's services and offer to provide or pass along info if > > someone > > contacts you directly. > > > > Unless you are a vendor trying to disguise yourself....or unless > > you're > > receiving some sort of "kickback" from this company by referring > > potential > > customers (i.e. Histonetters who have contacted you), why do you > > just not > > name the company so that all subscribers can benefit? > > I don't mean to offend you in any way, but why not share the > > company name > > and info with everyone? It would benefit the company as well as > > Histonet > > subscribers. > > > > Sincerely, > > Sherri > > > > > > >From: Justin Thomas > > >To: histonet@lists.utsouthwestern.edu > > >Subject: [Histonet] RE: Microtome knife sharpening service/ > > >Microwaveaccessories > > >Date: Tue, 4 Apr 2006 13:22:18 -0700 (PDT) > > > > > >Just to let everyone know: > > > > > > There is finally a company out there that sharpens microtome >knifes > > >(both stainless steel & tungsten) and manufactures plastic >microwave > > >accessories at a low price. I send all our knifes to them and I am > > >extremely satisfied with the customer service I receive, along > > with the > > >pricing. Also, they have made a number of accessories for us and > > I have > > >nothing but good words to say about them. The accessory's work > > well and > > >with no problem. The best thing about them is the company offers a > > >warranty with each accessory. If you wish, email me back with > > your contact > > >information and I will pass it on to the sharpening and MW accessory > > >company. > > > > > > Thanks, > > > Dr. Thomas > > > > > > > > >--------------------------------- > > >New Yahoo! Messenger with Voice. Call regular phones from your PC > > and save > > >big. > > >_______________________________________________ > > >Histonet mailing list > > >Histonet@lists.utsouthwestern.edu > > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > _________________________________________________________________ > > Don't just search. Find. Check out the new MSN Search! > > http://search.msn.click-url.com/go/onm00200636ave/direct/01/ > > > > > > _______________________________________________ > > Histonet mailing list > > Histonet@lists.utsouthwestern.edu > > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _________________________________________________________________ Don't just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Margaret.Perry <@t> sdstate.edu Wed Apr 5 15:17:50 2006 From: Margaret.Perry <@t> sdstate.edu (Perry, Margaret) Date: Wed Apr 5 15:18:03 2006 Subject: [Histonet] TRP-2 antibody Message-ID: Has anyone used the TRP-2 antibodies from Santa Cruz for detection of melanomas? I also found a DCT (dopachrome tautomerase) polyclonal from Gentaur and would like feedback if anyone has used it. Margaret Perry HT(ASCP) Vet Science South Dakota State University From pruegg <@t> ihctech.net Wed Apr 5 15:46:47 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Apr 5 15:47:01 2006 Subject: [Histonet] cardio myocyte histomorpometry Message-ID: <200604052046.k35KkjrF007129@chip.viawest.net> We want to measure the size of cardio myocytes. The investigator heard tell of a "Silver" stain used for this but he doesn't know what silver stain. What is recommended? This will be done using a computerized image analysis system. Thanks in advance, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. From HornHV <@t> archildrens.org Wed Apr 5 15:46:44 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Wed Apr 5 15:47:23 2006 Subject: [Histonet] Slide identifiers Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BE029@EMAIL.archildrens.org> This does not apply to slides/cassettes. It the initial specimen container of when the specimen is collected. hh -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of qbta@aol.com Sent: Wednesday, April 05, 2006 3:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide identifiers Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------------ The information contained in this message may be privileged and confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. ============================================================================== From JWEEMS <@t> sjha.org Wed Apr 5 15:50:27 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Apr 5 15:50:34 2006 Subject: [Histonet] Slide identifiers Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01D34F80@sjhaexc02.sjha.org> We use the case number and the name. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of qbta@aol.com Sent: Wednesday, April 05, 2006 4:02 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide identifiers Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From jkiernan <@t> uwo.ca Wed Apr 5 16:31:52 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Wed Apr 5 16:31:53 2006 Subject: [Histonet] Mayer's hematox References: <7bc655d477fe96232fcc967de2f2eea5@uqar.qc.ca> <4433F9D7.527A6ECF@uwo.ca> Message-ID: <44343748.1BAE6E1E@uwo.ca> You should have said! Its: Haematoxylin 1G Sodium iodate 0.2G Potassium alum 50G Citric acid 1G Chloral hydrate 50G Dist. water 1000ml Allow the first 3 items to dissolve in the water overnight. Add citric acid and chloral hydrate, then boil for 5 mins. Ready for use when cool. D & W call the chloral hydrate a "preservative," which is a statement I don't understand. Preserving what? This matter has been discussed on Histonet in the past (not by me), and the assertion has been made that Mayer's haemalum works just as well without the chloral hydrate. I can't remember who said that; I've used only the complete mixture myself. It would be worth searching in www.histosearch.com -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Julien Lambrey de Souza wrote: > > Thanks, > but actually I meant I needed the recipe for making Mayer's hematox > according to Drury and Wallington (We don't have the book here and > although I ordered it, I won't have it in time for the due dates...). > > Julien. > > Le 06-04-05, ? 13:09, John A. Kiernan a ?crit : > > > From the 4th edition (1967), p.127: > > > > "Stain for 5-10 minutes." > > -- > > ------------------------------- > > John A. Kiernan > > Department of Anatomy and Cell Biology > > The University of Western Ontario > > London, Canada N6A 5C1 > > kiernan[AT]uwo.ca > > http://publish.uwo.ca/~jkiernan/ > > http://instruct.uwo.ca/anatomy/530/index.htm > > _______________________________ > > Julien Lambrey de Souza wrote: > >> > >> Hello, > >> > >> Does anyone have the protocol for Mayer's hematox from Drury and > >> Wallington's book (Carleton's histological techniques)? I need to > >> compare with other protocols I have. > >> > >> Thanks' > >> > >> Julien Lambrey de Souza > >> Research assistant, Evolutionary biology > >> University of Quebec at Rimouski > >> > >> Tel: (418) 723-1986 #1714 > >> Fax: (418) 724-1849 > >> > >> _______________________________________________ > >> Histonet mailing list > >> Histonet@lists.utsouthwestern.edu > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From NKMitche <@t> ahs.llumc.edu Wed Apr 5 17:29:54 2006 From: NKMitche <@t> ahs.llumc.edu (Mitchell, Nancy K.) Date: Wed Apr 5 17:30:00 2006 Subject: [Histonet] procedure for calibrating the wattage on a microwave oven Message-ID: <8EB1459EA20E4E498AC2E05378F8F30A536EBD@mind.mc.ad.lluahsc.org> Does anyone have a procedure that they can share for calibrating the temp/wattage on a microwave oven? We use ours (house hold type-800 watt) for heating biological stains only-no processing. Thank you. Nancy Mitchell, HT ASCP Histology Supervisor Loma Linda Pathology Medical Group 11370 Anderson Street Ste 2960 Loma Linda, CA 92354 909-558-2012 FAX-909-796-3667 From pmcardle <@t> ebsciences.com Wed Apr 5 17:49:38 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Wed Apr 5 17:49:45 2006 Subject: [Histonet] procedure for calibrating the wattage on a microwave oven In-Reply-To: <8EB1459EA20E4E498AC2E05378F8F30A536EBD@mind.mc.ad.lluahsc.org> References: <8EB1459EA20E4E498AC2E05378F8F30A536EBD@mind.mc.ad.lluahsc.org> Message-ID: <44344982.4020507@ebsciences.com> Hi: First off, I'm a vendor, but here's some info: First of all, there is no way to "calibrate" either temperature or wattage on a household microwave without temperature feedback/control (i.e., temperature probe) and/or a means provided by the manufacturer to adjust/calibrate. However, one CAN do an output power test, and adjust your protocols accordingly; the link below outlines the method: http://www.ebsciences.com/papers/microwave_quality.htm Other material in our "library" section may be useful as well, plus our sheet of CAP-related recommendations has been well-received: http://www.ebsciences.com/pdf/EBS_CAP_RECOMMEND.pdf Best regards, Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson Mitchell, Nancy K. wrote: > Does anyone have a procedure that they can share for calibrating the temp/wattage on a microwave oven? > We use ours (house hold type-800 watt) for heating biological stains only-no processing. > Thank you. > Nancy Mitchell, HT ASCP > Histology Supervisor > Loma Linda Pathology Medical Group > 11370 Anderson Street Ste 2960 > Loma Linda, CA 92354 > 909-558-2012 FAX-909-796-3667 > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- From srodriguez <@t> phenopath.com Wed Apr 5 17:53:13 2006 From: srodriguez <@t> phenopath.com (Stephanie Rodriguez) Date: Wed Apr 5 17:53:34 2006 Subject: [Histonet] Re: PTH In-Reply-To: Message-ID: Hi Rich, We have been purchasing the same clone we used to get from Dako >From LabVision. (800-828-1628, labvision.com). We use citrate/heat Pretreatment, not enzyme, and Rat IgG with Vector Elite/DAB detection. Stephanie Rodriguez, BS, HTL(ASCP), QIHC Phenopath Laboratories Seattle, WA > Message: 1 > Date: Wed, 05 Apr 2006 10:27:03 -0400 > From: maramydave@aol.com > Subject: [Histonet] PTH > To: histonet@lists.utsouthwestern.edu > Message-ID: <8C826D4245C3947-1698-B98@FWM-D10.sysops.aol.com> > Content-Type: text/plain; charset="us-ascii"; format=flowed > > Rich > We had the same problem, Vector's antibody is good with PK. > Mary Helie > > > Message: 1 > Date: Tue, 04 Apr 2006 13:13:52 -0400 > From: "Richard Cartun" > Subject: [Histonet] PTH > To: > Message-ID: > Content-Type: text/plain; charset=US-ASCII > > Is anyone doing immunohistochemical staining for parathyroid hormone > (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose > antibody are you using? I just found out that DAKO no longer sells the > rat monoclonal antibody that we had been using. > > It would be nice if suppliers notified their customers before they > discontinue a product so that we can find a replacement before we run > out of reagent. > > Richard > > Richard W. Cartun, Ph.D. > Director, Immunopathology & Histology > Assistant Director, Anatomic Pathology > Hartford Hospital > 80 Seymour Street > Hartford, CT 06102 > (860) 545-1596 > (860) 545-0174 Fax > > > ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From liz <@t> premierlab.com Wed Apr 5 18:22:18 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Wed Apr 5 18:20:38 2006 Subject: [Histonet] cardio myocyte histomorpometry In-Reply-To: <200604052046.k35KkjrF007129@chip.viawest.net> Message-ID: <003001c65907$c96ffa80$0300a8c0@Chlipala> Patsy I believe most reference just measure the diameter of the myocyte through the center. A regular H&E should work. You can use an ocular micrometer or take a digital image and use a image analysis program. That's what we have done in the past. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, April 05, 2006 2:47 PM To: histonet@pathology.swmed.edu Cc: nmitchell@ihctech.net Subject: [Histonet] cardio myocyte histomorpometry We want to measure the size of cardio myocytes. The investigator heard tell of a "Silver" stain used for this but he doesn't know what silver stain. What is recommended? This will be done using a computerized image analysis system. Thanks in advance, Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________ NOD32 1.1474 (20060405) Information __________ This message was checked by NOD32 antivirus system. http://www.eset.com From jkiernan <@t> uwo.ca Wed Apr 5 23:43:56 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Wed Apr 5 23:42:58 2006 Subject: [Histonet] Sizes of cardiomyocytes Message-ID: <44349C8C.1090603@uwo.ca> Dear Patsy, Cardiac muscle fibres contain many nuclei and are branched, with partitions at the intercalated discs. Are your investigator's "cardiomyocytes" syncytia in the heart or objects in tissue culture? Does "size" mean area of a cultured cell, diameter of a sectioned fibre, or volume of a syncytial unit? You should tell the investigator to grow up, go to the library and then formulate a methodological question after reading at least 50 papers in the field. Feel free to pass on this advice. John Kiernan Anatomy, UWO London, Canada. ----------------------------------- Patsy ruegg wrote: > We want to measure the size of cardio myocytes. The investigator heard > tell of a "Silver" stain used for this but he doesn't know what silver > stain. What is recommended? This will be done using a computerized > image analysis system. Thanks in advance, Patsy > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 216 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > From Tony_Reilly <@t> health.qld.gov.au Thu Apr 6 00:52:25 2006 From: Tony_Reilly <@t> health.qld.gov.au (Anthony Reilly) Date: Thu Apr 6 00:54:11 2006 Subject: [Histonet] Mayer's hematox Message-ID: We have been using Mayer's in our lab for over 30 years. The mixture we use is similar to John Kiernan's except we use acetic acid and do not boil. A number of years ago we tried this mixture without the chloral hydrate due to the inconvenience of having to continually renew our licence to handle it.(chloral hydrate is a drug often prescribed for use as a sedative) There was no change in staining without chloral hydrate and as John states I query the 'preservative' tag. It certainly does not preserve the quality of the stain as we generally make large batches for use in the lab. We use it as a progressively for 1-2 minutes as a counterstain for IHC, PAS etc and regressively for H&E staining with a time of 4 minutes. Though as stated it can be applied for up to 10 minutes depending of differentiation used. regards Tony Reilly Chief Scientist Anatomical Pathology QHPS-Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au >>> "John A. Kiernan" 04/06/06 7:31 am >>> You should have said! Its: Haematoxylin 1G Sodium iodate 0.2G Potassium alum 50G Citric acid 1G Chloral hydrate 50G Dist. water 1000ml Allow the first 3 items to dissolve in the water overnight. Add citric acid and chloral hydrate, then boil for 5 mins. Ready for use when cool. D & W call the chloral hydrate a "preservative," which is a statement I don't understand. Preserving what? This matter has been discussed on Histonet in the past (not by me), and the assertion has been made that Mayer's haemalum works just as well without the chloral hydrate. I can't remember who said that; I've used only the complete mixture myself. It would be worth searching in www.histosearch.com -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Julien Lambrey de Souza wrote: > > Thanks, > but actually I meant I needed the recipe for making Mayer's hematox > according to Drury and Wallington (We don't have the book here and > although I ordered it, I won't have it in time for the due dates...). > > Julien. > > Le 06-04-05, ? 13:09, John A. Kiernan a ?crit : > > > From the 4th edition (1967), p.127: > > > > "Stain for 5-10 minutes." > > -- > > ------------------------------- > > John A. Kiernan > > Department of Anatomy and Cell Biology > > The University of Western Ontario > > London, Canada N6A 5C1 > > kiernan[AT]uwo.ca > > http://publish.uwo.ca/~jkiernan/ > > http://instruct.uwo.ca/anatomy/530/index.htm > > _______________________________ > > Julien Lambrey de Souza wrote: > >> > >> Hello, > >> > >> Does anyone have the protocol for Mayer's hematox from Drury and > >> Wallington's book (Carleton's histological techniques)? I need to > >> compare with other protocols I have. > >> > >> Thanks' > >> > >> Julien Lambrey de Souza > >> Research assistant, Evolutionary biology > >> University of Quebec at Rimouski > >> > >> Tel: (418) 723-1986 #1714 > >> Fax: (418) 724-1849 > >> > >> _______________________________________________ > >> Histonet mailing list > >> Histonet@lists.utsouthwestern.edu > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/ received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. **************************************************************** From L.Driessen <@t> orthop.umcn.nl Thu Apr 6 00:54:04 2006 From: L.Driessen <@t> orthop.umcn.nl (L.Driessen@orthop.umcn.nl) Date: Thu Apr 6 00:54:11 2006 Subject: [Histonet] Antibodies for human cartilage Message-ID: <5169A2936294864EACC9DD433D003E7408D479@umcnet30.umcn.nl> Hello, We're are looking for good antibodies that can be used on human cartilage. We already perform histochemic stainings like alcian blue, toluidin blue and safranin, but now we also want to do immuno on it. We already have an antibody for collagen type II that can be used, but we would like to know if there are other antobodies that can be used for human cartilage. We work on frozen sections and cell-cultures. Thank you L?on Driessen Orthopaedisch Research Lab UMC St. Radboudziekenhuis, Nijmegen 024-3615145/3614932 l.driessen@orthop.umcn.nl From mverdu <@t> histopat.es Thu Apr 6 01:02:04 2006 From: mverdu <@t> histopat.es (Montse verdu) Date: Thu Apr 6 01:02:29 2006 Subject: [Histonet] HYPOPHYSEAL HORMONES Message-ID: <001501c6593f$a7458280$3800a8c0@Histopat.com> Is anyone doing immunohistochemical staining for hypophyseal hormones > on formalin-fixed, paraffin-embedded tissue? If so, whose > antibody are you using? I just found out that DAKO no longer sells > the monoclonal antibodies that we had been using: ACTH (M350101), FSH (M350401), HGH (A057001), LH (M350201), TSH (M350301) and PRO (A056901). > > It would be nice if suppliers notified their customers before they > discontinue a product so that we can find a replacement before we run > out of reagent. Montse Verd? histopat LABORATORIS Tel. 932033000 Fax 932802160 mverdu@histopat.es From cpomajzl <@t> cpllabs.com Thu Apr 6 05:15:30 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Thu Apr 6 05:13:12 2006 Subject: [Histonet] It's no wonder... References: Message-ID: <002501c65963$2277d1b0$26fca8c0@CSP> I agree with this assessment. Healthy constructive criticism is how change is made... for the better. I welcome constructive comments that are well thought out and presented in a polite, professional manner. However, most people would rather complain than take the time and effort to provide an alternative solution. What I am referring to is the constant whining, bitching, "so and so said this" or "so and so did that" hen house cackling that goes on day after day. It seems as though a lot people cannot function without this soap opera drama. The point I am trying to make is that the focus appears to have gotten away from the patient and is more on the "what are you doing for me" from the histotechs. And yes... hospitals and lab directors are lining their pockets. They went to school and have the education that put them in that position. That is what they are getting paid to do. And you are getting paid too. If you don't think you are getting paid enough, then present the facts to your supervisor and ask for a raise. If you work hard, don't complain, and do a good job, you will most likely be rewarded for your efforts. On the other hand, if you are the proverbial thorn, don't expect much. This is Business 101. Nobody is going to look out for you but you. Just be professional. xoxoxo -Chris ----- Original Message ----- From: "Rittman, Barry R" To: ; Sent: Wednesday, April 05, 2006 12:54 PM Subject: RE: [Histonet] It's no wonder... I apologize, my wrist caught one of the computer buttons before I finished.... It is not unusual for people to vent their feeling regarding their job. A study several years ago concluded that people that complain are at least interested in their job. Apathy on the other hand gets us nowhere (as seen with our political system and lack of votes). Venting is I believe a healthy thing and prevents internal pressure building up, providing that there is a limit to venting. I think that it is difficult not to be frustrated when we see the valuable contribution that the profession makes and yet often sees little reward. We are all expected to do more work in the same time and many times the service/hospital etc. may benefit financially but this does not seem to trickle down to the professionals that are actually doping the work. Please do not think that this is unique to this profession. Just look at higher education where the student faculty ratio has been increasing for several year. I believe that we should channel this love of the profession into something positive. If we want to have better recognition, better pay and conditions it is up to us. We need to inform the general public, legislators etc. about what we do and its value to the communities we serve. So the bottom line is get it off your chest - you will feel a lot better but also get off your butt and write your comments to the above groups. Above all be have constructive suggestions. No good complaining unless you have constructive comments on how to improve the situation. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Robyn Vazquez Sent: Wednesday, April 05, 2006 11:55 AM To: cpomajzl@cpllabs.com; histonet@lists.utsouthwestern.edu; cking@rallansci.com Subject: RE: [Histonet] It's no wonder... I second it... Robyn >>> "King, Curtis - RAS" 4/5/2006 7:54 AM >>> Well said Chris. Curt -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Chris Pomajzl Sent: Wednesday, April 05, 2006 10:52 AM To: HISTONET Subject: [Histonet] It's no wonder... ...that Histology finds itself in this current state of anger, lethargy, and apathy. From the content posted over the last several weeks, it seems that a lot of people are very hypersensitive and hypercritical. I personally don't understand all of the hostility. I for one am very thankful for this profession and my position. We perform a very critical aspect of anatomic pathology, and I have learned a tremendous wealth of information since I have moved to the clinical world. I am very grateful for that. I can certainly get the fact that a lot of histotechs feel unappreciated, but let's be realistic. Most people do not know what histology is, and they do not know what we do. They do not know what to appreciate. Why can't you take personal satisfaction in hard work and a job well done? And not to mention, this is a job, it is a profession. This is not about me, it's not about you. It's about the patient! We have a responsibility to every specimen that comes through the door. And I treat every specimen as if it belongs to my wife, or my son, or a friend or family member. If you do your job well, without complaint, the accolades will come. Your supervisors will take notice, and you can be rewarded for that. I am so tired of the drama and the in-fighting within the histology community. Be thankful that you have a job. Be thankful that you have a job that pays fairly well. And for most, it did not even require a college degree. It was not long ago that I was making $7 an hour woking in veterinary medicine. I have come a long way since then, and I owe it to Histology. You make the choice to be happy, so why be miserable? I feel sorry for you. I just wish that people would stop complaining, stop being a part of the problem, and start being a part of the solution. Every day I have people come to me with a complaint. The first thing I ask them is what they propose we do about it. Invariably, they had not even considered how to improve the situation. Every one of us has a stake in this profession, and it will continue to spiral out of control until we change our mindset and adopt a new pardigm. Rant over. Thanks. I love you all. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@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 cdemarinis <@t> SARATOGACARE.ORG Thu Apr 6 06:34:14 2006 From: cdemarinis <@t> SARATOGACARE.ORG (Demarinis, Carolyn) Date: Thu Apr 6 06:35:21 2006 Subject: [Histonet] inventory Message-ID: We are a Meditech hospital and we use the Materials Management module for ordering. At the present time I keep a file of all supply orders for Anatomic Pathology on index cards. Is there software I can purchase to documents lab orders and manage inventory? From JosefaNava <@t> texashealth.org Thu Apr 6 07:21:31 2006 From: JosefaNava <@t> texashealth.org (Nava, Josefa) Date: Thu Apr 6 07:21:36 2006 Subject: [Histonet] suggestion for sections washing off Message-ID: <2C515C1049EAF5459EFD8C9B929078A419464E@phdex03.txhealth.org> Hello to Everyone, I am running my IHC using Ventana stainer . Sometimes a positive charged slide is not enough to hold the sections. Any suggestions that I can do to the positive charged slides? Will putting thin coat of Albumin to the positive charged slides do the trick? I heard that it will affect the positivity of the slide. Will it not affect the immuno staining? Any suggestion is very much appreciated. Thanks, Josie Nava 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 Barry.R.Rittman <@t> uth.tmc.edu Thu Apr 6 08:01:24 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Apr 6 08:02:16 2006 Subject: [Histonet] It's no wonder. Long reply Message-ID: I agree with many of the comments but you have to look at the big picture of what is going on in industry as a whole today. The concept of teamwork is a great one and one to which I fully subscribe. This is a symbiotic relationship, a two way street if you will in which everyone benefits. What has happened in most industries including our profession is that while "we are a team" or "we are a family" if the cutting edge phrase, the bottom line has become the major driving force with less and less regard of how we to get there. This has resulted in everyone having to do more. I am essentially a basic scientist and love teaching but as with most of my colleagues my teaching load has doubles over the past two years. I am fortunate in many ways in that my department chair has also given me some salary increase. However the student/faculty ratio is increasing in all of the institutes of higher education with little recourse for those who have an increased load. Administration in many places has increased dramatically. I believe that administration is necessary and some increase is necessitated by increasing federal and state regulations but it seems to go far beyond that. Over the past several years there has been a growing need for both parents to work and for an increasing number of single parent families. Thus priorities have changed. The need for extra income and for a desirable workplace is even more important. No one wants to go home late and do housework or to have to deal alone with a discipline problem with children. Fifty years ago work was the major force in people's lives. Today because of financial and other pressures this is no longer the case. Work may be important but it is no longer the number one priority in most people's lives. There is a life outside work. I think that most of us in this profession feel that we no longer have much input, nor do we often benefit appropriately from the current work situation. As to some previous comments that we are lucky to have a job. While we may be lucky to be employed, many of us are older than the hills and have extensive experience. The employer is in most cases also lucky to have people with our experience. It's a two way street. I am not sure that you realize the drastic shortage of professionals in Histotechnology and also of basic science teachers. I feel that histotechs while they love the service portion of their job also need to feel that they are valued. This is not necessarily in the form of huge salary increases but in the attitude of supervisors and administrators up to and including those at the top. Opportunities for training and for learning outside what in some instances is a narrow field of work is critical. It is however dangerous to generalize as some workplaces do an excellent job in this area. Let us have some input into how things are done. After all we are the troops in the trenches, while policy and other decisions are often made by generals sipping wine in a comfortable area. This is an easy task and fits into the concept of teamwork. PS I do not consider this bitching - wife told me not to use the term as it is gender specific. Barry From rjbuesa <@t> yahoo.com Thu Apr 6 08:21:49 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Apr 6 08:21:55 2006 Subject: [Histonet] procedure for calibrating the wattage on a microwave oven In-Reply-To: <8EB1459EA20E4E498AC2E05378F8F30A536EBD@mind.mc.ad.lluahsc.org> Message-ID: <20060406132149.31653.qmail@web61219.mail.yahoo.com> Nancy: I have published a paper on the subject. I am sending it to you directly. Ren? J. "Mitchell, Nancy K." wrote: Does anyone have a procedure that they can share for calibrating the temp/wattage on a microwave oven? We use ours (house hold type-800 watt) for heating biological stains only-no processing. Thank you. Nancy Mitchell, HT ASCP Histology Supervisor Loma Linda Pathology Medical Group 11370 Anderson Street Ste 2960 Loma Linda, CA 92354 909-558-2012 FAX-909-796-3667 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Apr 6 08:43:53 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Apr 6 08:43:19 2006 Subject: [Histonet] It's no wonder. Long reply Message-ID: Short reply: UK 'Techs' empathise with what you say but the only way we found 'our place in the Sun' was when the Royal Colleges brought a moratorium down on Medical Students at Uni. No medical students, no Pathologists, no-one to report cervical smears, no-one to do the grossing, etc. Suddenly Advanced Practitioners reporting cervical cytology, Transfusion Practitioners, Consultant Practitioners, etc. Nothing beats being in the right place at the right time holding the right 'ticket'. What worries me is that there will soon be a glut of Medical Students; I hope our Sun is not eclipsed. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "Following the line of least resistance makes both men and rivers crooked" This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Rittman, Barry R [mailto:Barry.R.Rittman@uth.tmc.edu] Sent: Thursday, April 06, 2006 2:01 PM To: Chris Pomajzl; HISTONET Subject: RE: [Histonet] It's no wonder. Long reply I agree with many of the comments but you have to look at the big picture of what is going on in industry as a whole today. The concept of teamwork is a great one and one to which I fully subscribe. This is a symbiotic relationship, a two way street if you will in which everyone benefits. What has happened in most industries including our profession is that while "we are a team" or "we are a family" if the cutting edge phrase, the bottom line has become the major driving force with less and less regard of how we to get there. This has resulted in everyone having to do more. I am essentially a basic scientist and love teaching but as with most of my colleagues my teaching load has doubles over the past two years. I am fortunate in many ways in that my department chair has also given me some salary increase. However the student/faculty ratio is increasing in all of the institutes of higher education with little recourse for those who have an increased load. Administration in many places has increased dramatically. I believe that administration is necessary and some increase is necessitated by increasing federal and state regulations but it seems to go far beyond that. Over the past several years there has been a growing need for both parents to work and for an increasing number of single parent families. Thus priorities have changed. The need for extra income and for a desirable workplace is even more important. No one wants to go home late and do housework or to have to deal alone with a discipline problem with children. Fifty years ago work was the major force in people's lives. Today because of financial and other pressures this is no longer the case. Work may be important but it is no longer the number one priority in most people's lives. There is a life outside work. I think that most of us in this profession feel that we no longer have much input, nor do we often benefit appropriately from the current work situation. As to some previous comments that we are lucky to have a job. While we may be lucky to be employed, many of us are older than the hills and have extensive experience. The employer is in most cases also lucky to have people with our experience. It's a two way street. I am not sure that you realize the drastic shortage of professionals in Histotechnology and also of basic science teachers. I feel that histotechs while they love the service portion of their job also need to feel that they are valued. This is not necessarily in the form of huge salary increases but in the attitude of supervisors and administrators up to and including those at the top. Opportunities for training and for learning outside what in some instances is a narrow field of work is critical. It is however dangerous to generalize as some workplaces do an excellent job in this area. Let us have some input into how things are done. After all we are the troops in the trenches, while policy and other decisions are often made by generals sipping wine in a comfortable area. This is an easy task and fits into the concept of teamwork. PS I do not consider this bitching - wife told me not to use the term as it is gender specific. Barry _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From ROrr <@t> enh.org Thu Apr 6 09:06:15 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Thu Apr 6 09:06:20 2006 Subject: [Histonet] RE: Histonet Digest, Vol 29, Issue 7 Message-ID: Annette, It would be good to have that vendor generated validation in writing. This is only my opinion, I don't really have a reference for this. On the installation, I would pick a broad sample of tests from your menu, (Maybe 10% -20% of the total?) And have them run (either you run them or get the TSR to run them :) on both instruments. so you would have at least something from your own validation process. I don't know off the top of my head if there is a specific protocol that must be adhered to, when installing new equipment. Choose antibodies with different HEIR, and Enzyme digestions, also. I would be interested to know what anyone else would do to validate in their lab, if there is a documented process. Hope this helps. Becky Orr CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 Message: 1 Date: Wed, 5 Apr 2006 07:05:58 -0400 From: "Featherstone, Annette" Subject: [Histonet] validation for CAP To: Message-ID: <9B4A77DF11463E4FB723D484214AE9BCA5515B@KALEXMB02.KaleidaHealth.org> Content-Type: text/plain; charset="iso-8859-1" We have just purchased 2 new Dako immuno stainers and I was wondering what CAP requirements are for validation. The company says that both instruments are validated against each other and therefore you can validate the antibodies on either stainer. I hope this is fact. Annette Featherstone HT/MLT MT(HEW) From gu.lang <@t> gmx.at Thu Apr 6 09:11:37 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Apr 6 09:11:44 2006 Subject: AW: [Histonet] suggestion for sections washing off In-Reply-To: <2C515C1049EAF5459EFD8C9B929078A419464E@phdex03.txhealth.org> Message-ID: <000301c65984$055d17e0$eeeea8c0@SERVER01> Josie, When we loose a section in the Ventana Benchmark it's mostly due to water under the tissue. Often the sections dry from the margins and in the middle a bit of water is trapped. The water prevents the tissue to attach the glass. Therefore I try shake off the water before. We put fatty slides in the 60 degree oven for 5 to 10 min before the run. I don't think that using an extra adhesive is a good idea, because the machine is adjusted to the superfrost slides. Perhaps it would lead to an uneven staining. Gudrun Lang Linz, Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Nava, Josefa Gesendet: Donnerstag, 06. April 2006 14:22 An: histonet@pathology.swmed.edu Betreff: [Histonet] suggestion for sections washing off Hello to Everyone, I am running my IHC using Ventana stainer . Sometimes a positive charged slide is not enough to hold the sections. Any suggestions that I can do to the positive charged slides? Will putting thin coat of Albumin to the positive charged slides do the trick? I heard that it will affect the positivity of the slide. Will it not affect the immuno staining? Any suggestion is very much appreciated. Thanks, Josie Nava 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 From jkiernan <@t> uwo.ca Thu Apr 6 10:37:46 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Thu Apr 6 10:37:48 2006 Subject: [Histonet] suggestion for sections washing off References: <2C515C1049EAF5459EFD8C9B929078A419464E@phdex03.txhealth.org> Message-ID: <443535CA.3B8C4E0E@uwo.ca> If you put a layer of protein on the slide, you'll cover up the positively charged glass surface, so you may as well use ordinary, less expensive slides. Albumen (from eggs) is a commonly used adhesive; I don't think albumin is often used for this purpose. John Kiernan Anatomy, UWO London, Canada. ----------------------- "Nava, Josefa" wrote: > > Hello to Everyone, > > I am running my IHC using Ventana stainer . Sometimes a positive > charged slide is not enough to hold the sections. Any suggestions that I > can do to the positive charged slides? > Will putting thin coat of Albumin to the positive charged slides do the > trick? I heard that it will affect the positivity of the slide. Will it > not affect the immuno staining? > Any suggestion is very much appreciated. > > > Thanks, > Josie Nava > > 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 From Janet.Bonner <@t> FLHOSP.ORG Thu Apr 6 11:27:36 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Thu Apr 6 11:28:21 2006 Subject: [Histonet] Slide identifiers References: <8C82702DF23DF9C-444-3514@mblk-d32.sysops.aol.com> Message-ID: We use the surgical number and the patient name on all slides. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of qbta@aol.com Sent: Wed 4/5/2006 4:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide identifiers Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Thu Apr 6 11:42:29 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Apr 6 11:42:33 2006 Subject: [Histonet] Slide identifiers In-Reply-To: Message-ID: <20060406164229.51947.qmail@web61218.mail.yahoo.com> We were going to do that but it was in conflict with patient confidentiality issues (anybody could see the patient's name) so we did not do it. The moment the specimen was received it was numbered and that number was used for blocks, slides, special procedures and all reports. It was unique since the year was also included as well as the type of specimen (S for surgicals, C for cytologies, L for reference specimens, A for autopsies). Ren? J. "Bonner, Janet" wrote: We use the surgical number and the patient name on all slides. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of qbta@aol.com Sent: Wed 4/5/2006 4:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide identifiers Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. From hymclab <@t> hyhc.com Thu Apr 6 12:00:42 2006 From: hymclab <@t> hyhc.com (hymclab) Date: Thu Apr 6 11:56:13 2006 Subject: [Histonet] Slide identifiers Message-ID: We just use the surgical number on slides. Dawn -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: Thursday, April 06, 2006 11:28 AM To: qbta@aol.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Slide identifiers We use the surgical number and the patient name on all slides. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of qbta@aol.com Sent: Wed 4/5/2006 4:01 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide identifiers Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. _______________________________________________ Histonet mailing list Histonet@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 bwhitaker <@t> brownpathology.com Thu Apr 6 12:02:04 2006 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Thu Apr 6 12:00:58 2006 Subject: [Histonet] Image disclaimer Message-ID: <000001c6599b$d48189e0$3601a8c0@brownpathology.net> Hi All, Is anyone out there using a disclaimer on reports with images that they would share with me? (such as: "Images in this report are for illustrative purposes only and are not meant to be diagnostic") If you use a disclaimer, who wrote it? Someone in pathology or the hospital's or lab's legal department? Thanks! Bonnie Whitaker Lab Manager Brown & Associates Medical Laboratories 8076 El Rio Houston, Texas 77054 713-741-6677 From rjbuesa <@t> yahoo.com Thu Apr 6 12:06:49 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Apr 6 12:06:53 2006 Subject: [Histonet] Image disclaimer In-Reply-To: <000001c6599b$d48189e0$3601a8c0@brownpathology.net> Message-ID: <20060406170649.50348.qmail@web61223.mail.yahoo.com> And what do you find wrong with yours? I think it serves its purpose and is short and to the point. I don't think you need another. Ren? J. Bonnie Whitaker wrote: Hi All, Is anyone out there using a disclaimer on reports with images that they would share with me? (such as: "Images in this report are for illustrative purposes only and are not meant to be diagnostic") If you use a disclaimer, who wrote it? Someone in pathology or the hospital's or lab's legal department? 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 --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From ja.mitchell <@t> hosp.wisc.edu Thu Apr 6 12:32:24 2006 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Thu Apr 6 12:31:42 2006 Subject: [Histonet] RE: Slide identifiers Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3192C1A10@UWHIS-XCHNG2.uwhis.hosp.wisc.edu> All of our slides are required per JCAHO regulations to have "2" patient identifiers. We use the assigned laboratory accession number and the patient last name. The JCAHO inspector that recently inspected my laboratory peeled off our slide labels on several cases to ensure that the slides themselves were labeled permanantly with marker/or etched with "2" patient identifiers. If you do not care to use a patient's last name as an identifier a patient's hospital medical record # or birthdate can be used along with the laboratory accession number. Using a patients last name should not be a breach of confidentiality - if HIPAA regulations are followed all and any patient identifying information is covered when taken out of the laboratory or when outside people are present in your lab. Jean Mitchell University of Wisconsin Hospital & Clinics Neuromuscular Laboratory Madison, WI -----Original Message----- From: Bonner, Janet [mailto:Janet.Bonner@FLHOSP.ORG] Sent: Thursday, April 06, 2006 11:28 AM Please help me to clarify the new regulation of JCAHO regarding the two identifiers, because I have been requested to use 2 identifiers on the slides. As I understand this subject, it will apply to the specimen collection container when it is submitted for processing, because the identification must be done in the presence of the patient. Slides identification is way pass this point! I appreciate any help. Thank you very much. Magaly Rojas,HTL(ASCP) Histopathology Manager Inova Fairfax Hospital Falls Church,Va. _______________________________________________ Histonet mailing list Histonet@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 victor <@t> pathology.washington.edu Thu Apr 6 12:31:38 2006 From: victor <@t> pathology.washington.edu (Victor Tobias) Date: Thu Apr 6 12:31:46 2006 Subject: [Histonet] Image disclaimer In-Reply-To: <20060406170649.50348.qmail@web61223.mail.yahoo.com> References: <20060406170649.50348.qmail@web61223.mail.yahoo.com> Message-ID: <4435507A.3010600@pathology.washington.edu> Has someone run into an issue by not having a disclaimer or is it just to CYA? Rene J Buesa wrote: >And what do you find wrong with yours? I think it serves its purpose and is short and to the point. I don't think you need another. > Ren? J. > >Bonnie Whitaker wrote: > Hi All, > >Is anyone out there using a disclaimer on reports with images that they >would share with me? (such as: "Images in this report are for illustrative >purposes only and are not meant to be diagnostic") If you use a disclaimer, >who wrote it? Someone in pathology or the hospital's or lab's legal >department? > >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 > > > > > >--------------------------------- >Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From Traczyk7 <@t> aol.com Thu Apr 6 12:49:55 2006 From: Traczyk7 <@t> aol.com (Traczyk7@aol.com) Date: Thu Apr 6 12:50:06 2006 Subject: [Histonet] Microwave Accessories Message-ID: <2e4.53ba60b.3166aec3@aol.com> I would love to hear from anyone who has gotten a response from Dr. Thomas about microwave accessories. I have emailed him (twice) for the contact information but have not received an answer. Perhaps as a vendor I am viewed as competition, who knows? As someone who is out and about in many facilities, I find it very useful to have as much information on where products are available as possible. I am often asked for products I don't carry or tips on technique. This list helps with many things I can't answer and if this accessory supplier is legitimate, then I would like that information for my files too. If you have the company's contact information, please respond on the list or to me directly at _Traczyk7@aol.com_ (mailto:Traczyk7@aol.com) . Thanks for your help. Dorothy Dorothy Murphy Traczyk National Sales Manager Hacker Instruments & Industries Inc. PO Box 1176 Winnsboro, SC 29180 1-800-442-2537 hackerlab@aol.com _www.hacker_ (http://www.hacker/) insruments.com From bwhitaker <@t> brownpathology.com Thu Apr 6 13:42:19 2006 From: bwhitaker <@t> brownpathology.com (Bonnie Whitaker) Date: Thu Apr 6 13:41:12 2006 Subject: [Histonet] Image disclaimer In-Reply-To: <4435507A.3010600@pathology.washington.edu> Message-ID: <001701c659a9$d5b76970$3601a8c0@brownpathology.net> We are in the preparatory stages of adding images to our reports, and had not considered a disclaimer until one of the pathologists mentioned it yesterday. It would be a CYA thing, and I'm not sure if it really IS necessary. Thanks, Bonnie Whitaker -----Original Message----- From: Victor Tobias [mailto:victor@pathology.washington.edu] Sent: Thursday, April 06, 2006 12:32 PM To: Rene J Buesa Cc: Bonnie Whitaker; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Image disclaimer Has someone run into an issue by not having a disclaimer or is it just to CYA? Rene J Buesa wrote: >And what do you find wrong with yours? I think it serves its purpose >and is short and to the point. I don't think you need another. > Ren? J. > >Bonnie Whitaker wrote: > Hi All, > >Is anyone out there using a disclaimer on reports with images that they >would share with me? (such as: "Images in this report are for >illustrative purposes only and are not meant to be diagnostic") If you >use a disclaimer, who wrote it? Someone in pathology or the hospital's >or lab's legal department? > >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 > > > > > >--------------------------------- >Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ >countries) for 2?/min or less. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. From Vickroy.Jim <@t> mhsil.com Thu Apr 6 15:19:10 2006 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Thu Apr 6 15:19:17 2006 Subject: [Histonet] P16 Clone E6H4 Message-ID: Does anyone know where we can purchase the P16 antibody (clone E6H4)? Our pathologist would like this clone only and has given us information about it. Unfortunately the company we have found that has the clone only sells it with a detection kit to use with their antibody. We would prefer an antibody we could use with our current Ventana system. Thanks James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. From JEllin <@t> yumaregional.org Thu Apr 6 15:34:35 2006 From: JEllin <@t> yumaregional.org (Jesus Ellin) Date: Thu Apr 6 15:35:24 2006 Subject: [Histonet] Image disclaimer Message-ID: Well this is an interesting topic. Why would you need a disclaimer for an image. It is there of esthetics, unless the physician is using it to demonstrate his or hers dx. Then I would say you would need to talk to your companies legal Department, but it does raise a flag to me. Jesus Ellin Yuma Regional Medical Center >>> "Victor Tobias" 04/06/06 10:31AM >>> Has someone run into an issue by not having a disclaimer or is it just to CYA? Rene J Buesa wrote: >And what do you find wrong with yours? I think it serves its purpose and is short and to the point. I don't think you need another. > Ren? J. > >Bonnie Whitaker wrote: > Hi All, > >Is anyone out there using a disclaimer on reports with images that they >would share with me? (such as: "Images in this report are for illustrative >purposes only and are not meant to be diagnostic") If you use a disclaimer, >who wrote it? Someone in pathology or the hospital's or lab's legal >department? > >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 > > > > > >--------------------------------- >Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LRaff <@t> lab.uropartners.com Thu Apr 6 15:36:11 2006 From: LRaff <@t> lab.uropartners.com (Lester Raff) Date: Thu Apr 6 15:36:14 2006 Subject: [Histonet] Technologist needed Message-ID: <5DA1CA5D0B98A84985B545A24423B822A7BB@UPLAB01.uplab.local> Hello all, Sorry to temporarily hijack the histology list, but I can't find a comparable cytology site. We are a small private histology lab outside Chicago that will be expanding into urine cytology and FISH testing. We are looking for an experienced tech, and are willing to pay for one. If any of you in the histo world have cytology friends across the hall who might be interested, please pass this on to them! Lester J. Raff, MD Medical Director UroPartners, LLC Laboratory ph: 708-486-0076 fax: 708-486-0080 Toll free: 866-818-3473 lraff@lab.uropartners.com From tpmorken <@t> labvision.com Thu Apr 6 16:13:58 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Thu Apr 6 16:14:06 2006 Subject: [Histonet] Image disclaimer Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D825@usca0082k08.labvision.apogent.com> There was a letter in the July 2005 CAP Today magazine in which the writer (commenting about an article "Digging its way in: lab digital imaging", May 2005 CAP Today) was concerned about being "second guessed" about a diagnosis if a picture representing a lesion is included on a report. Maybe that is where this idea about a disclaimer comes from. Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jesus Ellin Sent: Thursday, April 06, 2006 1:35 PM To: victor@pathology.washington.edu; rjbuesa@yahoo.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Image disclaimer Well this is an interesting topic. Why would you need a disclaimer for an image. It is there of esthetics, unless the physician is using it to demonstrate his or hers dx. Then I would say you would need to talk to your companies legal Department, but it does raise a flag to me. Jesus Ellin Yuma Regional Medical Center >>> "Victor Tobias" 04/06/06 10:31AM >>> >>> Has someone run into an issue by not having a disclaimer or is it just to CYA? Rene J Buesa wrote: >And what do you find wrong with yours? I think it serves its purpose >and is short and to the point. I don't think you need another. > Ren? J. > >Bonnie Whitaker wrote: > Hi All, > >Is anyone out there using a disclaimer on reports with images that they >would share with me? (such as: "Images in this report are for >illustrative purposes only and are not meant to be diagnostic") If you >use a disclaimer, who wrote it? Someone in pathology or the hospital's >or lab's legal department? > >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 > > > > > >--------------------------------- >Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ >countries) for 2?/min or less. >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > -- Victor Tobias Clinical Applications Analyst University of Washington Medical Center Dept of Pathology Room BB220 1959 NE Pacific Seattle, WA 98195 victor@pathology.washington.edu 206-598-2792 206-598-7659 Fax ================================================= Privileged, confidential or patient identifiable information may be contained in this message. This information is meant only for the use of the intended recipients. If you are not the intended recipient, or if the message has been addressed to you in error, do not read, disclose, reproduce, distribute, disseminate or otherwise use this transmission. Instead, please notify the sender by reply e-mail, and then destroy all copies of the message and any attachments. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Thu Apr 6 17:52:31 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Apr 6 17:52:38 2006 Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Message-ID: Including hydrochloric acid in your staining solution might help (0.25ml per 100ml). Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: Guillermo Palao [mailto:gpbnas@yahoo.es] Sent: Thursday, 6 April 2006 1:51 AM To: Tony Henwood; Histonet Subject: RE: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Thanks, you advice going straight from sirius red to 70% ethanol or maybe to 100% as suggested in other reply, but how do I get rid of the excess red stuff on the slide? Best regards, Guillermo Tony Henwood escribi?: Drop step 3 and 4 (picric acid is soluble in water) Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.eddu] On Behalf Of Guillermo Palao Sent: Wednesday, 5 April 2006 4:59 AM To: Histonet Subject: [Histonet] Sirius red with low contrast between cells cytoplasm andstained fibers Hello all: I would really appreciate any suggestions to improve sirius red staining of kidney samples. Although collagen fibers in tissue nicely stain red, cells cytoplasm do not stain yellow, but sort of pinkish, making it very difficult to separate from red-stained extracellular fibers for quantification purposes. My protocol is as follows: 1. Deparafinize and hydrate samples 2. Stain 1 h in 0.1% Sirius Red in saturated aqueous picric acid 3. Wash 2 min 0.01 N HCl 4. Rinse in water 5. Dehydrate and cover slides I have tried shorter staining times and rinsing with water before HCl washing without improving results. Any help would be greatly appreciated. Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** ________________________________ LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From AnthonyH <@t> chw.edu.au Thu Apr 6 19:01:15 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Thu Apr 6 19:01:35 2006 Subject: [Histonet] Mayer's hematox Message-ID: We have used Garvies with good results: Garvey's Modified Mayer's Haematoxylin Principle: This solution is a progressive haematoxylin that uses 10% less alum. Chloral hydrate, because of its toxicity, has been replaced with ethanol, a more effective penetrator and bacteriostat. Mercury oxide has been replaced with sodium iodate. Fixation: 10% buffered formalin. Microtomy: paraffin sections at 5um. Reagents: Dissolve 45g ammonium or potassium alum in 900ml distilled water with the aid of heat. Dissolve 2.5g Haematoxylin (CI 75290) in 100ml absolute ethanol. Combine above solutions and add 0.2g Sodium Iodate and 1g citric acid. Mix well. The solution is stable for several months. Procedure: 1. Dewax and hydrate sections. 2. Stain in modified Mayer's Haematoxylin 40secs. 3. Wash in warm water 5min. 4. If required, counterstain in Eosin solution. 5. Dehydrate, clear and mount. Reference: Garvie, W., (1991) J. Histotechn 14(3):164-165. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anthony Reilly Sent: Thursday, 6 April 2006 3:52 PM To: histonet@lists.utsouthwestern.edu; julien_lambreydesouza@UQAR.QC.CA; jkiernan@uwo.ca Subject: Re: [Histonet] Mayer's hematox We have been using Mayer's in our lab for over 30 years. The mixture we use is similar to John Kiernan's except we use acetic acid and do not boil. A number of years ago we tried this mixture without the chloral hydrate due to the inconvenience of having to continually renew our licence to handle it.(chloral hydrate is a drug often prescribed for use as a sedative) There was no change in staining without chloral hydrate and as John states I query the 'preservative' tag. It certainly does not preserve the quality of the stain as we generally make large batches for use in the lab. We use it as a progressively for 1-2 minutes as a counterstain for IHC, PAS etc and regressively for H&E staining with a time of 4 minutes. Though as stated it can be applied for up to 10 minutes depending of differentiation used. regards Tony Reilly Chief Scientist Anatomical Pathology QHPS-Prince Charles Hospital Rode rd Chermside Q 4032 Australia Ph: 07 3350 8543 Fax: 07 33508546 tony_reilly@health.qld.gov.au >>> "John A. Kiernan" 04/06/06 7:31 am >>> You should have said! Its: Haematoxylin 1G Sodium iodate 0.2G Potassium alum 50G Citric acid 1G Chloral hydrate 50G Dist. water 1000ml Allow the first 3 items to dissolve in the water overnight. Add citric acid and chloral hydrate, then boil for 5 mins. Ready for use when cool. D & W call the chloral hydrate a "preservative," which is a statement I don't understand. Preserving what? This matter has been discussed on Histonet in the past (not by me), and the assertion has been made that Mayer's haemalum works just as well without the chloral hydrate. I can't remember who said that; I've used only the complete mixture myself. It would be worth searching in www.histosearch.com -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Julien Lambrey de Souza wrote: > > Thanks, > but actually I meant I needed the recipe for making Mayer's hematox > according to Drury and Wallington (We don't have the book here and > although I ordered it, I won't have it in time for the due dates...). > > Julien. > > Le 06-04-05, ? 13:09, John A. Kiernan a ?crit : > > > From the 4th edition (1967), p.127: > > > > "Stain for 5-10 minutes." > > -- > > ------------------------------- > > John A. Kiernan > > Department of Anatomy and Cell Biology > > The University of Western Ontario > > London, Canada N6A 5C1 > > kiernan[AT]uwo.ca > > http://publish.uwo.ca/~jkiernan/ > > http://instruct.uwo.ca/anatomy/530/index.htm > > _______________________________ > > Julien Lambrey de Souza wrote: > >> > >> Hello, > >> > >> Does anyone have the protocol for Mayer's hematox from Drury and > >> Wallington's book (Carleton's histological techniques)? I need to > >> compare with other protocols I have. > >> > >> Thanks' > >> > >> Julien Lambrey de Souza > >> Research assistant, Evolutionary biology > >> University of Quebec at Rimouski > >> > >> Tel: (418) 723-1986 #1714 > >> Fax: (418) 724-1849 > >> > >> _______________________________________________ > >> Histonet mailing list > >> Histonet@lists.utsouthwestern.edu > >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***************************************************************** This email, including any attachments sent with it, is confidential and for the sole use of the intended recipient(s). This confidentiality is not waived or lost, if you receive it and you are not the intended recipient(s), or if it is transmitted/ received in error. Any unauthorised use, alteration, disclosure, distribution or review of this email is strictly prohibited. The information contained in this email, including any attachment sent with it, may be subject to a statutory duty of confidentiality if it relates to health service matters. If you are not the intended recipient(s), or if you have received this email in error, you are asked to immediately notify the sender by telephone collect on Australia +61 1800 198 175 or by return email. You should also delete this email, and any copies, from your computer system network and destroy any hard copies produced. If not an intended recipient of this email, you must not copy, distribute or take any action(s) that relies on it; any form of disclosure, modification, distribution and/or publication of this email is also prohibited. Although Queensland Health takes all reasonable steps to ensure this email does not contain malicious software, Queensland Health does not accept responsibility for the consequences if any person's computer inadvertently suffers any disruption to services, loss of information, harm or is infected with a virus, other malicious computer programme or code that may occur as a consequence of receiving this email. Unless stated otherwise, this email represents only the views of the sender and not the views of the Queensland Government. **************************************************************** _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From cbass <@t> bidmc.harvard.edu Fri Apr 7 00:19:36 2006 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Apr 7 00:19:42 2006 Subject: [Histonet] liver damage detection Message-ID: <7A675819-A8A7-4E80-BD0C-76202BE80378@bidmc.harvard.edu> Dear Histonetters, I am working on an experimental model of viral liver damage. Does anyone have suggestions as to how I should best proceed in establishing the type or extent of damage caused? I don't expect damage to the extent that the mice die or even show any overt signs of illness. There is no noticeable gross change in the liver. Ideally I would like to establish a biochemical marker of damage such as serum ALT as well as a morphological analysis. I figure an H&E stain would be enough. I would like to use NBF fixed tissue. Are there any suggestions as to the thickness of the liver sections? I can reliably section 30 um, anything thinner would require special arrangements. Any suggestions for a kit to measure ALT? A colleague suggested that I submit the samples to the hospital's diagnostic lab, but would they be able to detect mouse ALT? Any suggestions would be appreciated. I have never worked with liver damage, so I really don't know what to do here. Thanks in advance, Caroline From malek.j <@t> ghc.org Fri Apr 7 07:10:27 2006 From: malek.j <@t> ghc.org (Malek, Jack M) Date: Fri Apr 7 07:10:38 2006 Subject: [Histonet] Xylene Resistant Slide Labels Message-ID: <4B2B5C876118AC47998E76D07206505D0829B8@ex05.GHCMASTER.GHC.ORG> Can anyone recommend a smudge proof xylene resistant slide label? We're preparing to go-live with a new computer system and are printing barcodes on slides. It seems that they all smudge a little, some worse than others. Please reply by email or phone... Jack 206-326-3251 From akbitting <@t> geisinger.edu Fri Apr 7 09:07:29 2006 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Fri Apr 7 09:08:10 2006 Subject: [Histonet] Cyclin D1 Message-ID: Is anyone using BioCare's Cyclin D1 or Labvision's Cyclin D1,clone SP4 on the Ventana XT? Would you mind sharing your protocols with me? IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From Traczyk7 <@t> aol.com Fri Apr 7 09:15:27 2006 From: Traczyk7 <@t> aol.com (Traczyk7@aol.com) Date: Fri Apr 7 09:15:36 2006 Subject: [Histonet] Re: Histonet "Solicitation" Message-ID: <36d.155cbbe.3167cdff@aol.com> Thanks to everyone who sent me the contact information. Knowing what I do about Coleman Manufacturing makes me wonder who "Dr. Thomas" is. Are any of the histologists on this list familiar with this guy or where he works? I'd like to find out which microwave system his lab works with and how substitute accessories are working out over the long run. As a mw vendor, I know the time that may need to be invested in testing products. It can sometimes be weeks before a problem presents itself. Thanks again. Dorothy Murphy Traczyk Hacker Instruments & Industries Inc. From la.sebree <@t> hosp.wisc.edu Fri Apr 7 09:20:05 2006 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Fri Apr 7 09:19:24 2006 Subject: [Histonet] Cyclin D1 Message-ID: We've used both Angela but right now are using LabVision's at 1:100 for 32" at 37 degrees C, mCC2, amplification, AB block and HemII for 8". Works great for us even on bm bxs. Linda Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-3224 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Angela Bitting Sent: Friday, April 07, 2006 9:07 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Cyclin D1 Is anyone using BioCare's Cyclin D1 or Labvision's Cyclin D1,clone SP4 on the Ventana XT? Would you mind sharing your protocols with me? IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stefanie.werner <@t> gmx.de Fri Apr 7 09:30:26 2006 From: stefanie.werner <@t> gmx.de (Stefanie Werner) Date: Fri Apr 7 09:30:32 2006 Subject: [Histonet] toluidine blue staining Message-ID: <31129.1144420226@www052.gmx.net> Hello, i have a problem with my toluidine blue staining after radioactive in situ hybridization. After developing the slides i stain them with 0,2% toluidine blue. After staining I often see under the microscope a red background colour on the slide and in the tissue. Therefore I can't see my radioactive signal properly. Did anybody have the same problem and can help me? I don't know what to do, because it just appears sometimes, even when the procedure is the same. Thanks. Steffi -- Analog-/ISDN-Nutzer sparen mit GMX SmartSurfer bis zu 70%! Kostenlos downloaden: http://www.gmx.net/de/go/smartsurfer From ploykasek <@t> phenopath.com Fri Apr 7 10:19:31 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Apr 7 10:19:56 2006 Subject: [Histonet] P16 Clone E6H4 In-Reply-To: Message-ID: James, Dako sells that clone of antibody. Don't know about it's performance on the Ventana. Patti Loykasek > > > Does anyone know where we can purchase the P16 antibody (clone E6H4)? > > Our pathologist would like this clone only and has given us information > about it. Unfortunately the company we have found that has the clone > only sells it with a detection kit to use with their antibody. We would > prefer an antibody we could use with our current Ventana system. Thanks > > > > James R. Vickroy > > Supervisor - MMC Surgical Pathology > > 217-788-4046 > > > > > > This message (including any attachments) contains confidential information > intended for a > specific individual and purpose, and is protected by law. If you are not the > intended recipient, > you should delete this message. Any disclosure, copying, or distribution of > this message, or the > taking of any action based on it, is strictly prohibited. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From bharmala <@t> ucalgary.ca Fri Apr 7 10:39:14 2006 From: bharmala <@t> ucalgary.ca (Aamir Bharmal) Date: Fri Apr 7 10:39:22 2006 Subject: [Histonet] A quick note Message-ID: <1619.136.159.235.215.1144424354.squirrel@136.159.235.215> Sorry, I just noticed that I should not be including any photographs or attachments on sent e-mails. I have posted a copy of the images (within the Word document) entitled "Bonus Assignment 2006.doc" to www.histonet.org. Thanks again, Aamir Bharmal University of Calgary From bharmala <@t> ucalgary.ca Fri Apr 7 10:48:54 2006 From: bharmala <@t> ucalgary.ca (Aamir Bharmal) Date: Fri Apr 7 10:48:59 2006 Subject: [Histonet] Bonus Assignment Message-ID: <1726.136.159.235.215.1144424934.squirrel@136.159.235.215> Hello, I am an undergraduate student who has been given a bonus assignment for my pathology course. It is to identify what is in the following photographs. Just to let you know, I have been given permission to use any resource for identification. I have uploaded three photographs to www.histonet.org to a Microsoft Word document entitled "Bonus Assignment 2006.doc." Can someone help me identify what is being shown in the pictures? Thank you in advance for your help. Aamir Bharmal University of Calgary From rjbuesa <@t> yahoo.com Fri Apr 7 11:03:05 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Apr 7 11:03:09 2006 Subject: [Histonet] liver damage detection In-Reply-To: <7A675819-A8A7-4E80-BD0C-76202BE80378@bidmc.harvard.edu> Message-ID: <20060407160305.99222.qmail@web61219.mail.yahoo.com> Caroline: Just some points: 1- first you need to be very familiar and knowledgeable of mice liver normal histology; 2- I do not understand why you do think that H&E will be sufficient vwithout any previous idea of the type of damage you could expect to find. It is very likely that H&E will not be sufficient after all; 3- A 30 ?m section will be probably useless to detect perhaps subtle chages; you have to do those "special arrangements" and get thinner sections. 4- Finally, where is this virus supposed to "attack"? Cytoplasm, nuclei? Depending on where it is supposed to act is where you should try to look for changes and it will indicate the type of stain you will need. Just some thought that I hope will help you! Ren? J. Caroline Bass wrote: Dear Histonetters, I am working on an experimental model of viral liver damage. Does anyone have suggestions as to how I should best proceed in establishing the type or extent of damage caused? I don't expect damage to the extent that the mice die or even show any overt signs of illness. There is no noticeable gross change in the liver. Ideally I would like to establish a biochemical marker of damage such as serum ALT as well as a morphological analysis. I figure an H&E stain would be enough. I would like to use NBF fixed tissue. Are there any suggestions as to the thickness of the liver sections? I can reliably section 30 um, anything thinner would require special arrangements. Any suggestions for a kit to measure ALT? A colleague suggested that I submit the samples to the hospital's diagnostic lab, but would they be able to detect mouse ALT? Any suggestions would be appreciated. I have never worked with liver damage, so I really don't know what to do here. Thanks in advance, Caroline _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- How low will we go? Check out Yahoo! Messenger?s low PC-to-Phone call rates. From Bauer.Karen <@t> mayo.edu Fri Apr 7 11:57:23 2006 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Apr 7 11:57:33 2006 Subject: [Histonet] IHC Question... Message-ID: Hi to all. I recently stained a lymph node case and it has brought up an interesting question that I can't explain to our Pathologist. I'm hoping that someone out there can give me some suggestions. First of all, a little background... The lymph node was processed, by accident, on a "fast run" on our VIP Tissue Tek. The processing run was complete in about 3 and a half hours. (We thought there were just small biopsies on the run, but the lymph node cassette was probably thrown in the wrong basket by mistake.) Anyway, the lymph node tissue didn't turn out all that bad, but the fatty tissue did not process adequately (no surprise there!!). Since the lymph node tissue cut fairly well, the block was not run back and re-processed. H&E's were turned in and stains were ordered. Here's my problem: The CD3 and CD20 stained well and the lymph node tissue and controls looked as they should, but the LCA on the patient tissue did not stain at all. We put the patient tissue on a control slide, so the control and tissue are stained the same all the way through the run. The LCA control looked beautiful, but the lymph node tissue had no staining. How can you have positive staining for CD3 and CD20, but no staining for LCA on a lymph node? Since we use a tonsil for our control, we re-ran the LCA with the tonsil control, a different lymph node case and then the same lymph node case that we were staining. Again, the control looked great, but the lymph node did not stain again. The different lymph node case that was also run turned out just fine, so we knew it was tissue specific. We thought maybe this lack of staining was due to the insufficient processing, but then why did the CD3 and CD20 turn out so well? Shouldn't they have no staining also? I'm perplexed and was wondering if anyone else out there has had this happen to them. As always, thank you. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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 Heather.A.Harper <@t> pcola.med.navy.mil Fri Apr 7 11:58:26 2006 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Fri Apr 7 12:01:29 2006 Subject: [Histonet] Intestinal Polyps Message-ID: We do a lot of intestinal polyps and all of a sudden, there is chatter in the section. We are not doing anything different. I need to trouble shoot, so please give input. Could it be sitting in formalin too long? The only thing that did change is that Endoscopy would submit the polyps in Ultrum, a non-formadelhyde fixative. Now they have started submitting the tissue in formalin. This is the only thing that has changed. Our processing, cutting, staining is the same. Any input would be gladly appreciated. Heather A. Harper Supervisor of Histology Naval Hospital of Pensacola, FL From Bauer.Karen <@t> mayo.edu Fri Apr 7 12:01:07 2006 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Apr 7 12:02:54 2006 Subject: [Histonet] IHC Question... References: Message-ID: Oops! I forgot to mention that we use a BenchMark from Ventana for our IHC staining and we use prediluted antibodies. Karen ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bauer, Karen Sent: Fri 4/7/2006 11:57 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Question... Hi to all. I recently stained a lymph node case and it has brought up an interesting question that I can't explain to our Pathologist. I'm hoping that someone out there can give me some suggestions. First of all, a little background... The lymph node was processed, by accident, on a "fast run" on our VIP Tissue Tek. The processing run was complete in about 3 and a half hours. (We thought there were just small biopsies on the run, but the lymph node cassette was probably thrown in the wrong basket by mistake.) Anyway, the lymph node tissue didn't turn out all that bad, but the fatty tissue did not process adequately (no surprise there!!). Since the lymph node tissue cut fairly well, the block was not run back and re-processed. H&E's were turned in and stains were ordered. Here's my problem: The CD3 and CD20 stained well and the lymph node tissue and controls looked as they should, but the LCA on the patient tissue did not stain at all. We put the patient tissue on a control slide, so the control and tissue are stained the same all the way through the run. The LCA control looked beautiful, but the lymph node tissue had no staining. How can you have positive staining for CD3 and CD20, but no staining for LCA on a lymph node? Since we use a tonsil for our control, we re-ran the LCA with the tonsil control, a different lymph node case and then the same lymph node case that we were staining. Again, the control looked great, but the lymph node did not stain again. The different lymph node case that was also run turned out just fine, so we knew it was tissue specific. We thought maybe this lack of staining was due to the insufficient processing, but then why did the CD3 and CD20 turn out so well? Shouldn't they have no staining also? I'm perplexed and was wondering if anyone else out there has had this happen to them. As always, thank you. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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 ********************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 settembr <@t> umdnj.edu Fri Apr 7 12:04:50 2006 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Fri Apr 7 12:05:36 2006 Subject: [Histonet] IHC Question... Message-ID: How do you run IHC? By hand? Shandon coverplates? automation? Dana Settembre University Hospital-UMDNJ Newark, NJ >>> "Bauer, Karen" 04/07/06 12:57 PM >>> Hi to all. I recently stained a lymph node case and it has brought up an interesting question that I can't explain to our Pathologist. I'm hoping that someone out there can give me some suggestions. First of all, a little background... The lymph node was processed, by accident, on a "fast run" on our VIP Tissue Tek. The processing run was complete in about 3 and a half hours. (We thought there were just small biopsies on the run, but the lymph node cassette was probably thrown in the wrong basket by mistake.) Anyway, the lymph node tissue didn't turn out all that bad, but the fatty tissue did not process adequately (no surprise there!!). Since the lymph node tissue cut fairly well, the block was not run back and re-processed. H&E's were turned in and stains were ordered. Here's my problem: The CD3 and CD20 stained well and the lymph node tissue and controls looked as they should, but the LCA on the patient tissue did not stain at all. We put the patient tissue on a control slide, so the control and tissue are stained the same all the way through the run. The LCA control looked beautiful, but the lymph node tissue had no staining. How can you have positive staining for CD3 and CD20, but no staining for LCA on a lymph node? Since we use a tonsil for our control, we re-ran the LCA with the tonsil control, a different lymph node case and then the same lymph node case that we were staining. Again, the control looked great, but the lymph node did not stain again. The different lymph node case that was also run turned out just fine, so we knew it was tissue specific. We thought maybe this lack of staining was due to the insufficient processing, but then why did the CD3 and CD20 turn out so well? Shouldn't they have no staining also? I'm perplexed and was wondering if anyone else out there has had this happen to them. As always, thank you. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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 cpomajzl <@t> cpllabs.com Fri Apr 7 12:16:06 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Fri Apr 7 12:12:55 2006 Subject: [Histonet] Bonus Assignment References: <1726.136.159.235.215.1144424934.squirrel@136.159.235.215> Message-ID: <008e01c65a66$f5083e40$26fca8c0@CSP> If you're just looking for the answers, well that's not really doing any work - is it? I haven't seen the images, but my suggestion would be to do some of the research on your own. If you ask questions that show that you've at least done some investigating, I would be more inclined to help you out. I would also like to make sure that nobody just gives Aamir any answers. We would be doing a disservice to his education and our profession. Thanks. ----- Original Message ----- From: "Aamir Bharmal" To: Sent: Friday, April 07, 2006 10:48 AM Subject: [Histonet] Bonus Assignment Hello, I am an undergraduate student who has been given a bonus assignment for my pathology course. It is to identify what is in the following photographs. Just to let you know, I have been given permission to use any resource for identification. I have uploaded three photographs to www.histonet.org to a Microsoft Word document entitled "Bonus Assignment 2006.doc." Can someone help me identify what is being shown in the pictures? Thank you in advance for your help. Aamir Bharmal University of Calgary _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bharmala <@t> ucalgary.ca Fri Apr 7 12:36:56 2006 From: bharmala <@t> ucalgary.ca (Aamir Bharmal) Date: Fri Apr 7 12:37:39 2006 Subject: [Histonet] Bonus Assignment In-Reply-To: <008e01c65a66$f5083e40$26fca8c0@CSP> References: <1726.136.159.235.215.1144424934.squirrel@136.159.235.215> <008e01c65a66$f5083e40$26fca8c0@CSP> Message-ID: <1275.136.159.239.166.1144431416.squirrel@136.159.239.166> Hi Chris, I do understand your concern. I posted the photos in hopes of identifying what is actually going on, however, my job is then to write up a discussion of the pathophysiology of the given condition. I have only been provided with images (no supplementary information whatsover) and have thus been given no means of actually identifying what the images show. Thus, using the resources available to me, an online group of professionals, I hope to successfully identify what the specific condition in the images actually is. It is only once the condition is identified that I can actually discuss the pathophysiology and how it is associated with human disease. Given that I have had no lab training in the area, I personally felt this online group was a very helpful resource for identification and would thus give me the tools to continue my own individual research. Thanks, Aamir Bharmal > If you're just looking for the answers, well that's not really doing any > work - is it? > > I haven't seen the images, but my suggestion would be to do some of the > research on your own. If you ask questions that show that you've at least > done some investigating, I would be more inclined to help you out. > > I would also like to make sure that nobody just gives Aamir any answers. > We > would be doing a disservice to his education and our profession. > > Thanks. > > ----- Original Message ----- > From: "Aamir Bharmal" > To: > Sent: Friday, April 07, 2006 10:48 AM > Subject: [Histonet] Bonus Assignment > > > Hello, > > I am an undergraduate student who has been given a bonus assignment for > my > pathology course. It is to identify what is in the following photographs. > Just to let you know, I have been given permission to use any resource > for > identification. > > I have uploaded three photographs to www.histonet.org to a Microsoft Word > document entitled "Bonus Assignment 2006.doc." Can someone help me > identify what is being shown in the pictures? > > Thank you in advance for your help. > > Aamir Bharmal > University of Calgary > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From ROrr <@t> enh.org Fri Apr 7 13:19:57 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Apr 7 13:20:08 2006 Subject: [Histonet] IHC question Message-ID: Hi Karen, My experienced guess would be 2 things. I am suggesting this based on my experiences in the field troubleshooting just this type of issue, so I'm not really an expert. But, I would consider these things: 1) The CD 3 and CD 20 epitope sites may not need as much formalin fixation to stain appropriately. There may be some "wiggle room" in the titer of these antibodies. What is your pretreatment for all 3? Are they all the same? In my lab I run concentrates and my CD3 works best with Tris HIER and the CD 20 works best with citrate HIER. Back in the day, LCA didn't even NEED HEIR! Or: 2)You may have simply over done the HIER and caused a prozone effect. Because the LCA control worked that shows me that the Antibody itself is working appropriately. These immunogens are developed separately so even though LCA will stain the same sites as CD3 and CD20, they aren't "the same". This may not be a fact but comes more from my experience. I'm sure you could make the LCA work if you compensated for the variable that changed, in this case the formalin/processing time. Either lighten up on the hier or if you can, extend the incubation time. Or Change the concentration. Sometimes errors are made and you have make a change to the standardized protocol to compensate. Hope this helps Becky Orr CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- Message: 23 Date: Fri, 7 Apr 2006 11:57:23 -0500 From: "Bauer, Karen" Subject: [Histonet] IHC Question... To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi to all. I recently stained a lymph node case and it has brought up an interesting question that I can't explain to our Pathologist. I'm hoping that someone out there can give me some suggestions. First of all, a little background... The lymph node was processed, by accident, on a "fast run" on our VIP Tissue Tek. The processing run was complete in about 3 and a half hours. (We thought there were just small biopsies on the run, but the lymph node cassette was probably thrown in the wrong basket by mistake.) Anyway, the lymph node tissue didn't turn out all that bad, but the fatty tissue did not process adequately (no surprise there!!). Since the lymph node tissue cut fairly well, the block was not run back and re-processed. H&E's were turned in and stains were ordered. Here's my problem: The CD3 and CD20 stained well and the lymph node tissue and controls looked as they should, but the LCA on the patient tissue did not stain at all. We put the patient tissue on a control slide, so the control and tissue are stained the same all the way through the run. The LCA control looked beautiful, but the lymph node tissue had no staining. How can you have positive staining for CD3 and CD20, but no staining for LCA on a lymph node? Since we use a tonsil for our control, we re-ran the LCA with the tonsil control, a different lymph node case and then the same lymph node case that we were staining. Again, the control looked great, but the lymph node did not stain again. The different lymph node case that was also run turned out just fine, so we knew it was tissue specific. We thought maybe this lack of staining was due to the insufficient processing, but then why did the CD3 and CD20 turn out so well? Shouldn't they have no staining also? I'm perplexed and was wondering if anyone else out there has had this happen to them. As always, thank you. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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 End of Histonet Digest, Vol 29, Issue 11 **************************************** From rjbuesa <@t> yahoo.com Fri Apr 7 13:21:34 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Apr 7 13:21:39 2006 Subject: [Histonet] Bonus Assignment In-Reply-To: <1275.136.159.239.166.1144431416.squirrel@136.159.239.166> Message-ID: <20060407182134.86611.qmail@web61222.mail.yahoo.com> Aamir: You are about to be barbequed! The least that you can do is to, along with the images, provide YOUR interpretation. Don't fear saying something foolish because probably only few people will have an idea of what the images are. Come with something and ask the people who knows to add or express their ideas. Ren? J. Aamir Bharmal wrote: Hi Chris, I do understand your concern. I posted the photos in hopes of identifying what is actually going on, however, my job is then to write up a discussion of the pathophysiology of the given condition. I have only been provided with images (no supplementary information whatsover) and have thus been given no means of actually identifying what the images show. Thus, using the resources available to me, an online group of professionals, I hope to successfully identify what the specific condition in the images actually is. It is only once the condition is identified that I can actually discuss the pathophysiology and how it is associated with human disease. Given that I have had no lab training in the area, I personally felt this online group was a very helpful resource for identification and would thus give me the tools to continue my own individual research. Thanks, Aamir Bharmal > If you're just looking for the answers, well that's not really doing any > work - is it? > > I haven't seen the images, but my suggestion would be to do some of the > research on your own. If you ask questions that show that you've at least > done some investigating, I would be more inclined to help you out. > > I would also like to make sure that nobody just gives Aamir any answers. > We > would be doing a disservice to his education and our profession. > > Thanks. > > ----- Original Message ----- > From: "Aamir Bharmal" > To: > Sent: Friday, April 07, 2006 10:48 AM > Subject: [Histonet] Bonus Assignment > > > Hello, > > I am an undergraduate student who has been given a bonus assignment for > my > pathology course. It is to identify what is in the following photographs. > Just to let you know, I have been given permission to use any resource > for > identification. > > I have uploaded three photographs to www.histonet.org to a Microsoft Word > document entitled "Bonus Assignment 2006.doc." Can someone help me > identify what is being shown in the pictures? > > Thank you in advance for your help. > > Aamir Bharmal > University of Calgary > > > > _______________________________________________ > Histonet mailing list > Histonet@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 --------------------------------- Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1¢/min. From cpomajzl <@t> cpllabs.com Fri Apr 7 13:31:48 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Fri Apr 7 13:28:40 2006 Subject: [Histonet] Bonus Assignment References: <20060407182134.86611.qmail@web61222.mail.yahoo.com> Message-ID: <001101c65a71$88918400$26fca8c0@CSP> Mmmm... ...barbeque. ; ) "Momma's got the cornbread, Daddy's got the sauce." ----- Original Message ----- From: Rene J Buesa To: Aamir Bharmal ; Chris Pomajzl Cc: HISTONET Sent: Friday, April 07, 2006 1:21 PM Subject: Re: [Histonet] Bonus Assignment Aamir: You are about to be barbequed! The least that you can do is to, along with the images, provide YOUR interpretation. Don't fear saying something foolish because probably only few people will have an idea of what the images are. Come with something and ask the people who knows to add or express their ideas. Ren? J. Aamir Bharmal wrote: Hi Chris, I do understand your concern. I posted the photos in hopes of identifying what is actually going on, however, my job is then to write up a discussion of the pathophysiology of the given condition. I have only been provided with images (no supplementary information whatsover) and have thus been given no means of actually identifying what the images show. Thus, using the resources available to me, an online group of professionals, I hope to successfully identify what the specific condition in the images actually is. It is only once the condition is identified that I can actually discuss the pathophysiology and how it is associated with human disease. Given that I have had no lab training in the area, I personally felt this online group was a very helpful resource for identification and would thus give me the tools to continue my own individual research. Thanks, Aamir Bharmal > If you're just looking for the answers, well that's not really doing any > work - is it? > > I haven't seen the images, but my suggestion would be to do some of the > research on your own. If you ask questions that show that you've at least > done some investigating, I would be more inclined to help you out. > > I would also like to make sure that nobody just gives Aamir any answers. > We > would be doing a disservice to his education and our profession. > > Thanks. > > ----- Original Message ----- > From: "Aamir Bharmal" > To: > Sent: Friday, April 07, 2006 10:48 AM > Subject: [Histonet] Bonus Assignment > > > Hello, > > I am an undergraduate student who has been given a bonus assignment for > my > pathology course. It is to identify what is in the following photographs. > Just to let you know, I have been given permission to use any resource > for > identification. > > I have uploaded three photographs to www.histonet.org to a Microsoft Word > document entitled "Bonus Assignment 2006.doc." Can someone help me > identify what is being shown in the pictures? > > Thank you in advance for your help. > > Aamir Bharmal > University of Calgary > > > > _______________________________________________ > Histonet mailing list > Histonet@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 ------------------------------------------------------------------------------ Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1?/min. From Barb.Richmond <@t> mckennan.org Fri Apr 7 13:12:10 2006 From: Barb.Richmond <@t> mckennan.org (Barb Richmond) Date: Fri Apr 7 13:29:26 2006 Subject: [Histonet] ANP.28860 Message-ID: New CAP question: Are all containers used in the micorwave devices made from microwave-transparent material. I'm having trouble finding transparent plastic coplin jars. We have pyrex, but they break in the microwave. Any suggestions where we can purchase transparent plastic coplin jars?? ----------------------------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From gcallis <@t> montana.edu Fri Apr 7 13:54:52 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Apr 7 13:55:01 2006 Subject: [Histonet] ANP.28860 In-Reply-To: References: Message-ID: <6.0.0.22.1.20060407123732.01b8be28@gemini.msu.montana.edu> Evergreen Scientific 800-421-6261. They have two types - Staining dish made of TPX with a polypropylene 20-slide carrier, w/two lids - #258-4070-000 these are microwave compatible up to 5 minutes at a time. Hellendahl type (Coplin style, only square sided for 8 or more slides) made of TPX, #258-4088-000 these are microwave compatible up to 5 minutes at a time. Sometimes you can't find these on their website, but they are very nice people with very reasonable pricing and still have all items in stock. Be sure to ask for a hard copy catalog! Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From pmcardle <@t> ebsciences.com Fri Apr 7 13:59:40 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Fri Apr 7 13:59:54 2006 Subject: [Histonet] ANP.28860 In-Reply-To: References: Message-ID: <4436B69C.7060608@ebsciences.com> Hello: If you'll forgive a "vendor" response, we (Energy Beam Sciences) sell sets of microwave transparent Coplin jars: http://www.ebsstore.com/control/crosssell/~category_id=C/~product_id=H2504 Best regards, Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson Barb Richmond wrote: > New CAP question: Are all containers used in the micorwave devices made from microwave-transparent material. I'm having trouble finding transparent plastic coplin jars. We have pyrex, but they break in the microwave. Any suggestions where we can purchase transparent plastic coplin jars?? > > > > ----------------------------------------- > Confidentiality Notice: This e-mail message, including any > attachments, is for the sole use of the intended recipient(s) and > may contain confidential and privileged information. Any > unauthorized review, use, disclosure, or distribution is > prohibited. If you are not the intended recipient, please contact > the sender by reply e-mail and destroy all copies of the original > message. > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From sjchtascp <@t> yahoo.com Fri Apr 7 14:56:18 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Fri Apr 7 14:56:22 2006 Subject: [Histonet] histogel processing Message-ID: <20060407195618.62554.qmail@web38211.mail.mud.yahoo.com> I just got a sample of histogel and will be using it to FFPE cultured and stem cell bodies. Anyone have any suggestions as to the lenght of processing. Thanks, Steve --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. From liz <@t> premierlab.com Fri Apr 7 15:06:35 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Apr 7 15:04:44 2006 Subject: [Histonet] histogel processing In-Reply-To: <20060407195618.62554.qmail@web38211.mail.mud.yahoo.com> Message-ID: <000f01c65a7e$c58a4470$0300a8c0@Chlipala> Steve I have used histogel and for cell culture cells I always processed by hand 15 minutes per step. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Friday, April 07, 2006 1:56 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] histogel processing I just got a sample of histogel and will be using it to FFPE cultured and stem cell bodies. Anyone have any suggestions as to the lenght of processing. Thanks, Steve --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Bauer.Karen <@t> mayo.edu Fri Apr 7 15:01:24 2006 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Fri Apr 7 15:04:56 2006 Subject: [Histonet] IHC question References: Message-ID: Thanks for the reply Becky. We do not use any HIER on our CD3, CD20 or LCA antibodies. I didn't fool around with the LCA protocol, since the CD3 and CD20 worked just fine on the tissue. I'll have to do some "testing" with that. Thanks again, Karen ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Orr, Rebecca Sent: Fri 4/7/2006 1:19 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC question Hi Karen, My experienced guess would be 2 things. I am suggesting this based on my experiences in the field troubleshooting just this type of issue, so I'm not really an expert. But, I would consider these things: 1) The CD 3 and CD 20 epitope sites may not need as much formalin fixation to stain appropriately. There may be some "wiggle room" in the titer of these antibodies. What is your pretreatment for all 3? Are they all the same? In my lab I run concentrates and my CD3 works best with Tris HIER and the CD 20 works best with citrate HIER. Back in the day, LCA didn't even NEED HEIR! Or: 2)You may have simply over done the HIER and caused a prozone effect. Because the LCA control worked that shows me that the Antibody itself is working appropriately. These immunogens are developed separately so even though LCA will stain the same sites as CD3 and CD20, they aren't "the same". This may not be a fact but comes more from my experience. I'm sure you could make the LCA work if you compensated for the variable that changed, in this case the formalin/processing time. Either lighten up on the hier or if you can, extend the incubation time. Or Change the concentration. Sometimes errors are made and you have make a change to the standardized protocol to compensate. Hope this helps Becky Orr CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 -----Original Message----- Message: 23 Date: Fri, 7 Apr 2006 11:57:23 -0500 From: "Bauer, Karen" Subject: [Histonet] IHC Question... To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hi to all. I recently stained a lymph node case and it has brought up an interesting question that I can't explain to our Pathologist. I'm hoping that someone out there can give me some suggestions. First of all, a little background... The lymph node was processed, by accident, on a "fast run" on our VIP Tissue Tek. The processing run was complete in about 3 and a half hours. (We thought there were just small biopsies on the run, but the lymph node cassette was probably thrown in the wrong basket by mistake.) Anyway, the lymph node tissue didn't turn out all that bad, but the fatty tissue did not process adequately (no surprise there!!). Since the lymph node tissue cut fairly well, the block was not run back and re-processed. H&E's were turned in and stains were ordered. Here's my problem: The CD3 and CD20 stained well and the lymph node tissue and controls looked as they should, but the LCA on the patient tissue did not stain at all. We put the patient tissue on a control slide, so the control and tissue are stained the same all the way through the run. The LCA control looked beautiful, but the lymph node tissue had no staining. How can you have positive staining for CD3 and CD20, but no staining for LCA on a lymph node? Since we use a tonsil for our control, we re-ran the LCA with the tonsil control, a different lymph node case and then the same lymph node case that we were staining. Again, the control looked great, but the lymph node did not stain again. The different lymph node case that was also run turned out just fine, so we knew it was tissue specific. We thought maybe this lack of staining was due to the insufficient processing, but then why did the CD3 and CD20 turn out so well? Shouldn't they have no staining also? I'm perplexed and was wondering if anyone else out there has had this happen to them. As always, thank you. Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ********************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 End of Histonet Digest, Vol 29, Issue 11 **************************************** _______________________________________________ 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 rjbuesa <@t> yahoo.com Fri Apr 7 16:22:16 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Apr 7 16:22:21 2006 Subject: [Histonet] ANP.28860 In-Reply-To: Message-ID: <20060407212216.88169.qmail@web61211.mail.yahoo.com> Barb: Just to clarify: microwave-transparent does not mean "clear plastic", it means that the plastic does not absorb microwaves (MW) and, therefore, is considered as "transparent". Examples: Distilled water lets MW to travel only 3.4 cm in it, but those same MW can travel 400 meters in Styrofoam. Water, that is a transparent liquid, is almost 12 thousand times less "MW transparent" than Styrofoan, in spite of being an opaque material. You can have a black plastic container that is more "MW transparent" than water. That is what the CAP question is all about: Are you using "MW transparent" containers, i.e. Are you sure than the container is not interfering with the MW? Hope this will clarify your question. Ren? J. Barb Richmond wrote: New CAP question: Are all containers used in the micorwave devices made from microwave-transparent material. I'm having trouble finding transparent plastic coplin jars. We have pyrex, but they break in the microwave. Any suggestions where we can purchase transparent plastic coplin jars?? ----------------------------------------- Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From jqb7 <@t> cdc.gov Fri Apr 7 18:37:48 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Fri Apr 7 18:42:57 2006 Subject: [Histonet] histogel processing References: <20060407195618.62554.qmail@web38211.mail.mud.yahoo.com> Message-ID: We use it all the time and love it. After we prepare the pellet in histogel we put it on the processor for our normal overnight processing schedule. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Steven Coakley Sent: Fri 4/7/2006 3:56 PM To: Histonet@lists.utsouthwestern.edu Cc: Subject: [Histonet] histogel processing I just got a sample of histogel and will be using it to FFPE cultured and stem cell bodies. Anyone have any suggestions as to the lenght of processing. Thanks, Steve --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From barbwebb <@t> webtv.net Tue Apr 4 13:34:23 2006 From: barbwebb <@t> webtv.net (Barbara Webb) Date: Sun Apr 9 17:29:29 2006 Subject: [Histonet] (no subject) Message-ID: <20060404183423.2EB0CD864@smtpout-3201.bay.webtv.net> From lpwenk <@t> sbcglobal.net Sun Apr 9 17:33:49 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Sun Apr 9 17:35:20 2006 Subject: [Histonet] Health and Safety Guidelines for the Lab by L Montgomeryneeded In-Reply-To: Message-ID: <000501c65c25$ac141360$0a000a4b@HPPav2> When I studied for the SLS (Specialist in Laboratory Safety) exam a couple of years ago, I found the Hazmat Manual from Anatech to be very helpful. It's not on the ASCP BOR reading list for the SLS certification exam, but I think it should be. http://www.anatechltdusa.com/Hazmat.html Also, there is a self-assessment booklet from NSH on Laboratory Safety (#14). I was the editor of the booklet, published after I took the SLS exam. Wish I had done all the research and reading needed for this booklet, BEFORE I took the exam. http://www.nsh.org/education/materials.html Let me know what you think about these are references for taking the SLS exam. Good luck on the exam. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of jason.burrill@crl.com Sent: Wednesday, March 29, 2006 1:25 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Health and Safety Guidelines for the Lab by L Montgomeryneeded Hello all, I am in the process of taking the SLS (Specialist in Laboratory Safety) certification from ASCP and can not locate a new or used text of "Health and Safety Guidelines for the Laboratory" by Lynn Montgomery. I have searched the internet, Amazon and contacted ASCP with no luck (out of print), if anyone has any suggestions on where else I can look I would really appreciate it. Thanks in advance for your help. Jason Jason D. Burrill Manager, Histology Charles River Laboratories 251 Ballardvale Street Wilmington, MA 01887 ph: 978-658-6000 ext 1652 fax: 978-988-8793 E-mail: jason.burrill@crl.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From anh2006 <@t> med.cornell.edu Sun Apr 9 22:52:00 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Sun Apr 9 22:51:31 2006 Subject: [Histonet] Anti-mouse CD34 clone MEC14.7 Message-ID: Has anyone used this clone to stain mouse FFPE sections? If so can someone share their favorite protocol or stories of success? Thanks! Andrea -- From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Apr 10 03:14:36 2006 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Mon Apr 10 03:15:31 2006 Subject: [Histonet] P16 clone Message-ID: We are currently using the P16 from Ventana on the Benchmark XT with the protocol T-37 mCC1 32 mins and it works far better then the other clone we tried which was the Vision Biosystems NCL-P16-432 clone. I have found occasionally that certain clones work better with one system than another (manual v sequenza v Dako auto v Bond-max v Benchmark etc, etc) which may explain it. Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Apr 10 03:22:12 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Apr 10 03:21:36 2006 Subject: [Histonet] Chimerism Message-ID: I have a student that wants to follow a NVQ (National Vocational Qualification) and has chosen Chimerism (Genetic Mosaicism) about which I know nowt. Does anyone out there know anything about this subject or anyone who does? I've vainly been trying to find my feminine side and wonder if I've been looking in the wrong place; if this fascinating subject is to be believed it could, genetically, be anywhere . Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 "Following the line of least resistance makes both men and rivers crooked" This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation From Jacqueline.Malam <@t> rli.mbht.nhs.uk Mon Apr 10 03:59:55 2006 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Mon Apr 10 04:00:40 2006 Subject: [Histonet] Cyclin D1 Message-ID: We use Lab Vision's clone at 1/100 with a protocol of room temp, standard CC1 60 mins on the Benchmark XT (you could cut the time down which we will shortly do) and we are very pleased with it. Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From histology.bc <@t> shaw.ca Mon Apr 10 09:18:51 2006 From: histology.bc <@t> shaw.ca (Paul Bradbury) Date: Mon Apr 10 09:19:46 2006 Subject: [Histonet] Chimerism In-Reply-To: References: Message-ID: <443A694B.3030905@shaw.ca> Hi Kemlo, >The only times I have come across chimerism has been associated with blood transfusion. The patient presents with two different, but apparently coexisting, cell populations. So my best guess to look for further info would be the National Blood Transfusion Service, or a similar body. I believe this is an exceedingly rare condition, but certainly a fascinating one, should make an interesting thesis topic. > Paul Bradbury Kamloops, BC, Canada Kemlo Rogerson wrote: >I have a student that wants to follow a NVQ (National Vocational >Qualification) and has chosen Chimerism (Genetic Mosaicism) about which I >know nowt. Does anyone out there know anything about this subject or anyone >who does? > >I've vainly been trying to find my feminine side and wonder if I've been >looking in the wrong place; if this fascinating subject is to be believed it >could, genetically, be anywhere . > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > > > > > > > > From petepath <@t> yahoo.com Mon Apr 10 09:31:36 2006 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Mon Apr 10 09:31:43 2006 Subject: [Histonet] What is your favorite frozen section fixative Message-ID: <20060410143136.99818.qmail@web30413.mail.mud.yahoo.com> HI histonetters, I would like to do a comparison of the morphologic results using different fixatives for frozen section slides. I have always been happy with 95% ETOH. I know there are other good fixatives people use and I thought it would be useful to compare them. I will photograph and add these results to my web site tutorial ( http://pathologyinnovations.com/frozen_section_technique.htm ) and frozen section lecture ( NSH Region 1 April 28 & 29 and California Society of Histotechnology May 18-21). Please let me know what you find is the best fixative for frozen section morphology. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com From mgdelaware <@t> comcast.net Mon Apr 10 09:40:59 2006 From: mgdelaware <@t> comcast.net (Marian powers) Date: Mon Apr 10 09:39:17 2006 Subject: [Histonet] QIHC Message-ID: <000a01c65cac$c89201a0$be4dff45@MEGAN> Looking for a good IHC reference to study for the QIHC? Thanks in advance, Marian From Malcolm.McCallum <@t> tamut.edu Mon Apr 10 09:37:45 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Mon Apr 10 09:39:19 2006 Subject: [Histonet] Chimerism Message-ID: there was a good program on Discovery/science channel or Discovery health a while back. I do not recall if it was actually on fetus in feto (sp?), blood tests, or chimeraism. Hope that helps! Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Paul Bradbury Sent: Mon 4/10/2006 9:18 AM To: Kemlo Rogerson; HistoNet Server Subject: Re: [Histonet] Chimerism Hi Kemlo, >The only times I have come across chimerism has been associated with blood transfusion. The patient presents with two different, but apparently coexisting, cell populations. So my best guess to look for further info would be the National Blood Transfusion Service, or a similar body. I believe this is an exceedingly rare condition, but certainly a fascinating one, should make an interesting thesis topic. > Paul Bradbury Kamloops, BC, Canada Kemlo Rogerson wrote: >I have a student that wants to follow a NVQ (National Vocational >Qualification) and has chosen Chimerism (Genetic Mosaicism) about which I >know nowt. Does anyone out there know anything about this subject or anyone >who does? > >I've vainly been trying to find my feminine side and wonder if I've been >looking in the wrong place; if this fascinating subject is to be believed it >could, genetically, be anywhere . > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > > > > > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Apr 10 09:40:25 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Apr 10 09:40:41 2006 Subject: [Histonet] What is your favorite frozen section fixative Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176CA@lsexch.lsmaster.lifespan.org> For most immuno work, acetone or methanol. For strictly morphologic work, 10% formalin in 70% ethanol. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Stephen Peters M.D. > Sent: Monday, April 10, 2006 7:31 AM > To: Histonet@lists.utsouthwestern.edu > Subject: [Histonet] What is your favorite frozen section fixative > > HI histonetters, > > I would like to do a comparison of the morphologic results using > different fixatives > for frozen section slides. I have always been happy with 95% ETOH. I > know there are other good fixatives people use and I thought it would be > useful to compare them. I will photograph and add these results to my > web site tutorial ( > http://pathologyinnovations.com/frozen_section_technique.htm ) and frozen > section lecture ( NSH Region 1 April 28 & 29 and California Society > of Histotechnology May 18-21). > Please let me know what you find is the best fixative for frozen section > morphology. > > Stephen > > > > > > Stephen Peters M.D. > Vice Chairman of Pathology > Hackensack University Medical Center > 201 996 4836 > > Pathology Innovations, LLC > 410 Old Mill Lane, > Wyckoff, NJ 07481 > 201 847 7600 > www.pathologyinnovations.com > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From cpomajzl <@t> cpllabs.com Mon Apr 10 09:57:16 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Mon Apr 10 09:54:26 2006 Subject: [Histonet] Chimerism References: <443A694B.3030905@shaw.ca> Message-ID: <003b01c65caf$0fbe74d0$26fca8c0@CSP> I can tell you that there is much research being done using chimeric models. When I was working in research, we had mouse lines that were genetically engineered as chimeras. This particular lab was testing the role of heat-shock proteins during cardiac ischemia. I'm not sure if there was any interesting data obtained from the chimera, but the research possibilities are fascinating. Also, there was a TLC or Discovery special not long ago on Chimeras. It was an interesting program. It is a genetic anomaly that defies logic, but as Paul stated, it is an organism with two or more genetically distinct cell populations. Very neat stuff. ----- Original Message ----- From: "Paul Bradbury" To: "Kemlo Rogerson" ; "HistoNet Server" Sent: Monday, April 10, 2006 9:18 AM Subject: Re: [Histonet] Chimerism > Hi Kemlo, > > >The only times I have come across chimerism has been associated with blood transfusion. The patient presents with two different, but apparently coexisting, cell populations. So my best guess to look for further info would be the National Blood Transfusion Service, or a similar body. I believe this is an exceedingly rare condition, but certainly a fascinating one, should make an interesting thesis topic. > > > > Paul Bradbury > Kamloops, BC, Canada > > > Kemlo Rogerson wrote: > > >I have a student that wants to follow a NVQ (National Vocational > >Qualification) and has chosen Chimerism (Genetic Mosaicism) about which I > >know nowt. Does anyone out there know anything about this subject or anyone > >who does? > > > >I've vainly been trying to find my feminine side and wonder if I've been > >looking in the wrong place; if this fascinating subject is to be believed it > >could, genetically, be anywhere . > > > >Kemlo Rogerson > >Pathology Manager > >Ext 3311 > >DD 01934 647057 > >Mob 07749 754194 > > > > > > > > > > > > > > > > > > > > > > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Mon Apr 10 11:48:55 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Apr 10 11:49:00 2006 Subject: [Histonet] What is your favorite frozen section fixative In-Reply-To: <20060410143136.99818.qmail@web30413.mail.mud.yahoo.com> References: <20060410143136.99818.qmail@web30413.mail.mud.yahoo.com> Message-ID: <6.0.0.22.1.20060410104445.01b33270@gemini.msu.montana.edu> What is the fixation for? H&E or another stain for rapid diagnostic purposes? For H&E frozen sections, 95% ethanol works fine but we also use neutral buffered formalin. NBF, however, is not as FAST a fixative, but for morphology purposes, we like it better than 95% ethanol. For immunohistochemistry purposes, our fixation is MUCH different for frozen sections. At 08:31 AM 4/10/2006, you wrote: >HI histonetters, > > I would like to do a comparison of the morphologic results using > different fixatives > for frozen section slides. I have always been happy with 95% ETOH. I > know there are other good fixatives people use and I thought it would be > useful to compare them. I will photograph and add these results to my > web site tutorial > ( http://pathologyinnovations.com/frozen_section_technique.htm ) and > frozen section lecture ( NSH Region 1 April 28 & 29 and California Society > of Histotechnology May 18-21). > Please let me know what you find is the best fixative for frozen > section morphology. > > Stephen > > > > > >Stephen Peters M.D. >Vice Chairman of Pathology >Hackensack University Medical Center >201 996 4836 > >Pathology Innovations, LLC >410 Old Mill Lane, >Wyckoff, NJ 07481 >201 847 7600 >www.pathologyinnovations.com > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 406 994-4303 (FAX) From cornettl <@t> hotmail.com Mon Apr 10 11:49:58 2006 From: cornettl <@t> hotmail.com (Lorraine Cornett) Date: Mon Apr 10 11:50:07 2006 Subject: [Histonet] Feedback on Milestone Medical's RHS Series MicrowaveTissue Processor In-Reply-To: <8C8264C3B5C9264-1088-88C3@mblk-d50.sysops.aol.com> Message-ID: We are looking at this processor as well, please include me on the replies. Thanks Lorraine Cornett, HT (ASCP) Highlands Pathology Blue Ridge Division, Kingsport, TN 423 224-5793 fax 423 224-5349 >From: jenbug812@aol.com >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Feedback on Milestone Medical's RHS Series >MicrowaveTissue Processor >Date: Tue, 04 Apr 2006 18:14:10 -0400 > >Just wondering if anyone is currently using this tissue processor and if >so, can you provide any feedback, good or bad? My company is conducting >demos with this product and I would like to be prepared for the sales rep. >Thanks >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From Janet.Bonner <@t> flhosp.org Mon Apr 10 12:05:56 2006 From: Janet.Bonner <@t> flhosp.org (Bonner, Janet) Date: Mon Apr 10 12:07:28 2006 Subject: [Histonet] What is your favorite frozen section fixative References: <6.0.0.22.1.20060410104445.01b33270@gemini.msu.montana.edu> Message-ID: We use a mix of the two for FS fixative; 190cc 95% EtOH and 70cc NBF. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Gayle Callis Sent: Mon 4/10/2006 12:48 PM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] What is your favorite frozen section fixative What is the fixation for? H&E or another stain for rapid diagnostic purposes? For H&E frozen sections, 95% ethanol works fine but we also use neutral buffered formalin. NBF, however, is not as FAST a fixative, but for morphology purposes, we like it better than 95% ethanol. For immunohistochemistry purposes, our fixation is MUCH different for frozen sections. At 08:31 AM 4/10/2006, you wrote: >HI histonetters, > > I would like to do a comparison of the morphologic results using > different fixatives > for frozen section slides. I have always been happy with 95% ETOH. I > know there are other good fixatives people use and I thought it would be > useful to compare them. I will photograph and add these results to my > web site tutorial > ( http://pathologyinnovations.com/frozen_section_technique.htm ) and > frozen section lecture ( NSH Region 1 April 28 & 29 and California Society > of Histotechnology May 18-21). > Please let me know what you find is the best fixative for frozen > section morphology. > > Stephen > > > > > >Stephen Peters M.D. >Vice Chairman of Pathology >Hackensack University Medical Center >201 996 4836 > >Pathology Innovations, LLC >410 Old Mill Lane, >Wyckoff, NJ 07481 >201 847 7600 >www.pathologyinnovations.com > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 406 994-4303 (FAX) _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Mon Apr 10 12:46:51 2006 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Mon Apr 10 12:46:59 2006 Subject: [Histonet] substitute for Uranyl Nitrate in Snooks retic. method Message-ID: <4.3.2.7.2.20060410104128.025836e0@algranth.inbox.email.arizona.edu> I know this was discussed a while back and I have checked the archives where phosphomolybdic acid and zinc formalin is suggested instead of 1% Uranyl Nitrate. Is anyone out there doing this method and what are you using - if you are using the phosphomolybdic acid, what percentage are you using? Thanks. Andi ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From linhines <@t> yahoo.com Mon Apr 10 13:18:58 2006 From: linhines <@t> yahoo.com (Linda Hines) Date: Mon Apr 10 13:19:01 2006 Subject: [Histonet] Calibration of Thermometers Message-ID: <20060410181858.36822.qmail@web33601.mail.mud.yahoo.com> Hello, All Does anyone have a procedure for calibrating thermometers in the pathology lab? Do we need to purchase a certain thermometer for calibrations, any help is greatly appreciated. Linda Hines HT --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From germckeon <@t> excite.com Mon Apr 10 13:22:43 2006 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Mon Apr 10 13:22:49 2006 Subject: [Histonet] IHC staining for astrocytes Message-ID: <20060410182243.0A1EA3DD8@xprdmailfe12.nwk.excite.com> Hello Histonetters,I am in need of some information which is likely to be only found amongst people of your intellectual calibre (flattery gets you everywhere!).I am interested in immunostaining for glia, specifically astrocytes, in the brain and spinal column of the zebrafish.Would anyone know of any antibodies that would stain definitively for these cells?Hoping you can be of some help.Thanking you,Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From cgfields <@t> lexhealth.org Mon Apr 10 13:29:44 2006 From: cgfields <@t> lexhealth.org (Carole Fields) Date: Mon Apr 10 13:26:08 2006 Subject: [Histonet] substitute for Uranyl Nitrate in Snooks retic. met hod Message-ID: Hi Andi, I have a procedure that substitutes 2% Ferric Ammonium Sulfate for the 1% Uranyl ...we used it all the time in San Diego. I will attach the procedure. If anyone else needs a copy let me know. Carole Fields, HT,ASCP Pathology Supervisor Lexington Medical Center 2720 Sunset Blvd. W. Columbia, SC 29169 -----Original Message----- From: Andrea Grantham [mailto:algranth@u.arizona.edu] Sent: Monday, April 10, 2006 1:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] substitute for Uranyl Nitrate in Snooks retic. method I know this was discussed a while back and I have checked the archives where phosphomolybdic acid and zinc formalin is suggested instead of 1% Uranyl Nitrate. Is anyone out there doing this method and what are you using - if you are using the phosphomolybdic acid, what percentage are you using? Thanks. Andi ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________ 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 Ambergoetze <@t> aol.com Mon Apr 10 14:04:27 2006 From: Ambergoetze <@t> aol.com (Ambergoetze@aol.com) Date: Mon Apr 10 14:04:44 2006 Subject: [Histonet] Holzer stain for glial fibers Message-ID: <372.17e67b2.316c063b@aol.com> Hello everyone, I am doing research on the Holzer staining technique, I was wondering if anyone could tell me the functions of the following reagents: 0.5% Aqueous Phosphomolybdic Acid solution, Phosphomolybdic Alcohol solution, Absolute Alcohol Chloroform mixture, and Potassium Bromide solution. Thank you! From cfavara <@t> niaid.nih.gov Mon Apr 10 14:19:22 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Mon Apr 10 14:19:32 2006 Subject: [Histonet] IHC staining for astrocytes Message-ID: Astrocytes are stained quite nicely with GFAP. Oligo Olig-2 I do not know if they will cross with zebra fish that is out of my league by like 2000. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: germckeon@excite.com [mailto:germckeon@excite.com] Sent: Monday, April 10, 2006 11:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC staining for astrocytes Hello Histonetters,I am in need of some information which is likely to be only found amongst people of your intellectual calibre (flattery gets you everywhere!).I am interested in immunostaining for glia, specifically astrocytes, in the brain and spinal column of the zebrafish.Would anyone know of any antibodies that would stain definitively for these cells?Hoping you can be of some help.Thanking you,Gerald McKeon _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From germckeon <@t> excite.com Mon Apr 10 14:22:27 2006 From: germckeon <@t> excite.com (germckeon@excite.com) Date: Mon Apr 10 14:22:33 2006 Subject: [Histonet] IHC staining for astrocytes Message-ID: <20060410192227.A2F143DC5@xprdmailfe12.nwk.excite.com> Hello,I am using GFAP but want something which is more specific to astrocytes.Thanks,Gerald McKeon --- On Mon 04/10, Favara, Cynthia (NIH/NIAID) [E] < cfavara@niaid.nih.gov > wrote:From: Favara, Cynthia (NIH/NIAID) [E] [mailto: cfavara@niaid.nih.gov]To: germckeon@excite.com, histonet@lists.utsouthwestern.eduDate: Mon, 10 Apr 2006 15:19:22 -0400Subject: RE: [Histonet] IHC staining for astrocytesAstrocytes are stained quite nicely with GFAP. Oligo Olig-2 I do notknow if they will cross with zebra fish that is out of my league by like2000.cCynthia FavaraNIAID/NIH/RML/LPVD903 South 4th StreetHamilton, MT 59840406-363-9317Disclaimer: The information in this e-mail and any of its attachments isconfidential and may contain sensitive information. It should not beused by anyone who is not the original intended recipient. If you havereceived this e-mail in error please inform the sender and delete itfrom your mailbox or any other storage devices. National Institute ofAllergy and Infectious Diseases shall not accept liability for anystatements made that are sender's own and not expressly made on behalfof the NIAID by one of its representatives-----Original Message-----From: germckeon@excite.com [mailto:germckeon@excite.com] Sent: Monday, April 10, 2006 11:23 AMTo: histonet@lists.utsouthwestern.eduSubject: [Histonet] IHC staining for astrocytes Hello Histonetters,I am in need of some information which is likely tobe only found amongst people of your intellectual calibre (flattery getsyou everywhere!).I am interested in immunostaining for glia,specifically astrocytes, in the brain and spinal column of thezebrafish.Would anyone know of any antibodies that would staindefinitively for these cells?Hoping you can be of some help.Thankingyou,Gerald McKeon_______________________________________________Join Excite! - http://www.excite.comThe most personalized portal on the Web!_______________________________________________Histonet mailing listHistonet@lists.utsouthwestern.eduhttp://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Join Excite! - http://www.excite.com The most personalized portal on the Web! From Nadia.Gale <@t> vch.ca Mon Apr 10 15:08:33 2006 From: Nadia.Gale <@t> vch.ca (Gale, Nadia [NS]) Date: Mon Apr 10 15:09:06 2006 Subject: [Histonet] Slides compatible with Ventana Message-ID: We have been using a Ventana Benchmark XT for all of our immunostaining for one year. We have found the only slides that consistently retain all of the tissue are Fisherbrand Superfrost/Plus slides, Catalog No. 12-550-15. These slides may also be purchased with a "control box". Nadia Gale IHC Technologist Anatomical Pathology Laboratory Lions Gate Hospital 604-984-5802 nadia.gale@vch.ca From JPaulB42 <@t> aol.com Mon Apr 10 15:42:08 2006 From: JPaulB42 <@t> aol.com (JPaulB42@aol.com) Date: Mon Apr 10 15:42:18 2006 Subject: [Histonet] Feedback on Milestone Medical's RHS Series MicrowaveTissue Processor Message-ID: <21e.b47e9c8.316c1d20@aol.com> I just finished my first run on the Milestone RHS-110. Excellent results on large tissue samples. Uterus, colon, placenta, prostate, breast all fixed well, processed well with a small amount of shrinkage. Which will be fixed with a minor adjustment of the long program. The H&E staining turned out slightly or more than we are accustomed to but that too will easily be resolved. A few more runs to come shortly and we'll see where we go from here. James Bradford Orlando Regional Medical Center Orlando, FL 321-841-5498 From barry_m <@t> ozemail.com.au Mon Apr 10 16:02:06 2006 From: barry_m <@t> ozemail.com.au (Barry Madigan) Date: Mon Apr 10 16:02:14 2006 Subject: [Histonet] PTH In-Reply-To: Message-ID: <000001c65ce2$06f372a0$0201010a@WORKSTATION1> We do PTH on formalin fixed paraffin-embedded tissue. Novocastra antibody NCL-PTH-488 (clone 105G7) at a dilution of 1:1000 and using heat retrieval with a high pH EDTA solution. This is done on the BOND MAX Immunostainer. Barry Madigan Immunohistochemistry QHPS-Central Queensland Australia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Richard Cartun Sent: Wednesday, 5 April 2006 3:14 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PTH Is anyone doing immunohistochemical staining for parathyroid hormone (PTH) on formalin-fixed, paraffin-embedded tissue? If so, whose antibody are you using? I just found out that DAKO no longer sells the rat monoclonal antibody that we had been using. It would be nice if suppliers notified their customers before they discontinue a product so that we can find a replacement before we run out of reagent. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nfournier <@t> sasktel.net Mon Apr 10 17:36:34 2006 From: nfournier <@t> sasktel.net (N Fournier) Date: Mon Apr 10 17:36:38 2006 Subject: [Histonet] cryoprotection help rat brain Message-ID: <58ac66ec7321.443a8992@sasktel.net> Hi, I have been having some problems with our laboratory protocol for making up a viscuous solution for rat brain section. We use the protocol of Watson and colleagues published in 1986 in Peptides (page 155-59). The original protocol states to make 1 L cryoprotectant solution one needs 500 ml of PB, 300 g sucrose (30% w/v), 10 g polyvinylpyrrolidone (1% w/v), and 300 ml of ethylene glycol (30% v/v). I noticed that once I made up this solution and placed it in our -20 degree C freezer, the solution seemed to precipitate and become very viscous (strands of gelatinous material in the soluion). The solution looked clear before I placed it in the freezer. Has anyone encountered this before? Should I have let the solution stir overnight before placing in the fridge? Once the solution has been put in the fridge, taken out because of the observation above and restirred at room temperature, is it alright to return the solution bottle back to the fridge after that, or is the solution ruined. Lastly, can I transfer brain sections into a cryoprotectant solution at -20 degrees if they have been cut recently and stored in PB at 4 degrees C? I appreciate the help, Neil From jkiernan <@t> uwo.ca Mon Apr 10 17:40:50 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Mon Apr 10 17:40:53 2006 Subject: [Histonet] Holzer stain for glial fibers References: <372.17e67b2.316c063b@aol.com> Message-ID: <443ADEF2.7FA5A424@uwo.ca> I don't think there are answers, other than wild speculation, to your questions! A few statements can be made about the reagents, but they don't necessarily relate to Holzer's method. When a PMA solution is applied to a section its large anions stick to regions of protein that are permeable and rich in basic amino acids. (This happens in collagen, in trichrome methods.) PMA also makes insoluble pigment precipitates when mixed with cationic triphenylmethane dyes such as crystal violet. Non-aqueous PMA may have tissue affinity different from that of an aqueous solution. With the chemically related phosphotungstic acid, the solvent and other factors influence the tissue components that take up the metal when PTA is used as a contrast stain for EM (see Hayat 1993 Stains and Cytochemical Methods, various places in book). The Holzer method I'm now reading (in H. Cook's "Manual", 1974) does not have a reagent called phosphomolybdic alcohol, but the PMA is dissolved (0.5%) in 95% alcohol. I have no idea what the ethanol-chloroform mixture does. As the solvent for the dye, it might help to carry crystal violet into hydrophobic domains of the section. Clearly care is taken not to have any water in contact with the sections before, during and immediately after the staining in non-aqueous crystal violet. Cook suggests that the KBr might act as a trapping agen for previously bound dye. (This would be an action similar to the iodine-KI solution used in Gram staining.) The aniline-chloroform-ammonia mixture is used to differentiate the stain. Ordinarily you wouldn't expect a cationic dye like cresyl violet to be extracted by an alkaline solution. Possibly it extracts bound PMA, or dissolves a PMA-dye-bromide complex out of hydrophobic but not out of hydrophilic domains in the tissue. All guessing! There's plenty of room for benchtop testing of ideas with this one. It will be splendid if you come up with hard evidence for a possible mechanism for Holzer's stain. Several years ago I was talking with a pathologist who had tried to show that GFAP was the substrate for Holzer's method. He failed to do so, using methods equivalent to the ones he'd used to show that Bodian's protargol method demonstrated one of the neurofilament proteins. Nevertheless, it's difficult to believe that Holzer's method and the related Anderson's victoria blue staining anything other than GFAP; is there another major strctural protein in astrogliosis tissue? Good luck with your research. I'd recommend reading Holzer's 1921 paper; he must have developed the method on the basis of some sort of reasoning! I don't know of any published work on the mechanism of Holzer's methods for gliosis. Perhaps someone else will come up with something. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Ambergoetze@aol.com wrote: > > Hello everyone, > I am doing research on the Holzer staining technique, I was wondering if > anyone could tell me the functions of the following reagents: 0.5% Aqueous > Phosphomolybdic Acid solution, Phosphomolybdic Alcohol solution, Absolute > Alcohol Chloroform mixture, and Potassium Bromide solution. > > > Thank you! --------------- From cfranci <@t> rigel.com Mon Apr 10 19:17:09 2006 From: cfranci <@t> rigel.com (Christian Franci) Date: Mon Apr 10 19:17:21 2006 Subject: [Histonet] fixing tissues with zinc formalin Message-ID: <727df04461a9589535f3243dcf82c5ed@rigel.com> Hello histokids! I tried looking in the archives but there were too many past postings labeled "zinc formalin" that didn't address my questions so, I decided to post instead. is there one particular brand that works better or, is all commercially available zF about the same? Buffered better than not? for fixing tissues such as small tumors of no more than half a mL in volume, is there a good rule of thumb? (I know that for NBF if we pickle more than 8 hrs we can't retrieve any epitopes later on... does that happen as well with zinc formalin?) I seem to recall people mentioning that the zinc precipitaties? Does the fixed tissue need to be rinsed before putting it in the processor? any pointers or tricks are more than welcomed! Thanks in advance. Chris From jdmd77 <@t> hotmail.com Mon Apr 10 20:38:38 2006 From: jdmd77 <@t> hotmail.com (Julia Dahl) Date: Mon Apr 10 20:38:46 2006 Subject: [Histonet] Image disclaimer In-Reply-To: Message-ID: There was a recent article in the Journal Of Dermatology that covers this topic. >From: "Jesus Ellin" >To: victor@pathology.washington.edu,rjbuesa@yahoo.com >CC: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Image disclaimer >Date: Thu, 06 Apr 2006 13:34:35 -0700 >MIME-Version: 1.0 >Received: from swlx162.swmed.edu ([199.165.152.162]) by >bay0-mc3-f19.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.1830); Thu, 6 >Apr 2006 13:35:45 -0700 >Received: from localhost ([127.0.0.1] helo=swlx162.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34)id 1FRbCP-0000Q8-My; Thu, 06 Apr >2006 15:35:25 -0500 >Received: from [199.165.152.166] (helo=swlx166.swmed.edu)by >swlx162.swmed.edu with esmtp (Exim 4.34) id 1FRbCL-0000Pz-Ptfor >histonet@lists.utsouthwestern.edu; Thu, 06 Apr 2006 15:35:23 -0500 >Received: from twmms.carebridge.net ([139.177.224.67])by swlx166.swmed.edu >with esmtp (Exim 4.44) id 1FRbCJ-0008Ix-Cufor >histonet@lists.utsouthwestern.edu; Thu, 06 Apr 2006 15:35:21 -0500 >Received: from 198.212.202.20 by twmms.carebridge.net with SMTP >(SMTPRelay); Thu, 06 Apr 2006 16:35:09 -0400 >Received: from YRMCGW-Message_Server by FSGROUPWISE.yumaregional.orgwith >Novell_GroupWise; Thu, 06 Apr 2006 13:35:08 -0700 >X-Message-Info: FdnYIHvXcrINCkuGOOmAg97se0PzOHhwnIwi6tinmN8= >X-Server-Uuid: 8F3C3CFB-FA04-4E2B-BFC2-0E4F125CD055 >X-Mailer: Novell GroupWise 5.5.5 >X-TMWD-Spam-Summary: TS=20060406203511; SEV=2.0.1; >DFV=A2006040608;IFV=2.0.4,4.0-8; RPD=4.00.0004; >ENG=IBF;RPDID=303030312E30413039303230362E34343335374137372E303034372D412D;CAT=NONE; >CON=NONE >X-MMS-Spam-Filter-ID: A2006040608_4.00.0004_4.0-8 >X-WSS-ID: 682BA4F7XR8290257-01-01 >X-Scan-Signature: a1858e828dfa2042deb55ef2dd43bec4 >X-BeenThere: histonet@lists.utsouthwestern.edu >X-Mailman-Version: 2.1.5 >Precedence: list >List-Id: For the exchange of information pertaining to histotechnology >andrelated fields >List-Unsubscribe: >, > >List-Archive: >List-Post: >List-Help: >List-Subscribe: >, >Errors-To: histonet-bounces@lists.utsouthwestern.edu >X-Scan-Signature: 4fcdaf9e0a622c13a93ecd4a6192e06b >X-SA-Exim-Connect-IP: 127.0.0.1 >X-SA-Exim-Mail-From: histonet-bounces@lists.utsouthwestern.edu >X-Spam-Checker-Version: SpamAssassin 2.64 (2004-01-11) on swlx162.swmed.edu >X-Spam-Level: X-Spam-Status: No, hits=0.0 required=5.5 tests=none >autolearn=no version=2.64 >X-SA-Exim-Version: 4.0 (built Wed, 07 Apr 2004 12:14:41 -0500) >X-SA-Exim-Scanned: Yes (on swlx162.swmed.edu) >Return-Path: histonet-bounces@lists.utsouthwestern.edu >X-OriginalArrivalTime: 06 Apr 2006 20:35:45.0926 (UTC) >FILETIME=[AE91F260:01C659B9] > >Well this is an interesting topic. Why would you need a disclaimer for an >image. It is there of esthetics, unless the physician is using it to >demonstrate his or hers dx. Then I would say you would need to talk to >your companies legal Department, but it does raise a flag to me. > > >Jesus Ellin >Yuma Regional Medical Center > > >>> "Victor Tobias" 04/06/06 10:31AM >>> >Has someone run into an issue by not having a disclaimer or is it just >to CYA? > >Rene J Buesa wrote: > > >And what do you find wrong with yours? I think it serves its purpose and >is short and to the point. I don't think you need another. > > René J. > > > >Bonnie Whitaker wrote: > > Hi All, > > > >Is anyone out there using a disclaimer on reports with images that they > >would share with me? (such as: "Images in this report are for >illustrative > >purposes only and are not meant to be diagnostic") If you use a >disclaimer, > >who wrote it? Someone in pathology or the hospital's or lab's legal > >department? > > > >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 > > > > > > > > > > > >--------------------------------- > >Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ >countries) for 2¢/min or less. > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > > >-- >Victor Tobias >Clinical Applications Analyst >University of Washington Medical Center >Dept of Pathology Room BB220 >1959 NE Pacific >Seattle, WA 98195 >victor@pathology.washington.edu >206-598-2792 >206-598-7659 Fax >================================================= >Privileged, confidential or patient identifiable information may be >contained in this message. This information is meant only for the use >of the intended recipients. If you are not the intended recipient, or >if the message has been addressed to you in error, do not read, >disclose, reproduce, distribute, disseminate or otherwise use this >transmission. Instead, please notify the sender by reply e-mail, and >then destroy all copies of the message and any attachments. > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From AnthonyH <@t> chw.edu.au Mon Apr 10 18:07:19 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Mon Apr 10 22:22:21 2006 Subject: [Histonet] What is your favorite frozen section fixative Message-ID: Stephen, We routinely use methanol followed by a rapid H&E. It is important to immediately fix the slide and not allow any airdrying. 30-60 sec at room temp is enough. I will forward on to you some example pics on a separate email Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Stephen Peters M.D. Sent: Tuesday, 11 April 2006 12:32 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] What is your favorite frozen section fixative HI histonetters, I would like to do a comparison of the morphologic results using different fixatives for frozen section slides. I have always been happy with 95% ETOH. I know there are other good fixatives people use and I thought it would be useful to compare them. I will photograph and add these results to my web site tutorial ( http://pathologyinnovations.com/frozen_section_technique.htm ) and frozen section lecture ( NSH Region 1 April 28 & 29 and California Society of Histotechnology May 18-21). Please let me know what you find is the best fixative for frozen section morphology. Stephen Stephen Peters M.D. Vice Chairman of Pathology Hackensack University Medical Center 201 996 4836 Pathology Innovations, LLC 410 Old Mill Lane, Wyckoff, NJ 07481 201 847 7600 www.pathologyinnovations.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From AnthonyH <@t> chw.edu.au Mon Apr 10 17:55:30 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Mon Apr 10 22:22:42 2006 Subject: [Histonet] Chimerism Message-ID: I believe this is known as Proteus syndrome. The following is from a paper I am writing on the cytological findings of a hydrocele fluid from a boy with Proteus syndrome (I must pull my finger out and submit it). As an aside, it is believed that the "elephant man" suffered from this disorder. Proteus syndrome is a rare disorder and it estimated that there are only several hundred patients affected. It is postulated that the syndrome is caused by a post-zygotic mosaic alteration in a gene that is lethal in its non-mosaic state (4). Apart from connective tissue and epidermal nevi, disproportionate overgrowth of some bones and other malformations, bilateral ovarian cystadenomas are regarded as having diagnostic value in Proteus syndrome when occurring in the first two decades of life. Papillary lesions, resembling common epithelial ovarian tumours are rare in the testis with only a handful being described in pre-pubertal males (4). It has been suggested that these papillary lesions, so called serous papillary cystadenomas of low malignant potential, may prove to have the same diagnostic value in Proteus syndrome, as do bilateral cystic ovarian tumours (4). Some references follows: 3. Bale, P.M., Watson, G., Collins, F., (1993) "Pathology of Osseous and Genitourinary Lesions of Proteus Syndrome" Ped. Pathol 13:797-809. 4. Raju, R.R., Hart, W.R., Magnuson, D.K., Reid, J.R., Rogers, D.G., (2002) "Genital Tract Tumours in Proteus Syndrome: Report of a case of Bilateral Paraovarian Endometrioid Cystic Tumours of Borderline Malignancy and Review of the Literature" Mod Pathol 15(2):172-180. 5. McClure, R.F., Keeney, G.L., Sebo, T.J., Cheville, J.C., (2001) "Serous Borderline Tumour of the Paratestis: A Report of Seven Cases" Am J Surg Pathol 25(3):373-378. 6. Austin, P.F., Rink, R.C., Cain, M.P., Casale, A.J., Davis, M.M., Faught, P.R., (1998) "Testicular Serous Papillary Cystadenomatous Tumor of Low Malignant Potential in a Child" J Urol 160(6-I):2002-2003. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Paul Bradbury Sent: Tuesday, 11 April 2006 12:19 AM To: Kemlo Rogerson; HistoNet Server Subject: Re: [Histonet] Chimerism Hi Kemlo, >The only times I have come across chimerism has been associated with >blood transfusion. The patient presents with two different, but >apparently coexisting, cell populations. So my best guess to look for >further info would be the National Blood Transfusion Service, or a >similar body. I believe this is an exceedingly rare condition, but >certainly a fascinating one, should make an interesting thesis topic. > Paul Bradbury Kamloops, BC, Canada Kemlo Rogerson wrote: >I have a student that wants to follow a NVQ (National Vocational >Qualification) and has chosen Chimerism (Genetic Mosaicism) about which >I know nowt. Does anyone out there know anything about this subject or >anyone who does? > >I've vainly been trying to find my feminine side and wonder if I've >been looking in the wrong place; if this fascinating subject is to be >believed it could, genetically, be anywhere . > >Kemlo Rogerson >Pathology Manager >Ext 3311 >DD 01934 647057 >Mob 07749 754194 > > > > > > > > > > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From lpwenk <@t> sbcglobal.net Tue Apr 11 03:03:34 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Tue Apr 11 03:05:10 2006 Subject: [Histonet] QIHC In-Reply-To: <000a01c65cac$c89201a0$be4dff45@MEGAN> Message-ID: <001201c65d3e$6e67a6c0$3b2c1544@HPPav2> The Immunohistochemistry Resource Group of NSH has the about 4 books that they recommend: http://www.ihcrg.org/ Click on "Important change in the QIHC exam effective January 1, 2005. More... " in the middle of the page. For more books, click on "IHC References" on the left. Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marian powers Sent: Monday, April 10, 2006 10:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QIHC Looking for a good IHC reference to study for the QIHC? Thanks in advance, Marian _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From andrew.bond <@t> imperial.ac.uk Tue Apr 11 11:12:47 2006 From: andrew.bond <@t> imperial.ac.uk (Bond, Andrew) Date: Tue Apr 11 11:12:54 2006 Subject: [Histonet] Endothelial Cell Border stains Message-ID: <793D7D892DCFD441972F743EB40C9CE66EE965@icex3.ic.ac.uk> Hi, I am trying to look at changes in endothelial cell morphology in the rabbit thoracic aorta. I have managed to get Hautchen preparations to work fairly well, and now need to stain endothelial cell borders. Only problem is I am trying to carry out automated image analysis therefore need a very uniform and specific fluorescent stain, that leaves very clear lines without any cytoplasm/nuclear staining. I've been told silver nitrate staining would work but is very inconsistent. Does anyone have any suggestions? Thanks. Andrew Bond Imperial College London, UK From rjbuesa <@t> yahoo.com Tue Apr 11 11:42:56 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Apr 11 11:43:07 2006 Subject: [Histonet] Endothelial Cell Border stains In-Reply-To: <793D7D892DCFD441972F743EB40C9CE66EE965@icex3.ic.ac.uk> Message-ID: <20060411164256.15724.qmail@web61216.mail.yahoo.com> Andrew: Bensley & Bensley (Anat.Rec.,vol 64 pp 46,1935) descibe a method with silver chloride dichlorfluoresceinate. It requires to inject the animal intravenously with 0.8% aq. dichlorofluorescein until the animal becomes quite yellow. Kill the animal and immerse the targeted tissue in 10% aq. silver nitrate. Fix in 10% neutral formalin, dehydrate in ethanol, clear in iso-safrol, mount in balsam. The endothelial cells will be outlined in pink. Exposed to light silver becomes park brown to black with defined outlines. Silver inconsistencies are usually due to procedural inconsistencies. Hope this will help! Ren? J. "Bond, Andrew" wrote: Hi, I am trying to look at changes in endothelial cell morphology in the rabbit thoracic aorta. I have managed to get Hautchen preparations to work fairly well, and now need to stain endothelial cell borders. Only problem is I am trying to carry out automated image analysis therefore need a very uniform and specific fluorescent stain, that leaves very clear lines without any cytoplasm/nuclear staining. I've been told silver nitrate staining would work but is very inconsistent. Does anyone have any suggestions? Thanks. Andrew Bond Imperial College London, UK _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From sarah <@t> kidneybiopsy.com Tue Apr 11 12:25:49 2006 From: sarah <@t> kidneybiopsy.com (Sarah Holmes) Date: Tue Apr 11 12:07:25 2006 Subject: [Histonet] Dako unresponsive? Message-ID: <001501c65d8c$f9ee81c0$780a010a@wp.comcast.net> Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah From sjchtascp <@t> yahoo.com Tue Apr 11 12:12:26 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Tue Apr 11 12:12:34 2006 Subject: [Histonet] histogel Message-ID: <20060411171227.11273.qmail@web38206.mail.mud.yahoo.com> Ok, now I have successfull processed cells using histogel. Do I dry and stain the slides as usual? Steve --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From deb.vaneyck <@t> phci.org Tue Apr 11 12:21:23 2006 From: deb.vaneyck <@t> phci.org (Van Eyck, Deb) Date: Tue Apr 11 12:21:38 2006 Subject: [Histonet] Parathyroid hormone Message-ID: I agree with Richard-----it would be nice to know when you order a product that it is being discontinued. I trialed several PTH antibodies and I ended up liking the Novocastra PTH NCL-PTH-488 - it is research use only-------couldn't find a source for IVD PTH. Hope this helps! Deb Van Eyck Cytology/Immunohistochemistry Waukesha Memorial Hospital 725 American Avenue Waukesha,WI 53188 phone:262-928-2112 fax:262-928-4849 deb.vaneyck@phci.org 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. From jqb7 <@t> cdc.gov Tue Apr 11 12:22:10 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Tue Apr 11 12:25:00 2006 Subject: [Histonet] histogel Message-ID: Yep! Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Tuesday, April 11, 2006 1:12 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] histogel Ok, now I have successfull processed cells using histogel. Do I dry and stain the slides as usual? Steve --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cpomajzl <@t> cpllabs.com Tue Apr 11 12:36:02 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Tue Apr 11 12:33:20 2006 Subject: [Histonet] Dako unresponsive? References: <001501c65d8c$f9ee81c0$780a010a@wp.comcast.net> Message-ID: <003a01c65d8e$6a9e0d40$26fca8c0@CSP> Dako is switching over to a new business system. So you are going to have some trouble. Supposedly, they were going to be able to take calls as of yesterday. We placed an order at the end of March, and our supplies just arrived today. Good luck. ----- Original Message ----- From: "Sarah Holmes" To: Sent: Tuesday, April 11, 2006 12:25 PM Subject: [Histonet] Dako unresponsive? Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charlene.Henry <@t> STJUDE.ORG Tue Apr 11 12:38:57 2006 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Tue Apr 11 12:39:52 2006 Subject: [Histonet] CD61 Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1BEA@SJMEMXMB02.stjude.sjcrh.local> Dako has discontinued the CD61 (Dako M0753) antibody that we have been using for years and I was wondering if anyone knows of a good CD61 that I could try as a replacement? I have tried the Ventana CD61 (Y2/51) and Cell Marque CD61 (2f2) and they are not as good as our old antibody from Dako. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 From sa.drew <@t> hosp.wisc.edu Tue Apr 11 12:45:12 2006 From: sa.drew <@t> hosp.wisc.edu (Drew Sally A.) Date: Tue Apr 11 12:45:24 2006 Subject: [Histonet] CD61 Message-ID: We use the Cell Marque CD61 (clone 2f2) via Ventana, and it seems to work well for us with mild CC1 on our VMS instruments. Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Henry, Charlene Sent: Tuesday, April 11, 2006 12:39 PM To: histonet@pathology.swmed.edu Subject: [Histonet] CD61 Dako has discontinued the CD61 (Dako M0753) antibody that we have been using for years and I was wondering if anyone knows of a good CD61 that I could try as a replacement? I have tried the Ventana CD61 (Y2/51) and Cell Marque CD61 (2f2) and they are not as good as our old antibody from Dako. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From linhines <@t> yahoo.com Tue Apr 11 12:46:50 2006 From: linhines <@t> yahoo.com (Linda Hines) Date: Tue Apr 11 12:46:52 2006 Subject: [Histonet] Cell block prep Message-ID: <20060411174650.78180.qmail@web33603.mail.mud.yahoo.com> Hi! All, Thank-you for the responses regarding the thermometer's. Are any of you working with the Thin-prep and preparing cytology blocks upon completing the slide preparation? My question is how we prepare the cell blocks. Any information is greatly appreciated. We are a new lab, and this is the first time I have'nt made my own smears etc. Thank-you Linda Hines HT (ASCP) --------------------------------- Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1¢/min. From settembr <@t> umdnj.edu Tue Apr 11 12:46:17 2006 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Tue Apr 11 12:47:18 2006 Subject: [Histonet] Dako unresponsive? Message-ID: I was waiting too long on hold so I left a complete voicemail message as they suggested and received a call back that they got my message. The processed my order. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Sarah Holmes 04/11/06 1:25 PM >>> Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kmerriam2003 <@t> yahoo.com Tue Apr 11 12:53:08 2006 From: kmerriam2003 <@t> yahoo.com (Kim Merriam) Date: Tue Apr 11 12:53:12 2006 Subject: [Histonet] Sudan Red 7B staining procedure Message-ID: <20060411175308.21420.qmail@web50308.mail.yahoo.com> Hello, I have found a reference for staining fat with sudan red 7B, it calls for a "saturated" solution of sudan red 7B in 70% ethanol. Does anyone know what the saturation point of sudan red 7B is in 70% ethanol; a general guide would be very helpful! Also - has anyone used this procedure before and what were the results compared to a regular Oil Red O (both in isopropanol and/or propylene glycol). Thanks in advance, Kim Kim Merriam, MA, HT(ASCP) Novartis Cambridge, MA --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. From Lynne.Bell <@t> hitchcock.org Tue Apr 11 13:01:36 2006 From: Lynne.Bell <@t> hitchcock.org (Bell, Lynne) Date: Tue Apr 11 13:01:46 2006 Subject: [Histonet] Dako unresponsive? Message-ID: We are also having a major problem getting through to Dako. We were told a couple of our items are on backorder for a few weeks. I am planning to call our account rep and voice my displeasure. It would have been nice to have received some communication regarding this "changeover" and perhaps we could have upped our order before all hell broke loose. Lynne Lynne A. Bell, HT (ASCP) Laboratory Central Vermont Hospital P.O. Box 547 Barre, VT 05641 (802)371-4122 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Dana Settembre Sent: Tue 4/11/2006 1:46 PM To: Sarah Holmes; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Dako unresponsive? I was waiting too long on hold so I left a complete voicemail message as they suggested and received a call back that they got my message. The processed my order. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Sarah Holmes 04/11/06 1:25 PM >>> Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah _______________________________________________ Histonet mailing list Histonet@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 Apr 11 13:06:04 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Apr 11 13:06:16 2006 Subject: [Histonet] New edition of Polak and van Noorden immunohistochemistry book Message-ID: <6.0.0.22.1.20060411115552.01b3bc98@gemini.msu.montana.edu> If you are looking for a concise, introductory book to immunohistochemical staining, I strongly suggest the long awaited new edition of: Introduction to ImmunoCytochemistry, 3rd Edition, JM Polak and S van Noorden, ISBN 185996 208 4. A paperback with 179 pages. This new edition was easy to follow and had excellent basic information. The appendix was packed with Technical notes on buffers, blocks, chromogens, etc. It cost ~$61 from Amazon.com 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 406 994-4303 (FAX) From NKMitche <@t> ahs.llumc.edu Tue Apr 11 13:09:00 2006 From: NKMitche <@t> ahs.llumc.edu (Mitchell, Nancy K.) Date: Tue Apr 11 13:09:04 2006 Subject: [Histonet] CSH meeting in May-Costa Mesa Message-ID: <8EB1459EA20E4E498AC2E05378F8F30A536EDA@mind.mc.ad.lluahsc.org> If any of you California Histoechs are out there, I am having problems getting a copy of the workshops that are going to be available. Who should I contact? Nancy Mitchell, HT ASCP Histology Supervisor Loma Linda Pathology Medical Group 11370 Anderson Street Ste 2960 Loma Linda, CA 92354 909-558-2012 FAX-909-796-3667 From mlm11 <@t> cornell.edu Tue Apr 11 13:21:15 2006 From: mlm11 <@t> cornell.edu (Mary Lou Norman) Date: Tue Apr 11 13:21:19 2006 Subject: [Histonet] tendon processing/cutting Message-ID: <5.2.1.1.2.20060411140143.02cbf850@postoffice9.mail.cornell.edu> Thanks Elizabeth and Gayle for the tendon info. I've done the longer soaking which does help quite a bit. I've got big hunky horse pieces. My question today: After the slide has dried all night on the warming tray I still see white tissue, definitely not as opaque as I'd like. Any suggestions on making sure the tissue doesn't fall off? I'm just the cutter so I don't know what IHC is going to be done. Could I put them in the oven 60o? What's wrong with melting the paraffin? Stacy, can I put them in the oven? Thanks, Mary Lou Norman From tpmorken <@t> labvision.com Tue Apr 11 13:24:58 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Tue Apr 11 13:25:10 2006 Subject: [Histonet] CSH meeting in May-Costa Mesa Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D843@usca0082k08.labvision.apogent.com> Nancy, you can download the program and registration information from www.californiahistology.org Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mitchell, Nancy K. Sent: Tuesday, April 11, 2006 11:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CSH meeting in May-Costa Mesa If any of you California Histoechs are out there, I am having problems getting a copy of the workshops that are going to be available. Who should I contact? Nancy Mitchell, HT ASCP Histology Supervisor Loma Linda Pathology Medical Group 11370 Anderson Street Ste 2960 Loma Linda, CA 92354 909-558-2012 FAX-909-796-3667 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Tue Apr 11 13:59:07 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Tue Apr 11 13:59:14 2006 Subject: [Histonet] Sudan Red 7B staining procedure Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176CD@lsexch.lsmaster.lifespan.org> I don't have an exact figure, but according to one of my older texts (Histopathologic Technic and Practical Histochemistry by Lillie and Fullmer), 4.25 grams Sudan Red 7B will dissolve in 100 ml absolute ethanol. So the solubility in 70% ethanol should be substantially less than that. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kim > Merriam > Sent: Tuesday, April 11, 2006 10:53 AM > To: Histonet > Subject: [Histonet] Sudan Red 7B staining procedure > > Hello, > > I have found a reference for staining fat with sudan red 7B, it calls > for a "saturated" solution of sudan red 7B in 70% ethanol. Does anyone > know what the saturation point of sudan red 7B is in 70% ethanol; a > general guide would be very helpful! > > Also - has anyone used this procedure before and what were the results > compared to a regular Oil Red O (both in isopropanol and/or propylene > glycol). > > Thanks in advance, > Kim > > > Kim Merriam, MA, HT(ASCP) > Novartis > Cambridge, MA > > --------------------------------- > Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From dgaupp <@t> tulane.edu Tue Apr 11 14:22:03 2006 From: dgaupp <@t> tulane.edu (dgaupp@tulane.edu) Date: Tue Apr 11 14:22:11 2006 Subject: [Histonet] re-freezing frozen sections Message-ID: <1144783323.443c01db190b1@webmail.tulane.edu> Histonet: I have tissues(monkey brain) that were fixed(4%Paraformaldehyde), cryprotected(20% sucrose), and flash froze(-70C isopentane). Unfortunately, mothernature(hurricane katrina) came threw and destroyed everything. This tissues sat for 3-4 months at blistering temps in the same position(melted OCT) & some even had a surprise for me(fungus). When the University let us come back into our building, I rinsed off OCT and put the tissues in 10%NBF at room temp. The tissue was immersed for 3-4months in fixative. I rinsed the tissue and once again, flash froze one of the sections, and they cut horribly. I don't know what to do. I am now tempting to paraffin process. I prefer to do frozens because its easier to get almost every section of tissue(one sections I usually get 500-700 slides). As most of you know, brain likes to explode in water, so I can do a better job if they are frozen. Any help, I would appreciate! Dina Dina D. Gaupp, B.S., M.T. Medical Research Specialist Center for Gene Therapy Tulane University Health Sciences Center JBJ Bldg/Rm 658, SL-99 1430 Tulane Avenue New Orleans, LA 70112 Lab: 504.988.1194 Fax: 504.988.7710 Email: dgaupp@tulane.edu From cfavara <@t> niaid.nih.gov Tue Apr 11 16:43:52 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Tue Apr 11 16:43:57 2006 Subject: [Histonet] re-freezing frozen sections Message-ID: I have done some squirrel monkey brain in paraffin with good results - happy to help with processing schedules etc if you like. Let me know c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: dgaupp@tulane.edu [mailto:dgaupp@tulane.edu] Sent: Tuesday, April 11, 2006 12:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re-freezing frozen sections Histonet: I have tissues(monkey brain) that were fixed(4%Paraformaldehyde), cryprotected(20% sucrose), and flash froze(-70C isopentane). Unfortunately, mothernature(hurricane katrina) came threw and destroyed everything. This tissues sat for 3-4 months at blistering temps in the same position(melted OCT) & some even had a surprise for me(fungus). When the University let us come back into our building, I rinsed off OCT and put the tissues in 10%NBF at room temp. The tissue was immersed for 3-4months in fixative. I rinsed the tissue and once again, flash froze one of the sections, and they cut horribly. I don't know what to do. I am now tempting to paraffin process. I prefer to do frozens because its easier to get almost every section of tissue(one sections I usually get 500-700 slides). As most of you know, brain likes to explode in water, so I can do a better job if they are frozen. Any help, I would appreciate! Dina Dina D. Gaupp, B.S., M.T. Medical Research Specialist Center for Gene Therapy Tulane University Health Sciences Center JBJ Bldg/Rm 658, SL-99 1430 Tulane Avenue New Orleans, LA 70112 Lab: 504.988.1194 Fax: 504.988.7710 Email: dgaupp@tulane.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From abilger <@t> ptd.net Tue Apr 11 19:04:54 2006 From: abilger <@t> ptd.net (Andrea C. Bilger) Date: Tue Apr 11 19:04:25 2006 Subject: [Histonet] Milestone microwave processor Message-ID: <058f01c65dc4$baebb320$0300a8c0@andrea> We had the RHS I in our lab for a couple of days to test it out. It performed well. The PA's didn't change how they grossed the tissue. This was a big plus. No need for special tools and tiny sized tissues. You can program it to the largest size specimen being processed. Our lab is looking into getting at least one of the small units and we may go for the new Pathos they just came out with. The Pathos is larger and automatically changes the solutions. No need for manually changing them like you do with the RHS I. Now if we can just convince the top brass to give us the money for one of them. Andrea Bilger From ekukurt <@t> yahoo.com Wed Apr 12 09:40:41 2006 From: ekukurt <@t> yahoo.com (Emre Kukurt) Date: Wed Apr 12 09:40:46 2006 Subject: [Histonet] One question Message-ID: <20060412144041.65778.qmail@web31001.mail.mud.yahoo.com> Hello Histonetters, Im new here, not sure whether or no my problem appropriate for the list topic. I have treated low electric current on grafted plants. Then, they were sectioned, prepared and photographed either light or electron microscope. I need to show them to an expert on plant histology who can interpret the pictures in order to decide what (sub)celullar changes appeared before/after electric current applied. Can some please one help me ? Best regards - Emre __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Jessica.Vacca <@t> HCAhealthcare.com Wed Apr 12 10:08:51 2006 From: Jessica.Vacca <@t> HCAhealthcare.com (Vacca Jessica) Date: Wed Apr 12 10:08:55 2006 Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tek VIP 2000 Message-ID: <41E16A15CE78374EA45B57E0F94339B84B5ADC@ORLEV01.hca.corpad.net> Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax From KevinMcGovern <@t> catholichealth.net Wed Apr 12 10:26:06 2006 From: KevinMcGovern <@t> catholichealth.net (McGovern, Kevin) Date: Wed Apr 12 10:26:25 2006 Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tekVIP 2000 Message-ID: Jessica, You may need two boxes, depending on how the instruments signal they are in an alarm state. In many cases, they simply short together their output wires, in some, they send out 5VDC. We have a VIP E-150 with a home-brewed alarm box that works well. Your local biomed should be able to cook one up if he has the electrical diagrams for the units. It ain't rocket science! Good luck. Kevin Kevin S. McGovern, BMETII Good Samaritan Hospital Clinical Engineering Dept. 10 East 31st Street Kearney, NE 68836 Phone: (308) 865-7051 Fax: (308) 865-2914 e-Mail: kevinmcgovern@catholichealth.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vacca Jessica Sent: Wednesday, April 12, 2006 10:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tekVIP 2000 Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mari.ann.mailhiot <@t> leica-microsystems.com Wed Apr 12 11:04:46 2006 From: mari.ann.mailhiot <@t> leica-microsystems.com (mari.ann.mailhiot@leica-microsystems.com) Date: Wed Apr 12 11:05:03 2006 Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tek VIP 2000 In-Reply-To: <41E16A15CE78374EA45B57E0F94339B84B5ADC@ORLEV01.hca.corpad.net> Message-ID: Jessica I have asked Mike Gallo to give you a call in regards to your alarm for the Leica processor. However, your biomedical engineers should be able to wire the alarm for you. If you have questions just give me a call. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Vacca Jessica" To Sent by: histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] Auto dialers (remote alarms) for leica and tissue tek 04/12/2006 10:08 VIP 2000 AM Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax _______________________________________________ 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 pruegg <@t> ihctech.net Wed Apr 12 11:43:25 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Apr 12 11:43:29 2006 Subject: [Histonet] QIHC In-Reply-To: <000a01c65cac$c89201a0$be4dff45@MEGAN> Message-ID: <200604121643.k3CGhNka013874@chip.viawest.net> Marian you can join the NSH IHC Resource Group online at www.ihcrg.org We have a list of references and you can actually access the ws on taking the QIHC exam presented by myself and Ethel Macrea at the NSH S/C last year. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marian powers Sent: Monday, April 10, 2006 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QIHC Looking for a good IHC reference to study for the QIHC? Thanks in advance, Marian _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vic <@t> vetmed.wsu.edu Wed Apr 12 12:55:57 2006 From: vic <@t> vetmed.wsu.edu (Leyva-Grado, Victor) Date: Wed Apr 12 12:56:01 2006 Subject: [Histonet] Questions about the mouse olfactory bulb Message-ID: <34EFB08480241347BEBE1F095ABB96F401C024@cvm36.vetmed.wsu.edu> Hi everybody, I'm working with mouse olfactory bulb (OB) for DAB-IHC and Alexa fluor double labeling. I've found several difficulties and I'd like to have your input about them. We perfused the mice with 4% PFA using a syringe (first saline then PFA) and we found out that usually the brain is well perfused and fixed, but the OB not always gets well fixed. Any suggestions on how can we improve the OB fixation to conserve better the histology of the tissue? A bad fixation can result in unspecific DAB labeling? In the other hand, when we do the immunofluorescence, the region of the glomerular layer is always fill with unspecific dots of fluorescence. Is this "endogenous fluorescence" or is a consequence of bad perfusion/fixation? Thanks a lot for your comments, Victor Leyva Washington State University From carl.hobbs <@t> kcl.ac.uk Wed Apr 12 13:30:46 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Wed Apr 12 13:31:35 2006 Subject: [Histonet] RE: New book Message-ID: Interesting that you use the phrase "immunohistochemistry" , when the book you recommend uses "immunocytochemistry". I'd be grateful for clarification of terms, Gail. Best wishes, Carl From gcallis <@t> montana.edu Wed Apr 12 14:25:10 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Apr 12 14:25:23 2006 Subject: [Histonet] Polak and von Noorden book title correction Message-ID: <6.0.0.22.1.20060412132352.01b09ac0@gemini.msu.montana.edu> It was pointed out to me the I had typed Immunohistochemistry in title of the 3rd Edtion of their new book. Please note the correction: Introduction to Immunocytochemistry 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 406 994-4303 (FAX) From rodger_65489 <@t> yahoo.com Wed Apr 12 14:29:39 2006 From: rodger_65489 <@t> yahoo.com (Roger Smithwell) Date: Wed Apr 12 14:51:59 2006 Subject: [Histonet] Opinions on the RHS-1; RHS-2, and the TT Mega Message-ID: <20060412192939.75934.qmail@web38313.mail.mud.yahoo.com> To all: If anyone has any pro's or con's with these three units (RHS-1; RHS-2; TT Mega) it would be greatly appreciated! Thanks in advance, Rodger --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From vic <@t> vetmed.wsu.edu Wed Apr 12 15:58:02 2006 From: vic <@t> vetmed.wsu.edu (Leyva-Grado, Victor) Date: Wed Apr 12 15:58:08 2006 Subject: [Histonet] Some details about my questions on mouse olfactory bulb perfusion/fixation. Message-ID: <34EFB08480241347BEBE1F095ABB96F401C025@cvm36.vetmed.wsu.edu> Hi everybody, I received some input about my questions on OB (thanks), but as some of you noticed I didn't include in my previous mail some details in the procedure that are important. So, here I go again. After collecting the brain, we post-fix it in 4% PFA for 4 h at 4C and then we sink the brain in 20% sucrose (usually overnight). After sucrose, we froze the tissue with dry ice and then we store them at -80C until we used them. We cut serial sections in a microtome (in a cold platform, 30um) and then we split some sections for DAB staining and some sections for double labeling. For double labeling we tried adding the two Alexa fluorochromes at the same time or using an incubation with Streptavidin Alexa 568 against the first Ab and then Alexa 488 for the second antibody. In both cases we had the endogenous fluorescence particularly in the glomerular layer, which unfortunately is one of the regions of the OB that I'm most interested in. Thanks a lot, Victor H Leyva Washington State University From Jodiputnam <@t> aol.com Wed Apr 12 16:29:43 2006 From: Jodiputnam <@t> aol.com (Jodiputnam@aol.com) Date: Wed Apr 12 16:29:49 2006 Subject: [Histonet] (no subject) Message-ID: <27a.81a2bfc.316ecb47@aol.com> Hi everyone. I have a few questions for you immunofluorescence experts out there. I recently have started doing IF on derm cases. I have noticed that quite a few of my sections are washing off or folding. So far the doctor has been able to make a diagnosis on all of the them, but I would like to improve my methods. After I cut the frozens, I dip the slides in acetone and allow them to dry completely. I've noticed that the sections start coming off or folding during my PBS washes. I'm very gentle while rinsing but still have problems. Any tips would be very much appreciated. Its so frustrating to have beautiful sections at the cryostat and end up with very little to show for the finished product. Thanks!! Jodi From tkngflght <@t> yahoo.com Wed Apr 12 16:59:26 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Wed Apr 12 16:59:30 2006 Subject: [Histonet] (no subject) In-Reply-To: <27a.81a2bfc.316ecb47@aol.com> Message-ID: <20060412215926.68317.qmail@web50902.mail.yahoo.com> Hi Jodi- As most of my methods are proven by practice, not by science, please try this on an old block first: When you cut the section before dipping in a fixative, hold the slide's empty side against the back of your hand or arm (covered of course by a glove) to warm the OCT and 'melt' the section onto the charged slide. As soon as you see the OCT liquefy, drop it gently into the acetone. We've used 95% etOH as a fixative as well--seems to be a bit gentler on things that roll off...but again I caution--practice on a case that has been signed out as a parallel. Cheryl Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. From AnthonyH <@t> chw.edu.au Wed Apr 12 17:53:31 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Wed Apr 12 17:53:41 2006 Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tek VIP 2000 Message-ID: We have two Leica TP1020's and despite extensive investigation and trials have been unable to connect them to our external auto-diallers. I truly wish that Leica would at least consider our dilemma and suggest a solution. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of mari.ann.mailhiot@leica-microsystems.com Sent: Thursday, 13 April 2006 2:05 AM To: Vacca Jessica Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Auto dialers (remote alarms) for leica and tissue tek VIP 2000 Jessica I have asked Mike Gallo to give you a call in regards to your alarm for the Leica processor. However, your biomedical engineers should be able to wire the alarm for you. If you have questions just give me a call. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Vacca Jessica" To Sent by: histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] Auto dialers (remote alarms) for leica and tissue tek 04/12/2006 10:08 VIP 2000 AM Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax _______________________________________________ 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 ______________________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Wed Apr 12 19:42:14 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Wed Apr 12 19:42:16 2006 Subject: [Histonet] (no subject) References: <27a.81a2bfc.316ecb47@aol.com> Message-ID: <000f01c676ef$3b98f8a0$e8bd0b43@yourxhtr8hvc4p> Jodi, we cut ours on plus slides and let them air dry. We don't dip them in acetone. If they are to be stained the next day, we leave them in a refrigerator overnight. Hope this helps. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Wednesday, April 12, 2006 4:29 PM Subject: [Histonet] (no subject) > Hi everyone. I have a few questions for you immunofluorescence experts out > there. I recently have started doing IF on derm cases. I have noticed that > quite a few of my sections are washing off or folding. So far the doctor > has been > able to make a diagnosis on all of the them, but I would like to improve > my > methods. After I cut the frozens, I dip the slides in acetone and allow > them > to dry completely. I've noticed that the sections start coming off or > folding > during my PBS washes. I'm very gentle while rinsing but still have > problems. > Any tips would be very much appreciated. Its so frustrating to have > beautiful sections at the cryostat and end up with very little to show > for the > finished product. > > Thanks!! > Jodi > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > Internal Virus Database is out-of-date. > Checked by AVG Free Edition. > Version: 7.1.385 / Virus Database: 268.4.1/309 - Release Date: 4/11/2006 > > From Eric <@t> ategra.com Wed Apr 12 20:50:04 2006 From: Eric <@t> ategra.com (Eric Dye (ext 223)) Date: Wed Apr 12 20:38:23 2006 Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed- Immediate openings Message-ID: Fellow-Histonetters I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) Here are some of my Hottest Histology Jobs: 1. Ohio (Southwestern Ohio) (Full-time, Perm, Histo Manager) 2. Northern New Jersey (Full-time, Perm and/or Temp, Bench Histo Tech, 1st shift) 3. Rhode Island - Providence (Full-time, Perm, Bench Histo Tech, 2nd Shift) 4. Northern New Jersey (Full-time, Perm, Bench Histo Tech) 5. Ohio (Southwestern Ohio) (Full-time, Perm, Bench Histo Tech) 6. New York (Long Island) (Full-time, Perm, SUPERVISOR Histo Tech) 7. Virginia(South - Coastal) (Full-time, Perm, Bench Histo Tech) 8. Virginia - Metro DC (Full-time, Perm, Lead Histo Tech) 9. South Florida (Full-time, Perm, Supervisor Histo Tech) 10. South Florida (Full-time, Perm, Bench Histo Tech) 11. Mass (Boston) (Part-time, Perm, Bench Histo Tech) 12. West Viriginia (Full-time, Perm, Bench Histo Tech) 13. Texas (Austin area) (Full-time, Perm, Supervisor and Bench Histo Tech) The clients are currently interviewing Histology candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- Note: this message is intended for: Fellow-Histonetters at histonet@lists.utsouthwestern.edu ---------------------------------------------------------------- ---------------------------------------------------------------- If you received this by mistake, or if you wish not to hear from me, please shoot me a mail to let me know and I'll not mail you again. ---------------------------------------------------------------- From gu.lang <@t> gmx.at Thu Apr 13 02:45:03 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Apr 13 02:45:08 2006 Subject: [Histonet] AW: immunofluorescence In-Reply-To: <000f01c676ef$3b98f8a0$e8bd0b43@yourxhtr8hvc4p> Message-ID: <000101c65ece$2d3a57b0$eeeea8c0@SERVER01> Joe, How long is it possible to store the slides in the refrigerator before staining? One, two days? Do you store them in simple boxes, or with a desiccant? Gudrun Lang Linz, Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Joe Nocito Gesendet: Sonntag, 14. Mai 2006 02:42 An: Jodiputnam@aol.com; histonet@lists.utsouthwestern.edu Betreff: Re: [Histonet] (no subject) Jodi, we cut ours on plus slides and let them air dry. We don't dip them in acetone. If they are to be stained the next day, we leave them in a refrigerator overnight. Hope this helps. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Wednesday, April 12, 2006 4:29 PM Subject: [Histonet] (no subject) > Hi everyone. I have a few questions for you immunofluorescence experts out > there. I recently have started doing IF on derm cases. I have noticed that > quite a few of my sections are washing off or folding. So far the doctor > has been > able to make a diagnosis on all of the them, but I would like to improve > my > methods. After I cut the frozens, I dip the slides in acetone and allow > them > to dry completely. I've noticed that the sections start coming off or > folding > during my PBS washes. I'm very gentle while rinsing but still have > problems. > Any tips would be very much appreciated. Its so frustrating to have > beautiful sections at the cryostat and end up with very little to show > for the > finished product. > > Thanks!! > Jodi > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > Internal Virus Database is out-of-date. > Checked by AVG Free Edition. > Version: 7.1.385 / Virus Database: 268.4.1/309 - Release Date: 4/11/2006 > > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fferreir <@t> ibmc.up.pt Thu Apr 13 06:49:48 2006 From: fferreir <@t> ibmc.up.pt (Fatima Ferreira) Date: Thu Apr 13 07:09:53 2006 Subject: [Histonet] Murine PNS neurodegeneration markers Message-ID: <1144928988.443e3adc96b85@webmail.ibmc.up.pt> Hello Everyone! I'm looking for peripheral axonal degeneration marker antibodies that would work in a murine model. Any leads? Regards, Fatima IBMC-Porto Portugal ---------------------------------------------------------------- This message was sent using IMP, the Internet Messaging Program. From cpomajzl <@t> cpllabs.com Thu Apr 13 09:22:13 2006 From: cpomajzl <@t> cpllabs.com (Chris Pomajzl) Date: Thu Apr 13 09:34:38 2006 Subject: [Histonet] Immunofluorescence - CAP 21850 Message-ID: <001c01c65f05$ae2b5b30$26fca8c0@CSP> Revisiting this CAP question regarding appropriate controls for IF: I understand this question to read that internal controls can be used on renal bx's, but what about skin bx's? In speaking with one of our dermatopathologists, he has asked some colleagues, and they claim that a positive tissue control needs to be run for each antibody tested. Is this correct? And if so, how does one acquire positive tissue other than from a positive case? If we do not have any positive controls at the moment, how can we do IF's and be of positive/negative fluorescence? I am concerned because we have not been running positive controls for several years now. I know that this is a Phase II NEW question from CAP, so I want to make sure that we address the issue properly. Thanks, Chris From pmcardle <@t> ebsciences.com Thu Apr 13 09:53:02 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Thu Apr 13 09:53:23 2006 Subject: [Histonet] RE: New book In-Reply-To: References: Message-ID: <443E65CE.3090001@ebsciences.com> Hi: I believe the difference between the two terms is: Immunocytochemistry pertains to cultured cells (and possibly suspensions?), while immunohistochemistry pertains to sectioned tissue. (If I'm wrong, I'd appreciate someone setting me straight, too!) Best regards, Phil McArdle Carl Hobbs wrote: > Interesting that you use the phrase "immunohistochemistry" , when the book > you recommend uses "immunocytochemistry". I'd be grateful for clarification > of terms, Gail. > Best wishes, > Carl > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson From Rcartun <@t> harthosp.org Thu Apr 13 09:58:43 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Thu Apr 13 09:59:38 2006 Subject: [Histonet] Immunofluorescence - CAP 21850 Message-ID: We use frozen tonsil (readily available) as our positive control for immunofluorescence studies performed on kidney and skin. It will label for IgG, IgA, IgM, C3, kappa, lambda, and fibrinogen. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "Chris Pomajzl" 04/13/06 10:22AM >>> Revisiting this CAP question regarding appropriate controls for IF: I understand this question to read that internal controls can be used on renal bx's, but what about skin bx's? In speaking with one of our dermatopathologists, he has asked some colleagues, and they claim that a positive tissue control needs to be run for each antibody tested. Is this correct? And if so, how does one acquire positive tissue other than from a positive case? If we do not have any positive controls at the moment, how can we do IF's and be of positive/negative fluorescence? I am concerned because we have not been running positive controls for several years now. I know that this is a Phase II NEW question from CAP, so I want to make sure that we address the issue properly. Thanks, Chris _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From fredericoacazevedo <@t> gmail.com Thu Apr 13 10:20:45 2006 From: fredericoacazevedo <@t> gmail.com (frederico azevedo) Date: Thu Apr 13 10:20:49 2006 Subject: [Histonet] Need 2 digital articles. Message-ID: Hi Histonetters, I need desesperately to get two digital articles but I don?t have access to those journals. I think it?s faster to ask you all than the author. The articles are kind of old. Could someone please help me? Title: The fuzzy logic of visuomotor control Author: Prochazka A. Journal:* *Can J Physiol Pharmacol 1996 Apr;74(4):456-62. Title: Development of a fuzzy logic based system to monitor the electrical responses of nerve fiber. Authors: al-Holou N, Joo DS. Journal: Biomed Sci Instrum 1997;33:376-81. Thank you very much! -- Frederico A. C. Azevedo Dept. of Anatomy - Neuroplasticity UFRJ, Brasil From cfavara <@t> niaid.nih.gov Thu Apr 13 10:40:46 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Thu Apr 13 10:40:53 2006 Subject: [Histonet] Need 2 digital articles. Message-ID: You should be able to access the NIH Library in Bethesda and they can help you. That way all authors' rights will be honored. I have included the web site. http://nihlibrary.nih.gov/ Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: frederico azevedo [mailto:fredericoacazevedo@gmail.com] Sent: Thursday, April 13, 2006 8:21 AM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] Need 2 digital articles. Hi Histonetters, I need desesperately to get two digital articles but I don?t have access to those journals. I think it?s faster to ask you all than the author. The articles are kind of old. Could someone please help me? Title: The fuzzy logic of visuomotor control Author: Prochazka A. Journal:* *Can J Physiol Pharmacol 1996 Apr;74(4):456-62. Title: Development of a fuzzy logic based system to monitor the electrical responses of nerve fiber. Authors: al-Holou N, Joo DS. Journal: Biomed Sci Instrum 1997;33:376-81. Thank you very much! -- Frederico A. C. Azevedo Dept. of Anatomy - Neuroplasticity UFRJ, Brasil _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From lpjones <@t> srhs-pa.org Thu Apr 13 11:27:14 2006 From: lpjones <@t> srhs-pa.org (Jones, Laura) Date: Thu Apr 13 11:27:29 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: <8E7AD740937B954F947F0DB4467EFEE056E978@mail.srhs-pa.org> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? From BlazekL <@t> childrensdayton.org Thu Apr 13 11:37:42 2006 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Thu Apr 13 11:38:32 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RRA <@t> Stowers-Institute.org Thu Apr 13 11:39:51 2006 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Thu Apr 13 11:40:16 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: I love that show! Especially the big honk of tissue that was only enough for 2 tests! Rhonda Allen Stowers Institute 1000 E. 50th street Kansas City, Missouri 64110 rra@stowers-institute.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jones, Laura Sent: Thursday, April 13, 2006 11:27 AM To: Histonet (E-mail) Subject: [Histonet] Anyone catch "House" the other night? Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From steven.p.postl <@t> abbott.com Thu Apr 13 11:41:28 2006 From: steven.p.postl <@t> abbott.com (Steven P Postl) Date: Thu Apr 13 11:41:56 2006 Subject: [Histonet] Anyone catch "House" the other night? In-Reply-To: <8E7AD740937B954F947F0DB4467EFEE056E978@mail.srhs-pa.org> Message-ID: Great catch. I was thinking the same thing. It was the fastest CD68 stain ever done. Also, I never been to a hospital were the doctors do everything from MRI, cat scans, colonoscopy to brain surgery, histology, serology, immunochemistry. "Jones, Laura" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/13/2006 11:27 AM To "Histonet (E-mail)" cc Subject [Histonet] Anyone catch "House" the other night? Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From la.sebree <@t> hosp.wisc.edu Thu Apr 13 11:45:44 2006 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Thu Apr 13 11:45:54 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: Their "procedure" was so far removed from the real thing, I didn't even realize that it was an immuno procedure they were trying to portray! I still love the show even if those fellows can do every thing under the sun; the production budget must not allow for actors portraying technologists! Linda Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-3224 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jones, Laura Sent: Thursday, April 13, 2006 11:27 AM To: Histonet (E-mail) Subject: [Histonet] Anyone catch "House" the other night? Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Thu Apr 13 11:34:30 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Apr 13 11:46:46 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: I saw that, too. But honestly, isn't it more fun to watch them do it their way? Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jones, Laura Sent: Thursday, April 13, 2006 12:27 PM To: Histonet (E-mail) Subject: [Histonet] Anyone catch "House" the other night? Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Barry.R.Rittman <@t> uth.tmc.edu Thu Apr 13 11:52:31 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Thu Apr 13 11:52:34 2006 Subject: [Histonet] RE: New book Message-ID: Both terms are derived from Immunochemistry - the chemistry of immunological phenomena. Immunocytochemistry has to do with the study of cell components by immunological methods often antibodies. Immunohistochemistry is much broader and is the microscopic localization of specific antigens in tissues usually by the use of antibodies. While these are the usual definitions, we also usually include other reactions such as lectin binding. In some instances the terms overlap in their usage. Both are often applied when immunochemistry is used on cells in culture, smears, on tissue sections and on blocks of tissue etc. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Phil McArdle Sent: Thursday, April 13, 2006 9:53 AM To: Carl Hobbs; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] RE: New book Hi: I believe the difference between the two terms is: Immunocytochemistry pertains to cultured cells (and possibly suspensions?), while immunohistochemistry pertains to sectioned tissue. (If I'm wrong, I'd appreciate someone setting me straight, too!) Best regards, Phil McArdle Carl Hobbs wrote: > Interesting that you use the phrase "immunohistochemistry" , when the book > you recommend uses "immunocytochemistry". I'd be grateful for clarification > of terms, Gail. > Best wishes, > Carl > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Jackie.O'Connor <@t> abbott.com Thu Apr 13 11:54:56 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Thu Apr 13 11:55:23 2006 Subject: [Histonet] Anyone catch "House" the other night? In-Reply-To: Message-ID: That's because they don't have any nurses, either. If someone can get me Dr. House's phone number, tell him to call me. Jackie O' Steven P Postl Sent by: histonet-bounces@lists.utsouthwestern.edu 04/13/2006 11:41 AM To "Jones, Laura" cc "Histonet \(E-mail\)" , histonet-bounces@lists.utsouthwestern.edu Subject Re: [Histonet] Anyone catch "House" the other night? Great catch. I was thinking the same thing. It was the fastest CD68 stain ever done. Also, I never been to a hospital were the doctors do everything from MRI, cat scans, colonoscopy to brain surgery, histology, serology, immunochemistry. "Jones, Laura" Sent by: histonet-bounces@lists.utsouthwestern.edu 04/13/2006 11:27 AM To "Histonet (E-mail)" cc Subject [Histonet] Anyone catch "House" the other night? Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@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 mtitford <@t> aol.com Thu Apr 13 11:56:25 2006 From: mtitford <@t> aol.com (mtitford@aol.com) Date: Thu Apr 13 11:56:32 2006 Subject: [Histonet] Another "House" fan. Message-ID: <8C82D32557D6E00-C74-215C@MBLK-M36.sysops.aol.com> I love that show, especially when he is rude to everyone and never has to apologize! One day I must sit down with a calculator and figure out how large the bill is for one of his patients. Let me see: four doctors for one patient, numerous exotic tests (sometimes even a transplant "rushed through" the committee), house calls to patients homes, etc. Then there is that glass fronted office.... Strangly though many of the medical predicaments Dr House and his staff face really do exist. That show must have good writers. Mike Titford USA Pathology Mobile AL From jqb7 <@t> cdc.gov Thu Apr 13 12:13:26 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Apr 13 12:13:53 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: Hey, they stretched it to 3...give them credit! Jeanine -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Allen, Rhonda Sent: Thursday, April 13, 2006 12:40 PM To: Jones, Laura; Histonet (E-mail) Subject: RE: [Histonet] Anyone catch "House" the other night? I love that show! Especially the big honk of tissue that was only enough for 2 tests! Rhonda Allen Stowers Institute 1000 E. 50th street Kansas City, Missouri 64110 rra@stowers-institute.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jones, Laura Sent: Thursday, April 13, 2006 11:27 AM To: Histonet (E-mail) Subject: [Histonet] Anyone catch "House" the other night? Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From carl.hobbs <@t> kcl.ac.uk Thu Apr 13 12:25:27 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Thu Apr 13 12:25:38 2006 Subject: [Histonet] re: Some details about my questions on mouse olfactory Message-ID: Just to confirm: do you get the same "endog. pattern" when using DAB-based immunolocalisation? If not, and they are normal tissues, one should not get those "unspecific dots", UNLESS they are due to aggregates of the fluorochrome-labelled secondary reagent. A pic would be good. Try posting here http://www.immunoportal.com/index.php as well? Also, are you using free-floating sections? Depending on the requirements, I will take a perfused-fixed brain and and cut it parasaggitally.Then further fix, as you do. However, if I'm interested in the OB only, I'll cut them off after disection and fix separately. Given that your procedure is as you state, I can't see a problem with "inadequate " fixation. Carl From gcallis <@t> montana.edu Thu Apr 13 12:52:34 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Apr 13 12:52:46 2006 Subject: [Histonet] re: Some details about my questions on mouse olfactory In-Reply-To: References: Message-ID: <6.0.0.22.1.20060413114328.01b3ab68@gemini.msu.montana.edu> A few more thoughts. One way to get rid of unwanted fluorescent aggregates is to spin down a diluted antibody-fluorophore just before application. We call this annoying problem "glowing garbage" although most of the time it is not fine punctate dots, but bigger clumps of stuff. Also, if one uses Strepavidin-Alexa dye conjugates and the tissue is notorious (or you want to just make sure) for containing endogenous biotin, a Strepavidin or avidin biotin block step should be added. Vector is one source for both kits. Molecular Probes advised never diluting Strepavidin-Alexa conjugates with normal serum in the buffer, the SA can bind to any endogenous biotin found in the normal serum. A buffer containing Tween 20 was acceptable. At 11:25 AM 4/13/2006, you wrote: >Just to confirm: do you get the same "endog. pattern" when using DAB-based >immunolocalisation? If not, and they are normal tissues, one should not get >those "unspecific dots", UNLESS they are due to aggregates of the >fluorochrome-labelled secondary reagent. >A pic would be good. > Try posting here http://www.immunoportal.com/index.php as well? >Also, are you using free-floating sections? >Depending on the requirements, I will take a perfused-fixed brain and and >cut it parasaggitally.Then further fix, as you do. However, if I'm >interested in the OB only, I'll cut them off after disection and fix >separately. Given that your procedure is as you state, I can't see a problem >with "inadequate " fixation. >Carl > > >_______________________________________________ >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 406 994-4303 (FAX) From POWELL_SA <@t> Mercer.edu Thu Apr 13 13:40:59 2006 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Thu Apr 13 13:41:40 2006 Subject: [Histonet] Anyone catch "House" the other night? In-Reply-To: Message-ID: <01M17O621QEE8X2034@Macon2.Mercer.edu> Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@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 c.m.vanderloos <@t> amc.uva.nl Thu Apr 13 13:59:56 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Apr 13 14:00:23 2006 Subject: [Histonet] RE: Polak and von Noorden book title correction Message-ID: <14a2f414e8b6.14e8b614a2f4@amc.uva.nl> Hi all, I don't think that Gayle is the one who should apologize for her "mistake" with respect to the terms "immunohistochemistry" and "immunocytochemistry". In fact she was right. I had the honour to discuss this over with Susan van Noorden in London a couple of years ago. She agreed with me that those two terms are not always used in the right way. Especially after she saw our very different immunoenzymatic staining results of anti-IFNgamma on intact cells and tissue sections from embedded cells (that was my topic of the NSH international lecture in 2001 - Charlotte. NC). To my opinion it's a bit missed chance that the authors continue with the term "immunocytochemistry" where mostly tissue work is concerned. I totally agree with Phil McArdle that "immunohistochemistry" stands for immunoenzymatic staining of tissue sections and "immunocytochemistry" for the immunoenzymatic staining of cell specimens. Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Wed, 12 Apr 2006 13:25:10 -0600 From: Gayle Callis Subject: [Histonet] Polak and von Noorden book title correction To: Histonet@lists.utsouthwestern.edu It was pointed out to me the I had typed Immunohistochemistry in title of the 3rd Edtion of their new book. Please note the correction: Introduction to Immunocytochemistry 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 406 994-4303 (FAX) From dmarsha3 <@t> utmem.edu Thu Apr 13 14:01:06 2006 From: dmarsha3 <@t> utmem.edu (Dana Marshall) Date: Thu Apr 13 14:01:13 2006 Subject: [Histonet] Anyone catch "House" the other night? References: <01M17O621QEE8X2034@Macon2.Mercer.edu> Message-ID: <001501c65f2c$9e1788a0$f623c084@DanaM> one time on The X-files Scully had 4 hours until a meeting and she needed to prove she was infected with the alien virus at the meeting. She took her own blood, purified lymphocytes, extracted DNA, digested the DNA, ran a gel, transferred the gel to a filter to which she hybridized a "really hot probe" that she had also managed to synthesize/purify (not clear) and label while the gel was running and transferring, then washed the blot and exposed it to film and got to the meeting with the data. I am still trying to recover from that bastardization of science (and that has been quite a few years now) so in the meantime I simply watch House because of Hugh Laurie (I digress from this digression but the actor who plays his friend Wilson doesn't get the kudos he deserves for playing his character) although one of my colleagues is totally torn up over the fact that House uses his cane in the wrong hand ;-P whew! ok, back to reality. dm ----- Original Message ----- From: "Shirley Powell" To: "'Linda Blazek'" ; ; Sent: Thursday, April 13, 2006 1:40 PM Subject: RE: [Histonet] Anyone catch "House" the other night? > > Turn off the TV get in the Jacuzzi, have a glass of wine and relax. > Work is work, TV is TV and they get paid a lot more than we do for doing > histology wrong. Go figure. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda > Blazek > Sent: Thursday, April 13, 2006 11:38 AM > To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org > Subject: Re: [Histonet] Anyone catch "House" the other night? > > My husband turned to me as said "I thought you said doing immunos was a > complicted thing." > >>>> "Jones, Laura" 4/13/2006 12:27 PM >>> > > Dr. House and his colleagues were doing immuno on Tuesday night! > Procedure: > > Take large chunk of heart tissue which has not been frozen or in fixative, > but in a petri dish in the fridge, and flop it on to your microscope > slide. > Put the slide with the large chunk of tissue on it under the scope. Add > "one microlitre of immunoperoxidase" to the tissue. Look into scope. If > it > turns red, it's positive. > > I wish our immuno were that easy and quick. Get a Histo consultant on > staff, please! :o) > > P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, > and then test it all? In evening gowns and tuxes? > > _______________________________________________ > Histonet mailing list > Histonet@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 LRaff <@t> lab.uropartners.com Thu Apr 13 14:05:38 2006 From: LRaff <@t> lab.uropartners.com (Lester Raff) Date: Thu Apr 13 14:05:44 2006 Subject: [Histonet] Tissue transfer Message-ID: <5DA1CA5D0B98A84985B545A24423B822A7E7@UPLAB01.uplab.local> I have been reading about a procedure for transferring small fragments of tissue from stained slides to charged slides for IHC. The procedure calls for Mount-Quick mounting medium. I have found this on the web, but don't know whether the procedure replies to the aqueous based or solvent based product. 1. Have people used a transfer procedure like this effectively? 2. Which Mount-Quick is used? 3. Would anyone like to share a their protocol? Thanks Lester J. Raff, MD Medical Director UroPartners, LLC Laboratory ph: 708-486-0076 fax: 708-486-0080 From NKMitche <@t> ahs.llumc.edu Thu Apr 13 15:07:27 2006 From: NKMitche <@t> ahs.llumc.edu (Mitchell, Nancy K.) Date: Thu Apr 13 15:07:33 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: <8EB1459EA20E4E498AC2E05378F8F30A536EEB@mind.mc.ad.lluahsc.org> TV does have its "moments" though. I personally would not mind working with a pathologist that looks like: George Clooney, Brad Pitt, Keanu Reeves, etc. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Shirley Powell Sent: Thursday, April 13, 2006 11:41 To: 'Linda Blazek'; Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: RE: [Histonet] Anyone catch "House" the other night? Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality 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 jqb7 <@t> cdc.gov Thu Apr 13 15:11:03 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Thu Apr 13 15:11:14 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: I have my pitcher of Stoli Dolis in the refrig as we speak....... Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Shirley Powell Sent: Thursday, April 13, 2006 2:41 PM To: 'Linda Blazek'; Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: RE: [Histonet] Anyone catch "House" the other night? Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@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 POWELL_SA <@t> Mercer.edu Thu Apr 13 15:48:37 2006 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Thu Apr 13 15:49:10 2006 Subject: [Histonet] Anyone catch "House" the other night? In-Reply-To: <8EB1459EA20E4E498AC2E05378F8F30A536EEB@mind.mc.ad.lluahsc.org> Message-ID: <01M17SMAXV7K8X23YZ@Macon2.Mercer.edu> You do have a point. -----Original Message----- From: Mitchell, Nancy K. [mailto:NKMitche@ahs.llumc.edu] Sent: Thursday, April 13, 2006 3:07 PM To: Shirley Powell; Linda Blazek; Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: RE: [Histonet] Anyone catch "House" the other night? TV does have its "moments" though. I personally would not mind working with a pathologist that looks like: George Clooney, Brad Pitt, Keanu Reeves, etc. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Shirley Powell Sent: Thursday, April 13, 2006 11:41 To: 'Linda Blazek'; Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: RE: [Histonet] Anyone catch "House" the other night? Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality 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 cjkelley <@t> cmh.edu Thu Apr 13 16:18:05 2006 From: cjkelley <@t> cmh.edu (Kelley, Cynthia,) Date: Thu Apr 13 16:18:17 2006 Subject: [Histonet] Job Posting Message-ID: We currently have an opening for a registered Histology Technician in our laboratory. This is a unique full time day shift position that will rotate in all areas of Histology including routine procedures, IHC, and some specimen grossing. If you are interested in this great opportunity please log on to the childrens-mercy.org website and apply on-line. Cynthia Kelley Laboratory Services Manager Children's Mercy Hospital & Clinics 2401 Gillham Road Kansas City, MO 64108 816.234.3023 office 816.821.9224 pager From bjdewe <@t> aol.com Thu Apr 13 17:09:03 2006 From: bjdewe <@t> aol.com (bjdewe@aol.com) Date: Thu Apr 13 17:09:10 2006 Subject: [Histonet] CD31 Message-ID: <8C82D5E024966D9-1CBC-803@MBLK-M39.sysops.aol.com> Can anyone share their protocol for doing IHC on rate tissue. I have been going nuts trying to get it to work and I need some advice... I have been using Santa Cruz Bio catalogue # SC-1506... Cheers, Lorie Compassion, in which all ethics must take root, can only attain its full breadth and depth if it embraces all living creatures and does not limit itself to human kind". ~ Albert Einstein ******************************************************************* Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s)and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ******************************************************************* From ploykasek <@t> phenopath.com Thu Apr 13 17:38:03 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Thu Apr 13 17:39:29 2006 Subject: [Histonet] Cdx antibody Message-ID: Hi all. I was wondering if anyone can recommend an alternate source for antibody Cdx2 clone CDX2-88. We are currently buying this antibody from Biogenex. The price has just increased exponentially. I'll bite the bullet if I have to, but would prefer another source. Thank you. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From liz <@t> premierlab.com Thu Apr 13 16:01:20 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Thu Apr 13 19:48:47 2006 Subject: [Histonet] BD hamster anti-mouse UC3-10A6 Message-ID: <000601c65f3d$6a4b4780$0300a8c0@Chlipala> Has anyone used this antibody it from BD Biosciences on FFPE tissue. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From Malcolm.McCallum <@t> tamut.edu Thu Apr 13 19:56:49 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Thu Apr 13 19:58:09 2006 Subject: [Histonet] New Herpetology Journal Message-ID: While this may not fit histology directly, I figured some on here might be interested. We announce the release of the new journal "Herpetological Conservation and Biology." This journal is dedicated to the conservation, natural history, and ecology of amphibians and reptiles. The journal has no page charges and no download fees and is run by a group of 50 scientists from around the world. HCB follow ICZN guidelines for deposition of permanent copies of the journal and has been carefully orchestrated to qualify for inclusion in the ISI journal impact ratings. The journal is/will be indexed by biosys, google scholar, medline, and others. Please check out the HCB webpage www.herpconbio.org I hope you enjoy perusing the new journal's website and consider submitting an article Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html From lr_hsvlle2 <@t> yahoo.com.au Thu Apr 13 21:09:28 2006 From: lr_hsvlle2 <@t> yahoo.com.au (Lorraine Rolston) Date: Thu Apr 13 21:09:37 2006 Subject: [Histonet] Histology learning & teaching resource. Message-ID: <20060414020928.57692.qmail@web30503.mail.mud.yahoo.com> This site is certainly worth a look! http://www.visualhistology.com/Visual_Histology_Atlas/ Cheers, Lorraine Send instant messages to your online friends http://au.messenger.yahoo.com From Malcolm.McCallum <@t> tamut.edu Thu Apr 13 21:26:04 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Thu Apr 13 21:26:20 2006 Subject: [Histonet] Histology learning & teaching resource. Message-ID: That is a GREAT SITE!!!!!! :) Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Lorraine Rolston Sent: Thu 4/13/2006 9:09 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Histology learning & teaching resource. This site is certainly worth a look! http://www.visualhistology.com/Visual_Histology_Atlas/ Cheers, Lorraine Send instant messages to your online friends http://au.messenger.yahoo.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mbmphoto <@t> gmail.com Thu Apr 13 22:53:08 2006 From: mbmphoto <@t> gmail.com (Maria Mejia) Date: Thu Apr 13 22:52:36 2006 Subject: [Histonet] Reichert Jung Histocut 820 II rotary microtome Message-ID: I have a Reichert Jung rotary microtome that needs servicing & possible repair. Can anyone recommend a service company or someone in the Bay Area (San Francisco) that can provide this? I would greatly appreciate any input. Yours Maria Bartola Mejia University of California San Francisco (UCSF) Department of Neurosurgery San Francisco, CA. 94103 From marjoh3 <@t> telus.net Fri Apr 14 06:51:07 2006 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Fri Apr 14 06:51:05 2006 Subject: [Histonet] Slide Mailers Message-ID: <001401c65fb9$b78cbc00$6401a8c0@VALUED20606295> Hi Histonetters, I have to send wet-Permount, H&E slides to one of my clients in another city, by courier.Turn-around time is critical. I've tried the plastic slide mailers and the cardboard ones, but the wet coverslips stick to the contacted surfaces. Does anyone have a special slide mailer or technique you can share with me? Thank you in advance. Marilyn Johnson Edmonton, Alberta, Canada From Barry.R.Rittman <@t> uth.tmc.edu Fri Apr 14 07:12:46 2006 From: Barry.R.Rittman <@t> uth.tmc.edu (Rittman, Barry R) Date: Fri Apr 14 07:14:47 2006 Subject: [Histonet] Slide Mailers Message-ID: Marilyn Just a suggestion - early in the morning and I've only had one cup of coffee. How about wrapping the slides in Teflon tape? Permount will not stick to this. Can get this in the plumbing section of hardware stores and some have this in different widths. Barry -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marilyn Johnson Sent: Friday, April 14, 2006 6:51 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Slide Mailers Hi Histonetters, I have to send wet-Permount, H&E slides to one of my clients in another city, by courier.Turn-around time is critical. I've tried the plastic slide mailers and the cardboard ones, but the wet coverslips stick to the contacted surfaces. Does anyone have a special slide mailer or technique you can share with me? Thank you in advance. Marilyn Johnson Edmonton, Alberta, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Don.Birgerson <@t> leica-microsystems.com Fri Apr 14 07:19:02 2006 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Fri Apr 14 07:19:12 2006 Subject: [Histonet] Reichert Jung Histocut 820 II rotary microtome In-Reply-To: Message-ID: Hi Maria, Reichert Jung is now called Leica-Microsystems. Service can be arranged by phoning "service dispatch." (1-800-248-0223 8:00 to 5:00 CDT) Best regards, Don Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 Maria Mejia To Sent by: histonet@lists.utsouthwestern.edu histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] Reichert Jung Histocut 820 II rotary microtome 04/13/2006 10:53 PM I have a Reichert Jung rotary microtome that needs servicing & possible repair. Can anyone recommend a service company or someone in the Bay Area (San Francisco) that can provide this? I would greatly appreciate any input. Yours Maria Bartola Mejia University of California San Francisco (UCSF) Department of Neurosurgery San Francisco, CA. 94103 _______________________________________________ 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 robert.fisher <@t> fda.hhs.gov Fri Apr 14 08:05:50 2006 From: robert.fisher <@t> fda.hhs.gov (Fisher, Robert) Date: Fri Apr 14 08:06:04 2006 Subject: [Histonet] IHC for NK cells in Balb/c mice? Message-ID: <2185E187D9585E40B796B256F5AA60B60CC14CE5@cbsms05> Can anyone suggest a protocol for staining NK cells in formalin-fixed, paraffin-embedded Balb/c mouse tissue? It is my understanding that the NK1.1 antibody only works in BL/6 background mice (but I'd love to be corrected on this point!). Thanks, Robert Robert W. Fisher, Ph.D. Staff Fellow FDA/CBER/OBRR/DH/LPD 301-496-2579 Robert.Fisher@fda.hhs.gov From Yves.Heremans <@t> vub.ac.be Fri Apr 14 08:22:25 2006 From: Yves.Heremans <@t> vub.ac.be (Yves Heremans) Date: Fri Apr 14 08:22:34 2006 Subject: [Histonet] Microscopy Today January 2006 Message-ID: Dear Histonetters, I am desperately trying to get hold of the January 2006 issue of Microscopy Today (I am interested in the section on mounting media and antifade reagents). If someone would be so kind to send me this issue, it would be very much appreciated. Yves From hts <@t> gator.net Fri Apr 14 09:12:44 2006 From: hts <@t> gator.net (hts@gator.net) Date: Fri Apr 14 09:12:49 2006 Subject: [Histonet] HT position in Gainesville, Florida Message-ID: <1432.65.81.70.231.1145023964.squirrel@webmail.gru.net> Lead Histotechnician position open in Gainesville, Florida. Come work in a Histotech owned lab. We are looking for a tech with excellent microtomy and organizational skills. Works directly with Lab Manager and owner. We perform routine and IHC histology for human diagnostics, biotech research, QA for biotech companies, and are CLIA and GLP Compliant. Excellent working environment, M-F, day shift, no weekends. HT(ASCP) required. Prefer 5 + years of experience. Florida licensure now only requires an HT(ASCP) license to apply. Competetive salary, good benefit package. Come live in the sunshine!! We did! Gainesville is located in North Central Florida, a short drive to the east and west beaches of Florida. University of Florida is located here, as well as many outdoor activities. Great for the single person or a family. A town of great diversity. Contact Beverly Robinson HT(ASCP) (352) 338-0045 From funderwood <@t> mcohio.org Fri Apr 14 09:37:42 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Fri Apr 14 09:38:09 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: I think there was an episode where they used a jacuzzi to perform HIER. >>> Shirley Powell 04/13/06 02:40PM >>> Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@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 RRA <@t> Stowers-Institute.org Fri Apr 14 09:41:37 2006 From: RRA <@t> Stowers-Institute.org (Allen, Rhonda) Date: Fri Apr 14 09:41:54 2006 Subject: [Histonet] Anyone catch "House" the other night? Message-ID: That sounds like a new discovery! We should try it. Rhonda Allen BA HT(ASCP)HTL, QIHC Histotechnology Specialist II Stowers Institute 1000 E. 50th Street Kansas City, MO 64110 816-926-4305 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Friday, April 14, 2006 9:38 AM To: BlazekL@childrensdayton.org; Histonet@lists.utsouthwestern.edu; POWELL_SA@Mercer.edu; lpjones@srhs-pa.org Subject: RE: [Histonet] Anyone catch "House" the other night? I think there was an episode where they used a jacuzzi to perform HIER. >>> Shirley Powell 04/13/06 02:40PM >>> Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From liz <@t> premierlab.com Fri Apr 14 10:01:20 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Apr 14 10:01:28 2006 Subject: [Histonet] detecting mouse tissue in porcine implants Message-ID: <000001c65fd4$4a26f390$0300a8c0@Chlipala> Hello everyone I have a question that I think might have been brought up before but I've searched the archives and I can't seem to find anything. Is there an immunomarker that will detect all mouse cells. I'm working with contructs that have been implanted into mice these constructs are seeded with epitheial cells from a different species (porcine), unfortunately most cytokeratin antibodies seem to cross react with mouse cytokeratin also and I need to prove that the cells within the contruct are not mouse cells, is there a way I can do this? thanks in advance Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 From BlazekL <@t> childrensdayton.org Fri Apr 14 10:13:14 2006 From: BlazekL <@t> childrensdayton.org (Linda Blazek) Date: Fri Apr 14 10:14:22 2006 Subject: [Histonet] Job opening Message-ID: Dear all, There is a current job opening here at Children's Medical Center, Dayton, OH as today is my last day. These are a really great bunch of people to work with and a great place to work. If you are interested go to the web site listed below and follow the links to job openings. Linda http://www.childrensdayton.org/ Linda Blazek, HT (ASCP) Department of Pathology Children's Medical Center Dayton, Ohio 45404 (937) 641-3358 fax (937)641-5482 blazekl@childrensdayton.org From koellinr <@t> amgen.com Fri Apr 14 10:43:12 2006 From: koellinr <@t> amgen.com (Koelling, Ray) Date: Fri Apr 14 10:43:26 2006 Subject: [Histonet] IHC for NK cells in Balb/c mice? Message-ID: <16834C6DFFA6004C88DE4507FB8AE54403947CA7@wa-mb4-sea.amgen.com> Robert, you are correct NK1.1 works in BL/6 backround (and a few others like NZB) but NOT in Balb/c, DBA, etc. Clone 2B4 is nice but similar MHC restriction to NK1.1 and is a mouse so you'd have mouse-on-mouse issues. Might try clone DX-5 which is a rat IgM and which is not restricted by haplotype and works in Balb/c mice. But have used these in flow and not in IHC so if you try, I'd stick with frozen or something like Zinc/tris fixative. And you probably know, there is no known NK cell only marker known so far. So in IHC you'd be picking up subpopulations of other cells (like small set of T's). By flow we can gate out through dual and triple lables to see only NK's. Ray Raymond Koelling Pathology Research Scientist Amgen Corp. Seattle, WA -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Fisher, Robert Sent: Friday, April 14, 2006 6:06 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] IHC for NK cells in Balb/c mice? Can anyone suggest a protocol for staining NK cells in formalin-fixed, paraffin-embedded Balb/c mouse tissue? It is my understanding that the NK1.1 antibody only works in BL/6 background mice (but I'd love to be corrected on this point!). Thanks, Robert Robert W. Fisher, Ph.D. Staff Fellow FDA/CBER/OBRR/DH/LPD 301-496-2579 Robert.Fisher@fda.hhs.gov From PMonfils <@t> Lifespan.org Fri Apr 14 11:18:05 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Fri Apr 14 11:18:10 2006 Subject: [Histonet] Tissue transfer Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176D2@lsexch.lsmaster.lifespan.org> I'm not familiar with Mount-Quick, but years ago I used to occasionally lift sections from slides, using a product called Pro-Texx. Here's the protocol I used: Remove coverslip from slide by immersion in xylene. One hour additional time in xylene, to ensure removal of mounting medium. While slide is damp with xylene, place a drop of Permount on any section that is to remain on the slide, and a drop of Pro-Texx on the section that is to be lifted, being careful the two media do not mix. Do not spread the media thin. You want a thick layer. Allow to dry thoroughly, using gentle heat (low setting of a slide warmer). Alternatively they can dry overnight at room temperature. Immerse slide in distilled water until the Pro-Texx loosens from the slide (the Permount will not). With forceps, peel the layer of Pro-Texx, containing the tissue section, off the slide. I did this many years ago and we did not have charged slides at that time, so I would place the Pro-Texx film, tissue side down, onto a slide smeared with a thin layer of albumen, lay it flat on the same warm surface, cover it with a small piece of polyethylene film, and place a flat weight on top of it to ensure good contact between tissue and slide until dry. Remove Pro-Texx by immersion in xylene. Presumably using charged slides might facilitate this final step. If the process you are doing is similar to this, then obviously you would want the solvent-based medium. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of > Lester Raff > Sent: Thursday, April 13, 2006 12:05 PM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Tissue transfer > > I have been reading about a procedure for transferring small fragments > of tissue from stained slides to charged slides for IHC. The procedure > calls for Mount-Quick mounting medium. I have found this on the web, > but don't know whether the procedure replies to the aqueous based or > solvent based product. > > > > 1. Have people used a transfer procedure like this effectively? > 2. Which Mount-Quick is used? > 3. Would anyone like to share a their protocol? > > > > Thanks > > > > Lester J. Raff, MD > Medical Director > UroPartners, LLC Laboratory > ph: 708-486-0076 > fax: 708-486-0080 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > From arrrid702 <@t> skmc.gov.ae Fri Apr 14 11:49:13 2006 From: arrrid702 <@t> skmc.gov.ae (Ridhwaan Arries) Date: Fri Apr 14 11:49:13 2006 Subject: [Histonet] RE: Histonet Digest, Vol 29, Issue 15 Message-ID: HENRY CHARLENE TRY NOVOCASTRA CD61. I HAVE JUST OPTIMISED IT FOR OUR LAB AND IT WORKS GREAT!!! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Wednesday, April 12, 2006 9:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, Issue 15 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. Dako unresponsive? (Sarah Holmes) 2. histogel (Steven Coakley) 3. Parathyroid hormone (Van Eyck, Deb) 4. RE: histogel (Bartlett, Jeanine) 5. Re: Dako unresponsive? (Chris Pomajzl) 6. CD61 (Henry, Charlene) 7. RE: CD61 (Drew Sally A.) 8. Cell block prep (Linda Hines) 9. Re: Dako unresponsive? (Dana Settembre) 10. Sudan Red 7B staining procedure (Kim Merriam) 11. RE: Dako unresponsive? (Bell, Lynne) 12. New edition of Polak and van Noorden immunohistochemistry book (Gayle Callis) 13. CSH meeting in May-Costa Mesa (Mitchell, Nancy K.) 14. tendon processing/cutting (Mary Lou Norman) 15. RE: CSH meeting in May-Costa Mesa (Morken, Tim - Labvision) 16. RE: Sudan Red 7B staining procedure (Monfils, Paul) 17. re-freezing frozen sections (dgaupp@tulane.edu) 18. RE: re-freezing frozen sections (Favara, Cynthia (NIH/NIAID) [E]) 19. Milestone microwave processor (Andrea C. Bilger) 20. One question (Emre Kukurt) 21. Auto dialers (remote alarms) for leica and tissue tek VIP 2000 (Vacca Jessica) 22. RE: Auto dialers (remote alarms) for leica and tissue tekVIP 2000 (McGovern, Kevin) 23. Re: Auto dialers (remote alarms) for leica and tissue tek VIP 2000 (mari.ann.mailhiot@leica-microsystems.com) 24. RE: QIHC (Patsy Ruegg) ---------------------------------------------------------------------- Message: 1 Date: Tue, 11 Apr 2006 12:25:49 -0500 From: "Sarah Holmes" Subject: [Histonet] Dako unresponsive? To: Message-ID: <001501c65d8c$f9ee81c0$780a010a@wp.comcast.net> Content-Type: text/plain; charset="iso-8859-1" Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah ------------------------------ Message: 2 Date: Tue, 11 Apr 2006 10:12:26 -0700 (PDT) From: Steven Coakley Subject: [Histonet] histogel To: Histonet@lists.utsouthwestern.edu Message-ID: <20060411171227.11273.qmail@web38206.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Ok, now I have successfull processed cells using histogel. Do I dry and stain the slides as usual? Steve --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. ------------------------------ Message: 3 Date: Tue, 11 Apr 2006 12:21:23 -0500 From: "Van Eyck, Deb" Subject: [Histonet] Parathyroid hormone To: "' histonet@lists.utsouthwestern.edu'" Message-ID: Content-Type: text/plain; charset="iso-8859-1" I agree with Richard-----it would be nice to know when you order a product that it is being discontinued. I trialed several PTH antibodies and I ended up liking the Novocastra PTH NCL-PTH-488 - it is research use only-------couldn't find a source for IVD PTH. Hope this helps! Deb Van Eyck Cytology/Immunohistochemistry Waukesha Memorial Hospital 725 American Avenue Waukesha,WI 53188 phone:262-928-2112 fax:262-928-4849 deb.vaneyck@phci.org 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. ------------------------------ Message: 4 Date: Tue, 11 Apr 2006 13:22:10 -0400 From: "Bartlett, Jeanine" Subject: RE: [Histonet] histogel To: "Steven Coakley" , Message-ID: Content-Type: text/plain; charset="iso-8859-1" Yep! Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Steven Coakley Sent: Tuesday, April 11, 2006 1:12 PM To: Histonet@lists.utsouthwestern.edu Subject: [Histonet] histogel Ok, now I have successfull processed cells using histogel. Do I dry and stain the slides as usual? Steve --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 5 Date: Tue, 11 Apr 2006 12:36:02 -0500 From: "Chris Pomajzl" Subject: Re: [Histonet] Dako unresponsive? To: "Sarah Holmes" , "HISTONET" Message-ID: <003a01c65d8e$6a9e0d40$26fca8c0@CSP> Content-Type: text/plain; charset="iso-8859-1" Dako is switching over to a new business system. So you are going to have some trouble. Supposedly, they were going to be able to take calls as of yesterday. We placed an order at the end of March, and our supplies just arrived today. Good luck. ----- Original Message ----- From: "Sarah Holmes" To: Sent: Tuesday, April 11, 2006 12:25 PM Subject: [Histonet] Dako unresponsive? Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Tue, 11 Apr 2006 12:38:57 -0500 From: "Henry, Charlene" Subject: [Histonet] CD61 To: Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1BEA@SJMEMXMB02.stjude.sjcrh.local> Content-Type: text/plain; charset="us-ascii" Dako has discontinued the CD61 (Dako M0753) antibody that we have been using for years and I was wondering if anyone knows of a good CD61 that I could try as a replacement? I have tried the Ventana CD61 (Y2/51) and Cell Marque CD61 (2f2) and they are not as good as our old antibody from Dako. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 ------------------------------ Message: 7 Date: Tue, 11 Apr 2006 12:45:12 -0500 From: "Drew Sally A." Subject: RE: [Histonet] CD61 To: "Histonet" Message-ID: Content-Type: text/plain; charset="us-ascii" We use the Cell Marque CD61 (clone 2f2) via Ventana, and it seems to work well for us with mild CC1 on our VMS instruments. Sally Ann Drew, MT(ASCP) IHC/ISH Laboratory University of Wisconsin Hosp. & Clinics Madison, WI 53792 (608)265-6596 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Henry, Charlene Sent: Tuesday, April 11, 2006 12:39 PM To: histonet@pathology.swmed.edu Subject: [Histonet] CD61 Dako has discontinued the CD61 (Dako M0753) antibody that we have been using for years and I was wondering if anyone knows of a good CD61 that I could try as a replacement? I have tried the Ventana CD61 (Y2/51) and Cell Marque CD61 (2f2) and they are not as good as our old antibody from Dako. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 8 Date: Tue, 11 Apr 2006 10:46:50 -0700 (PDT) From: Linda Hines Subject: [Histonet] Cell block prep To: histonet Message-ID: <20060411174650.78180.qmail@web33603.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hi! All, Thank-you for the responses regarding the thermometer's. Are any of you working with the Thin-prep and preparing cytology blocks upon completing the slide preparation? My question is how we prepare the cell blocks. Any information is greatly appreciated. We are a new lab, and this is the first time I have'nt made my own smears etc. Thank-you Linda Hines HT (ASCP) --------------------------------- Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1¢/min. ------------------------------ Message: 9 Date: Tue, 11 Apr 2006 13:46:17 -0400 From: "Dana Settembre" Subject: Re: [Histonet] Dako unresponsive? To: "Sarah Holmes" , Message-ID: Content-Type: text/plain; charset=US-ASCII I was waiting too long on hold so I left a complete voicemail message as they suggested and received a call back that they got my message. The processed my order. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Sarah Holmes 04/11/06 1:25 PM >>> Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 10 Date: Tue, 11 Apr 2006 10:53:08 -0700 (PDT) From: Kim Merriam Subject: [Histonet] Sudan Red 7B staining procedure To: Histonet Message-ID: <20060411175308.21420.qmail@web50308.mail.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello, I have found a reference for staining fat with sudan red 7B, it calls for a "saturated" solution of sudan red 7B in 70% ethanol. Does anyone know what the saturation point of sudan red 7B is in 70% ethanol; a general guide would be very helpful! Also - has anyone used this procedure before and what were the results compared to a regular Oil Red O (both in isopropanol and/or propylene glycol). Thanks in advance, Kim Kim Merriam, MA, HT(ASCP) Novartis Cambridge, MA --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. ------------------------------ Message: 11 Date: Tue, 11 Apr 2006 14:01:36 -0400 From: "Bell, Lynne" Subject: RE: [Histonet] Dako unresponsive? To: "Dana Settembre" , "Sarah Holmes" , Message-ID: We are also having a major problem getting through to Dako. We were told a couple of our items are on backorder for a few weeks. I am planning to call our account rep and voice my displeasure. It would have been nice to have received some communication regarding this "changeover" and perhaps we could have upped our order before all hell broke loose. Lynne Lynne A. Bell, HT (ASCP) Laboratory Central Vermont Hospital P.O. Box 547 Barre, VT 05641 (802)371-4122 ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Dana Settembre Sent: Tue 4/11/2006 1:46 PM To: Sarah Holmes; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Dako unresponsive? I was waiting too long on hold so I left a complete voicemail message as they suggested and received a call back that they got my message. The processed my order. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> Sarah Holmes 04/11/06 1:25 PM >>> Is anyone else having difficulty getting through to place orders with Dako? Thanks, Sarah _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 12 Date: Tue, 11 Apr 2006 12:06:04 -0600 From: Gayle Callis Subject: [Histonet] New edition of Polak and van Noorden immunohistochemistry book To: Histonet@lists.utsouthwestern.edu Message-ID: <6.0.0.22.1.20060411115552.01b3bc98@gemini.msu.montana.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed If you are looking for a concise, introductory book to immunohistochemical staining, I strongly suggest the long awaited new edition of: Introduction to ImmunoCytochemistry, 3rd Edition, JM Polak and S van Noorden, ISBN 185996 208 4. A paperback with 179 pages. This new edition was easy to follow and had excellent basic information. The appendix was packed with Technical notes on buffers, blocks, chromogens, etc. It cost ~$61 from Amazon.com 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 406 994-4303 (FAX) ------------------------------ Message: 13 Date: Tue, 11 Apr 2006 11:09:00 -0700 From: "Mitchell, Nancy K." Subject: [Histonet] CSH meeting in May-Costa Mesa To: Message-ID: <8EB1459EA20E4E498AC2E05378F8F30A536EDA@mind.mc.ad.lluahsc.org> Content-Type: text/plain; charset="iso-8859-1" If any of you California Histoechs are out there, I am having problems getting a copy of the workshops that are going to be available. Who should I contact? Nancy Mitchell, HT ASCP Histology Supervisor Loma Linda Pathology Medical Group 11370 Anderson Street Ste 2960 Loma Linda, CA 92354 909-558-2012 FAX-909-796-3667 ------------------------------ Message: 14 Date: Tue, 11 Apr 2006 14:21:15 -0400 From: Mary Lou Norman Subject: [Histonet] tendon processing/cutting To: Histonet@lists.utsouthwestern.edu, Stacy.Semevolos@oregonstate.edu Message-ID: <5.2.1.1.2.20060411140143.02cbf850@postoffice9.mail.cornell.edu> Content-Type: text/plain; charset="us-ascii"; format=flowed Thanks Elizabeth and Gayle for the tendon info. I've done the longer soaking which does help quite a bit. I've got big hunky horse pieces. My question today: After the slide has dried all night on the warming tray I still see white tissue, definitely not as opaque as I'd like. Any suggestions on making sure the tissue doesn't fall off? I'm just the cutter so I don't know what IHC is going to be done. Could I put them in the oven 60o? What's wrong with melting the paraffin? Stacy, can I put them in the oven? Thanks, Mary Lou Norman ------------------------------ Message: 15 Date: Tue, 11 Apr 2006 14:24:58 -0400 From: "Morken, Tim - Labvision" Subject: RE: [Histonet] CSH meeting in May-Costa Mesa To: histonet@lists.utsouthwestern.edu Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D843@usca0082k08.labvision.apogent.com> Content-Type: text/plain Nancy, you can download the program and registration information from www.californiahistology.org Tim Morken -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Mitchell, Nancy K. Sent: Tuesday, April 11, 2006 11:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] CSH meeting in May-Costa Mesa If any of you California Histoechs are out there, I am having problems getting a copy of the workshops that are going to be available. Who should I contact? Nancy Mitchell, HT ASCP Histology Supervisor Loma Linda Pathology Medical Group 11370 Anderson Street Ste 2960 Loma Linda, CA 92354 909-558-2012 FAX-909-796-3667 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 16 Date: Tue, 11 Apr 2006 14:59:07 -0400 From: "Monfils, Paul" Subject: RE: [Histonet] Sudan Red 7B staining procedure To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176CD@lsexch.lsmaster.lifespan.org> Content-Type: text/plain; charset="iso-8859-1" I don't have an exact figure, but according to one of my older texts (Histopathologic Technic and Practical Histochemistry by Lillie and Fullmer), 4.25 grams Sudan Red 7B will dissolve in 100 ml absolute ethanol. So the solubility in 70% ethanol should be substantially less than that. > ---------- > From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kim > Merriam > Sent: Tuesday, April 11, 2006 10:53 AM > To: Histonet > Subject: [Histonet] Sudan Red 7B staining procedure > > Hello, > > I have found a reference for staining fat with sudan red 7B, it calls > for a "saturated" solution of sudan red 7B in 70% ethanol. Does anyone > know what the saturation point of sudan red 7B is in 70% ethanol; a > general guide would be very helpful! > > Also - has anyone used this procedure before and what were the results > compared to a regular Oil Red O (both in isopropanol and/or propylene > glycol). > > Thanks in advance, > Kim > > > Kim Merriam, MA, HT(ASCP) > Novartis > Cambridge, MA > > --------------------------------- > Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > ------------------------------ Message: 17 Date: Tue, 11 Apr 2006 14:22:03 -0500 From: dgaupp@tulane.edu Subject: [Histonet] re-freezing frozen sections To: histonet@lists.utsouthwestern.edu Message-ID: <1144783323.443c01db190b1@webmail.tulane.edu> Content-Type: text/plain; charset=ISO-8859-1 Histonet: I have tissues(monkey brain) that were fixed(4%Paraformaldehyde), cryprotected(20% sucrose), and flash froze(-70C isopentane). Unfortunately, mothernature(hurricane katrina) came threw and destroyed everything. This tissues sat for 3-4 months at blistering temps in the same position(melted OCT) & some even had a surprise for me(fungus). When the University let us come back into our building, I rinsed off OCT and put the tissues in 10%NBF at room temp. The tissue was immersed for 3-4months in fixative. I rinsed the tissue and once again, flash froze one of the sections, and they cut horribly. I don't know what to do. I am now tempting to paraffin process. I prefer to do frozens because its easier to get almost every section of tissue(one sections I usually get 500-700 slides). As most of you know, brain likes to explode in water, so I can do a better job if they are frozen. Any help, I would appreciate! Dina Dina D. Gaupp, B.S., M.T. Medical Research Specialist Center for Gene Therapy Tulane University Health Sciences Center JBJ Bldg/Rm 658, SL-99 1430 Tulane Avenue New Orleans, LA 70112 Lab: 504.988.1194 Fax: 504.988.7710 Email: dgaupp@tulane.edu ------------------------------ Message: 18 Date: Tue, 11 Apr 2006 17:43:52 -0400 From: "Favara, Cynthia \(NIH/NIAID\) [E]" Subject: RE: [Histonet] re-freezing frozen sections To: , Message-ID: Content-Type: text/plain; charset="us-ascii" I have done some squirrel monkey brain in paraffin with good results - happy to help with processing schedules etc if you like. Let me know c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: dgaupp@tulane.edu [mailto:dgaupp@tulane.edu] Sent: Tuesday, April 11, 2006 12:22 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] re-freezing frozen sections Histonet: I have tissues(monkey brain) that were fixed(4%Paraformaldehyde), cryprotected(20% sucrose), and flash froze(-70C isopentane). Unfortunately, mothernature(hurricane katrina) came threw and destroyed everything. This tissues sat for 3-4 months at blistering temps in the same position(melted OCT) & some even had a surprise for me(fungus). When the University let us come back into our building, I rinsed off OCT and put the tissues in 10%NBF at room temp. The tissue was immersed for 3-4months in fixative. I rinsed the tissue and once again, flash froze one of the sections, and they cut horribly. I don't know what to do. I am now tempting to paraffin process. I prefer to do frozens because its easier to get almost every section of tissue(one sections I usually get 500-700 slides). As most of you know, brain likes to explode in water, so I can do a better job if they are frozen. Any help, I would appreciate! Dina Dina D. Gaupp, B.S., M.T. Medical Research Specialist Center for Gene Therapy Tulane University Health Sciences Center JBJ Bldg/Rm 658, SL-99 1430 Tulane Avenue New Orleans, LA 70112 Lab: 504.988.1194 Fax: 504.988.7710 Email: dgaupp@tulane.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 19 Date: Tue, 11 Apr 2006 20:04:54 -0400 From: "Andrea C. Bilger" Subject: [Histonet] Milestone microwave processor To: Message-ID: <058f01c65dc4$baebb320$0300a8c0@andrea> Content-Type: text/plain; charset="iso-8859-1" We had the RHS I in our lab for a couple of days to test it out. It performed well. The PA's didn't change how they grossed the tissue. This was a big plus. No need for special tools and tiny sized tissues. You can program it to the largest size specimen being processed. Our lab is looking into getting at least one of the small units and we may go for the new Pathos they just came out with. The Pathos is larger and automatically changes the solutions. No need for manually changing them like you do with the RHS I. Now if we can just convince the top brass to give us the money for one of them. Andrea Bilger ------------------------------ Message: 20 Date: Wed, 12 Apr 2006 07:40:41 -0700 (PDT) From: Emre Kukurt Subject: [Histonet] One question To: histonet@lists.utsouthwestern.edu Message-ID: <20060412144041.65778.qmail@web31001.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Hello Histonetters, Im new here, not sure whether or no my problem appropriate for the list topic. I have treated low electric current on grafted plants. Then, they were sectioned, prepared and photographed either light or electron microscope. I need to show them to an expert on plant histology who can interpret the pictures in order to decide what (sub)celullar changes appeared before/after electric current applied. Can some please one help me ? Best regards - Emre __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com ------------------------------ Message: 21 Date: Wed, 12 Apr 2006 11:08:51 -0400 From: "Vacca Jessica" Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tek VIP 2000 To: Message-ID: <41E16A15CE78374EA45B57E0F94339B84B5ADC@ORLEV01.hca.corpad.net> Content-Type: text/plain; charset="iso-8859-1" Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax ------------------------------ Message: 22 Date: Wed, 12 Apr 2006 10:26:06 -0500 From: "McGovern, Kevin" Subject: RE: [Histonet] Auto dialers (remote alarms) for leica and tissue tekVIP 2000 To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=iso-8859-1 Jessica, You may need two boxes, depending on how the instruments signal they are in an alarm state. In many cases, they simply short together their output wires, in some, they send out 5VDC. We have a VIP E-150 with a home-brewed alarm box that works well. Your local biomed should be able to cook one up if he has the electrical diagrams for the units. It ain't rocket science! Good luck. Kevin Kevin S. McGovern, BMETII Good Samaritan Hospital Clinical Engineering Dept. 10 East 31st Street Kearney, NE 68836 Phone: (308) 865-7051 Fax: (308) 865-2914 e-Mail: kevinmcgovern@catholichealth.net -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Vacca Jessica Sent: Wednesday, April 12, 2006 10:09 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Auto dialers (remote alarms) for leica and tissue tekVIP 2000 Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 23 Date: Wed, 12 Apr 2006 11:04:46 -0500 From: mari.ann.mailhiot@leica-microsystems.com Subject: Re: [Histonet] Auto dialers (remote alarms) for leica and tissue tek VIP 2000 To: "Vacca Jessica" Cc: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain; charset=US-ASCII Jessica I have asked Mike Gallo to give you a call in regards to your alarm for the Leica processor. However, your biomedical engineers should be able to wire the alarm for you. If you have questions just give me a call. Best Regards Mari Ann Mailhiot BA HT ASCP Application Specialist Leica Technical Assistance Center 800 248 0123 x7267 847 236 3063 fax mari.ann.mailhiot@leica-microsystems.com www.leica-microsystems.com "Vacca Jessica" To Sent by: histonet-bounces@ cc lists.utsouthwest ern.edu Subject [Histonet] Auto dialers (remote alarms) for leica and tissue tek 04/12/2006 10:08 VIP 2000 AM Another day has gone by with the machine alarming over the weekend and nobody knew until it was too late. Does anyone know about the remote alarms for both of these types of processors? Do I need 2 separate ones or can biomed hook up 1 remote to 2 machines? Where can I find this product? Save a histotech from another Manic Monday! Thanks Jessica Vacca Histology Supervisor Brandon Regional Hospital 119 Oakfield Dr Brandon FL 33511 (813) 681-5551 ext 2454 (813) 571-5193 (813) 681-5169 Fax _______________________________________________ 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 ______________________________________________________________________ ------------------------------ Message: 24 Date: Wed, 12 Apr 2006 10:43:25 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] QIHC To: "'Marian powers'" , Message-ID: <200604121643.k3CGhNka013874@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Marian you can join the NSH IHC Resource Group online at www.ihcrg.org We have a list of references and you can actually access the ws on taking the QIHC exam presented by myself and Ethel Macrea at the NSH S/C last year. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Marian powers Sent: Monday, April 10, 2006 7:41 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] QIHC Looking for a good IHC reference to study for the QIHC? Thanks in advance, Marian _______________________________________________ Histonet mailing list Histonet@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 29, Issue 15 **************************************** From Charlene.Henry <@t> STJUDE.ORG Fri Apr 14 13:19:41 2006 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Apr 14 13:19:48 2006 Subject: [Histonet] Anyone catch "House" the other night?. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1BF6@SJMEMXMB02.stjude.sjcrh.local> Hey! It is the right temperature so perhaps we should all budget for a Jacuzzi next year. Charlene -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Fred Underwood Sent: Friday, April 14, 2006 9:38 AM To: BlazekL@childrensdayton.org; Histonet@lists.utsouthwestern.edu; POWELL_SA@Mercer.edu; lpjones@srhs-pa.org Subject: RE: [Histonet] Anyone catch "House" the other night?. . I think there was an episode where they used a jacuzzi to perform HIER. >>> Shirley Powell 04/13/06 02:40PM >>> Turn off the TV get in the Jacuzzi, have a glass of wine and relax. Work is work, TV is TV and they get paid a lot more than we do for doing histology wrong. Go figure. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Linda Blazek Sent: Thursday, April 13, 2006 11:38 AM To: Histonet@lists.utsouthwestern.edu; lpjones@srhs-pa.org Subject: Re: [Histonet] Anyone catch "House" the other night? My husband turned to me as said "I thought you said doing immunos was a complicted thing." >>> "Jones, Laura" 4/13/2006 12:27 PM >>> Dr. House and his colleagues were doing immuno on Tuesday night! Procedure: Take large chunk of heart tissue which has not been frozen or in fixative, but in a petri dish in the fridge, and flop it on to your microscope slide. Put the slide with the large chunk of tissue on it under the scope. Add "one microlitre of immunoperoxidase" to the tissue. Look into scope. If it turns red, it's positive. I wish our immuno were that easy and quick. Get a Histo consultant on staff, please! :o) P.S. And how about those doctors who do biopsies, ultrasounds, draw blood, and then test it all? In evening gowns and tuxes? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rachael_emerson <@t> urmc.rochester.edu Fri Apr 14 13:33:26 2006 From: rachael_emerson <@t> urmc.rochester.edu (Rachael Emerson) Date: Fri Apr 14 13:33:42 2006 Subject: [Histonet] Wright-giemsa Message-ID: Hello. Does anyone have a protocol that they would like to share for doing a manual Wright-Giemsa stain on cytospins? Thanks! Rachael -- Rachael L. Emerson Center for Pediatric Biomedical Research University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 From gsennello <@t> osip.com Fri Apr 14 13:57:14 2006 From: gsennello <@t> osip.com (Sennello, Gina) Date: Fri Apr 14 13:56:40 2006 Subject: [Histonet] CD 34 Message-ID: I was wondering if any one has a good protocol for doing CD34 on mouse brains? Thank you, Gina Sennello OSIP Boulder CO From ROrr <@t> enh.org Fri Apr 14 13:56:37 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Fri Apr 14 13:56:42 2006 Subject: [Histonet] IHC Tech Spec position Message-ID: Hi Histonetters, Evanston Northwestern Healthcare is looking for a Technical Specialist to over see Immunohistochemistry procedures. This position will be responsible for developing, modifying and implementing new techniques pertaining to special staining procedures. Requirements include: registration of HT/HTL (with 3-5 years full-time experience with immunoperoxidase procedures and equipment), *HTL (ASCP) or Immunohistochemistry qualifications and *QIHC (ASCP) certification preferred. This is a great opportunity to work in a progressive environment with a great staff. Please go to www.enh.org/careers then reference job number 020081. I'll be away until April 21, but you can contact Nikole Wagner in HR at 847-570-1484 or Linda Delk, Anatomic Pathology Manager at 847-570-1433 with any questions. Becky Orr CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 From Charlene.Henry <@t> STJUDE.ORG Fri Apr 14 14:07:54 2006 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Apr 14 14:07:58 2006 Subject: [Histonet] IHC Tech Spec position. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1BF8@SJMEMXMB02.stjude.sjcrh.local> Where is Evanston Northwestern Healthcare located? Charlene -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, April 14, 2006 1:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Tech Spec position. . Hi Histonetters, Evanston Northwestern Healthcare is looking for a Technical Specialist to over see Immunohistochemistry procedures. This position will be responsible for developing, modifying and implementing new techniques pertaining to special staining procedures. Requirements include: registration of HT/HTL (with 3-5 years full-time experience with immunoperoxidase procedures and equipment), *HTL (ASCP) or Immunohistochemistry qualifications and *QIHC (ASCP) certification preferred. This is a great opportunity to work in a progressive environment with a great staff. Please go to www.enh.org/careers then reference job number 020081. I'll be away until April 21, but you can contact Nikole Wagner in HR at 847-570-1484 or Linda Delk, Anatomic Pathology Manager at 847-570-1433 with any questions. Becky Orr CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Nalini.Makhijani <@t> va.gov Fri Apr 14 14:18:10 2006 From: Nalini.Makhijani <@t> va.gov (Makhijani, Nalini S) Date: Fri Apr 14 14:18:17 2006 Subject: [Histonet] PE Tumor tissue; Transwell embedding Message-ID: <715FA772CEA81A47B1A762AB6E45123AE254B8@VHAV22MSGA3.v22.med.va.gov> Hi Histonetters, I have 2 questions: 1. Does anyone have an opinion on the correct disposal method for paraffin embedded tumor tissue? 2. Does anyone have experience with paraffin embedding, sectioning and H&E staining of Transwell membrane inserts, either Corning Costar, or B-D? I've been following the protocol from the Corning website, using Citrisolv from Fisher instead of Histoclear. The sections of the membranes are thin lines about 3mm long. After about 2 min in Citrisolv the sections look so clear that it's not clear if they are on or not. If they are, they are usually lost in the first absolute alcohol. After staining, nothing is visible. Do these membranes, without cells, stain or remain transparent? Are 3mm sections too small to remain on Superfrost Plus slides? Should the slides be subbed? Does anyone else have a problem getting transwells and cells to stick to the slides? Thanks, Nalini VAMC, West Los Angeles From liz <@t> premierlab.com Fri Apr 14 14:50:52 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Fri Apr 14 14:51:02 2006 Subject: [Histonet] PE Tumor tissue; Transwell embedding In-Reply-To: <715FA772CEA81A47B1A762AB6E45123AE254B8@VHAV22MSGA3.v22.med.va.gov> Message-ID: <000901c65ffc$bddcf690$0300a8c0@Chlipala> Nalini We have processed millipore membrane inserts. We just processed them routinely to paraffin. We wrote a article for Sakura Histologic about it. The link is below: http://www.sakura-americas.com/histologic/pdf/05_may.pdf Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Makhijani, Nalini S Sent: Friday, April 14, 2006 1:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] PE Tumor tissue; Transwell embedding Hi Histonetters, I have 2 questions: 1. Does anyone have an opinion on the correct disposal method for paraffin embedded tumor tissue? 2. Does anyone have experience with paraffin embedding, sectioning and H&E staining of Transwell membrane inserts, either Corning Costar, or B-D? I've been following the protocol from the Corning website, using Citrisolv from Fisher instead of Histoclear. The sections of the membranes are thin lines about 3mm long. After about 2 min in Citrisolv the sections look so clear that it's not clear if they are on or not. If they are, they are usually lost in the first absolute alcohol. After staining, nothing is visible. Do these membranes, without cells, stain or remain transparent? Are 3mm sections too small to remain on Superfrost Plus slides? Should the slides be subbed? Does anyone else have a problem getting transwells and cells to stick to the slides? Thanks, Nalini VAMC, West Los Angeles _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________ NOD32 1.1489 (20060414) Information __________ This message was checked by NOD32 antivirus system. http://www.eset.com From amosbrooks <@t> gmail.com Fri Apr 14 14:12:57 2006 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Fri Apr 14 15:07:12 2006 Subject: [Histonet] Re: Histonet Digest, Vol 29, Issue 17 In-Reply-To: <443fba00.29c144c4.016e.06ebSMTPIN_ADDED@mx.gmail.com> References: <443fba00.29c144c4.016e.06ebSMTPIN_ADDED@mx.gmail.com> Message-ID: <582736990604141212x51f7e864sd42d92360d019356@mail.gmail.com> Marylin, Try popping them into a 40 to 60 deg. oven for a while just before sending them. The Permount will dry and won't stick to everything. It shouldn't take long to dry them out. Good luck, Amos Message: 19 > Date: Fri, 14 Apr 2006 05:51:07 -0600 > From: "Marilyn Johnson" > Subject: [Histonet] Slide Mailers > To: > Message-ID: <001401c65fb9$b78cbc00$6401a8c0@VALUED20606295> > Content-Type: text/plain; charset="iso-8859-1" > > Hi Histonetters, > I have to send wet-Permount, H&E slides to one of my clients in another > city, > by courier.Turn-around time is critical. I've tried the plastic slide > mailers and > the cardboard ones, but the wet coverslips stick to the contacted > surfaces. > Does anyone have a special slide mailer or technique you can share with > me? > Thank you in advance. > > Marilyn Johnson > Edmonton, Alberta, Canada From Rcartun <@t> harthosp.org Fri Apr 14 15:48:24 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Fri Apr 14 15:49:03 2006 Subject: [Histonet] CK5/6 - nuclear staining Message-ID: Has anyone observed "nuclear" staining with Dako's cytokeratin 5/6 monoclonal antibody? I think it may represent an artifact from the heat retrieval, but I wanted to query the Histonet membership in case others have seen this too. Thank you. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax From bob-meyer <@t> northwestern.edu Fri Apr 14 15:49:09 2006 From: bob-meyer <@t> northwestern.edu (bob-meyer@northwestern.edu) Date: Fri Apr 14 15:49:13 2006 Subject: [Histonet] IHC Tech Spec position. . Message-ID: <20060414204909.95A5035C6E@casbah.it.northwestern.edu> It is in Evanston, IL. A suburb located north of Chicago. Bob Meyer, HTL(ASCP),QIHC Sr. Research Technologist Northwestern University Pathology Core Facility Chicago, IL 312-908-5546 ==============Original message text=============== On Fri, 14 Apr 2006 7:07:54 pm +0000 "Henry, Charlene" wrote: Where is Evanston Northwestern Healthcare located? Charlene -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Orr, Rebecca Sent: Friday, April 14, 2006 1:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] IHC Tech Spec position. . Hi Histonetters, Evanston Northwestern Healthcare is looking for a Technical Specialist to over see Immunohistochemistry procedures. This position will be responsible for developing, modifying and implementing new techniques pertaining to special staining procedures. Requirements include: registration of HT/HTL (with 3-5 years full-time experience with immunoperoxidase procedures and equipment), *HTL (ASCP) or Immunohistochemistry qualifications and *QIHC (ASCP) certification preferred. This is a great opportunity to work in a progressive environment with a great staff. Please go to www.enh.org/careers then reference job number 020081. I'll be away until April 21, but you can contact Nikole Wagner in HR at 847-570-1484 or Linda Delk, Anatomic Pathology Manager at 847-570-1433 with any questions. Becky Orr CLA,HT(ASCP) Assistant Manager, Anatomic Pathology Evanston Northwestern Healthcare 847-570-2771 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ===========End of original message text=========== From bhewlett <@t> cogeco.ca Fri Apr 14 16:22:17 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Apr 14 16:22:23 2006 Subject: [Histonet] CK5/6 - nuclear staining References: Message-ID: <000901c66009$822801f0$6500a8c0@mainbox> Hi Richard, The only time I have observed "nuclear" immunoreactivity with CK5/6 was in a poorly fixed specimen. When the aggressive HIER (pressure cooker) was replaced by more gentle steaming, the reaction disappeared. This type of artifactual reaction can even rear it's ugly head in well fixed tissues with some antibodies. Best regards, Bryan ----- Original Message ----- From: "Richard Cartun" To: Sent: Friday, April 14, 2006 4:48 PM Subject: [Histonet] CK5/6 - nuclear staining > Has anyone observed "nuclear" staining with Dako's cytokeratin 5/6 > monoclonal antibody? I think it may represent an artifact from the heat > retrieval, but I wanted to query the Histonet membership in case others > have seen this too. Thank you. > > Richard > > Richard W. Cartun, Ph.D. > Director, Immunopathology & Histology > Assistant Director, Anatomic Pathology > Hartford Hospital > 80 Seymour Street > Hartford, CT 06102 > (860) 545-1596 > (860) 545-0174 Fax > > > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From JPaulB42 <@t> aol.com Fri Apr 14 16:41:46 2006 From: JPaulB42 <@t> aol.com (JPaulB42@aol.com) Date: Fri Apr 14 16:42:01 2006 Subject: [Histonet] Milestone Processor Run2 Message-ID: <389.8502e3.3171711a@aol.com> Hey Histonetters, I have now completed a my second run on the Milestone Microwave Processor 110. This time it was just biopsies and cell blocks. Both processed well on the short cycle. The only problem this time was the staining again. The nuclear detail was fine for the pathologist that looked at the slides but the cytoplasmic stain was inconsistant. Some were pale, some were slightly heavy. The cytologists that examined the cell blocks were pleased and surprised that it only took 70 minutes for processing. I'm starting to do parallel runs from here on out. So far the pathologists that have seen the work are okay with the results. More to come as this process continues. James Bradford Orlando Regional Medical Center Orlando, FL 321-841-5498 From mhwac <@t> yahoo.com Sun Apr 16 22:05:13 2006 From: mhwac <@t> yahoo.com (mh how) Date: Sun Apr 16 22:05:23 2006 Subject: [Histonet] Hacker MARS Message-ID: <20060417030513.78542.qmail@web53007.mail.yahoo.com> Hi Histonetters, I am just wondering whether anyone has any experience on the HACKER MARS, microwave tissue processor ? Thanks ! Shannon --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From akbitting <@t> geisinger.edu Mon Apr 17 07:13:31 2006 From: akbitting <@t> geisinger.edu (Angela Bitting) Date: Mon Apr 17 07:14:00 2006 Subject: [Histonet] EBV (EBER) and HPV Message-ID: Hi Histofriends, I'm looking for EBER positive and HPV ( high risk) positive tissue slides to validate my probes for CISH. We only have a few pos blocks and we've exhausted most of them. If anyone can share a couple slides with me that would be sooooo excellent! I'll send you my Fed Ex acct number. Thanks as always, Ang Angela Bitting, HT(ASCP) Technical Specialist, Histology Geisinger Medical Center 100 N Academy Ave. MC 23-00 Danville, PA 17822 phone 570-214-9634 fax 570-271-5916 IMPORTANT WARNING: The information in this message (and the documents attached to it, if any) is confidential and may be legally privileged. It is intended solely for the addressee. Access to this message by anyone else is unauthorized. If you are not the intended recipient, any disclosure, copying, distribution or any action taken, or omitted to be taken, in reliance on it is prohibited and may be unlawful. If you have received this message in error, please delete all electronic copies of this message (and the documents attached to it, if any), destroy any hard copies you may have created and notify me immediately by replying to this email. Thank you. From fawn <@t> cs.cmu.edu Mon Apr 17 08:02:34 2006 From: fawn <@t> cs.cmu.edu (Fawn Jones) Date: Mon Apr 17 08:02:43 2006 Subject: [Histonet] Help with pricing Message-ID: <444391EA.5040602@cs.cmu.edu> Hi histonetters, I was wondering if anyone could help me figure out the cost and turn around time of doing some plastic slides. It invoves embedding 2 whole mice in MMA and sectioning sagittally through the mice to look at the spine along with an implant. I appreciate any suggestions and comments. Fawn Jones From squadeer <@t> utmb.edu Mon Apr 17 10:17:04 2006 From: squadeer <@t> utmb.edu (Quadeer, Shahnaz S.) Date: Mon Apr 17 10:17:11 2006 Subject: [Histonet] RE-unscribe Message-ID: <13362C82244EEA4B9BD90AC1D90E5C1BEAFE4B@EXCH2K4.utmb.edu> Unscirbe From Tracey.Lenek <@t> CLS.ab.ca Mon Apr 17 10:30:00 2006 From: Tracey.Lenek <@t> CLS.ab.ca (Tracey.Lenek@CLS.ab.ca) Date: Mon Apr 17 10:30:10 2006 Subject: [Histonet] P16 Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE02C1D5F9@mail1.calgary.com> Hi, We recently started using Neomarkers P16 - clone 16P07. We are finding increased cytoplasmic staining making nuclear interpretation difficult. We are staining on the Ventana Nexes using their IVIEW DAB detection kit with HIER using a steamer. Has anyone experienced the same or different staining pattern with this antibody? Thanks Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify the sender immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From rodger_65489 <@t> yahoo.com Mon Apr 17 10:47:53 2006 From: rodger_65489 <@t> yahoo.com (Roger Smithwell) Date: Mon Apr 17 10:47:58 2006 Subject: [Histonet] RHS-1 and MicroMed T/T Mega Message-ID: <20060417154753.79318.qmail@web38304.mail.mud.yahoo.com> If you are familiar with using either of these units, or currently process with these units please let me know you're opinions (good or bad). We are going to be in the market for possibly both units soon. Thanks in advance to all members who respond. Rodger --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. From naje1972 <@t> yahoo.com Mon Apr 17 13:02:07 2006 From: naje1972 <@t> yahoo.com (cynthia haynes) Date: Mon Apr 17 13:02:39 2006 Subject: [Histonet] special stain with a immunohistochemical stain Message-ID: <20060417180207.48347.qmail@web33005.mail.mud.yahoo.com> Hello fellow Histonetters A researcher gave me a DAB stained slides. He would like for me to take the cover glass off and stain the slide with LUXOL FAST BLUE. My question is when you want to combine a special stain with immunohistochemical stains, which should you do first? The special stain or the immuno stain? Thanks in advance. Cynthia Haynes H.T. From settembr <@t> umdnj.edu Mon Apr 17 13:43:15 2006 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Mon Apr 17 13:44:18 2006 Subject: [Histonet] special stain with a immunohistochemical stain Message-ID: Good question, Cynthia. I have only run immunos on slides that have had special stains run on them already. They work. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> cynthia haynes 04/17/06 2:02 PM >>> Hello fellow Histonetters A researcher gave me a DAB stained slides. He would like for me to take the cover glass off and stain the slide with LUXOL FAST BLUE. My question is when you want to combine a special stain with immunohistochemical stains, which should you do first? The special stain or the immuno stain? Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From PMonfils <@t> Lifespan.org Mon Apr 17 13:56:10 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Mon Apr 17 13:56:17 2006 Subject: [Histonet] special stain with a immunohistochemical stain Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176D7@lsexch.lsmaster.lifespan.org> The immuno stain should ordinarily be done first, as it is practically impossible to remove the DAB staining histochemically, while it is very possible to remove or otherwise alter some of the histochemical dyes while doing the immuno stain. Just last week I did some trichromes on previously-stained immuno slides. The result was very nice. They had stained a marker in mouse heart, and then wanted to know if it was associated with areas of fibrosis. Wherever areas of DAB staining occurred, they were surrounded by the blue-stained collagen fibers. From liz <@t> premierlab.com Mon Apr 17 13:57:43 2006 From: liz <@t> premierlab.com (Elizabeth Chlipala) Date: Mon Apr 17 13:58:03 2006 Subject: [Histonet] special stain with a immunohistochemical stain In-Reply-To: Message-ID: <000001c66250$cf101040$0300a8c0@Chlipala> Cynthia DAB as a chromagen is very stable, we run the immuno first and then the special stain. We have done this with DAB and then Acid fast, there is also a nice Histologic article on this type of dual staining, I think it was on T cells and PAS in kidney samples. Liz Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC Manager Premier Laboratory, LLC P.O. Box 18592 Boulder, Colorado 80308 Office: (303) 735-5001 Fax: (303) 735-3540 liz@premierlab.com www.premierlab.com Ship to Address: Premier Laboratory University of Colorado MCDB, Room A3B40 Boulder, Colorado 80309 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Dana Settembre Sent: Monday, April 17, 2006 12:43 PM To: histonet@pathology.swmed.edu; cynthia haynes Subject: Re: [Histonet] special stain with a immunohistochemical stain Good question, Cynthia. I have only run immunos on slides that have had special stains run on them already. They work. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> cynthia haynes 04/17/06 2:02 PM >>> Hello fellow Histonetters A researcher gave me a DAB stained slides. He would like for me to take the cover glass off and stain the slide with LUXOL FAST BLUE. My question is when you want to combine a special stain with immunohistochemical stains, which should you do first? The special stain or the immuno stain? Thanks in advance. Cynthia Haynes H.T. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet __________ NOD32 1.1492 (20060416) Information __________ This message was checked by NOD32 antivirus system. http://www.eset.com From sluhisto <@t> yahoo.com Mon Apr 17 14:48:38 2006 From: sluhisto <@t> yahoo.com (Histology SLU) Date: Mon Apr 17 14:48:54 2006 Subject: [Histonet] Cryostat Message-ID: <20060417194838.81812.qmail@web51001.mail.yahoo.com> Hello All: We are looking to demo for a new cryostat. What is everyone using and what are your thoughts on them? Thanks bunches. Susan --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From RJLevier <@t> LancasterGeneral.org Mon Apr 17 14:55:01 2006 From: RJLevier <@t> LancasterGeneral.org (LeVier, Rebecca J) Date: Mon Apr 17 14:55:06 2006 Subject: [Histonet] Microtome Repair Message-ID: <0FDBF29200C637468CCC1F9E7E85A3D2D6DE4A@MAIL-LR.lha.org> Hey Histonetters, I work in South Central Pennsylvania and I was wondering if anyone knew of a good place to get service for microtomes? If they offer any other services could you please let me know what all they offer if you know. And do you like the service that you receive? Thank you in advance for the help. Becky Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. From pmcardle <@t> ebsciences.com Mon Apr 17 15:10:02 2006 From: pmcardle <@t> ebsciences.com (Phil McArdle) Date: Mon Apr 17 15:10:14 2006 Subject: [Histonet] Microtome Repair In-Reply-To: <0FDBF29200C637468CCC1F9E7E85A3D2D6DE4A@MAIL-LR.lha.org> References: <0FDBF29200C637468CCC1F9E7E85A3D2D6DE4A@MAIL-LR.lha.org> Message-ID: <4443F61A.5050208@ebsciences.com> Hello: Please forgive a "vendor" reply, but if you're looking specifically for JB-4 repair, we do it (we were the last company with manufacturing rights to this model). Best regards, Phil McArdle -- Phil McArdle Microwave Product Manager Energy Beam Sciences, Inc. Tel: 800.992.9037 x 341 Cell: 860.597.6796 Fax: 860.653.0422 PMcardle@ebsciences.com www.ebsciences.com "ADDING BRILLIANCE TO YOUR VISION" "I hate quotations. Tell me what you know." Ralph Waldo Emerson LeVier, Rebecca J wrote: > Hey Histonetters, > > I work in South Central Pennsylvania and I was wondering if anyone knew > of a good place to get service for microtomes? If they offer any other > services could you please let me know what all they offer if you know. > And do you like the service that you receive? > > Thank you in advance for the help. > > Becky > > > Confidentiality Notice: This e-mail message, including any > attachments, is for the sole use of intended recipient(s) and may > contain confidential and privileged information. Any unauthorized > review, use, disclosure or distribution is prohibited. If you are not > the intended recipient, please contact the sender by reply e-mail and > destroy all copies of the original message. > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From sheris <@t> MIT.EDU Mon Apr 17 15:39:04 2006 From: sheris <@t> MIT.EDU (Sheri L Simmons) Date: Mon Apr 17 15:39:17 2006 Subject: [Histonet] beginner question about methacrylate resins and nucleic acid ISH Message-ID: Hi, I'm a newcomer to the list and the world of histochemistry in general. I'm hoping that someone can help me with a few questions relating to targeting bacterial RNA in embedded thick sections (very thick- about a mm) with fluorescent methods. The goal is to detect species-specific oligonucleotide probes to the bacterial 16s rRNA with indirect CARD-FISH. Searching the list archives, it seems that although glycol methacrylate (GMA) and methyl methacrylate (MMA) resins are both used for histochemistry, MMA is generally preferred because it can be removed from sections with acetone. Is this your experience? Do you know of any direct comparisons of GMA and MMA for nucleic acid detection with in situ hybridization? Are there any special concerns in terms of the resin type or protocol relating to the use of nucleic acid probes rather than antibodies? Finally, which of the many MMA kits out there would you recommend? I've seen references to Technovit 9100, but this is significantly more expensive than a generic Methyl Methacrylate/Butyl Methacrylate kit from Electron Microscopy Sciences. Is there a reason to prefer one over the other? Thanks, Sheri --------------------------------------------------- Sheri Simmons Geomicrobiology Group MS #52, WHOI, Woods Hole MA 02543 http://www.whoi.edu/science/MCG/edwards/sherisimmons From brett_connolly <@t> merck.com Mon Apr 17 16:04:26 2006 From: brett_connolly <@t> merck.com (Connolly, Brett M) Date: Mon Apr 17 16:05:10 2006 Subject: [Histonet] Job opportunity Message-ID: <355C35514FEAC9458F75947F5270974D67CE35@usctmx1103.merck.com> Histonetter's We are looking for a histologist with good IHC experience to join our department. If you are interested or have any questions feel free to drop me an e-mail along with your CV. You can also visit the site listed below. Brett Brett M. Connolly, Ph.D. Merck & Co., Inc. MRL, Imaging Research WP-44K PO Box 4 West Point, PA 19486 PH 215-652-2501 fax. 215-993-6803 e-mail. brett_connolly@merck.com Job Description The Department of Imaging at the Merck Research Laboratories in West Point, PA is seeking an in vitro histologist to join the group. The successful candidate will be expected to work as part of a team using cutting edge imaging technologies to develop and evaluate non clinical models to support drug discovery projects. Experience with a variety of histological methodologies such as specimen fixation, embedding, sectioning, immunocytochemistry and immunofluorescence, and the ability to work effectively both independently and in a team environment is essential. Practical expertise with image analysis systems would be advantageous. Under general scientific supervision, the successful candidate will be primarily responsible for the generation and analysis of experimental data. Qualifications Qualifications for this position include a BS or MS degree in Biological science and ASCP certification as a histotechnician (HT) or histotechnologist (HTL) is required. The qualified individual must be highly motivated, conscientious, meticulous with expertise in immunohistological techniques as evidenced by several years of laboratory experience. She/he also needs to be computer literate, have excellent organizational and communication skills and enjoy working in dynamic teams. To be considered for this position, please visit our career site at www.merck.com/careers to create a profile and submit your CV for PHA000612. ------------------------------------------------------------------------------ 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 conniegrubaugh <@t> hotmail.com Mon Apr 17 18:45:55 2006 From: conniegrubaugh <@t> hotmail.com (connie grubaugh) Date: Mon Apr 17 18:46:02 2006 Subject: [Histonet] Job opening in Las Vegas Message-ID: Job opening in Las Vegas at Laboratory Medicine Consultants. Must be able to work independantly. Hours are from 10 pm till 6:30 am. Must have ASCP or be eligible. Contact Brett Brunson at 702-732-3441 No head hunters. From amosbrooks <@t> gmail.com Mon Apr 17 18:46:59 2006 From: amosbrooks <@t> gmail.com (Amos Brooks) Date: Mon Apr 17 18:47:05 2006 Subject: [Histonet] Spelling Hall of Fame? Message-ID: <582736990604171646o7f683622oc6aea64272baf538@mail.gmail.com> What are the chances of someone keeping a list of great mis-spellings of subscribe & unsubscribe? I think this one would make a great entry! Really, not to be nit-picky but this one was really funny. Amos Message: 4 > Date: Mon, 17 Apr 2006 10:17:04 -0500 > Subject: [Histonet] RE-unscribe > To: > Message-ID: <13362C82244EEA4B9BD90AC1D90E5C1BEAFE4B@EXCH2K4.utmb.edu> > Content-Type: text/plain; charset="us-ascii" > > Unscirbe > From b_pritt <@t> yahoo.com Mon Apr 17 20:19:36 2006 From: b_pritt <@t> yahoo.com (Bobbi Pritt) Date: Mon Apr 17 20:19:40 2006 Subject: [Histonet] Gomori silver versus Warthin-Starry Message-ID: <20060418011936.84272.qmail@web33202.mail.mud.yahoo.com> Could someone explain the difference between silver stains for fungi and those for spirochetes? I've been told that silver-based fungal stains (such as GMS) do not significantly deposit silver onto the fungal elements, therefore, one can accurately measure their size using such a preparation. On the other hand, I've been told that the silver stains for spirochetes (eg. Steiner, Warthin-Starry) are "deposition" stains, which make the organisms appear larger and therefore, more easily identifiable. Presumably, you would not want to measure size of organisms using this stain. Is all of this true, and can someone explain the differing mechanisms to me? Thank you, Bobbi Pritt --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. From jqb7 <@t> cdc.gov Tue Apr 18 05:13:22 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Tue Apr 18 05:13:30 2006 Subject: [Histonet] Gomori silver versus Warthin-Starry Message-ID: I am not an expert but the way I understand it is this: According to Grocott and Luna, the polysaccharides in the fungal cell wall are oxidized to aldehydes by the chromic acid. Only substances that possess a large quantity of polysaccharides (fungal cell wall, glycogen and mucins) remain reactive with the methenamine silver, reducing it to metallic silver. Most all of the silver spirochete techniques follow the principle that spirochetes are argryophilic (they have the ability to bind silver ions from a solution) but do not have the ability to reduce the silver to a visible metallic form. A chemical reducer is used for that purpose. Based on this chemical property it stands to reason that these organisms would appear larger than they actually are. Hope that helps. Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bobbi Pritt Sent: Monday, April 17, 2006 9:20 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Gomori silver versus Warthin-Starry Could someone explain the difference between silver stains for fungi and those for spirochetes? I've been told that silver-based fungal stains (such as GMS) do not significantly deposit silver onto the fungal elements, therefore, one can accurately measure their size using such a preparation. On the other hand, I've been told that the silver stains for spirochetes (eg. Steiner, Warthin-Starry) are "deposition" stains, which make the organisms appear larger and therefore, more easily identifiable. Presumably, you would not want to measure size of organisms using this stain. Is all of this true, and can someone explain the differing mechanisms to me? Thank you, Bobbi Pritt --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From mikael.niku <@t> helsinki.fi Tue Apr 18 07:21:43 2006 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Tue Apr 18 07:21:55 2006 Subject: [Histonet] Large vessel endothelium problem Message-ID: <4444D9D7.2060405@helsinki.fi> Hello! I need to do some immunostaining of large vessel endothelium (paraformaldehyde-fixed paraffin material). The problem is, the endothelium is easily stripped off and lost, apparently in early steps of sample processing. Any tips how to keep the stuff intact? With best regards, Mikael -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// From LRaff <@t> lab.uropartners.com Tue Apr 18 07:50:47 2006 From: LRaff <@t> lab.uropartners.com (Lester Raff) Date: Tue Apr 18 07:50:51 2006 Subject: [Histonet] VIP fixative Message-ID: <5DA1CA5D0B98A84985B545A24423B822A7FF@UPLAB01.uplab.local> Hi all: We process only small biopsies and have a new Sakura Tissue Tek VIP 5 processor. At the advice of the rep we have switched from formalin to Tissue Tek VIP Fixative for processing. In my opinion, this has made the tissues more brittle and has also had a negative impact on IHC. Has anyone else noticed a problem like this? Thanks, Lester J. Raff, MD Medical Director UroPartners, LLC Laboratory ph: 708-486-0076 fax: 708-486-0080 From petepath <@t> yahoo.com Tue Apr 18 08:32:14 2006 From: petepath <@t> yahoo.com (Stephen Peters M.D.) Date: Tue Apr 18 08:32:20 2006 Subject: [Histonet] Cryostat Message-ID: <20060418133214.79575.qmail@web30405.mail.mud.yahoo.com> Hi Susan, I posted this eply to a similar question a year ago. My feelings still hold. We have had 4 Leica 1850s at HUMC in a very busy surg path practice for about 10 years. I originally chose this cryostat, because it was the most comfortable and easy for me to cut using brush technique. My hands were not twisted in a painful knot. I felt the stage was the most accessable for retrieving. I also found the controls very easy and in no time I could function at my usual pace. It seemed to have a very solid microtome and was considerably more spacious than our old tissue teks. I compared the Leica to the other brands at this clinical level and found this to be my choice for all of the above reasons. I am very happy with the choice I made. They have been reliable work horses and have required very little more than routine maintenence. I have been quite satisfied with the quality of the preparations I am capable of making and I find the work space accomodating. You have 360 degree chuck rotation and a single knob for X-Y axis adjustment. I have kept up with most of the new designs of the other brands. I would still buy another 1850 tomorrow. Trust me none of them are perfect, or even close, but of what is out there this is my choice. I must mention that I do have a relationship with Leica for a about two years. They have been helping me promote an embedding system that I developed while practicing in these cryostats. I thought about whether to write this at all, but everything I have to say came long before. I have had great success in a very busy practice with these cryostats I am happy to recommend them. Stephen From HornHV <@t> archildrens.org Tue Apr 18 08:59:04 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Tue Apr 18 08:59:52 2006 Subject: [Histonet] Cryostat Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BE046@EMAIL.archildrens.org> We have a Leica 1800 and a 1850. We love both of them. They are reliable, user friendly and the cryostat I have ever used. We have had very few problems with them and Leica is responsive to our calls. Great cryostat, Good service. Hazel Horn ------------------------------------------------------------------------------ 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 gcallis <@t> montana.edu Tue Apr 18 09:40:37 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Apr 18 09:40:57 2006 Subject: [Histonet] Large vessel endothelium problem In-Reply-To: <4444D9D7.2060405@helsinki.fi> References: <4444D9D7.2060405@helsinki.fi> Message-ID: <6.0.0.22.1.20060418083554.01b56650@gemini.msu.montana.edu> You did not indicate how you do the paraformaldehyde fixation? Immersion or perfusion? Vascular perfusion followed by immersion for a few hours may help your problem - to ensure the tissue is fixed from the inside out and in particular the lumens of your large blood (?) vessels. You did not give details on what you do with the vessels (opened, etc, stretched out) during fixation or processing? At 06:21 AM 4/18/2006, you wrote: >Hello! > >I need to do some immunostaining of large vessel endothelium >(paraformaldehyde-fixed paraffin material). >The problem is, the endothelium is easily stripped off and lost, >apparently in early steps of sample processing. >Any tips how to keep the stuff intact? > >With best regards, >Mikael > >-- >//////////////////////////////////////////////////////////// > > Mikael Niku URL: www.helsinki.fi/~mniku/ > University of Helsinki Dept. Basic Veterinary Sciences > > - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? > Minusta se olisi erinomainen > ajatus! > - Gandhi > >//////////////////////////////////////////////////////////// > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 406 994-4303 (FAX) From TillRenee <@t> uams.edu Tue Apr 18 10:25:38 2006 From: TillRenee <@t> uams.edu (Till, Renee) Date: Tue Apr 18 10:26:05 2006 Subject: [Histonet] lendrums' phloxine/tartrazine Message-ID: <11F927674DEBDC43B960809A7403C5D2C04149@MAILPED.ad.uams.edu> Does anyone have experience with this stain? I have been trying it for a few weeks now on mouse jejunum. Tissues from some of my animals have wonderfully stained Paneth cells, while on others the Paneth cells are the first to be de-stained by the tartrazine. And these are all mouse jejunum from mice of the same age. Some do have a gene knockout, but I have not noticed the problem being only in the knockouts or only in the wild type, I am having problems with some from both. Any ideas? I have tried adjusting the time of the phloxine solution, but so far this hasn't helped any. Neither has making fresh (how long are these stains good for anyway?). Maybe there was more autolysis before fixation? I don't know. The times with the tartrazine solution have varied great on all the ones that have stained well. Anywhere from 5 minutes to over an hour. Renee' Till, HT Arkansas Childrens Nutrition Center 1120 Marshall Little Rock, AR 72202 ================================================================================================ Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. ================================================================================================ From scott9379 <@t> gmail.com Tue Apr 18 11:04:36 2006 From: scott9379 <@t> gmail.com (Scott Parker) Date: Tue Apr 18 11:04:41 2006 Subject: [Histonet] mouse spleen confocal Message-ID: Dear all, Can anybody help me with some antibody staining I have been doing on mouse spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) ( 53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately all we see is vast amounts of auto-fluorescence making antibody detection impossible. We really need to get this sorted because ultimatley we'd like to have GFP and two different secondaries (labelling 3 things in total). Our protocol is basically: Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and stored at -20. When ready for use slides are warmed to room temp and fixed in -20 C acetone for 3 mins then re-hydrated in PBS for 5 mins. Sections are allowed to dry and blocked for 30 min in 10% normal mouse serum in PBS (blocking buffer). Sections are stained in blocking buffer, washed and then stained with secondary (also in blocking buffer). We wash in PBS with 0.1% tween. Having done some searches on histonet I see that tween should probably not be used. Furthermore I have tried fixing in 3.2 paraformaldehyde with the same result. Sections that are treated as above without antibody look exactly the same as sections treated with antibody when viewed under the confocal. Basically lots of auto-fluorescence nicely labelling all the cells. Any help or protocols would be glady appreciated. Scott -- Scott Parker, Ph.D. School of Medicine St. Louis University USA From jcline <@t> wchsys.org Tue Apr 18 11:47:35 2006 From: jcline <@t> wchsys.org (Joyce Cline) Date: Tue Apr 18 11:47:52 2006 Subject: [Histonet] RE: RHS-1 and MicroMed T/T Mega/Rodger In-Reply-To: <20060417154753.79318.qmail@web38304.mail.mud.yahoo.com> Message-ID: <000001c66307$cc7ddf80$1d2a14ac@wchsys.org> I have the T/T Mega, I use it up to three times a day. Our pathologists like the way it processes biopsies, rather than the overnight processor. I process GI's, cervical, vocal cord, liver (wonderful results), lung and bladder. Your tissue ideally should be 2mm, some GI polyps have to be bisected. Large polyps are not put in. I use reagent alcohol before the JFC solution for all runs. Specimens have to be in 10% buffered formalin for a minimum of 2 hours. ************************************************************************ ** If you are familiar with using either of these units, or currently process with these units please let me know you're opinions (good or bad). We are going to be in the market for possibly both units soon. Thanks in advance to all members who respond. Rodger ***** 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 cfavara <@t> niaid.nih.gov Tue Apr 18 12:15:00 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Tue Apr 18 12:15:06 2006 Subject: [Histonet] CD-3 woes Message-ID: I have been using Dako CD-3 on murine formalin fixed paraffin embedded tissue and am experiencing some variation in staining. My protocol: pretreatment with Citrate pH 6.0 @120C for 5 minutes under pressure, an overnight incubation of the primary 1:1500 @4C, followed by 30 minute incubation with Vector biotinylated anti Rabbit IgG 1:250, 30 minutes Biogenex SS Streptavidin all steps following primary are done on the Ventana Nexus @RT using their AEC. I used to do the stain reliably but have a new lot and reliability is not optimal. I use a mouse spleen and brain with T-cell infiltration as positive controls and a NL brain as a negative control. Sometimes the stain is so crisp and other times it is muddy with a decrease in the number of cells stained. I have checked lot numbers on all regents and they are the same. It has occurred to me that doing the peroxidase block following the primary could be problematic however I do not see the same problem with other antibodies treated the same way and seem to recall that peroxidase block is done by many following the primary incubation. I have tried different buffers for pretreatment and also PK. Any suggestions/ commiserations would be welcome. Thanks, c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives From cfavara <@t> niaid.nih.gov Tue Apr 18 12:16:30 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Tue Apr 18 12:16:38 2006 Subject: [Histonet] mouse spleen confocal Message-ID: Hopefully Gayle Callis will reply she is the guru, she has written extensively and I am sure you can find information if you do a search of the archives. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Scott Parker [mailto:scott9379@gmail.com] Sent: Tuesday, April 18, 2006 9:05 AM To: histonet Subject: [Histonet] mouse spleen confocal Dear all, Can anybody help me with some antibody staining I have been doing on mouse spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) ( 53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately all we see is vast amounts of auto-fluorescence making antibody detection impossible. We really need to get this sorted because ultimatley we'd like to have GFP and two different secondaries (labelling 3 things in total). Our protocol is basically: Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and stored at -20. When ready for use slides are warmed to room temp and fixed in -20 C acetone for 3 mins then re-hydrated in PBS for 5 mins. Sections are allowed to dry and blocked for 30 min in 10% normal mouse serum in PBS (blocking buffer). Sections are stained in blocking buffer, washed and then stained with secondary (also in blocking buffer). We wash in PBS with 0.1% tween. Having done some searches on histonet I see that tween should probably not be used. Furthermore I have tried fixing in 3.2 paraformaldehyde with the same result. Sections that are treated as above without antibody look exactly the same as sections treated with antibody when viewed under the confocal. Basically lots of auto-fluorescence nicely labelling all the cells. Any help or protocols would be glady appreciated. Scott -- Scott Parker, Ph.D. School of Medicine St. Louis University USA _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gagnone <@t> KGH.KARI.NET Tue Apr 18 12:23:28 2006 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Tue Apr 18 12:23:33 2006 Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions Message-ID: Hello Listers, We currently do immunofluorescence for IgG, IgA, IgM, C3, C1q, Fib., Kappa, Lambda, and C4d on renal biopsies, and the first four on skin biopsies. We are using Dako Antibody Diluting Buffer and mostly Dako FITC antisera. We currently make up about 1 ml of each, and the dilutions last us between 1 and 3 weeks. We would like to keep them longer if possible, making them up in larger quantities, possibly with the goal of transferring them to dispensers on our Ventana XT's. ] Is there anyone doing the same immunofluorescence panel and dilutions, and if so, how long are you able to use your dilutions once made? Thanks for your assistance, Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada From gcallis <@t> montana.edu Tue Apr 18 12:34:56 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Apr 18 12:35:10 2006 Subject: [Histonet] mouse spleen confocal In-Reply-To: References: Message-ID: <6.0.0.22.1.20060418110943.01b45748@gemini.msu.montana.edu> Scott, Is your secondary adsorbed to mouse tissue? You blocking serum should be matched to the host of the secondary and you can add 1 to 2% mouse serum to this normal serum block AND use this NSB as the diluent of the secondary. The secondary should be F(ab')2 frag of IgG to not bind to fc receptors on your tissues. If your secondary is made in rabbit, then you may get background aka autofluorescence problems when the rabbit IgG sticks to everything. Did you do a dilution panel on your primary starting at 10 ug/ml? You did not give a concentration in ug/ml. I suggest you try this method to get rid of secondary entirely. NSB 30 min Strepavidin/biotin block (kit from Vector) kit instructions Biotinylated Rat anti Mouse CD8 (Ly3.2) diluted in the normal serum block I just described incubated for 30 min. CD8 is an antibody that requires a higher concentration at times - so do a dilution panel. Immunologist here indicated it was probably due to low affinity. Come back with Strepavidin-Alexa 633 diluted 1:1000 in buffer with Tween (yes, you can use it but we like a lower concentration!) Just to let you know, the GFP will probably NOT survive the acetone fixation, a common problem and CD8 will NOT survive formalin or paraformaldehyde fixation. GFP is nicely lit up by using an AntiGFP antibody. Rockland has an excellent Goat antiGFP for this purpose and we come back with donkey antiGoat f(ab')2 frag of IgG, adsorbed to mouse, a FITC conjugate - you will have to work faster with this one OR you can conjugate a favorite secondary with Alexa 488 per Molecular Probes kit - easy to do. We use Tween 20 for all our double immunofluorescence staining, just use 0.025%, very dilute to keep frozen sections intact. An excellent fixation protocol for murine CD markers for IFA work (including CLSM) is air dry your sections OVERNIGHT at RT, fix in 75% acetone/25% absolute ethanol for 5 min at RT the next day, go directly to buffer, do NOT AIR DRY AGAIN after fixation. You may be surprised at quality of CD8 staining but beware, GFP does NOT survive this solvent fixation. t 10:04 AM 4/18/2006, you wrote: >Dear all, >Can anybody help me with some antibody staining I have been doing on mouse >spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) ( >53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately all >we see is vast amounts of auto-fluorescence making antibody detection >impossible. We really need to get this sorted because ultimatley we'd like >to have GFP and two different secondaries (labelling 3 things in total). >Our protocol is basically: > >Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and >stored at -20. >When ready for use slides are warmed to room temp and fixed in -20 C acetone >for 3 mins then re-hydrated in PBS for 5 mins. >Sections are allowed to dry and blocked for 30 min in 10% normal mouse serum >in PBS (blocking buffer). >Sections are stained in blocking buffer, washed and then stained with >secondary (also in blocking buffer). We wash in PBS with 0.1% tween. >Having done some searches on histonet I see that tween should probably not >be used. Furthermore I have tried fixing in 3.2 paraformaldehyde with the >same result. >Sections that are treated as above without antibody look exactly the same as >sections treated with antibody when viewed under the confocal. Basically >lots of auto-fluorescence nicely labelling all the cells. >Any help or protocols would be glady appreciated. >Scott > >-- >Scott Parker, Ph.D. >School of Medicine >St. Louis University >USA >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet Gayle Callis MT,HT,HTL(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University - Bozeman PO Box 173610 Bozeman MT 59717-3610 406 994-6367 406 994-4303 (FAX) From cfavara <@t> niaid.nih.gov Tue Apr 18 12:52:16 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Tue Apr 18 12:52:20 2006 Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions Message-ID: You might want to check for yourself. Make up the amount you would like to use and run a control slide weekly until signal fades. I would make sure all other lot numbers are the same. I would think that time exposed to light may be a significant factor. Just a thought. c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Gagnon, Eric [mailto:gagnone@KGH.KARI.NET] Sent: Tuesday, April 18, 2006 10:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions Hello Listers, We currently do immunofluorescence for IgG, IgA, IgM, C3, C1q, Fib., Kappa, Lambda, and C4d on renal biopsies, and the first four on skin biopsies. We are using Dako Antibody Diluting Buffer and mostly Dako FITC antisera. We currently make up about 1 ml of each, and the dilutions last us between 1 and 3 weeks. We would like to keep them longer if possible, making them up in larger quantities, possibly with the goal of transferring them to dispensers on our Ventana XT's. ] Is there anyone doing the same immunofluorescence panel and dilutions, and if so, how long are you able to use your dilutions once made? Thanks for your assistance, Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Tue Apr 18 13:08:51 2006 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Tue Apr 18 13:08:58 2006 Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions Message-ID: Eric, I do all of the sam FITC antibodies for kidney here (all from Dako except for my C4d which is from Biogenesis). I make up 16ml. at a time and they have worked for me for upwards of 2-3 months with no pathologist complaints. I keep them in dark brown bottles and am very careful to minimize any other light exposure. I also make sure to keep them cool. Good Luck, Glen Dawson BS, HT & QIHC (ASCP) IHC Manager Milwaukee, WI -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Gagnon, Eric Sent: Tuesday, April 18, 2006 11:23 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions Hello Listers, We currently do immunofluorescence for IgG, IgA, IgM, C3, C1q, Fib., Kappa, Lambda, and C4d on renal biopsies, and the first four on skin biopsies. We are using Dako Antibody Diluting Buffer and mostly Dako FITC antisera. We currently make up about 1 ml of each, and the dilutions last us between 1 and 3 weeks. We would like to keep them longer if possible, making them up in larger quantities, possibly with the goal of transferring them to dispensers on our Ventana XT's. ] Is there anyone doing the same immunofluorescence panel and dilutions, and if so, how long are you able to use your dilutions once made? Thanks for your assistance, Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Tue Apr 18 13:10:06 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Tue Apr 18 13:09:02 2006 Subject: [Histonet] lendrums' phloxine/tartrazine In-Reply-To: <11F927674DEBDC43B960809A7403C5D2C04149@MAILPED.ad.uams.edu> References: <11F927674DEBDC43B960809A7403C5D2C04149@MAILPED.ad.uams.edu> Message-ID: <44452B7E.4020609@uwo.ca> Lendrum's original paper explains the method very thoroughly and has exact instructions for making adjustments to the colour scheme. The reference is LENDRUM, A. C. 1947. The phloxin-tartrazine method as a general histological stain and for the demonstration of inclusion bodies. Journal of Pathology and Bacteriology 59, 399-404. I strongly recommend reading this. Regarding stability, the phloxin B solution can be kept for one year; the saturated solution of tartrazine in ethylene glycol monoethyl ether is stable for years. It collects phloxin B with repeated re-use. A more recent paper in this field is CLARK, G. 1979. Displacement. Stain Technology 54: 111-119. Clark investigated the method and came up with an improved version using two different dyes. I've seen only one published reference to the Clark's method; perhaps most users are happy with Lendrum's. John Kiernan Anatomy, UWO London, Canada ____________________________ "Till, Renee" wrote: > > Does anyone have experience with this stain? I have been trying it for a > few weeks now on mouse jejunum. Tissues from some of my animals have > wonderfully stained Paneth cells, while on others the Paneth cells are > the first to be de-stained by the tartrazine. And these are all mouse > jejunum from mice of the same age. Some do have a gene knockout, but I > have not noticed the problem being only in the knockouts or only in the > wild type, I am having problems with some from both. Any ideas? I have > tried adjusting the time of the phloxine solution, but so far this > hasn't helped any. Neither has making fresh (how long are these stains > good for anyway?). Maybe there was more autolysis before fixation? I > don't know. The times with the tartrazine solution have varied great on > all the ones that have stained well. Anywhere from 5 minutes to over an > hour. > > > > > > Renee' Till, HT > > > > Arkansas Childrens Nutrition Center > > 1120 Marshall > > Little Rock, AR > > 72202 > > > > > > ================================================================================================ > > Confidentiality Notice: This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original message. > ================================================================================================ > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From scott9379 <@t> gmail.com Tue Apr 18 13:48:01 2006 From: scott9379 <@t> gmail.com (Scott Parker) Date: Tue Apr 18 13:48:06 2006 Subject: [Histonet] mouse spleen confocal In-Reply-To: <6.0.0.22.1.20060418110943.01b45748@gemini.msu.montana.edu> References: <6.0.0.22.1.20060418110943.01b45748@gemini.msu.montana.edu> Message-ID: Gayle, Thanks for the tips. We haven't even got to the stage where we dilute primaries. With or without antibodies added i see exactly the same thing. Just lots of auto-fluorescence! I'm happy to try the tips you gave me, but to be honest my primary concern at this time is to try to get a section that isn't completely red. Any ideas on that? Scott On 18/04/06, Gayle Callis wrote: > > Scott, > > Is your secondary adsorbed to mouse tissue? You blocking serum should be > matched to the host of the secondary and you can add 1 to 2% mouse serum > to > this normal serum block AND use this NSB as the diluent of the > secondary. The secondary should be F(ab')2 frag of IgG to not bind to fc > receptors on your tissues. If your secondary is made in rabbit, then you > may get background aka autofluorescence problems when the rabbit IgG > sticks > to everything. > > Did you do a dilution panel on your primary starting at 10 ug/ml? You did > not give a concentration in ug/ml. > > I suggest you try this method to get rid of secondary entirely. > > NSB 30 min > Strepavidin/biotin block (kit from Vector) kit instructions > > Biotinylated Rat anti Mouse CD8 (Ly3.2) diluted in the normal serum block > I > just described incubated for 30 min. CD8 is an antibody that requires a > higher concentration at times - so do a dilution panel. Immunologist here > indicated it was probably due to low affinity. > > Come back with Strepavidin-Alexa 633 diluted 1:1000 in buffer with Tween > (yes, you can use it but we like a lower concentration!) > > Just to let you know, the GFP will probably NOT survive the acetone > fixation, a common problem and CD8 will NOT survive formalin or > paraformaldehyde fixation. GFP is nicely lit up by using an AntiGFP > antibody. Rockland has an excellent Goat antiGFP for this purpose and we > come back with donkey antiGoat f(ab')2 frag of IgG, adsorbed to mouse, a > FITC conjugate - you will have to work faster with this one OR you can > conjugate a favorite secondary with Alexa 488 per Molecular Probes kit - > easy to do. > > We use Tween 20 for all our double immunofluorescence staining, just use > 0.025%, very dilute to keep frozen sections intact. > > An excellent fixation protocol for murine CD markers for IFA work > (including CLSM) is air dry your sections OVERNIGHT at RT, fix in 75% > acetone/25% absolute ethanol for 5 min at RT the next day, go directly to > buffer, do NOT AIR DRY AGAIN after fixation. You may be surprised at > quality of CD8 staining but beware, GFP does NOT survive this solvent > fixation. > > > > t 10:04 AM 4/18/2006, you wrote: > >Dear all, > >Can anybody help me with some antibody staining I have been doing on > mouse > >spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) > ( > >53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately > all > >we see is vast amounts of auto-fluorescence making antibody detection > >impossible. We really need to get this sorted because ultimatley we'd > like > >to have GFP and two different secondaries (labelling 3 things in total). > >Our protocol is basically: > > > >Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and > >stored at -20. > >When ready for use slides are warmed to room temp and fixed in -20 C > acetone > >for 3 mins then re-hydrated in PBS for 5 mins. > >Sections are allowed to dry and blocked for 30 min in 10% normal mouse > serum > >in PBS (blocking buffer). > >Sections are stained in blocking buffer, washed and then stained with > >secondary (also in blocking buffer). We wash in PBS with 0.1% tween. > >Having done some searches on histonet I see that tween should probably > not > >be used. Furthermore I have tried fixing in 3.2 paraformaldehyde with the > >same result. > >Sections that are treated as above without antibody look exactly the same > as > >sections treated with antibody when viewed under the confocal. Basically > >lots of auto-fluorescence nicely labelling all the cells. > >Any help or protocols would be glady appreciated. > >Scott > > > >-- > >Scott Parker, Ph.D. > >School of Medicine > >St. Louis University > >USA > >_______________________________________________ > >Histonet mailing list > >Histonet@lists.utsouthwestern.edu > >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > Gayle Callis > MT,HT,HTL(ASCP) > Research Histopathology Supervisor > Veterinary Molecular Biology > Montana State University - Bozeman > PO Box 173610 > Bozeman MT 59717-3610 > 406 994-6367 > 406 994-4303 (FAX) > > > -- Scott Parker, Ph.D. School of Medicine St. Louis University USA From jcline <@t> wchsys.org Tue Apr 18 15:14:46 2006 From: jcline <@t> wchsys.org (Joyce Cline) Date: Tue Apr 18 15:14:58 2006 Subject: [Histonet] RE: RHS-1 and MicroMed T/T Mega/Jeanine In-Reply-To: Message-ID: <000001c66324$be2472b0$1d2a14ac@wchsys.org> Jeanine, I use the microwave for both. I use a fixation program to fix my sentinel nodes and the breast tissue. I still hold the breast tissue overnight but it processes the next day much better. I also will fix the colon cases if they have not been in formalin long enough. The fixation takes an hour at 40 degrees. I have found a company that makes the plastic parts for the microwave. I have received a lid for my Mega T/T and find that it works the same as the manufacturer's pieces. The company is Coleman Manufacturing & Design. LLC. they are in Winnsboro, SC (they make the racks, stir plates, carrier and lids) ***** 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 squadeer <@t> utmb.edu Tue Apr 18 16:50:49 2006 From: squadeer <@t> utmb.edu (Quadeer, Shahnaz S.) Date: Tue Apr 18 16:50:54 2006 Subject: [Histonet] Re- Unscri be Message-ID: <13362C82244EEA4B9BD90AC1D90E5C1BEAFE50@EXCH2K4.utmb.edu> Please unscribe From kzhong888 <@t> yahoo.com Tue Apr 18 19:32:27 2006 From: kzhong888 <@t> yahoo.com (dfs dsaf) Date: Tue Apr 18 19:32:32 2006 Subject: [Histonet] IEC CTD harris cryostat Message-ID: <20060419003227.51473.qmail@web52902.mail.yahoo.com> Dear histonet subscribers: I am look for a very old IEC cryostat. Model CTD-harris or similar model. It is the small ice cream cart type. If you have one to sell or know someone please email me at Kzhong888@yahoo.com. Any condition with or without microtome. regards kirk Zhong histo tech --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From sohail_e <@t> yahoo.com Tue Apr 18 21:00:05 2006 From: sohail_e <@t> yahoo.com (sohail ejaz) Date: Tue Apr 18 21:00:13 2006 Subject: [Histonet] Over lapped flouresence in double staining Message-ID: <20060419020006.35066.qmail@web30615.mail.mud.yahoo.com> I am doing double staining of endothelial cells and smooth muscles of the blood vessels with vWF and SMA. While doing double staining I have encountered overlapping of both antibodies and I cant see a clear differentiation between the fluorescence of these two antibodies. Could you please guide me how to over come this problem. Thanks Dr. Sohail --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From BMolinari <@t> heart.thi.tmc.edu Wed Apr 19 05:33:46 2006 From: BMolinari <@t> heart.thi.tmc.edu (Molinari, Betsy) Date: Wed Apr 19 05:35:09 2006 Subject: [Histonet] Cryostat Message-ID: I have an 1800 and I agree with Hazel. Reliable and good tech support. Betsy Molinari HT (ASCP) Texas Heart Institute Cardiovascular Pathology 6770 Bertner Ave. Houston,TX 77030 832-355-6524 832-355-6812 (fax) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Horn, Hazel V Sent: Tuesday, April 18, 2006 8:59 AM To: Stephen Peters M.D.; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Cryostat We have a Leica 1800 and a 1850. We love both of them. They are reliable, user friendly and the cryostat I have ever used. We have had very few problems with them and Leica is responsive to our calls. Great cryostat, Good service. Hazel Horn ------------------------------------------------------------------------ ------ 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. ======================================================================== ====== _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From nancy.troiano <@t> yale.edu Wed Apr 19 06:53:58 2006 From: nancy.troiano <@t> yale.edu (Nancy W. Troiano) Date: Wed Apr 19 06:53:53 2006 Subject: [Histonet] beginner questions on methacrylate resins Message-ID: <5.2.1.1.2.20060419075141.00c44b98@email.med.yale.edu> Hi Sherri - we have extensive experience with methacrylate resins and prefer to make our own MMA , however we do use a kit for GMA. I am not familiar with your particular staining technique as we work primarily with bone specimens, but you certainly can contact me if you have any questions about embedding methods/applications. From jqb7 <@t> cdc.gov Wed Apr 19 09:01:12 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine) Date: Wed Apr 19 09:06:15 2006 Subject: [Histonet] M. leprae control slides Message-ID: Hello all: Can anyone give me a source of M. leprae controls? Wet tissue, paraffin blocks, slides, etc. Thanks, Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov From tkngflght <@t> yahoo.com Wed Apr 19 10:13:52 2006 From: tkngflght <@t> yahoo.com (Cheryl) Date: Wed Apr 19 10:13:57 2006 Subject: [Histonet] Microwave transparent plastics: Coleman Mfg contact information In-Reply-To: Message-ID: <20060419151352.620.qmail@web50906.mail.yahoo.com> Hi All~ Seems like the Coleman Manufacturing folks have several products we need. They make microwave transparent plastics in standard forms and custom designs (racks, vessels, etc), and are just getting into the AP Clinical and Research markets (us!). They have been in industrial applications so they aren't a new company, just new to lab. I talked with Jay & Jeremy Coleman a couple of times--they can make almost anything and their prices are--from what he quoted--reasonable (less costly than similar products from Miles and Sakura, and include a warranty.) He said they can provide documentation that their products are 'transparent' to cover the new CAP ANP regulation. They don't know AP/Med lab very well but have been responsive and willing to learn to be able to service our needs. Jeremy & Jay -- Coleman Manufacturing Phone 803.633.2124 Fax 803.635.9401 coleman_manufacturing@yahoo.com web site under construction--info to follow. They've worked with several labs to test their products with good results. No, I'm not getting a kickback--just doing my best to help as others have helped me :) Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. From jason.burrill <@t> crl.com Wed Apr 19 10:19:45 2006 From: jason.burrill <@t> crl.com (jason.burrill@crl.com) Date: Wed Apr 19 10:20:08 2006 Subject: [Histonet] Bouin's alternative/substitution question Message-ID: We are removing Bouin's fixative from our laboratory and we are looking for an alternative. Has anyone had experience in using the Bouin's Fixative Substitution distributed by IMEB or something similar from another vendor? From reviewing the Histonet archives I see that most people use Davidson's fixative but I just wanted to see if anyone had any personal experience with these substitutions. Thanks in advance, Jason Jason D. Burrill Manager, Histology Charles River Laboratories 251 Ballardvale Street Wilmington, MA 01887 ph: 978-658-6000 ext 1652 fax: 978-988-8793 E-mail: jason.burrill@crl.com From gcallis <@t> montana.edu Wed Apr 19 11:02:17 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Apr 19 11:02:29 2006 Subject: [Histonet] Re: Mouse lungs OCT and curling In-Reply-To: <46a3be380604181724q17ed252eic030749c4fe88ce6@mail.gmail.co m> References: <46a3be380604181724q17ed252eic030749c4fe88ce6@mail.gmail.com> Message-ID: <6.0.0.22.1.20060419095730.01b64be0@gemini.msu.montana.edu> Carment, How thick are you cutting? What is the temperature of the block and knife? Do you use a brush or antiroll device? How do you pick up the section? Give me more details about HOW you do your cryosectioning, what kind of cryostat, high or low profile blades, etc - then I will comment. We never get curled lung sections, they are flat. At 06:24 PM 4/18/2006, you wrote: >I was reading your advice about filling the lungs with OCT to maintain >their structure. I was wondering if you have a trick to keep them from >curling when they are sectioned on the cryostat? Thanks, Carmen Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From pruegg <@t> ihctech.net Wed Apr 19 13:48:50 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Apr 19 13:48:58 2006 Subject: [Histonet] dvaneyck IHCRG Membership Application Form Message-ID: <200604191848.k3JImnka016661@chip.viawest.net> verify NSH membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Wednesday, April 19, 2006 11:30 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: Van Eyck First_Name: Debra NSH_Member: Yes Employer: WAukesha Memorial Hospital Address: 725 American Avenue City: Waukesha State: WI Zip: 53188 Province: Country: USA Phone: 262-928-2112 Ext: Fax: 262-928-4849 Email: deb.vaneyck@phci.org Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Cytospins: Yes Smears_Touch_Preps: Yes Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: Yes IF: IHC_Other: ISH: Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: next 6 months Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 207.67.109.90 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 5.1; SV1; .NET CLR 1.1.4322) Other_Tech From gagnone <@t> KGH.KARI.NET Wed Apr 19 14:28:04 2006 From: gagnone <@t> KGH.KARI.NET (Gagnon, Eric) Date: Wed Apr 19 14:28:15 2006 Subject: [Histonet] RE: Expiry Period - Immunofluorescence Dilutions Message-ID: Cynthia, Glen and Gudrun, thanks for your input on making up extending the life of antisera-FITC dilutions for our immunofluorescence. Excellent answers, much appreciated. Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada From sjchtascp <@t> yahoo.com Wed Apr 19 14:49:18 2006 From: sjchtascp <@t> yahoo.com (Steven Coakley) Date: Wed Apr 19 14:49:22 2006 Subject: [Histonet] histogel cryosections Message-ID: <20060419194918.72148.qmail@web38208.mail.mud.yahoo.com> Has anyone ever used histogel/oct for FS. I've been asked to attempt cryosectioning cultured embryo bodies fr IHC. Steve --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day From EJones <@t> Ventanamed.com Wed Apr 19 14:51:21 2006 From: EJones <@t> Ventanamed.com (Emma JONES) Date: Wed Apr 19 14:51:25 2006 Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) Message-ID: Hi Eric,=20 Have you considered asking your Ventana rep for information on their = FITC reagents. The expiry dates on those are far longer than any you = will get from making your own up. Approximately 12 months + regards Emma Jones Today's Topics: 1. CD-3 woes (Favara, Cynthia (NIH/NIAID) [E]) 2. RE: mouse spleen confocal (Favara, Cynthia (NIH/NIAID) [E]) 3. Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) 4. Re: mouse spleen confocal (Gayle Callis) 5. RE: Expiry Period - Immunofluorescence Dilutions (Favara, Cynthia (NIH/NIAID) [E]) 6. RE: Expiry Period - Immunofluorescence Dilutions (Dawson, Glen) 7. Re: lendrums' phloxine/tartrazine (John Kiernan) 8. Re: mouse spleen confocal (Scott Parker) 9. RE: RE: RHS-1 and MicroMed T/T Mega/Jeanine (Joyce Cline) 10. Re- Unscri be (Quadeer, Shahnaz S.) 11. IEC CTD harris cryostat (dfs dsaf) 12. Over lapped flouresence in double staining (sohail ejaz) 13. RE: Cryostat (Molinari, Betsy) 14. beginner questions on methacrylate resins (Nancy W. Troiano) 15. M. leprae control slides (Bartlett, Jeanine) 16. Microwave transparent plastics: Coleman Mfg contact information (Cheryl) 17. Bouin's alternative/substitution question (jason.burrill@crl.com) 18. Re: Mouse lungs OCT and curling (Gayle Callis) ---------------------------------------------------------------------- =20 Message: 3 Date: Tue, 18 Apr 2006 13:23:28 -0400 From: "Gagnon, Eric" Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions To: Message-ID: Content-Type: text/plain; charset=3D"iso-8859-1" =20 =20 Hello Listers, =20 We currently do immunofluorescence for IgG, IgA, IgM, C3, C1q, Fib., = Kappa, Lambda, and C4d on renal biopsies, and the first four on skin = biopsies. We are using Dako Antibody Diluting Buffer and mostly Dako = FITC antisera. =20 We currently make up about 1 ml of each, and the dilutions last us = between 1 and 3 weeks. We would like to keep them longer if possible, = making them up in larger quantities, possibly with the goal of = transferring them to dispensers on our Ventana XT's. ] Is there anyone doing the same immunofluorescence panel and dilutions, = and if so, how long are you able to use your dilutions once made? =20 Thanks for your assistance, =20 Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada --=20 No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.2/314 - Release Date: = 16/04/2006 =20 From pruegg <@t> ihctech.net Wed Apr 19 14:53:46 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Wed Apr 19 14:53:58 2006 Subject: [Histonet] dvaneyck IHCRG Membership Application Form In-Reply-To: <200604191848.k3JImnka016661@chip.viawest.net> Message-ID: <200604191953.k3JJrjka001675@chip.viawest.net> Sorry, I meant to send this to NSH, it comes up as histo on my server and I hit the wrong button. Disregard this message to verify NSH membership to HISTONET Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, April 19, 2006 11:49 AM To: histonet@pathology.swmed.edu Subject: [Histonet] dvaneyck IHCRG Membership Application Form verify NSH membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Wednesday, April 19, 2006 11:30 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: Van Eyck First_Name: Debra NSH_Member: Yes Employer: WAukesha Memorial Hospital Address: 725 American Avenue City: Waukesha State: WI Zip: 53188 Province: Country: USA Phone: 262-928-2112 Ext: Fax: 262-928-4849 Email: deb.vaneyck@phci.org Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Cytospins: Yes Smears_Touch_Preps: Yes Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: Yes IF: IHC_Other: ISH: Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: next 6 months Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 207.67.109.90 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 5.1; SV1; .NET CLR 1.1.4322) Other_Tech _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From LGaliotto <@t> nch.org Wed Apr 19 14:58:11 2006 From: LGaliotto <@t> nch.org (Galiotto, Laura) Date: Wed Apr 19 14:58:17 2006 Subject: [Histonet] bone marrow Fe stains Message-ID: <270614B321ACB44D8C1D91F4F921FDC3036CBCBE@NCH01EX02.nch.org> I have a problem with bone marrow aspirate controls. The problem is that we are limited to the amount of controls we can collect. Even though the tissue control is working if the aspirate control (which are difficult to obtain) does not work we are being asked to repeat the stain until we find a aspirate control that is acceptable. Since I can not guarantee the aspirate control contains an adequate amount of Fe, I will be wasting time and reagents. Can anyone give feedback on how they process stains on bone marrow's, do you only use a tissue control, does anyone know where bone marrow aspirate controls -positive for Iron -can be purchased commercially? Laura Galiotto, HT (ASCP) Histology Facilitator Northwest Community Hospital 800 West Central Road Arlington Heights, Il 60005 847-618-6190 From anh2006 <@t> med.cornell.edu Wed Apr 19 18:56:24 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Wed Apr 19 18:55:50 2006 Subject: [Histonet] Mouse Cytokeratins Message-ID: Hi All, What antibodies are people using to stain for cytokeratins in mouse tissue - frozen or paraffin. Thanks in advance, Andrea -- From gmondragon <@t> gsopath.com Thu Apr 20 01:18:58 2006 From: gmondragon <@t> gsopath.com (Gus Mondragon) Date: Thu Apr 20 01:19:11 2006 Subject: [Histonet] CBG Recycler Message-ID: <03B63DE44B5C014D84F9A9D8A968B5174C4D60@lithium.corp.gsopath.com> Hello Histonetters; I have a CBG 5 gal. Alcohol or Xylene recycler for sale that we no longer use. Good efficient unit is 4 yrs old in excellent condition, trouble/maintenance free. I'd be glad to offer good suggestions on recycling program. If you have any available Leica electric or manual microtomes, let's talk. Gus Mondragon (336) 387-2536; Greensboro NC From c.m.vanderloos <@t> amc.uva.nl Thu Apr 20 01:51:11 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Apr 20 01:51:21 2006 Subject: [Histonet] RE: CD-3 woes (not any longer!) Message-ID: <3263403287ca.3287ca326340@amc.uva.nl> Cynthia, We were not that successful either with the Dako rabbit CD3 antibody on mouse tissue. We changed to the LabVision CD3 rabbit monoclonal (clone SP7) and this worked very well on human, rat and mouse tissues (both cryo and FFPE). E.g. for staining FFPE: HIER with Tris-EDTA pH9.0, CD3 1:1000 (60 min, RT), polymer anti-rabbit/HRP (30 min, RT), DAB+ (5 min, RT). Hope this helps, Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 18 Apr 2006 13:15:00 -0400 From: "Favara, Cynthia \(NIH/NIAID\) [E]" Subject: [Histonet] CD-3 woes To: I have been using Dako CD-3 on murine formalin fixed paraffin embedded tissue and am experiencing some variation in staining. My protocol: pretreatment with Citrate pH 6.0 @120C for 5 minutes under pressure, an overnight incubation of the primary 1:1500 @4C, followed by 30 minute incubation with Vector biotinylated anti Rabbit IgG 1:250, 30 minutes Biogenex SS Streptavidin all steps following primary are done on the Ventana Nexus @RT using their AEC. I used to do the stain reliably but have a new lot and reliability is not optimal. I use a mouse spleen and brain with T-cell infiltration as positive controls and a NL brain as a negative control. Sometimes the stain is so crisp and other times it is muddy with a decrease in the number of cells st! ained. I From c.m.vanderloos <@t> amc.uva.nl Thu Apr 20 02:05:26 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Apr 20 02:05:33 2006 Subject: [Histonet] RE: mouse spleen confocal Message-ID: <3294ac32b9ea.32b9ea3294ac@amc.uva.nl> Dear Scott, You didn't tell us what detection system was used. Your story sounds reminds me to our first experiments with streptavidin-based detection on mouse spleen when we were confronted with heavy endogenous biotin! For staining mouse spleen, either avoid a streptavidin-based detection system or try to block endogenous biotin as Gayle suggested. Please realize that if the above is true and/or Gayle's is right with her suggestion concerning the adsorption of the anti-rat reagent, the signal you observe is no autofluorescence but "unwanted" specific fluorescence! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands From: Scott Parker [[1]mailto:scott9379@gmail.com] Sent: Tuesday, April 18, 2006 9:05 AM To: histonet Subject: [Histonet] mouse spleen confocal Dear all, Can anybody help me with some antibody staining I have been doing on mouse spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) ( 53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately all we see is vast amounts of auto-fluorescence making antibody detection impossible. We really need to get this sorted because ultimatley we'd like to have GFP and two different secondaries (labelling 3 things in total). Our protocol is basically: Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and stored at -20. When ready for use slides are warmed to room temp and fixed in -20 C acetone for 3 mins then re-hydrated in PBS for 5 mins. Secti! ons are a References 1. javascript:main.compose('new', 't=scott9379@gmail.com') From c.m.vanderloos <@t> amc.uva.nl Thu Apr 20 02:16:29 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Thu Apr 20 02:16:49 2006 Subject: [Histonet] RE: Over lapped flouresence in double staining Message-ID: <50578550cc68.50cc68505785@amc.uva.nl> Dear Dr. Sohail, You didn't write us what detection system was used for this double staining. The fact you observed overlap of SMA and vWF (which isn't very likely indeed) makes your detection system a bit suspect. To my opinion the most straightforward (and safe) ways to doublestain these antibodies are: A: cocktail of SMA (mouse) + vWF (rabbit), followed by a secondary cocktail composed of goat anti-mouse/FLUO-1 + goat anti-rabbit/FLUO-2 B: cocktail of SMA, clone 1A4 (mouse IgG2a) + vWF , clone F8/86 (mouse IgG1), followed by a secondary cocktail composed of goat anti-mouse IgG2a/FLUO-1 + goat anti-mouse IgG1/FLUO-2 Lots of success! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 18 Apr 2006 19:00:05 -0700 (PDT) From: sohail ejaz Subject: [Histonet] Over lapped flouresence in double staining To: [1]histonet@lists.utsouthwestern.edu I am doing double staining of endothelial cells and smooth muscles of the blood vessels with vWF and SMA. While doing double staining I have encountered overlapping of both antibodies and I cant see a clear differentiation between the fluorescence of these two antibodies. Could you please guide me how to over come this problem. Thanks Dr. Sohail References 1. mailto:histonet@lists.utsouthwestern.edu From mikael.niku <@t> helsinki.fi Thu Apr 20 02:36:15 2006 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Thu Apr 20 02:36:24 2006 Subject: [Histonet] Large vessel endothelium problem In-Reply-To: <6.0.0.22.1.20060418083554.01b56650@gemini.msu.montana.edu> References: <4444D9D7.2060405@helsinki.fi> <6.0.0.22.1.20060418083554.01b56650@gemini.msu.montana.edu> Message-ID: <444739EF.3020701@helsinki.fi> Hello again. Sorry for a bit unclearly formatted question. More specifically, I'm doing samples of bovine blood vessels, obtained from a slaughterhouse. So perfusion is unfortunately out of question :) And this is about REALLY big vessels, such as the bovine aorta. So the samples are bits (about 1 cm2) of vessel wall cut off and placed in PFA for overnight at +4C, then processed for paraffin sections in a routine manner. And yes, this is primarily about blood vessels, not lymphatics. Regards, Mikael Gayle Callis wrote: > You did not indicate how you do the paraformaldehyde fixation? > Immersion or perfusion? > > Vascular perfusion followed by immersion for a few hours may help your > problem - to ensure the tissue is fixed from the inside out and in > particular the lumens of your large blood (?) vessels. > > You did not give details on what you do with the vessels (opened, etc, > stretched out) during fixation or processing? -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// From kemlo.rogerson <@t> waht.swest.nhs.uk Thu Apr 20 08:17:57 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Thu Apr 20 08:17:18 2006 Subject: [Histonet] Diff Quick Message-ID: We run a one-step FNAC breast clinic and have been using Diff-Quick as our stain; I personally don't like it as I prefer Paps but there you are. We had a recent FNAC that showed histiocytes in the tail of the prep but little else. As you know the bubbly cells tend to migrate to the edges as they are lighter but what we initially failed to notice was that in amongst the blood were poorly stained clumps of cells. After restain with a 'proper' Romanovsky stain they magically appeared and were benign. I shudder to think what would have happened if they had not been spotted and had been malignant. The problem I guess was the blood and together with the slow drying of the smear. Diff-Quick is fast but can have this effect, does anyone have a technique that is nearly as fast as Diff Quick but more robust in these cases? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Through return to simple living comes control of desires. In control of desires stillness is attained. In stillness the world is restored. -- Lao Tzu This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation From jcline <@t> wchsys.org Thu Apr 20 12:41:36 2006 From: jcline <@t> wchsys.org (Joyce Cline) Date: Thu Apr 20 12:41:51 2006 Subject: [Histonet] bone marrow Fe stains In-Reply-To: <270614B321ACB44D8C1D91F4F921FDC3036CBCBE@NCH01EX02.nch.org> Message-ID: <000001c664a1$acd6fa40$1d2a14ac@wchsys.org> I use only purchased iron controls from H.O.M.E. I use the same control for the aspirate block, the core block and the smears. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Galiotto, Laura Sent: Wednesday, April 19, 2006 3:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone marrow Fe stains I have a problem with bone marrow aspirate controls. The problem is that we are limited to the amount of controls we can collect. Even though the tissue control is working if the aspirate control (which are difficult to obtain) does not work we are being asked to repeat the stain until we find a aspirate control that is acceptable. Since I can not guarantee the aspirate control contains an adequate amount of Fe, I will be wasting time and reagents. Can anyone give feedback on how they process stains on bone marrow's, do you only use a tissue control, does anyone know where bone marrow aspirate controls -positive for Iron -can be purchased commercially? Laura Galiotto, HT (ASCP) Histology Facilitator Northwest Community Hospital 800 West Central Road Arlington Heights, Il 60005 847-618-6190 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From Lori.Disher <@t> HCAhealthcare.com Thu Apr 20 12:41:55 2006 From: Lori.Disher <@t> HCAhealthcare.com (Disher Lori) Date: Thu Apr 20 12:42:01 2006 Subject: [Histonet] RE: Bone marrow FE stains Message-ID: <095327C7CDBDF64B9E9728A54799091E1A9798@ORLEV03.hca.corpad.net> Laura, I make two drip slides when assisting in doing a bone marrow. After they dry we do an FE stain on one of them. If the drip is FE positive (as long as it is a good strong positive) I will save the 2nd drip slide for a control and we do not have to do the FE on the bx and clot the next day. Hope this helps. Lori Disher lori.disher@hcahealthcare.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, April 20, 2006 1:07 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, 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. dvaneyck IHCRG Membership Application Form (Patsy Ruegg) 2. RE: Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) 3. histogel cryosections (Steven Coakley) 4. Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) (Emma JONES) 5. RE: dvaneyck IHCRG Membership Application Form (Patsy Ruegg) 6. bone marrow Fe stains (Galiotto, Laura) 7. Mouse Cytokeratins (Andrea T. Hooper) 8. CBG Recycler (Gus Mondragon) 9. RE: CD-3 woes (not any longer!) (C.M. van der Loos) 10. RE: mouse spleen confocal (C.M. van der Loos) 11. RE: Over lapped flouresence in double staining (C.M. van der Loos) 12. Re: Large vessel endothelium problem (Mikael Niku) 13. Diff Quick (Kemlo Rogerson) ---------------------------------------------------------------------- Message: 1 Date: Wed, 19 Apr 2006 12:48:50 -0600 From: "Patsy Ruegg" Subject: [Histonet] dvaneyck IHCRG Membership Application Form To: Message-ID: <200604191848.k3JImnka016661@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" verify NSH membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Wednesday, April 19, 2006 11:30 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: Van Eyck First_Name: Debra NSH_Member: Yes Employer: WAukesha Memorial Hospital Address: 725 American Avenue City: Waukesha State: WI Zip: 53188 Province: Country: USA Phone: 262-928-2112 Ext: Fax: 262-928-4849 Email: deb.vaneyck@phci.org Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Cytospins: Yes Smears_Touch_Preps: Yes Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: Yes IF: IHC_Other: ISH: Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: next 6 months Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 207.67.109.90 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 5.1; SV1; .NET CLR 1.1.4322) Other_Tech ------------------------------ Message: 2 Date: Wed, 19 Apr 2006 15:28:04 -0400 From: "Gagnon, Eric" Subject: [Histonet] RE: Expiry Period - Immunofluorescence Dilutions To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Cynthia, Glen and Gudrun, thanks for your input on making up extending the life of antisera-FITC dilutions for our immunofluorescence. Excellent answers, much appreciated. Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada ------------------------------ Message: 3 Date: Wed, 19 Apr 2006 12:49:18 -0700 (PDT) From: Steven Coakley Subject: [Histonet] histogel cryosections To: Histonet@lists.utsouthwestern.edu Message-ID: <20060419194918.72148.qmail@web38208.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Has anyone ever used histogel/oct for FS. I've been asked to attempt cryosectioning cultured embryo bodies fr IHC. Steve --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day ------------------------------ Message: 4 Date: Wed, 19 Apr 2006 21:51:21 +0200 From: "Emma JONES" Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) To: Message-ID: Content-Type: text/plain; charset="windows-1250" Hi Eric, Have you considered asking your Ventana rep for information on their FITC reagents. The expiry dates on those are far longer than any you will get from making your own up. Approximately 12 months + regards Emma Jones Today's Topics: 1. CD-3 woes (Favara, Cynthia (NIH/NIAID) [E]) 2. RE: mouse spleen confocal (Favara, Cynthia (NIH/NIAID) [E]) 3. Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) 4. Re: mouse spleen confocal (Gayle Callis) 5. RE: Expiry Period - Immunofluorescence Dilutions (Favara, Cynthia (NIH/NIAID) [E]) 6. RE: Expiry Period - Immunofluorescence Dilutions (Dawson, Glen) 7. Re: lendrums' phloxine/tartrazine (John Kiernan) 8. Re: mouse spleen confocal (Scott Parker) 9. RE: RE: RHS-1 and MicroMed T/T Mega/Jeanine (Joyce Cline) 10. Re- Unscri be (Quadeer, Shahnaz S.) 11. IEC CTD harris cryostat (dfs dsaf) 12. Over lapped flouresence in double staining (sohail ejaz) 13. RE: Cryostat (Molinari, Betsy) 14. beginner questions on methacrylate resins (Nancy W. Troiano) 15. M. leprae control slides (Bartlett, Jeanine) 16. Microwave transparent plastics: Coleman Mfg contact information (Cheryl) 17. Bouin's alternative/substitution question (jason.burrill@crl.com) 18. Re: Mouse lungs OCT and curling (Gayle Callis) ---------------------------------------------------------------------- Message: 3 Date: Tue, 18 Apr 2006 13:23:28 -0400 From: "Gagnon, Eric" Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello Listers, We currently do immunofluorescence for IgG, IgA, IgM, C3, C1q, Fib., Kappa, Lambda, and C4d on renal biopsies, and the first four on skin biopsies. We are using Dako Antibody Diluting Buffer and mostly Dako FITC antisera. We currently make up about 1 ml of each, and the dilutions last us between 1 and 3 weeks. We would like to keep them longer if possible, making them up in larger quantities, possibly with the goal of transferring them to dispensers on our Ventana XT's. ] Is there anyone doing the same immunofluorescence panel and dilutions, and if so, how long are you able to use your dilutions once made? Thanks for your assistance, Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada -- No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.2/314 - Release Date: 16/04/2006 ------------------------------ Message: 5 Date: Wed, 19 Apr 2006 13:53:46 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] dvaneyck IHCRG Membership Application Form To: "'Patsy Ruegg'" , Message-ID: <200604191953.k3JJrjka001675@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Sorry, I meant to send this to NSH, it comes up as histo on my server and I hit the wrong button. Disregard this message to verify NSH membership to HISTONET Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, April 19, 2006 11:49 AM To: histonet@pathology.swmed.edu Subject: [Histonet] dvaneyck IHCRG Membership Application Form verify NSH membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Wednesday, April 19, 2006 11:30 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: Van Eyck First_Name: Debra NSH_Member: Yes Employer: WAukesha Memorial Hospital Address: 725 American Avenue City: Waukesha State: WI Zip: 53188 Province: Country: USA Phone: 262-928-2112 Ext: Fax: 262-928-4849 Email: deb.vaneyck@phci.org Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Cytospins: Yes Smears_Touch_Preps: Yes Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: Yes IF: IHC_Other: ISH: Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: next 6 months Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 207.67.109.90 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 5.1; SV1; .NET CLR 1.1.4322) Other_Tech _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 19 Apr 2006 14:58:11 -0500 From: "Galiotto, Laura" Subject: [Histonet] bone marrow Fe stains To: Message-ID: <270614B321ACB44D8C1D91F4F921FDC3036CBCBE@NCH01EX02.nch.org> Content-Type: text/plain; charset="iso-8859-1" I have a problem with bone marrow aspirate controls. The problem is that we are limited to the amount of controls we can collect. Even though the tissue control is working if the aspirate control (which are difficult to obtain) does not work we are being asked to repeat the stain until we find a aspirate control that is acceptable. Since I can not guarantee the aspirate control contains an adequate amount of Fe, I will be wasting time and reagents. Can anyone give feedback on how they process stains on bone marrow's, do you only use a tissue control, does anyone know where bone marrow aspirate controls -positive for Iron -can be purchased commercially? Laura Galiotto, HT (ASCP) Histology Facilitator Northwest Community Hospital 800 West Central Road Arlington Heights, Il 60005 847-618-6190 ------------------------------ Message: 7 Date: Wed, 19 Apr 2006 19:56:24 -0400 From: "Andrea T. Hooper" Subject: [Histonet] Mouse Cytokeratins To: Histonet Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi All, What antibodies are people using to stain for cytokeratins in mouse tissue - frozen or paraffin. Thanks in advance, Andrea -- ------------------------------ Message: 8 Date: Thu, 20 Apr 2006 02:18:58 -0400 From: "Gus Mondragon" Subject: [Histonet] CBG Recycler To: Message-ID: <03B63DE44B5C014D84F9A9D8A968B5174C4D60@lithium.corp.gsopath.com> Content-Type: text/plain; charset="us-ascii" Hello Histonetters; I have a CBG 5 gal. Alcohol or Xylene recycler for sale that we no longer use. Good efficient unit is 4 yrs old in excellent condition, trouble/maintenance free. I'd be glad to offer good suggestions on recycling program. If you have any available Leica electric or manual microtomes, let's talk. Gus Mondragon (336) 387-2536; Greensboro NC ------------------------------ Message: 9 Date: Thu, 20 Apr 2006 08:51:11 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: CD-3 woes (not any longer!) To: Histonet@lists.utsouthwestern.edu Message-ID: <3263403287ca.3287ca326340@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Cynthia, We were not that successful either with the Dako rabbit CD3 antibody on mouse tissue. We changed to the LabVision CD3 rabbit monoclonal (clone SP7) and this worked very well on human, rat and mouse tissues (both cryo and FFPE). E.g. for staining FFPE: HIER with Tris-EDTA pH9.0, CD3 1:1000 (60 min, RT), polymer anti-rabbit/HRP (30 min, RT), DAB+ (5 min, RT). Hope this helps, Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 18 Apr 2006 13:15:00 -0400 From: "Favara, Cynthia \(NIH/NIAID\) [E]" Subject: [Histonet] CD-3 woes To: I have been using Dako CD-3 on murine formalin fixed paraffin embedded tissue and am experiencing some variation in staining. My protocol: pretreatment with Citrate pH 6.0 @120C for 5 minutes under pressure, an overnight incubation of the primary 1:1500 @4C, followed by 30 minute incubation with Vector biotinylated anti Rabbit IgG 1:250, 30 minutes Biogenex SS Streptavidin all steps following primary are done on the Ventana Nexus @RT using their AEC. I used to do the stain reliably but have a new lot and reliability is not optimal. I use a mouse spleen and brain with T-cell infiltration as positive controls and a NL brain as a negative control. Sometimes the stain is so crisp and other times it is muddy with a decrease in the number of cells st! ained. I ------------------------------ Message: 10 Date: Thu, 20 Apr 2006 09:05:26 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: mouse spleen confocal To: Histonet@lists.utsouthwestern.edu Message-ID: <3294ac32b9ea.32b9ea3294ac@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Dear Scott, You didn't tell us what detection system was used. Your story sounds reminds me to our first experiments with streptavidin-based detection on mouse spleen when we were confronted with heavy endogenous biotin! For staining mouse spleen, either avoid a streptavidin-based detection system or try to block endogenous biotin as Gayle suggested. Please realize that if the above is true and/or Gayle's is right with her suggestion concerning the adsorption of the anti-rat reagent, the signal you observe is no autofluorescence but "unwanted" specific fluorescence! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands From: Scott Parker [[1]mailto:scott9379@gmail.com] Sent: Tuesday, April 18, 2006 9:05 AM To: histonet Subject: [Histonet] mouse spleen confocal Dear all, Can anybody help me with some antibody staining I have been doing on mouse spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) ( 53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately all we see is vast amounts of auto-fluorescence making antibody detection impossible. We really need to get this sorted because ultimatley we'd like to have GFP and two different secondaries (labelling 3 things in total). Our protocol is basically: Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and stored at -20. When ready for use slides are warmed to room temp and fixed in -20 C acetone for 3 mins then re-hydrated in PBS for 5 mins. Secti! ons are a References 1. javascript:main.compose('new', 't=scott9379@gmail.com') ------------------------------ Message: 11 Date: Thu, 20 Apr 2006 09:16:29 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: Over lapped flouresence in double staining To: histonet@lists.utsouthwestern.edu Cc: sohail_e@yahoo.com Message-ID: <50578550cc68.50cc68505785@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Dear Dr. Sohail, You didn't write us what detection system was used for this double staining. The fact you observed overlap of SMA and vWF (which isn't very likely indeed) makes your detection system a bit suspect. To my opinion the most straightforward (and safe) ways to doublestain these antibodies are: A: cocktail of SMA (mouse) + vWF (rabbit), followed by a secondary cocktail composed of goat anti-mouse/FLUO-1 + goat anti-rabbit/FLUO-2 B: cocktail of SMA, clone 1A4 (mouse IgG2a) + vWF , clone F8/86 (mouse IgG1), followed by a secondary cocktail composed of goat anti-mouse IgG2a/FLUO-1 + goat anti-mouse IgG1/FLUO-2 Lots of success! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 18 Apr 2006 19:00:05 -0700 (PDT) From: sohail ejaz Subject: [Histonet] Over lapped flouresence in double staining To: [1]histonet@lists.utsouthwestern.edu I am doing double staining of endothelial cells and smooth muscles of the blood vessels with vWF and SMA. While doing double staining I have encountered overlapping of both antibodies and I cant see a clear differentiation between the fluorescence of these two antibodies. Could you please guide me how to over come this problem. Thanks Dr. Sohail References 1. mailto:histonet@lists.utsouthwestern.edu ------------------------------ Message: 12 Date: Thu, 20 Apr 2006 10:36:15 +0300 From: Mikael Niku Subject: Re: [Histonet] Large vessel endothelium problem To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu Message-ID: <444739EF.3020701@helsinki.fi> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello again. Sorry for a bit unclearly formatted question. More specifically, I'm doing samples of bovine blood vessels, obtained from a slaughterhouse. So perfusion is unfortunately out of question :) And this is about REALLY big vessels, such as the bovine aorta. So the samples are bits (about 1 cm2) of vessel wall cut off and placed in PFA for overnight at +4C, then processed for paraffin sections in a routine manner. And yes, this is primarily about blood vessels, not lymphatics. Regards, Mikael Gayle Callis wrote: > You did not indicate how you do the paraformaldehyde fixation? > Immersion or perfusion? > > Vascular perfusion followed by immersion for a few hours may help your > problem - to ensure the tissue is fixed from the inside out and in > particular the lumens of your large blood (?) vessels. > > You did not give details on what you do with the vessels (opened, etc, > stretched out) during fixation or processing? -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// ------------------------------ Message: 13 Date: Thu, 20 Apr 2006 14:17:57 +0100 From: Kemlo Rogerson Subject: [Histonet] Diff Quick To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain We run a one-step FNAC breast clinic and have been using Diff-Quick as our stain; I personally don't like it as I prefer Paps but there you are. We had a recent FNAC that showed histiocytes in the tail of the prep but little else. As you know the bubbly cells tend to migrate to the edges as they are lighter but what we initially failed to notice was that in amongst the blood were poorly stained clumps of cells. After restain with a 'proper' Romanovsky stain they magically appeared and were benign. I shudder to think what would have happened if they had not been spotted and had been malignant. The problem I guess was the blood and together with the slow drying of the smear. Diff-Quick is fast but can have this effect, does anyone have a technique that is nearly as fast as Diff Quick but more robust in these cases? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Through return to simple living comes control of desires. In control of desires stillness is attained. In stillness the world is restored. -- Lao Tzu This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 23 **************************************** From detmar <@t> mshri.on.ca Thu Apr 20 12:49:13 2006 From: detmar <@t> mshri.on.ca (Jacqui Detmar) Date: Thu Apr 20 12:49:18 2006 Subject: [Histonet] cresyl violet on mouse placentae Message-ID: Hi all. I need some help interpreting my results. I have tried (twice) posting a picture on the histonet.org site, but I think my emails are going off into cyberspace. Anyway, I treated female mice with polycyclic aromatic hydrocarbons (PAHs; BaP and DMBA) for 6 weeks, then waited one week and mated the mice. When I collected the placentae and fetuses at d15.5 (mouse gestation is approx. 19 days), the fetuses were growth-restricted, but the placental weight was the same as controls. I wanted to see if there were any mast cells in the placenta, so I applied a cresyl violet stain I had downloaded from the internet (0.5% CV in 1% glycerol). I didn't see any mast cells, but what I *did* see were numerous rust-coloured deposits in the labyrinth (site of fetal-maternal exchange) of the treated placentae. I showed my pictures to a veterinary pathologist and he thought they might be hemosiderin or an iron-based deposit. When I did a Perl's Prussian Blue and a Turnbull reaction on the sections, I didn't see increased iron deposits in the tissue. Could somebody help me out with this? If you need/want pictures, please let me know. The .tif file is about 280 kb. Thanks, Jacqui Detmar Samuel Lunenfeld Research Institute, Mount Sinai Hospital. Toronto, Ontario, Canada From jcline <@t> wchsys.org Thu Apr 20 12:53:26 2006 From: jcline <@t> wchsys.org (Joyce Cline) Date: Thu Apr 20 12:53:37 2006 Subject: [Histonet] Microwave plastic parts for T/T Mega or RHS Message-ID: <000101c664a3$533c5eb0$1d2a14ac@wchsys.org> I have found the lid I received from Coleman Manuf. worked just as well as the "official" part. The prices are very good, I probably will get more parts from them. ***** 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 gcallis <@t> montana.edu Thu Apr 20 13:35:48 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Thu Apr 20 13:36:04 2006 Subject: [Histonet] Large vessel endothelium problem In-Reply-To: <444739EF.3020701@helsinki.fi> References: <4444D9D7.2060405@helsinki.fi> <6.0.0.22.1.20060418083554.01b56650@gemini.msu.montana.edu> <444739EF.3020701@helsinki.fi> Message-ID: <6.0.0.22.1.20060420122104.01b97bc8@gemini.msu.montana.edu> Mickael, Next question: Are you there when they slaughter of the animal, then collect the blood vessel as soon as possible after death? Or do you get the tissue after many hours of it sitting around in a slaughter house? If not, you may be dealing with autolyzed tissue and poorly preserved epithelium which may be sloughing off before you ever get a sample as per your assessment of "early steps of sample process." You had written: I need to do some immunostaining of large vessel endothelium (paraformaldehyde-fixed paraffin material). The problem is, the endothelium is easily stripped off and lost, apparently in early steps of sample process. This problem could be solved if you are there to collect and fix the tissue immediately after death of the bovine. I suspect this problem could occur due to any delay in opening the vessel and delay of fixation after the death of the animal. Be generous with the amount of fixative you use for the size of the tissue. At 01:36 AM 4/20/2006, you wrote: >Hello again. Sorry for a bit unclearly formatted question. > >More specifically, I'm doing samples of bovine blood vessels, obtained >from a slaughterhouse. So perfusion is unfortunately out of question :) >And this is about REALLY big vessels, such as the bovine aorta. So the >samples are bits (about 1 cm2) of vessel wall cut off and placed in PFA >for overnight at +4C, then processed for paraffin sections in a routine manner. > >And yes, this is primarily about blood vessels, not lymphatics. > >Regards, >Mikael 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 406 994-4303 (FAX) From rodger_65489 <@t> yahoo.com Thu Apr 20 14:02:05 2006 From: rodger_65489 <@t> yahoo.com (Roger Smithwell) Date: Thu Apr 20 14:02:11 2006 Subject: [Histonet] Microwave transparent plastics: Coleman Mfg contact information In-Reply-To: <20060419151352.620.qmail@web50906.mail.yahoo.com> Message-ID: <20060420190205.85448.qmail@web38302.mail.mud.yahoo.com> thanks for the contact info for the Coleman company; customer service is great. Cheryl wrote: Hi All~ Seems like the Coleman Manufacturing folks have several products we need. They make microwave transparent plastics in standard forms and custom designs (racks, vessels, etc), and are just getting into the AP Clinical and Research markets (us!). They have been in industrial applications so they aren't a new company, just new to lab. I talked with Jay & Jeremy Coleman a couple of times--they can make almost anything and their prices are--from what he quoted--reasonable (less costly than similar products from Miles and Sakura, and include a warranty.) He said they can provide documentation that their products are 'transparent' to cover the new CAP ANP regulation. They don't know AP/Med lab very well but have been responsive and willing to learn to be able to service our needs. Jeremy & Jay -- Coleman Manufacturing Phone 803.633.2124 Fax 803.635.9401 coleman_manufacturing@yahoo.com web site under construction--info to follow. They've worked with several labs to test their products with good results. No, I'm not getting a kickback--just doing my best to help as others have helped me :) Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day From kegraves <@t> UTMB.EDU Thu Apr 20 13:59:25 2006 From: kegraves <@t> UTMB.EDU (Graves, Kerry) Date: Thu Apr 20 14:03:42 2006 Subject: [Histonet] unsubscribe Message-ID: <649CE9B357B1174DB9FEE750C1369FC325AAC0@EXCH2K2.utmb.edu> From jestrong <@t> comcast.net Thu Apr 20 14:31:06 2006 From: jestrong <@t> comcast.net (Jes Strong) Date: Thu Apr 20 14:31:27 2006 Subject: [Histonet] Microwave transparent plastics: Coleman Mfg contactinformation In-Reply-To: <20060420190205.85448.qmail@web38302.mail.mud.yahoo.com> Message-ID: <000001c664b0$f8dad330$7400a8c0@Jes> Dear Histonetters, >From the early days of the Histonet, there has always been a policy (It seems to me that used to be enforced) that the Histonet was to be used for the exchange of ideas and peer to peer support, NOT commercial promotion. Having been a vendor to the Histology community for many years, I have always respected that principle and have only contacted members directly and discreetly when appropriate. Unfortunately these days there are a number of vendors, whose names do not need to be mentioned as you all know who they are, who feel that the Histonet is their personal, free, platform for self promotion. In a free market, the best form of censure and control is through the withholding of support from those who abuse the system. I would hope that those of you who feel the same way about this wonderful forum will not support the vendors who promote their products or services through the Histonet, regardless of the company or product; and help to put an end to commercialization of the Histonet. Jes Strong -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roger Smithwell Sent: Thursday, April 20, 2006 2:02 PM To: Cheryl; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microwave transparent plastics: Coleman Mfg contactinformation thanks for the contact info for the Coleman company; customer service is great. Cheryl wrote: Hi All~ Seems like the Coleman Manufacturing folks have several products we need. They make microwave transparent plastics in standard forms and custom designs (racks, vessels, etc), and are just getting into the AP Clinical and Research markets (us!). They have been in industrial applications so they aren't a new company, just new to lab. I talked with Jay & Jeremy Coleman a couple of times--they can make almost anything and their prices are--from what he quoted--reasonable (less costly than similar products from Miles and Sakura, and include a warranty.) He said they can provide documentation that their products are 'transparent' to cover the new CAP ANP regulation. They don't know AP/Med lab very well but have been responsive and willing to learn to be able to service our needs. Jeremy & Jay -- Coleman Manufacturing Phone 803.633.2124 Fax 803.635.9401 coleman_manufacturing@yahoo.com web site under construction--info to follow. They've worked with several labs to test their products with good results. No, I'm not getting a kickback--just doing my best to help as others have helped me :) Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DeBrosse_Beatrice <@t> Allergan.com Thu Apr 20 14:55:24 2006 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Thu Apr 20 14:56:45 2006 Subject: [Histonet] Microwave transparent plastics: Coleman Mfg contactinformation Message-ID: I have talked to Justin from Coleman Manufacturing before. They do have excellent customer service! Beatrice DeBrosse-Serra HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Roger Smithwell Sent: Thursday, April 20, 2006 12:02 PM To: Cheryl; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Microwave transparent plastics: Coleman Mfg contactinformation thanks for the contact info for the Coleman company; customer service is great. Cheryl wrote: Hi All~ Seems like the Coleman Manufacturing folks have several products we need. They make microwave transparent plastics in standard forms and custom designs (racks, vessels, etc), and are just getting into the AP Clinical and Research markets (us!). They have been in industrial applications so they aren't a new company, just new to lab. I talked with Jay & Jeremy Coleman a couple of times--they can make almost anything and their prices are--from what he quoted--reasonable (less costly than similar products from Miles and Sakura, and include a warranty.) He said they can provide documentation that their products are 'transparent' to cover the new CAP ANP regulation. They don't know AP/Med lab very well but have been responsive and willing to learn to be able to service our needs. Jeremy & Jay -- Coleman Manufacturing Phone 803.633.2124 Fax 803.635.9401 coleman_manufacturing@yahoo.com web site under construction--info to follow. They've worked with several labs to test their products with good results. No, I'm not getting a kickback--just doing my best to help as others have helped me :) Cheryl Kerry, HT(ASCP) Full Staff Inc. Staffing the AP Lab by helping one Tech at a time. 281.883.7704 c 281.852.9457 o admin@fullstaff.org Sign up for the FREE monthly newsletter AP News--updates, tricks of the trade and current issues for Anatomic Pathology Clinical Labs. Send a 'subscribe' request to APNews@fullstaff.org. Please include your name and specialty in the body of the email. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From RCHIOVETTI <@t> aol.com Thu Apr 20 15:01:05 2006 From: RCHIOVETTI <@t> aol.com (RCHIOVETTI@aol.com) Date: Thu Apr 20 15:01:11 2006 Subject: [Histonet] Microwave transparent plastics: Coleman Mfg contactinformation Message-ID: <370.2d53088.31794281@aol.com> I couldn't agree more with Jes. In fact, I've been following this thread with some interest as well. Several of the posts and requests for input have come from folks who have yahoo! email addresses...nothing wrong with that in itself, in fact I have a yahoo addy too. It's a good way to maintain anonymity, but on a listserver like this, it's impossible to tell what's an honest request or honest feedback, and what's a bogus, thinly veiled advertisement. I've been a Histonet member since day one, and please excuse me if I'm wrong, but I smell "bogus" in this whole thread, probably from the original poster who started it. Folks, even if this thread is honest and well-intentioned, you should always, always, *always* check with the manufacturer before using third-party parts in your expensive microwave processor! I'm sure there are warranty issues involved. Now getting off my soapbox... Bob Robert (Bob) Chiovetti, Ph.D. Owner The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Member, Arizona Small Business Association - ASBA (asba.com) Robert (Bob) Chiovetti, Ph.D. Owner The Microscope Works Arizona's Microscopy Resource 132 North Elster Drive Tucson, AZ 85710-3212 USA Tel./Fax 520-546-4986 Manufacturers' Representative Member, Arizona Small Business Association - ASBA From eca9 <@t> georgetown.edu Thu Apr 20 15:39:10 2006 From: eca9 <@t> georgetown.edu (Eva C Andersson) Date: Thu Apr 20 15:39:24 2006 Subject: [Histonet] precipitaion of DAB on top of tissue Message-ID: <26b23b269215.26921526b23b@imap.georgetown.edu> Hello Everyone, I have encountered a very annoying problem. During my DAB step it is precipitating on top of the tissue. At first it seems like black dots floating around on top of the tissue but now it has started attaching itself and it won't go away with washing. There is nothing forming in the tube the DAB is prepared in but only once it has been placed on the tissue. It happens regardless of if I do HRP, LSAB or ABC. I have also tried three different DAB kits and checked the pH of my TBST wash buffer. The only things that are the same for all the times this has happened is the H2O2 from Dako, Protein Block from Biogenex, TBST from DAKO and Antibody diluent with background reducing component from Dako. Would appreciate any suggestions you may have. I have also asked the people who cut my section and they say they have changed nothing in their procedures. And it happens with totally different tissues (kidney, skin). Thank you all in advance for your help, Eva Andersson Research assistant Georgetown University From gatorjanelle <@t> yahoo.com Thu Apr 20 21:21:29 2006 From: gatorjanelle <@t> yahoo.com (Janelle Novak) Date: Thu Apr 20 21:21:37 2006 Subject: [Histonet] Ki67 protocol--frozen sections Message-ID: <000601c664ea$4d2b0e30$4e02a8c0@Dell2005> Does anyone have any good recommendations for a Ki67 staining protocol for feline frozen tissue sections? Thank you, Janelle Novak From Michael.Rice <@t> holy-cross.com Fri Apr 21 07:27:03 2006 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Apr 21 07:27:28 2006 Subject: [Histonet] Meditech pathology module Message-ID: <3BC92F29BE821745AB15E04C98EE028D6D3921@HCH2KMAIL.holy-cross.com> Hi All, We are currently being asked to convert from Copath to Meditech as our pathology module. Does anyone out there have experience with them. So far every thing that I have seen shows it to be inferior to Copath as well as being extremely difficult to build as we in the department are responsible for that. Any and all opinions are welcomed. Thanks mike Michael Rice CT.HT(ASCP) Supervisor Of Pathology Holy Cross Hospital Ft Lauderdale, Fl 33308 954.776.3070 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of histonet-request@lists.utsouthwestern.edu Sent: Thursday, April 20, 2006 1:05 PM To: histonet@lists.utsouthwestern.edu Subject: Histonet Digest, Vol 29, 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. dvaneyck IHCRG Membership Application Form (Patsy Ruegg) 2. RE: Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) 3. histogel cryosections (Steven Coakley) 4. Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) (Emma JONES) 5. RE: dvaneyck IHCRG Membership Application Form (Patsy Ruegg) 6. bone marrow Fe stains (Galiotto, Laura) 7. Mouse Cytokeratins (Andrea T. Hooper) 8. CBG Recycler (Gus Mondragon) 9. RE: CD-3 woes (not any longer!) (C.M. van der Loos) 10. RE: mouse spleen confocal (C.M. van der Loos) 11. RE: Over lapped flouresence in double staining (C.M. van der Loos) 12. Re: Large vessel endothelium problem (Mikael Niku) 13. Diff Quick (Kemlo Rogerson) ---------------------------------------------------------------------- Message: 1 Date: Wed, 19 Apr 2006 12:48:50 -0600 From: "Patsy Ruegg" Subject: [Histonet] dvaneyck IHCRG Membership Application Form To: Message-ID: <200604191848.k3JImnka016661@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" verify NSH membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Wednesday, April 19, 2006 11:30 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: Van Eyck First_Name: Debra NSH_Member: Yes Employer: WAukesha Memorial Hospital Address: 725 American Avenue City: Waukesha State: WI Zip: 53188 Province: Country: USA Phone: 262-928-2112 Ext: Fax: 262-928-4849 Email: deb.vaneyck@phci.org Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Cytospins: Yes Smears_Touch_Preps: Yes Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: Yes IF: IHC_Other: ISH: Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: next 6 months Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 207.67.109.90 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 5.1; SV1; .NET CLR 1.1.4322) Other_Tech ------------------------------ Message: 2 Date: Wed, 19 Apr 2006 15:28:04 -0400 From: "Gagnon, Eric" Subject: [Histonet] RE: Expiry Period - Immunofluorescence Dilutions To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Cynthia, Glen and Gudrun, thanks for your input on making up extending the life of antisera-FITC dilutions for our immunofluorescence. Excellent answers, much appreciated. Eric Gagnon MLT Histology Laboratory Kingston General Hospital Kingston, Ontario, Canada ------------------------------ Message: 3 Date: Wed, 19 Apr 2006 12:49:18 -0700 (PDT) From: Steven Coakley Subject: [Histonet] histogel cryosections To: Histonet@lists.utsouthwestern.edu Message-ID: <20060419194918.72148.qmail@web38208.mail.mud.yahoo.com> Content-Type: text/plain; charset=iso-8859-1 Has anyone ever used histogel/oct for FS. I've been asked to attempt cryosectioning cultured embryo bodies fr IHC. Steve --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day ------------------------------ Message: 4 Date: Wed, 19 Apr 2006 21:51:21 +0200 From: "Emma JONES" Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) To: Message-ID: Content-Type: text/plain; charset="windows-1250" Hi Eric, Have you considered asking your Ventana rep for information on their FITC reagents. The expiry dates on those are far longer than any you will get from making your own up. Approximately 12 months + regards Emma Jones Today's Topics: 1. CD-3 woes (Favara, Cynthia (NIH/NIAID) [E]) 2. RE: mouse spleen confocal (Favara, Cynthia (NIH/NIAID) [E]) 3. Expiry Period - Immunofluorescence Dilutions (Gagnon, Eric) 4. Re: mouse spleen confocal (Gayle Callis) 5. RE: Expiry Period - Immunofluorescence Dilutions (Favara, Cynthia (NIH/NIAID) [E]) 6. RE: Expiry Period - Immunofluorescence Dilutions (Dawson, Glen) 7. Re: lendrums' phloxine/tartrazine (John Kiernan) 8. Re: mouse spleen confocal (Scott Parker) 9. RE: RE: RHS-1 and MicroMed T/T Mega/Jeanine (Joyce Cline) 10. Re- Unscri be (Quadeer, Shahnaz S.) 11. IEC CTD harris cryostat (dfs dsaf) 12. Over lapped flouresence in double staining (sohail ejaz) 13. RE: Cryostat (Molinari, Betsy) 14. beginner questions on methacrylate resins (Nancy W. Troiano) 15. M. leprae control slides (Bartlett, Jeanine) 16. Microwave transparent plastics: Coleman Mfg contact information (Cheryl) 17. Bouin's alternative/substitution question (jason.burrill@crl.com) 18. Re: Mouse lungs OCT and curling (Gayle Callis) ---------------------------------------------------------------------- Message: 3 Date: Tue, 18 Apr 2006 13:23:28 -0400 From: "Gagnon, Eric" Subject: [Histonet] Expiry Period - Immunofluorescence Dilutions To: Message-ID: Content-Type: text/plain; charset="iso-8859-1" Hello Listers, We currently do immunofluorescence for IgG, IgA, IgM, C3, C1q, Fib., Kappa, Lambda, and C4d on renal biopsies, and the first four on skin biopsies. We are using Dako Antibody Diluting Buffer and mostly Dako FITC antisera. We currently make up about 1 ml of each, and the dilutions last us between 1 and 3 weeks. We would like to keep them longer if possible, making them up in larger quantities, possibly with the goal of transferring them to dispensers on our Ventana XT's. ] Is there anyone doing the same immunofluorescence panel and dilutions, and if so, how long are you able to use your dilutions once made? Thanks for your assistance, Eric Gagnon MLT Histology Laboratory Kingston General Hospital, Kingston, Ontario, Canada -- No virus found in this outgoing message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.2/314 - Release Date: 16/04/2006 ------------------------------ Message: 5 Date: Wed, 19 Apr 2006 13:53:46 -0600 From: "Patsy Ruegg" Subject: RE: [Histonet] dvaneyck IHCRG Membership Application Form To: "'Patsy Ruegg'" , Message-ID: <200604191953.k3JJrjka001675@chip.viawest.net> Content-Type: text/plain; charset="us-ascii" Sorry, I meant to send this to NSH, it comes up as histo on my server and I hit the wrong button. Disregard this message to verify NSH membership to HISTONET Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Patsy Ruegg Sent: Wednesday, April 19, 2006 11:49 AM To: histonet@pathology.swmed.edu Subject: [Histonet] dvaneyck IHCRG Membership Application Form verify NSH membership please Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 216 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. _____ From: webmaster@neo.agsci.colostate.edu [mailto:webmaster@neo.agsci.colostate.edu] Sent: Wednesday, April 19, 2006 11:30 AM To: pruegg@ihctech.net Subject: IHCRG Membership Application Form _____ Last_Name: Van Eyck First_Name: Debra NSH_Member: Yes Employer: WAukesha Memorial Hospital Address: 725 American Avenue City: Waukesha State: WI Zip: 53188 Province: Country: USA Phone: 262-928-2112 Ext: Fax: 262-928-4849 Email: deb.vaneyck@phci.org Surgical_Pathology: Yes Hematopathology: Orthopedic: Veterinary: Immunology: Plastics: Research: Paraffin: Yes Frozen: Cytospins: Yes Smears_Touch_Preps: Yes Plastics_sample: Sample_Other: PAP: APAAP: ABC_HRP: Yes ABC_AP: Yes SA_HRP: Yes SA_AP: Yes IF: IHC_Other: ISH: Gels: PCR: Mol_Other: Automated_IHC: Yes Company: Dako IHC_Qualification: No Date_Taken_Exam: Like_To_Take_Exam: Yes Expected_Date: next 6 months Need_Tissues: No Tissue_Needed: Need_Reagents: No Reagents_Needed: Can_Provide_Tissues: No Tissues_Provided: Can_Provide_Reagents: No Reagents_Provided: Form: Submit Remote Name: 207.67.109.90 HTTP User Agent: Mozilla/4.0 (compatible; MSIE 6.0; Windows NT 5.1; SV1; .NET CLR 1.1.4322) Other_Tech _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------ Message: 6 Date: Wed, 19 Apr 2006 14:58:11 -0500 From: "Galiotto, Laura" Subject: [Histonet] bone marrow Fe stains To: Message-ID: <270614B321ACB44D8C1D91F4F921FDC3036CBCBE@NCH01EX02.nch.org> Content-Type: text/plain; charset="iso-8859-1" I have a problem with bone marrow aspirate controls. The problem is that we are limited to the amount of controls we can collect. Even though the tissue control is working if the aspirate control (which are difficult to obtain) does not work we are being asked to repeat the stain until we find a aspirate control that is acceptable. Since I can not guarantee the aspirate control contains an adequate amount of Fe, I will be wasting time and reagents. Can anyone give feedback on how they process stains on bone marrow's, do you only use a tissue control, does anyone know where bone marrow aspirate controls -positive for Iron -can be purchased commercially? Laura Galiotto, HT (ASCP) Histology Facilitator Northwest Community Hospital 800 West Central Road Arlington Heights, Il 60005 847-618-6190 ------------------------------ Message: 7 Date: Wed, 19 Apr 2006 19:56:24 -0400 From: "Andrea T. Hooper" Subject: [Histonet] Mouse Cytokeratins To: Histonet Message-ID: Content-Type: text/plain; charset=us-ascii; format=flowed Hi All, What antibodies are people using to stain for cytokeratins in mouse tissue - frozen or paraffin. Thanks in advance, Andrea -- ------------------------------ Message: 8 Date: Thu, 20 Apr 2006 02:18:58 -0400 From: "Gus Mondragon" Subject: [Histonet] CBG Recycler To: Message-ID: <03B63DE44B5C014D84F9A9D8A968B5174C4D60@lithium.corp.gsopath.com> Content-Type: text/plain; charset="us-ascii" Hello Histonetters; I have a CBG 5 gal. Alcohol or Xylene recycler for sale that we no longer use. Good efficient unit is 4 yrs old in excellent condition, trouble/maintenance free. I'd be glad to offer good suggestions on recycling program. If you have any available Leica electric or manual microtomes, let's talk. Gus Mondragon (336) 387-2536; Greensboro NC ------------------------------ Message: 9 Date: Thu, 20 Apr 2006 08:51:11 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: CD-3 woes (not any longer!) To: Histonet@lists.utsouthwestern.edu Message-ID: <3263403287ca.3287ca326340@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Cynthia, We were not that successful either with the Dako rabbit CD3 antibody on mouse tissue. We changed to the LabVision CD3 rabbit monoclonal (clone SP7) and this worked very well on human, rat and mouse tissues (both cryo and FFPE). E.g. for staining FFPE: HIER with Tris-EDTA pH9.0, CD3 1:1000 (60 min, RT), polymer anti-rabbit/HRP (30 min, RT), DAB+ (5 min, RT). Hope this helps, Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 18 Apr 2006 13:15:00 -0400 From: "Favara, Cynthia \(NIH/NIAID\) [E]" Subject: [Histonet] CD-3 woes To: I have been using Dako CD-3 on murine formalin fixed paraffin embedded tissue and am experiencing some variation in staining. My protocol: pretreatment with Citrate pH 6.0 @120C for 5 minutes under pressure, an overnight incubation of the primary 1:1500 @4C, followed by 30 minute incubation with Vector biotinylated anti Rabbit IgG 1:250, 30 minutes Biogenex SS Streptavidin all steps following primary are done on the Ventana Nexus @RT using their AEC. I used to do the stain reliably but have a new lot and reliability is not optimal. I use a mouse spleen and brain with T-cell infiltration as positive controls and a NL brain as a negative control. Sometimes the stain is so crisp and other times it is muddy with a decrease in the number of cells st! ained. I ------------------------------ Message: 10 Date: Thu, 20 Apr 2006 09:05:26 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: mouse spleen confocal To: Histonet@lists.utsouthwestern.edu Message-ID: <3294ac32b9ea.32b9ea3294ac@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Dear Scott, You didn't tell us what detection system was used. Your story sounds reminds me to our first experiments with streptavidin-based detection on mouse spleen when we were confronted with heavy endogenous biotin! For staining mouse spleen, either avoid a streptavidin-based detection system or try to block endogenous biotin as Gayle suggested. Please realize that if the above is true and/or Gayle's is right with her suggestion concerning the adsorption of the anti-rat reagent, the signal you observe is no autofluorescence but "unwanted" specific fluorescence! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands From: Scott Parker [[1]mailto:scott9379@gmail.com] Sent: Tuesday, April 18, 2006 9:05 AM To: histonet Subject: [Histonet] mouse spleen confocal Dear all, Can anybody help me with some antibody staining I have been doing on mouse spleen? We are trying to detect CD8 in 5 um sections using CD8b.2 (ly-3.2) ( 53-5.8) with secondary Alexa Fluor 633 from mol. probes. Unfortunately all we see is vast amounts of auto-fluorescence making antibody detection impossible. We really need to get this sorted because ultimatley we'd like to have GFP and two different secondaries (labelling 3 things in total). Our protocol is basically: Spleens are removed to OCT and frozen in isopentane, sectioned (5 um) and stored at -20. When ready for use slides are warmed to room temp and fixed in -20 C acetone for 3 mins then re-hydrated in PBS for 5 mins. Secti! ons are a References 1. javascript:main.compose('new', 't=scott9379@gmail.com') ------------------------------ Message: 11 Date: Thu, 20 Apr 2006 09:16:29 +0200 From: "C.M. van der Loos" Subject: [Histonet] RE: Over lapped flouresence in double staining To: histonet@lists.utsouthwestern.edu Cc: sohail_e@yahoo.com Message-ID: <50578550cc68.50cc68505785@amc.uva.nl> Content-Type: text/plain; charset="us-ascii" Dear Dr. Sohail, You didn't write us what detection system was used for this double staining. The fact you observed overlap of SMA and vWF (which isn't very likely indeed) makes your detection system a bit suspect. To my opinion the most straightforward (and safe) ways to doublestain these antibodies are: A: cocktail of SMA (mouse) + vWF (rabbit), followed by a secondary cocktail composed of goat anti-mouse/FLUO-1 + goat anti-rabbit/FLUO-2 B: cocktail of SMA, clone 1A4 (mouse IgG2a) + vWF , clone F8/86 (mouse IgG1), followed by a secondary cocktail composed of goat anti-mouse IgG2a/FLUO-1 + goat anti-mouse IgG1/FLUO-2 Lots of success! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Tue, 18 Apr 2006 19:00:05 -0700 (PDT) From: sohail ejaz Subject: [Histonet] Over lapped flouresence in double staining To: [1]histonet@lists.utsouthwestern.edu I am doing double staining of endothelial cells and smooth muscles of the blood vessels with vWF and SMA. While doing double staining I have encountered overlapping of both antibodies and I cant see a clear differentiation between the fluorescence of these two antibodies. Could you please guide me how to over come this problem. Thanks Dr. Sohail References 1. mailto:histonet@lists.utsouthwestern.edu ------------------------------ Message: 12 Date: Thu, 20 Apr 2006 10:36:15 +0300 From: Mikael Niku Subject: Re: [Histonet] Large vessel endothelium problem To: Gayle Callis Cc: Histonet@lists.utsouthwestern.edu Message-ID: <444739EF.3020701@helsinki.fi> Content-Type: text/plain; charset=ISO-8859-1; format=flowed Hello again. Sorry for a bit unclearly formatted question. More specifically, I'm doing samples of bovine blood vessels, obtained from a slaughterhouse. So perfusion is unfortunately out of question :) And this is about REALLY big vessels, such as the bovine aorta. So the samples are bits (about 1 cm2) of vessel wall cut off and placed in PFA for overnight at +4C, then processed for paraffin sections in a routine manner. And yes, this is primarily about blood vessels, not lymphatics. Regards, Mikael Gayle Callis wrote: > You did not indicate how you do the paraformaldehyde fixation? > Immersion or perfusion? > > Vascular perfusion followed by immersion for a few hours may help your > problem - to ensure the tissue is fixed from the inside out and in > particular the lumens of your large blood (?) vessels. > > You did not give details on what you do with the vessels (opened, etc, > stretched out) during fixation or processing? -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// ------------------------------ Message: 13 Date: Thu, 20 Apr 2006 14:17:57 +0100 From: Kemlo Rogerson Subject: [Histonet] Diff Quick To: histonet@lists.utsouthwestern.edu Message-ID: Content-Type: text/plain We run a one-step FNAC breast clinic and have been using Diff-Quick as our stain; I personally don't like it as I prefer Paps but there you are. We had a recent FNAC that showed histiocytes in the tail of the prep but little else. As you know the bubbly cells tend to migrate to the edges as they are lighter but what we initially failed to notice was that in amongst the blood were poorly stained clumps of cells. After restain with a 'proper' Romanovsky stain they magically appeared and were benign. I shudder to think what would have happened if they had not been spotted and had been malignant. The problem I guess was the blood and together with the slow drying of the smear. Diff-Quick is fast but can have this effect, does anyone have a technique that is nearly as fast as Diff Quick but more robust in these cases? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Through return to simple living comes control of desires. In control of desires stillness is attained. In stillness the world is restored. -- Lao Tzu This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation ------------------------------ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet End of Histonet Digest, Vol 29, Issue 23 **************************************** ====================== Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ====================== From snyman <@t> embl.de Fri Apr 21 08:47:46 2006 From: snyman <@t> embl.de (snyman@embl.de) Date: Fri Apr 21 08:47:54 2006 Subject: [Histonet] (no subject) Message-ID: <20060421154746.tri5eqxobe8sk0wg@webmail.embl.de> Dear all We are currently investigating the presence and function of mushroom bodies in polycheates and would like to try two techniques. The one is Casons modified trichrome and the other Feulgens reaction using Schiffs reagent. However our library here only has books from the 80's that do not have these 2 techniques and the internet is proving to be a right jungle . If anyone could assist me in anyway I would greatly appreciate it. I would also welcome any suggestions for other techniques similar to these ones Our samples are mostly cryosections but we are also able to produce wax sections or use whole fixed embryos. Sincerely Yours, Heidi From aaljabari <@t> zahrawi.com Fri Apr 21 09:07:18 2006 From: aaljabari <@t> zahrawi.com (Alaa Al-Jabari) Date: Fri Apr 21 09:08:03 2006 Subject: [Histonet] Histology Statistics Message-ID: Dear All, I am interested in finding the web sites that can provide me with all sort of statistics pertaining to Histology such as world wide number of slides, growth rate, number of centers, cancer rate increase, ..etc. so I would apreciate it if any can help out with a web site or an agency for such information . Thank you in advance . Best Regards, Alaa El-Jabari __________________________________________________________________________________________________________________________________________________________________________________________________ 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 named addressee you should not disseminate, distribute or copy this e-mail. Scanned By AL Zahrawi Group Mail Scan. Al-ZAHRAWI Group IT Dep. _________________________________________________________________________________________________________________________________________________________________ From stefanie.werner <@t> gmx.de Fri Apr 21 09:08:27 2006 From: stefanie.werner <@t> gmx.de (Stefanie Werner) Date: Fri Apr 21 09:08:37 2006 Subject: [Histonet] toluidine blue staining problem Message-ID: <1512.1145628507@www069.gmx.net> Hello, i have a problem with toluidine-blue staining on paraffin sections after radioactive in situ hybridization. After developing the slides I stain them with 0,2% toluidine blue. The procedure is as follows: 2 minutes 0,2% toluidine-blue (with or without 1% Borax) rinse in tap water at least 1 minute 30% ethanol (denatured) at least 1 minute 50% ethanol (denatured) at least 1 minute 75% ethanol (denatured) at least 1 minute 95% ethanol (denatured) at least 1 minute 100% ethanol (denatured) at least 1 minute 100% ethanol (denatured) at least 10 minutes Xylol at least 10 minutes Xylol The slides are covered using DPX. Staining solutions were made in either tap or distilled water. After staining, I often see a red background colour on the slide and in the tissue under the microscope. Therefore I can't see my radioactive signal properly. Did anybody have the same problem or has a suggestion to solve the problem? I also suggested, if the Kodak photo emulsion is the problem, but we always use the same emulsion. I don't know what to do, because it just appears sometimes, even when the procedure is the same. Thank you very much in advance. Stefanie Werner -- GMX Produkte empfehlen und ganz einfach Geld verdienen! Satte Provisionen für GMX Partner: http://www.gmx.net/de/go/partner From PIXLEYSK <@t> UCMAIL.UC.EDU Fri Apr 21 09:17:49 2006 From: PIXLEYSK <@t> UCMAIL.UC.EDU (Pixley, Sarah (pixleysk)) Date: Fri Apr 21 09:17:54 2006 Subject: [Histonet] Re: DAB precipitate Message-ID: Dear Eva: I would like to hear any answers to this also. We always have a DAB precipitate. To avoid it, we make the DAB and add the nickel sulfate and H2O2 immediately before the reaction and do the reaction quickly. (Using nickel in the solution increases the precipitate significantly.) The longer you wait, the greater the precipitation. If I am doing a lot of samples, I set half the DAB aside and change solutions in the middle (add the nickel sulfate and H2O2). Otherwise, I don't know any other tricks. One other phenomenon I have seen is after freezing DAB. We make up our DAB solution in phosphate buffer (without H2O2 or nickel) and freeze the leftover. That works very well as long as you freeze in glass (i.e. new glass scintillation vials). If you freeze in some plastics, I have seen a precipitate upon thawing. And the glass has to be very clean--it cannot be something that was used before for DAB and then bleached and cleaned. Even if you clean the glass with a good cleaner like Chromerge or Contrad, there can be enough bleach leached into the glass to inactivate the DAB. Good luck and I hope to hear some interesting solutions to your problem! Sarah Pixley From dusko.trajkovic <@t> pfizer.com Fri Apr 21 09:31:47 2006 From: dusko.trajkovic <@t> pfizer.com (Trajkovic, Dusko) Date: Fri Apr 21 09:32:09 2006 Subject: [Histonet] Re: DAB precipitate Message-ID: <3AD0BD3142459B4E9B12CBEAFF2B89B202A94FE0@lajamrexm01.amer.pfizer.com> Is all of this aggravation worth the extra cost that you would dish out for a commercially prepared DAB? I have used DAB from DAKO, Zymed, KPL, Biocare, Invitrogen, and Vector, (there are many more out there)and never had to deal with the issues that you are describing. Most of these DAB reagents that I mentioned can be prepared in volumes as small as 1 ml, and Invitrogen DAB is ready to use, so you can use as little as 100ul. How much simpler can it get? As my good friend Dr. Laura always says at the end of her major station breaks: Now go and do the right thing! Thank you and have a great weekend. To anyone that is celebrating a special day this Sunday, have a nice and enjoyable holiday. Dusko Trajkovic -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pixley, Sarah (pixleysk) Sent: Friday, April 21, 2006 7:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: DAB precipitate Dear Eva: I would like to hear any answers to this also. We always have a DAB precipitate. To avoid it, we make the DAB and add the nickel sulfate and H2O2 immediately before the reaction and do the reaction quickly. (Using nickel in the solution increases the precipitate significantly.) The longer you wait, the greater the precipitation. If I am doing a lot of samples, I set half the DAB aside and change solutions in the middle (add the nickel sulfate and H2O2). Otherwise, I don't know any other tricks. One other phenomenon I have seen is after freezing DAB. We make up our DAB solution in phosphate buffer (without H2O2 or nickel) and freeze the leftover. That works very well as long as you freeze in glass (i.e. new glass scintillation vials). If you freeze in some plastics, I have seen a precipitate upon thawing. And the glass has to be very clean--it cannot be something that was used before for DAB and then bleached and cleaned. Even if you clean the glass with a good cleaner like Chromerge or Contrad, there can be enough bleach leached into the glass to inactivate the DAB. Good luck and I hope to hear some interesting solutions to your problem! Sarah Pixley _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ---------------------------------------------------------------------- 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 c.m.vanderloos <@t> amc.uva.nl Fri Apr 21 09:33:58 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Apr 21 09:34:09 2006 Subject: [Histonet] RE: Ki67 protocol--frozen sections Message-ID: <527233529aac.529aac527233@amc.uva.nl> Dear Janelle, For staining of Ki67 we like to fix human cryostat tissues sections with Zamboni's or buffered formalin. This results into a sharper nuclear staining than acetone-fixation. Just any polymer/HRP visualization and DAB+ as chromogen will do. As for a Ki67 antibody working on feline tissues I would recommend you to test the SP6 clone (rabbit monoclonal) from LabVision or Vector. In our hands this clone already worked for a lot of different species. Perhaps you can expand the list with feline... Good luck! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Thu, 20 Apr 2006 22:21:29 -0400 From: "Janelle Novak" Subject: [Histonet] Ki67 protocol--frozen sections To: [1]histonet@lists.utsouthwestern.edu Does anyone have any good recommendations for a Ki67 staining protocol for feline frozen tissue sections? Thank you, Janelle Novak References 1. mailto:histonet@lists.utsouthwestern.edu From pruegg <@t> ihctech.net Fri Apr 21 11:14:18 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Apr 21 11:14:20 2006 Subject: [Histonet] Re: DAB precipitate In-Reply-To: Message-ID: <200604211614.k3LGEEka023031@chip.viawest.net> I thought DAB should not be frozen???? Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pixley, Sarah (pixleysk) Sent: Friday, April 21, 2006 7:18 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Re: DAB precipitate Dear Eva: I would like to hear any answers to this also. We always have a DAB precipitate. To avoid it, we make the DAB and add the nickel sulfate and H2O2 immediately before the reaction and do the reaction quickly. (Using nickel in the solution increases the precipitate significantly.) The longer you wait, the greater the precipitation. If I am doing a lot of samples, I set half the DAB aside and change solutions in the middle (add the nickel sulfate and H2O2). Otherwise, I don't know any other tricks. One other phenomenon I have seen is after freezing DAB. We make up our DAB solution in phosphate buffer (without H2O2 or nickel) and freeze the leftover. That works very well as long as you freeze in glass (i.e. new glass scintillation vials). If you freeze in some plastics, I have seen a precipitate upon thawing. And the glass has to be very clean--it cannot be something that was used before for DAB and then bleached and cleaned. Even if you clean the glass with a good cleaner like Chromerge or Contrad, there can be enough bleach leached into the glass to inactivate the DAB. Good luck and I hope to hear some interesting solutions to your problem! Sarah Pixley _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From cbass <@t> bidmc.harvard.edu Fri Apr 21 11:19:07 2006 From: cbass <@t> bidmc.harvard.edu (Caroline Bass) Date: Fri Apr 21 11:17:08 2006 Subject: [Histonet] fluorescent microscope problem Message-ID: <44911709-363F-45C3-B9B5-6BD5BB6B98A0@bidmc.harvard.edu> Hello, I am having a problem with my fluorescent microscope. It is an old NIkon Diaphot 300 inverted. I don't expect much from this scope, but it is usually good enough for a quick evaluation. However, since it is used by numerous people it is constantly being thrown out of whack. Today I have encountered a new problem. When I switch to fluorescence there is a considerable amount of white light coming through. I can even make out the outline of non-fluorescent cells when I should only be able to see fluorescence. Any ideas as to what may be causing this? I have tried everything I can think of. Thanks, Caroline From bhewlett <@t> cogeco.ca Fri Apr 21 11:25:46 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Fri Apr 21 11:25:49 2006 Subject: [Histonet] Re: DAB precipitate References: <200604211614.k3LGEEka023031@chip.viawest.net> Message-ID: <002201c66560$3ee1f7a0$6500a8c0@mainbox> Hi Patsy, I have frozen aliquots of DAB for years without any problem! Bryan ----- Original Message ----- From: "Patsy Ruegg" To: "'Pixley, Sarah (pixleysk)'" ; Sent: Friday, April 21, 2006 12:14 PM Subject: RE: [Histonet] Re: DAB precipitate >I thought DAB should not be frozen???? > Patsy > > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 215 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > > This email is confidential and intended solely for the use of the > Person(s) > ('the intended recipient') to whom it was addressed. Any views or opinions > presented are solely those of the author. It may contain information that > is > privileged & confidential within the meaning of applicable law. > Accordingly > any dissemination, distribution, copying, or other use of this message, or > any of its contents, by any person other than the intended recipient may > constitute a breach of civil or criminal law and is strictly prohibited. > If > you are NOT the intended recipient please contact the sender and dispose > of > this e-mail as soon as possible. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pixley, > Sarah (pixleysk) > Sent: Friday, April 21, 2006 7:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: DAB precipitate > > Dear Eva: > I would like to hear any answers to this also. We always have a DAB > precipitate. To avoid it, we make the DAB and add the nickel sulfate and > H2O2 immediately before the reaction and do the reaction quickly. (Using > nickel in the solution increases the precipitate significantly.) The > longer > you wait, the greater the precipitation. If I am doing a lot of samples, I > set half the DAB aside and change solutions in the middle (add the nickel > sulfate and H2O2). Otherwise, I don't know any other tricks. > > One other phenomenon I have seen is after freezing DAB. We make up our DAB > solution in phosphate buffer (without H2O2 or nickel) and freeze the > leftover. That works very well as long as you freeze in glass (i.e. new > glass scintillation vials). If you freeze in some plastics, I have seen a > precipitate upon thawing. And the glass has to be very clean--it cannot be > something that was used before for DAB and then bleached and cleaned. Even > if you clean the glass with a good cleaner like Chromerge or Contrad, > there > can be enough bleach leached into the glass to inactivate the DAB. > > Good luck and I hope to hear some interesting solutions to your problem! > Sarah Pixley > > > _______________________________________________ > Histonet mailing list > Histonet@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 Fri Apr 21 11:40:03 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Apr 21 11:40:20 2006 Subject: [Histonet] Re: DAB precipitate In-Reply-To: <200604211614.k3LGEEka023031@chip.viawest.net> References: <200604211614.k3LGEEka023031@chip.viawest.net> Message-ID: <6.0.0.22.1.20060421103632.01b48b50@gemini.msu.montana.edu> With Pierce DAB two liquid component kits, the DAB portion is kept in a freezer, although it does not freeze due to the solvent it is dissolved in, and there was and probably still is a ready to use DAB from Research Genetics(? and under a new name (?) was kept in a freezer and taken out for use. At 10:14 AM 4/21/2006, you wrote: >I thought DAB should not be frozen???? >Patsy > > >Patsy Ruegg, HT(ASCP)QIHC >IHCtech, LLC >Fitzsimmons BioScience Park >12635 Montview Blvd. Suite 215 >Aurora, CO 80010 >P-720-859-4060 >F-720-859-4110 >wk email pruegg@ihctech.net >web site www.ihctech.net > > >This email is confidential and intended solely for the use of the Person(s) >('the intended recipient') to whom it was addressed. Any views or opinions >presented are solely those of the author. It may contain information that is >privileged & confidential within the meaning of applicable law. Accordingly >any dissemination, distribution, copying, or other use of this message, or >any of its contents, by any person other than the intended recipient may >constitute a breach of civil or criminal law and is strictly prohibited. If >you are NOT the intended recipient please contact the sender and dispose of >this e-mail as soon as possible. > > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Pixley, >Sarah (pixleysk) >Sent: Friday, April 21, 2006 7:18 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] Re: DAB precipitate > >Dear Eva: >I would like to hear any answers to this also. We always have a DAB >precipitate. To avoid it, we make the DAB and add the nickel sulfate and >H2O2 immediately before the reaction and do the reaction quickly. (Using >nickel in the solution increases the precipitate significantly.) The longer >you wait, the greater the precipitation. If I am doing a lot of samples, I >set half the DAB aside and change solutions in the middle (add the nickel >sulfate and H2O2). Otherwise, I don't know any other tricks. > >One other phenomenon I have seen is after freezing DAB. We make up our DAB >solution in phosphate buffer (without H2O2 or nickel) and freeze the >leftover. That works very well as long as you freeze in glass (i.e. new >glass scintillation vials). If you freeze in some plastics, I have seen a >precipitate upon thawing. And the glass has to be very clean--it cannot be >something that was used before for DAB and then bleached and cleaned. Even >if you clean the glass with a good cleaner like Chromerge or Contrad, there >can be enough bleach leached into the glass to inactivate the DAB. > >Good luck and I hope to hear some interesting solutions to your problem! >Sarah Pixley > > >_______________________________________________ >Histonet mailing list >Histonet@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 406 994-4303 (FAX) From kegraves <@t> UTMB.EDU Fri Apr 21 11:53:01 2006 From: kegraves <@t> UTMB.EDU (Graves, Kerry) Date: Fri Apr 21 11:53:05 2006 Subject: [Histonet] unsubscribe Message-ID: <649CE9B357B1174DB9FEE750C1369FC325AAC5@EXCH2K2.utmb.edu> From pruegg <@t> ihctech.net Fri Apr 21 12:07:33 2006 From: pruegg <@t> ihctech.net (Patsy Ruegg) Date: Fri Apr 21 12:07:37 2006 Subject: [Histonet] Re: DAB precipitate In-Reply-To: <002201c66560$3ee1f7a0$6500a8c0@mainbox> Message-ID: <200604211707.k3LH7Tka005894@chip.viawest.net> Oh yea, sorry I had a brain fart, I actually use the Research Genetics ready to use DAB that is in glycerine or something so it doesn't actually freeze. Patsy Patsy Ruegg, HT(ASCP)QIHC IHCtech, LLC Fitzsimmons BioScience Park 12635 Montview Blvd. Suite 215 Aurora, CO 80010 P-720-859-4060 F-720-859-4110 wk email pruegg@ihctech.net web site www.ihctech.net This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible. -----Original Message----- From: Bryan Hewlett [mailto:bhewlett@cogeco.ca] Sent: Friday, April 21, 2006 9:26 AM To: Patsy Ruegg; 'Pixley, Sarah (pixleysk)'; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Re: DAB precipitate Hi Patsy, I have frozen aliquots of DAB for years without any problem! Bryan ----- Original Message ----- From: "Patsy Ruegg" To: "'Pixley, Sarah (pixleysk)'" ; Sent: Friday, April 21, 2006 12:14 PM Subject: RE: [Histonet] Re: DAB precipitate >I thought DAB should not be frozen???? > Patsy > > > Patsy Ruegg, HT(ASCP)QIHC > IHCtech, LLC > Fitzsimmons BioScience Park > 12635 Montview Blvd. Suite 215 > Aurora, CO 80010 > P-720-859-4060 > F-720-859-4110 > wk email pruegg@ihctech.net > web site www.ihctech.net > > > This email is confidential and intended solely for the use of the > Person(s) > ('the intended recipient') to whom it was addressed. Any views or > opinions presented are solely those of the author. It may contain > information that is privileged & confidential within the meaning of > applicable law. > Accordingly > any dissemination, distribution, copying, or other use of this > message, or any of its contents, by any person other than the intended > recipient may constitute a breach of civil or criminal law and is strictly prohibited. > If > you are NOT the intended recipient please contact the sender and > dispose of this e-mail as soon as possible. > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > Pixley, Sarah (pixleysk) > Sent: Friday, April 21, 2006 7:18 AM > To: histonet@lists.utsouthwestern.edu > Subject: [Histonet] Re: DAB precipitate > > Dear Eva: > I would like to hear any answers to this also. We always have a DAB > precipitate. To avoid it, we make the DAB and add the nickel sulfate > and > H2O2 immediately before the reaction and do the reaction quickly. > (Using nickel in the solution increases the precipitate > significantly.) The longer you wait, the greater the precipitation. If > I am doing a lot of samples, I set half the DAB aside and change > solutions in the middle (add the nickel sulfate and H2O2). Otherwise, > I don't know any other tricks. > > One other phenomenon I have seen is after freezing DAB. We make up our > DAB solution in phosphate buffer (without H2O2 or nickel) and freeze > the leftover. That works very well as long as you freeze in glass > (i.e. new glass scintillation vials). If you freeze in some plastics, > I have seen a precipitate upon thawing. And the glass has to be very > clean--it cannot be something that was used before for DAB and then > bleached and cleaned. Even if you clean the glass with a good cleaner > like Chromerge or Contrad, there can be enough bleach leached into the > glass to inactivate the DAB. > > Good luck and I hope to hear some interesting solutions to your problem! > Sarah Pixley > > > _______________________________________________ > Histonet mailing list > Histonet@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 Wanda.Smith <@t> HCAhealthcare.com Fri Apr 21 12:28:22 2006 From: Wanda.Smith <@t> HCAhealthcare.com (Smith Wanda) Date: Fri Apr 21 12:28:27 2006 Subject: [Histonet] Giving Specimens to Patients Message-ID: Happy Friday Histonet, We have aways given gallstones and foreign bodies to patients if they request them, however, I have a patient that wants his rib back that was removed in surgery. It just looks like a rib fracture, no atypia. Our Pathologist don't recommend this, but it is a call from the hospital legal department and I can't get in touch with them today. What is the practice everyone uses for giving/not giving patient's their specimens in formalin? Thanks, Wanda From juan.gutierrez <@t> christushealth.org Fri Apr 21 12:43:43 2006 From: juan.gutierrez <@t> christushealth.org (GUTIERREZ, JUAN) Date: Fri Apr 21 12:43:49 2006 Subject: [Histonet] Giving Specimens to Patients Message-ID: Check with tour state Health Dept. Here in Texas we can only give back teeth. Anything else has to be released to a licensed funeral home. Or you can also release tissues for research purposes, but again to bona fide institutions. Gone are the days when you could release gallstones, tonsils, hardware, fingers, flashlights, batteries and some small appliances. Hope this helps. Juan C. Gutierrez, HT(ASCP) Histology Laboratory Supervisor (210)704-2533 My opinions are my own and do not reflect those of my employer. Long live free speech! -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Smith Wanda Sent: Friday, April 21, 2006 12:28 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Giving Specimens to Patients Happy Friday Histonet, We have aways given gallstones and foreign bodies to patients if they request them, however, I have a patient that wants his rib back that was removed in surgery. It just looks like a rib fracture, no atypia. Our Pathologist don't recommend this, but it is a call from the hospital legal department and I can't get in touch with them today. What is the practice everyone uses for giving/not giving patient's their specimens in formalin? Thanks, Wanda _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From GDawson <@t> dynacaremilwaukee.com Fri Apr 21 12:52:22 2006 From: GDawson <@t> dynacaremilwaukee.com (Dawson, Glen) Date: Fri Apr 21 12:52:29 2006 Subject: [Histonet] FW: Help! Message-ID: -----Original Message----- From: Bobrowitz, Carol [mailto:carolb@phys.mcw.edu] Sent: Friday, April 21, 2006 11:50 AM To: Dawson, Glen Subject: Help! Glen, I would like to send the question below to the histonet but am having problems. Could you please post it for me. Have a great vacation. Thanks, Carol -----Original Message----- From: Bobrowitz, Carol Sent: Friday, April 21, 2006 11:44 AM To: HISTONET (E-mail) Subject: repeating AR heat treatment Hello, I have already stained FFPE rat kidney's using FITC with a 2 hour heat treatment in citrate buffer ph 6.0. (the staining was great) I am now going to restain the same rat kidney slides with an additional primary using DAB as the chromagen. This primary also requires heat treatment. Am I correct in my thinking that I should repeat the heat treatment again due to the fact that the epitope (binding) sites have already closed after accepting the application of the 1st primary? Any help will be appreciated. Thank you in advance. Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology 414-456-8179 cbobrowi@mcw.edu From failm <@t> musc.edu Fri Apr 21 13:10:03 2006 From: failm <@t> musc.edu (Mildred Fail) Date: Fri Apr 21 13:10:47 2006 Subject: [Histonet] bar code system for Ab storage Message-ID: Good afternoon We are looking for a bar code system for storage of antibodies in the refrigerator and a minus 70 freezer. We need cryogenic labels that fit on concentrated antibody vials. Does anyone have a bar code sytem in place for tracking immunohistochemical reagents, in particular antibodies? Thank you in advance for your help Rena Fail Lead Tech IHC/SS Medical University of SC Rena Fail From jkiernan <@t> uwo.ca Fri Apr 21 13:16:54 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Fri Apr 21 13:16:55 2006 Subject: [Histonet] toluidine blue staining problem References: <1512.1145628507@www069.gmx.net> Message-ID: <44492196.3D1F5AAD@uwo.ca> You are using a touidine blue recipe for staining semi-thin plastic-embedded sections, with which the idea is to stain everything - hence the alkaline solution of a cationic dye. To use toluidine blue as a counterstain, you want it to stain cell nuclei and not much else, and not very strongly - especially if your silver grains are over nuclei. So use the dye at pH 3.5 to 4.0, and rinse in water adjusted to the same pH. If the staining is just right, dehydrate in 3 X 100% alcohol. If staining is too strong, remove some dye with 50-70-% alcohol. Next time, stain at a lower pH. When I did autoradiography I preferred a red nuclear stain to a blue one. Brazalum was very good; you make it up like Mayer's haemalum, but with brazilin instead of haematoxylin. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ Stefanie Werner wrote: > > Hello, > > i have a problem with toluidine-blue staining on paraffin sections after > radioactive in situ hybridization. After developing the slides I stain > them with 0,2% toluidine blue. > The procedure is as follows: > > 2 minutes 0,2% toluidine-blue (with or without 1% Borax) > rinse in tap water > at least 1 minute 30% ethanol (denatured) > at least 1 minute 50% ethanol (denatured) > at least 1 minute 75% ethanol (denatured) > at least 1 minute 95% ethanol (denatured) > at least 1 minute 100% ethanol (denatured) > at least 1 minute 100% ethanol (denatured) > > at least 10 minutes Xylol > at least 10 minutes Xylol > > The slides are covered using DPX. > > Staining solutions were made in either tap or distilled water. > > After staining, I often see a red background colour on the slide and in > the tissue under the microscope. Therefore I can't see my > radioactive signal properly. > > Did anybody have the same problem or has a suggestion to solve the > problem? > > I also suggested, if the Kodak photo emulsion is the problem, but we > always use the same emulsion. > I don't know what to do, because it just appears sometimes, even > when the procedure is the same. > > Thank you very much in advance. > > Stefanie Werner > > -- > GMX Produkte empfehlen und ganz einfach Geld verdienen! > Satte Provisionen f?r GMX Partner: http://www.gmx.net/de/go/partner From bill501 <@t> mindspring.com Fri Apr 21 13:36:30 2006 From: bill501 <@t> mindspring.com (Bill Blank) Date: Fri Apr 21 13:36:38 2006 Subject: [Histonet] Giving Specimens to Patients In-Reply-To: References: Message-ID: Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if >they request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call >from the hospital legal department and I can't get in touch with >them today. What is the practice everyone uses for giving/not >giving patient's their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board From ynwang <@t> u.washington.edu Fri Apr 21 13:43:20 2006 From: ynwang <@t> u.washington.edu (Yak-Nam Wang) Date: Fri Apr 21 13:41:54 2006 Subject: [Histonet] Tissue-Tek Paraform Sectionable Cassette System Message-ID: Hello All, Has anyone tried the Tissue-Tek Paraform Sectionable Cassette System? If you have can you share your experiences please. Has anyone had trouble with the posts in the orientation cassette in terms of being in the section when staining and photographing? Thanks Yak-Nam Wang From gcallis <@t> montana.edu Fri Apr 21 13:58:35 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Apr 21 13:58:41 2006 Subject: [Histonet] Colocalization of different fluorophores Message-ID: <6.0.0.22.1.20060421125549.01b3e078@gemini.msu.montana.edu> I presented this question to Molecular Probes Technical services: What kind of co-localization color do we expect to see with antibody staining using Alexa 350 and FITC or Alexa 488, or Alexa 350 and rhodamine (either TRITC or RRX) or Alexa 555? We know that colocalization of FITC or Alexa 488 with Alexa 555 or Rhodamine is yellow - Their answer was inconclusive: "Thank you for contacting Molecular Probes Technical Assistance. We have not determine the color of the fluorescence from multiple dyes co-localizing. I would guess that the Alexa Fluor 350 and Alexa Fluor 488 would give a teal color and the Alexa Fluor 350 and Rhodamine would give a purplish color. " If anyone has done this, I would appreciate the "non-guess" response. 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 406 994-4303 (FAX) From rjbuesa <@t> yahoo.com Fri Apr 21 14:12:23 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Apr 21 14:12:26 2006 Subject: [Histonet] Giving Specimens to Patients In-Reply-To: Message-ID: <20060421191223.50146.qmail@web61220.mail.yahoo.com> To avoid getting in trouble, explain the situation to the patient and wait until you get in touch with your legal department. They will not accept not being available as an excuse if something "develops" from your action. Ren? J. Smith Wanda wrote: Happy Friday Histonet, We have aways given gallstones and foreign bodies to patients if they request them, however, I have a patient that wants his rib back that was removed in surgery. It just looks like a rib fracture, no atypia. Our Pathologist don't recommend this, but it is a call from the hospital legal department and I can't get in touch with them today. What is the practice everyone uses for giving/not giving patient's their specimens in formalin? Thanks, Wanda _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Celebrate Earth Day everyday! Discover 10 things you can do to help slow climate change. Yahoo! Earth Day From Jackie.O'Connor <@t> abbott.com Fri Apr 21 13:59:36 2006 From: Jackie.O'Connor <@t> abbott.com (Jackie M O'Connor) Date: Fri Apr 21 14:14:43 2006 Subject: [Histonet] Giving Specimens to Patients In-Reply-To: Message-ID: Sorry, Bill - I think that once he signed all the papers before surgery, he gives the hospital permission to 'dispose of' tissues removed in surgery. I think that makes his rib the hospital's property. We once had a guy ask for his toe bones back after he lost part of his foot in a motorcycle accident. He wanted to make a necklace. The pathologist said 'no'. End of story, unless we get into all the patients whose husbands ask for the placenta after childbirth. Jackie O' Bill Blank Sent by: histonet-bounces@lists.utsouthwestern.edu 04/21/2006 01:36 PM To "Smith Wanda" , cc Subject Re: [Histonet] Giving Specimens to Patients Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if >they request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call >from the hospital legal department and I can't get in touch with >them today. What is the practice everyone uses for giving/not >giving patient's their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Apr 21 14:18:23 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Apr 21 14:18:33 2006 Subject: Thank you!!! RE: [Histonet] Colocalization of different fluorophores In-Reply-To: References: Message-ID: <6.0.0.22.1.20060421131717.01b13c88@gemini.msu.montana.edu> Surita, Thank you, and I am forwarding your message to the technical person at Molecular Probes so they do not have to guess anymore!!! Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From gcallis <@t> montana.edu Fri Apr 21 14:31:42 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Apr 21 14:31:49 2006 Subject: [Histonet] Colocalization of different fluorophores In-Reply-To: <8ACFEE6579A6F04894933C3775C13D2D36F2A0@NTRSEVS30002.s3.ms. unilever.com> References: <8ACFEE6579A6F04894933C3775C13D2D36F2A0@NTRSEVS30002.s3.ms.unilever.com> Message-ID: <6.0.0.22.1.20060421132758.01b13148@gemini.msu.montana.edu> Manoj, It was a private answer to me and just what I needed to know. I am happy to share her comments. Enjoy! *************************************************************************************************************************** She wrote: We constantly do four color imaging work for various PIs here at the Buck. I can tell you with certainty the following results: When staining with Alexa 350, 488, 555, and 647 (which we always have trouble picking a color when looking at coloc of all four...) 488 and 555 = yellow 350 and 555 = purple 350 and 488 = we call cyan 350 and 488 and 555 = white adding the fourth 647 becomes a mess as you can imagine, but we always psuedocolor it various colors and analyze the coloc individually (for example make 647 'green' and compare with 555, and 350; then make it 'red' and compare to 488 and 350, etc.) before making any conclusions of all four combined. Now that I think about it, to avoid strange color combos, you could always psuedocolor the channels red and green for any alexa dye and keep track of the changes, this way you are always reporting the coloc as yellow (of course this would only work if you are looking at two proteins at a time I guess) Hope this helps, happy staining Surita ****************************************************************************************************************************** >Gayle Callis HTL, HT, MT(ASCP) >Research Histopathology Supervisor >Veterinary Molecular Biology >Montana State University >Bozeman MT 59717 From ploykasek <@t> phenopath.com Fri Apr 21 14:32:03 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Apr 21 14:32:26 2006 Subject: [Histonet] FW: Help! In-Reply-To: Message-ID: I don't have experience with animal tissue, but can give my experiences with human tissue. Every once in a while we will be asked to use a slide that has already been heat pretreated & the slide is either a negative IHC control or the original antibody was not positive. In that situation, we remove the coverslip, rehydrate the slide, and DO NOT repeat the heat retrieval. We have had positive staining with this method, but I think the staining is not as crisp as using a fresh cut section. Perhaps you can test this on a positive control slide before using your precious tissue. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > > -----Original Message----- > From: Bobrowitz, Carol [mailto:carolb@phys.mcw.edu] > Sent: Friday, April 21, 2006 11:50 AM > To: Dawson, Glen > Subject: Help! > > > > Glen, > > I would like to send the question below to the histonet but am having > problems. > Could you please post it for me. > > Have a great vacation. > > Thanks, > > Carol > > -----Original Message----- > From: Bobrowitz, Carol > Sent: Friday, April 21, 2006 11:44 AM > To: HISTONET (E-mail) > Subject: repeating AR heat treatment > > Hello, > > I have already stained FFPE rat kidney's using FITC with a 2 hour heat > treatment in citrate buffer ph 6.0. (the staining was great) > > I am now going to restain the same rat kidney slides with an additional > primary using DAB as the chromagen. This primary also requires heat > treatment. > > Am I correct in my thinking that I should repeat the heat treatment again due > to the fact that the epitope (binding) sites have already closed after > accepting the application of the 1st primary? > > Any help will be appreciated. Thank you in advance. > > Carol Ann Bobrowitz > Medical College of Wisconsin > Department of Physiology > 414-456-8179 > cbobrowi@mcw.edu > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From Sjohnso616 <@t> aol.com Fri Apr 21 15:04:48 2006 From: Sjohnso616 <@t> aol.com (Sjohnso616@aol.com) Date: Fri Apr 21 15:04:58 2006 Subject: [Histonet] Respirator fit test Message-ID: <289.90e8193.317a94e0@aol.com> Hello All, Would anyone be willing to share your laboratory procedure for respirator fit testing? Thanks in advance. Saundra Johnson Sarasota Pathology From Charles.Embrey <@t> carle.com Fri Apr 21 15:41:27 2006 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Fri Apr 21 15:42:02 2006 Subject: [Histonet] Giving Specimens to Patients Message-ID: I agree. Give the patient his rib. The only thing I would insist on is keeping it two weeks after sign-out to fulfill CAP requirements and do replace the formalin with ETOH. The only things we won't return directly to the patient here are amputated limbs. These must be released to a funeral home. Charles Embrey PA(ASCP) www.GreyRealm.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bill Blank Sent: Friday, April 21, 2006 1:37 PM To: Smith Wanda; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Giving Specimens to Patients Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if >they request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call >from the hospital legal department and I can't get in touch with >them today. What is the practice everyone uses for giving/not >giving patient's their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charlene.Henry <@t> STJUDE.ORG Fri Apr 21 16:23:55 2006 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Fri Apr 21 16:24:01 2006 Subject: [Histonet] Giving Specimens to Patients. . Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1C11@SJMEMXMB02.stjude.sjcrh.local> I would suggest waiting for an answer from a representative of your legal department. People do love lawsuits. Charlene -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bill Blank Sent: Friday, April 21, 2006 1:37 PM To: Smith Wanda; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Giving Specimens to Patients. . Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if they >request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call from >the hospital legal department and I can't get in touch with them today. >What is the practice everyone uses for giving/not giving patient's >their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From sohail_e <@t> yahoo.com Fri Apr 21 20:17:45 2006 From: sohail_e <@t> yahoo.com (sohail ejaz) Date: Fri Apr 21 20:17:50 2006 Subject: [Histonet] 3D software for histology Message-ID: <20060422011745.94473.qmail@web30609.mail.mud.yahoo.com> Hello every one I am interested in getting 3D pictures form my histology sections (serial sections). Can anyone guide or recommend me any good software that can produce a combined 3D pictures of my serial sections. Thanks Dr. Sohail --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From gu.lang <@t> gmx.at Sat Apr 22 02:48:00 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Sat Apr 22 02:48:01 2006 Subject: AW: [Histonet] Giving Specimens to Patients In-Reply-To: Message-ID: <000001c665e1$141ed610$eeeea8c0@SERVER01> Recently we had the case, that a woman wanted to get her abortion-tissue. You know what that looks like in a glass of formalin. It was not allowed and I think it was the right decision. In Germany there was the case, that a woman wanted to bury the hard of her husband in his grave. It was a long fight at court. At last she got right and her soul got peace. Gudrun Lang Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Jackie M O'Connor Gesendet: Freitag, 21. April 2006 21:00 An: Bill Blank Cc: histonet@lists.utsouthwestern.edu; Smith Wanda; histonet-bounces@lists.utsouthwestern.edu Betreff: Re: [Histonet] Giving Specimens to Patients Sorry, Bill - I think that once he signed all the papers before surgery, he gives the hospital permission to 'dispose of' tissues removed in surgery. I think that makes his rib the hospital's property. We once had a guy ask for his toe bones back after he lost part of his foot in a motorcycle accident. He wanted to make a necklace. The pathologist said 'no'. End of story, unless we get into all the patients whose husbands ask for the placenta after childbirth. Jackie O' Bill Blank Sent by: histonet-bounces@lists.utsouthwestern.edu 04/21/2006 01:36 PM To "Smith Wanda" , cc Subject Re: [Histonet] Giving Specimens to Patients Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if they >request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call from >the hospital legal department and I can't get in touch with them today. >What is the practice everyone uses for giving/not giving patient's >their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@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 djamesnz <@t> orcon.net.nz Sat Apr 22 04:26:54 2006 From: djamesnz <@t> orcon.net.nz (Darren James) Date: Sat Apr 22 04:27:10 2006 Subject: [Histonet] Giving Specimens to Patients In-Reply-To: <000001c665e1$141ed610$eeeea8c0@SERVER01> Message-ID: <000001c665ee$e57e7690$0301010a@DAZZA> In my neck of the woods the patient retains the right to request the return of any and all tissue, including blocks once a report has been issued. They sign a waiver and presto....after all, it is from their body. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Saturday, 22 April 2006 7:48 p.m. To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Giving Specimens to Patients Recently we had the case, that a woman wanted to get her abortion-tissue. You know what that looks like in a glass of formalin. It was not allowed and I think it was the right decision. In Germany there was the case, that a woman wanted to bury the hard of her husband in his grave. It was a long fight at court. At last she got right and her soul got peace. Gudrun Lang Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Jackie M O'Connor Gesendet: Freitag, 21. April 2006 21:00 An: Bill Blank Cc: histonet@lists.utsouthwestern.edu; Smith Wanda; histonet-bounces@lists.utsouthwestern.edu Betreff: Re: [Histonet] Giving Specimens to Patients Sorry, Bill - I think that once he signed all the papers before surgery, he gives the hospital permission to 'dispose of' tissues removed in surgery. I think that makes his rib the hospital's property. We once had a guy ask for his toe bones back after he lost part of his foot in a motorcycle accident. He wanted to make a necklace. The pathologist said 'no'. End of story, unless we get into all the patients whose husbands ask for the placenta after childbirth. Jackie O' Bill Blank Sent by: histonet-bounces@lists.utsouthwestern.edu 04/21/2006 01:36 PM To "Smith Wanda" , cc Subject Re: [Histonet] Giving Specimens to Patients Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if they >request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call from >the hospital legal department and I can't get in touch with them today. >What is the practice everyone uses for giving/not giving patient's >their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@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 bill501 <@t> mindspring.com Sat Apr 22 09:24:53 2006 From: bill501 <@t> mindspring.com (Bill Blank) Date: Sat Apr 22 09:25:07 2006 Subject: AW: [Histonet] Giving Specimens to Patients In-Reply-To: <000001c665e1$141ed610$eeeea8c0@SERVER01> References: <000001c665e1$141ed610$eeeea8c0@SERVER01> Message-ID: This sort of thing bothers me, because it is becoming the rule in the US as regulators and lawyers take away the ability to exercise professional judgement. IMO, given there are no public health concerns, I do not think this is the business of any government. And it is the Pathologist who should decide if there are public health concerns (like a tubercular lung, eg) in each specific case, not lawyers or bureaucrats as they lack the appropriate expertise. If someone wants to bury abortus tissue or limbs, that is their business. As a physician and a humanitarian, I would help make it safe and as visually appealing as possible for them. Bill At 9:48 AM +0200 4/22/06, Gudrun Lang wrote: > Recently we had the case, that a woman wanted to get her abortion-tissue. >You know what that looks like in a glass of formalin. It was not allowed and >I think it was the right decision. > >In Germany there was the case, that a woman wanted to bury the hard of her >husband in his grave. It was a long fight at court. At last she got right >and her soul got peace. -- May the Gods protect me from all the humans who want to protect me. From Malcolm.McCallum <@t> tamut.edu Sat Apr 22 09:42:34 2006 From: Malcolm.McCallum <@t> tamut.edu (Malcolm McCallum) Date: Sat Apr 22 09:44:41 2006 Subject: AW: [Histonet] Giving Specimens to Patients Message-ID: I agree with Bill Blank. All that need be done is dump the materials in formalin to preserve it. My guess is that this rule exists to reduce tracking requirements to save money rather than anything to do with public health. VISIT THE JOURNAL HERPETOLOGICAL CONSERVATION AND BIOLOGY www.herpconbio.org Malcolm L. McCallum Assistant Professor Department of Biological Sciences Texas A&M University Texarkana 2600 Robison Rd. Texarkana, TX 75501 O: 1-903-233-3134 H: 1-903-791-3843 Homepage: https://www.eagle.tamut.edu/faculty/mmccallum/index.html ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Bill Blank Sent: Sat 4/22/2006 9:24 AM To: histonet@lists.utsouthwestern.edu Subject: ~Re: AW: [Histonet] Giving Specimens to Patients This sort of thing bothers me, because it is becoming the rule in the US as regulators and lawyers take away the ability to exercise professional judgement. IMO, given there are no public health concerns, I do not think this is the business of any government. And it is the Pathologist who should decide if there are public health concerns (like a tubercular lung, eg) in each specific case, not lawyers or bureaucrats as they lack the appropriate expertise. If someone wants to bury abortus tissue or limbs, that is their business. As a physician and a humanitarian, I would help make it safe and as visually appealing as possible for them. Bill At 9:48 AM +0200 4/22/06, Gudrun Lang wrote: > Recently we had the case, that a woman wanted to get her abortion-tissue. >You know what that looks like in a glass of formalin. It was not allowed and >I think it was the right decision. > >In Germany there was the case, that a woman wanted to bury the hard of her >husband in his grave. It was a long fight at court. At last she got right >and her soul got peace. -- May the Gods protect me from all the humans who want to protect me. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From brian.chelack <@t> usask.ca Sat Apr 22 10:12:31 2006 From: brian.chelack <@t> usask.ca (Brian Chelack) Date: Sat Apr 22 10:11:30 2006 Subject: [Histonet] DAB (to freeze or not to Freeze) Message-ID: <000601c6661f$2daf4b80$0f13e980@PDS04> Our lab freezes down aliquots of DAB concentrate (typically a 50 mg/ml concentrate containing 0.1% Brij-35 and Tween -20 is frozen in 100 uL volumes in tiny eppendorf style plastic vials). We do this to minimize the handling and thus potential for contact with the DAB. A single 1 gram vial of dab is added to 20 mls of buffer in a closed 50 ml tube and mixed until dissolved. We run the concentrate through 0.45 um syringe filter prior to freezing to remove any insolubles. We thaw the vial when required, add it to 10 mls of buffer add the H2O2 and use. Occasionally we see a bit of ppt. when the concentrate is thawed, but this dissolves again easily when the concentrate is added to the buffer. The frozen concentrate is stable for at least 6 months, and it is convenient. The small volume of concentrate thaws in just a minute or so. My experience with DAB has been that the DAB sold by different manufacturers has slight impurity differences. This provides for different shades of brown. Most of them seem to work. Personally I don't think it matters which one you choose, as long as you like the colour. I've done the Ni and even Co, and they seem a bit more sensitive, although this might just be due to the different coloured reaction product, but really haven't been worth the bother. I've always found they also increase the overall background in a stain and the important part of the chromagen development is the ratio of the "signal" you want to the "noise" you don't want. If you need more sensitivity, generally the best way to achieve it is through your amplification method. When it comes to chromagens; keep it simple and keep it consistent, otherwise you run the risk of driving the slide evaluator nuts with the constant day to day variations We have been very happy with the DAB we purchase from EMS. It has been very consistent through numerous purchases over the past almost 20 years (man, I've been doing this way too long.) Brian Chelack Prairie Diagnostic Services Saskatoon, SK From carl.hobbs <@t> kcl.ac.uk Sat Apr 22 11:46:28 2006 From: carl.hobbs <@t> kcl.ac.uk (Carl Hobbs) Date: Sat Apr 22 11:47:01 2006 Subject: [Histonet] Re: DAB precipitate Message-ID: I routinely make up my DAB powder in water and aliquot and freeze in 1.5ml plastic m'fuge tubes..I never have any problems with it. Please see here http://www.immunoportal.com/ for my method. Look under IHC methods...."DAB". As I routinely use between 10 and 24 slides per run( manual) I always develope my racked slides in a 200ml working solution of DAB, to minimise contact. Pics of the plastic staining dishes on the Immunoportal site, too. carl From BFicher <@t> chomp.org Sat Apr 22 12:47:39 2006 From: BFicher <@t> chomp.org (Fischer, R. B) Date: Sat Apr 22 12:47:48 2006 Subject: [Histonet] job opening in Monterey,Ca. Message-ID: We are currently seeking an ASCP registered HT. or HTL.for our state of the art full service anatomic pathology laboratory. Candidates must have 3+ years experience with embedding, cutting, and staining (both routine and special stains), including processing of non-gyn cytology specimens. We prefer experience in immunohistochemistry staining procedures. We offer a complete benefit package, and relocation assistance is available.Minimum 2 years experience in clinical area or equivalent in specialty areas.Ability to operate Microsoft Office programs a help... R. Brian Fischer Histology Senior Tech Community Hospital of the Monterey Peninsula Monterey, Ca. 93942 831-625-4791 fax: 831-658-3683 From iatomatt <@t> yahoo.co.uk Sat Apr 22 19:12:57 2006 From: iatomatt <@t> yahoo.co.uk (MattG) Date: Sat Apr 22 19:13:01 2006 Subject: [Histonet] Impending CPA inspection Message-ID: <001201c6666a$ae9191e0$6501a8c0@acer311vpbceh0> Hi Histonetters, One primarily for the UK lot, but comments from all are welcome. We are about to have a CPA inspection, and a preliminary inspection by a haematologist has raised two queries. 1. We keep our solvents in a proper cabinet, but small quantities of various grades and flammable reagents are in bottles on the special stains shelf. It has been suggested that all flammable liquids should be kept in the cabinet. This seems like overkill to me, I have worked in other labs that passed CPA inspection when there were similar amounts out on shelves. Do any of you know if there are any guidelines on what is permissible? 2. We're a low volume lab, so we keep our specimen pots in a metal cabinet, much like a flammable cabinet, prior to them being disposed of. It has been suggested that we should use a special formalin specimen storage cabinet, which has ventilation. The formalin vapour level readings are very low, well below guideline levels. Is there any info about what sort of storage arrangements CPA consider acceptable? Any advice gratefully received, Matt ___________________________________________________________ To help you stay safe and secure online, we've developed the all new Yahoo! Security Centre. http://uk.security.yahoo.com From lhotaks <@t> mcmaster.ca Sat Apr 22 21:43:56 2006 From: lhotaks <@t> mcmaster.ca (Sarka Lhotak) Date: Sat Apr 22 21:44:01 2006 Subject: [Histonet] colocalization of fluorescence In-Reply-To: <200604221707.k3MH7aQL001356@coriana6.cis.mcmaster.ca> Message-ID: Hello Netters, Sorry if I misunderstood this thread but it is not making much sense. You will not be colocalizing colours, as in mixing the colors. You will see and image each color separately through a different filter. You might then superimpose the images on top of each other but it is not necessary. It is usual to publish separate colors side by side. The colors you will see in the microscope depend on your emission filters. If you have a long pass filter you will see both red and green with 488 excitation. With band filters you should only see one color at a time. Hope this helps, Sarka Lhotak McMaster University, Hamilton, Ontario, Canada From vanann702 <@t> skmc.gov.ae Sun Apr 23 03:04:02 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Sun Apr 23 03:03:07 2006 Subject: [Histonet] disclaimer Message-ID: Hello Histonetters Do you include any form of disclaimer at the end of you Path reports? Is this on every report or just on those with IHC or Cytology? Is it mandatory? Where would I find this information? Cheers Anne Anne S van Binsbegen Anatomical Pathology Laboratory Shaikh Khalifa Medical City PO BOX 51900 Abu Dhabu, U.A.E. ph: +971 2 6102695 mob:+971 50 3162804 fax +971 2 6104989 dance like no one is watching Sheikh Khalifa Medical City (SKMC) Disclaimer Notice: This mail message (including any attachments) is intended only for the designated recipient only and may contain privileged, proprietary, or otherwise private information. If you are not the addressee you must not copy, distribute, disclose or use any of the information in it. If you have received it in error, please notify the sender immediately and delete the original. From Xorren <@t> aol.com Sun Apr 23 07:56:57 2006 From: Xorren <@t> aol.com (Xorren@aol.com) Date: Sun Apr 23 07:57:05 2006 Subject: [Histonet] Re: Histonet Digest, Vol 29, Issue 26 Message-ID: <2e7.57e6f7d.317cd399@aol.com> HISTO CYTO JOBS IN NYC OR LONG ISLAND. Hello hisotnetters. Does anyone know of any positions in histolgy or cytoprep and stainging? I am a certified histotech with 12 years experience. Any help is very welcome. please email me at Xorren@aol.com Have a great week! From AnthonyH <@t> chw.edu.au Sun Apr 23 17:59:33 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Apr 23 17:59:46 2006 Subject: [Histonet] Diff Quick Message-ID: A Hair dryer helps with the drying. Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Kemlo Rogerson Sent: Thursday, 20 April 2006 11:18 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Diff Quick We run a one-step FNAC breast clinic and have been using Diff-Quick as our stain; I personally don't like it as I prefer Paps but there you are. We had a recent FNAC that showed histiocytes in the tail of the prep but little else. As you know the bubbly cells tend to migrate to the edges as they are lighter but what we initially failed to notice was that in amongst the blood were poorly stained clumps of cells. After restain with a 'proper' Romanovsky stain they magically appeared and were benign. I shudder to think what would have happened if they had not been spotted and had been malignant. The problem I guess was the blood and together with the slow drying of the smear. Diff-Quick is fast but can have this effect, does anyone have a technique that is nearly as fast as Diff Quick but more robust in these cases? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Through return to simple living comes control of desires. In control of desires stillness is attained. In stillness the world is restored. -- Lao Tzu This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. ********************************************************************** From mbmphoto <@t> gmail.com Mon Apr 24 01:10:17 2006 From: mbmphoto <@t> gmail.com (Maria Mejia) Date: Mon Apr 24 01:09:40 2006 Subject: [Histonet] searching for great sliding microtome Message-ID: I would like very much to hear from those folks who can recommend a very good USER FRIENDLY sliding microtome for large primate brain blocks for frozen sections cut at 40 microns. We are working with large & thick brain blocks measuring 2 inches by 2.5 inches. Yes, vendors are also welcome. In addition, I'm very keen on hearing from (anyone) who has a working AO sliding microtome & would like to get rid of it. I'm thinking of the cast iron type. I look forward to hearing from you. Any suggestions, & tips will be greatly appreciated. Yours Maria Bartola Mejia Department of Neurosurgery University of California San Francisco (UCSF) San Francisco, CA 94103 From abright <@t> brightinstruments.com Mon Apr 24 05:07:33 2006 From: abright <@t> brightinstruments.com (Alan Bright) Date: Mon Apr 24 04:57:29 2006 Subject: [Histonet] searching for great sliding microtome Message-ID: Dear Maria, We manufacture the Bright 8000 Retracting Base Sledge Microtome ( cast Iron type) that fits your criteria. Also to complement this microtome for your application we manufacture a 8000-208-02 Cryostage Freezing Stage for specimen sizes up to 130 X 90 mm, with a temperature range down to -30?C. For larger specimen sizes 250 X 110 mm, we also manufacture an 8250 cryostat with an anti-roll plate option, with a temperature range down to -35?C & a motorized drive system. These items can be viewed on our website. 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 Skype User ID: dazzle0 -----Original Message----- From: Maria Mejia [mailto:mbmphoto@gmail.com] Sent: 24 April 2006 07:10 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] searching for great sliding microtome I would like very much to hear from those folks who can recommend a very good USER FRIENDLY sliding microtome for large primate brain blocks for frozen sections cut at 40 microns. We are working with large & thick brain blocks measuring 2 inches by 2.5 inches. Yes, vendors are also welcome. In addition, I'm very keen on hearing from (anyone) who has a working AO sliding microtome & would like to get rid of it. I'm thinking of the cast iron type. I look forward to hearing from you. Any suggestions, & tips will be greatly appreciated. Yours Maria Bartola Mejia Department of Neurosurgery University of California San Francisco (UCSF) San Francisco, CA 94103 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Heather.A.Harper <@t> pcola.med.navy.mil Mon Apr 24 05:40:24 2006 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Mon Apr 24 05:43:53 2006 Subject: [Histonet] Bone decalcification Message-ID: I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL From vanann702 <@t> skmc.gov.ae Mon Apr 24 06:03:36 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Mon Apr 24 06:02:37 2006 Subject: [Histonet] Bone decalcification Message-ID: Even in darkest Africa....we were taught to 'Always fix first' - properly -then decal!!! Annieinarabia -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 2:40 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Apr 24 06:10:20 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Apr 24 06:09:39 2006 Subject: [Histonet] Bone decalcification Message-ID: If you decalc the bone then don't bother to fix it as you have probably ruined the morphology. Very odd school of thought that. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Heather.A.Harper@pcola.med.navy.mil [mailto:Heather.A.Harper@pcola.med.navy.mil] Sent: Monday, April 24, 2006 11:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Tiffany.L.Sheffield <@t> uth.tmc.edu Mon Apr 24 06:40:23 2006 From: Tiffany.L.Sheffield <@t> uth.tmc.edu (Sheffield, Tiffany L) Date: Mon Apr 24 06:40:09 2006 Subject: [Histonet] Bone decalcification Message-ID: One of the most important things I learned back in the day was the importance of choosing the appropriate fixative. In your case I would always fix the tissue/bone first before decal. Hope this helps :-) ************************************************** Tiffany Sheffield-Lopez,BS,HT(ASCP) Supervisor, Bone Histomorphometry & Biomaterials Lab Department of Orthopaedic Surgery The University of Texas Houston Health Science Center 6431 Fannin, Suite 6.144 MSB Houston, TX 77030 713-500-6803 Wk 713-500-0729 Fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Mon Apr 24 06:45:51 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Mon Apr 24 06:46:20 2006 Subject: [Histonet] Bone decalcification References: Message-ID: <83AACDB0810528418AA106F9AE9B7F7E01825452@sjhaexc02.sjha.org> I was always taught to fix first. Joyce -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu on behalf of Heather.A.Harper@pcola.med.navy.mil Sent: Mon 4/24/2006 6:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From DDDeltour <@t> mar.med.navy.mil Mon Apr 24 06:58:19 2006 From: DDDeltour <@t> mar.med.navy.mil (Deltour, Douglas D. (HM2)) Date: Mon Apr 24 06:58:56 2006 Subject: [Histonet] Bone decalcification Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE52@marxchg03.mar.med.navy.mil> Heather, Fixation is the key to a good section of bone. Surgipath makes a two step decal system. Decalcifier 1 is a combined fixative and decalcifying agent. Decalcifier 2 is rapid decalcification. We use this system and have had no problems. I can give you the catalog # if you need it. Good luck. Douglas Deltour HT(ASCP) -----Original Message----- From: Heather.A.Harper@pcola.med.navy.mil [mailto:Heather.A.Harper@pcola.med.navy.mil] Sent: Monday, April 24, 2006 6:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gillian.2.brown <@t> GSK.COM Mon Apr 24 07:08:19 2006 From: gillian.2.brown <@t> GSK.COM (gillian.2.brown@GSK.COM) Date: Mon Apr 24 07:07:10 2006 Subject: [Histonet] Bone decalcification In-Reply-To: Message-ID: Heather, do you not have any text books? As you have already seen the majority of answers are saying fixation is of prime importance if you wish to observe the cellular components of the bone using microscopy. Reason you want to decalcify is presumably to be able to cut nice thin paraffin wax sections? No point in doing that if you've already ruined the morphology by not fixing appropriately. Don't know how anyone could 'learn this the hard way' do people really walk in to labs and make it up as they go along? Gill Brown Heather.A.Harper@pcola.med.navy.mil Sent by: histonet-bounces@lists.utsouthwestern.edu 24-Apr-2006 11:40 To histonet@lists.utsouthwestern.edu cc Subject [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From caron_fournier <@t> yahoo.ca Mon Apr 24 08:50:21 2006 From: caron_fournier <@t> yahoo.ca (caron fournier) Date: Mon Apr 24 08:50:28 2006 Subject: [Histonet] (no subject) Message-ID: <20060424135022.75759.qmail@web35409.mail.mud.yahoo.com> Co-localization of proteins is no a simple as a color change. You can get a color change in an section if the proteins are above and below each other in the section and still not occupying the same space. There is a very complicated formula(s) that can be used when calculating cooficients of co-localization and it is much more complicated than a color change. You should be using image analysis software and doing calculations of these co-efficients to get a true account of what is really happening. Check out http://www.mediacy.com/index.aspx?page=FT_Colocalization Caron Fournier, BSc, R.T. Department of Orthopaedics, Division of Orthopaedic Engineering Research, U.B.C. __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From Charles.Embrey <@t> carle.com Mon Apr 24 08:23:47 2006 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Mon Apr 24 08:53:19 2006 Subject: [Histonet] Bone decalcification Message-ID: I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Charles.Embrey <@t> carle.com Mon Apr 24 08:36:37 2006 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Mon Apr 24 08:53:23 2006 Subject: [Histonet] Giving Specimens to Patients Message-ID: The POC issue is a very tricky and emotional issue. We had a law passed here in Illinois a few years ago to protect the Mother's rights concerning Products of Conception. Every woman that miscarriages in Illinois, regardless of gestational age, has the right to have a funeral home pick up the tissue for burial. The law came about after a nurse lost her baby at gestational age less than 20 weeks and she immediately asked that the abortus be saved for burial. When the funeral home came for the tissue it was discovered that it had been thrown out with the other tissue waste in pathology. Charles Embrey Jr. PA(ASCP) www.greyrealm.com -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Gudrun Lang Sent: Saturday, April 22, 2006 2:48 AM To: Histonetliste (Histonetliste) Subject: AW: [Histonet] Giving Specimens to Patients Recently we had the case, that a woman wanted to get her abortion-tissue. You know what that looks like in a glass of formalin. It was not allowed and I think it was the right decision. In Germany there was the case, that a woman wanted to bury the hard of her husband in his grave. It was a long fight at court. At last she got right and her soul got peace. Gudrun Lang Austria -----Urspr?ngliche Nachricht----- Von: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Im Auftrag von Jackie M O'Connor Gesendet: Freitag, 21. April 2006 21:00 An: Bill Blank Cc: histonet@lists.utsouthwestern.edu; Smith Wanda; histonet-bounces@lists.utsouthwestern.edu Betreff: Re: [Histonet] Giving Specimens to Patients Sorry, Bill - I think that once he signed all the papers before surgery, he gives the hospital permission to 'dispose of' tissues removed in surgery. I think that makes his rib the hospital's property. We once had a guy ask for his toe bones back after he lost part of his foot in a motorcycle accident. He wanted to make a necklace. The pathologist said 'no'. End of story, unless we get into all the patients whose husbands ask for the placenta after childbirth. Jackie O' Bill Blank Sent by: histonet-bounces@lists.utsouthwestern.edu 04/21/2006 01:36 PM To "Smith Wanda" , cc Subject Re: [Histonet] Giving Specimens to Patients Regardless of silly laws. It is HIS rib. I give it to them, but usually replace the formalin with ETOH. Bill At 12:28 PM -0500 4/21/06, Smith Wanda wrote: >We have aways given gallstones and foreign bodies to patients if they >request them, however, I have a patient that wants his rib back >that was removed in surgery. It just looks like a rib fracture, >no atypia. Our Pathologist don't recommend this, but it is a call from >the hospital legal department and I can't get in touch with them today. >What is the practice everyone uses for giving/not giving patient's >their specimens in formalin? -- _____________________________ Bill Blank http://kernunnos.com (Celtic studies and numismatics) OBOD's Message board: http://www.druidry.org/board _______________________________________________ Histonet mailing list Histonet@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 carolb <@t> phys.mcw.edu Fri Apr 21 11:29:51 2006 From: carolb <@t> phys.mcw.edu (Bobrowitz, Carol) Date: Mon Apr 24 09:06:39 2006 Subject: [Histonet] repeating AR heat treatment Message-ID: <8F78639AC56F4143B267FE5F5A1B92C80E42D3@guyton.phys.mcw.edu> I have already stained FFPE rat kidney's using FITC with a 2 hour heat treatment in citrate buffer ph 6.0. (the staining was great) I am now going to restain the same rat kidney slides with an additional primary using DAB as the chromagen. This primary also requires heat treatment. Am I correct in my thinking that I should repeat the heat treatment again due to the fact that the epitope (binding) sites have already closed after accepting the application of the 1st primary? Any help will be appreciated. Thank you in advance. Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology 414-456-8179 cbobrowi@mcw.edu From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Apr 24 09:14:11 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Apr 24 09:13:34 2006 Subject: [Histonet] Bone decalcification Message-ID: Does Pensacola make you burp if you drink it too fast? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 2:24 PM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From DDDeltour <@t> mar.med.navy.mil Mon Apr 24 09:20:51 2006 From: DDDeltour <@t> mar.med.navy.mil (Deltour, Douglas D. (HM2)) Date: Mon Apr 24 09:21:31 2006 Subject: [Histonet] Bone decalcification Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE55@marxchg03.mar.med.navy.mil> "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Apr 24 09:23:44 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Apr 24 09:23:50 2006 Subject: [Histonet] repeating AR heat treatment In-Reply-To: <8F78639AC56F4143B267FE5F5A1B92C80E42D3@guyton.phys.mcw.edu> Message-ID: <20060424142344.75024.qmail@web61220.mail.yahoo.com> Carol: HIER (Heat Induced Epitope Retrieval) treatment is to "undo" the formaldehyde crosslinking of proteins. The application of the first Ab was specifically binded to their antigenic sites and did nothing to other antigenic sites. HIER eliminated crosslinkage to all antigenic sites. If your new primary targets the same antigen as your FITC probably it will not be able to link unless you eliminate that FITC conjugated Ab; they are competing for the same site. I don't think that new HIER session will solve this problem.Perhaps immersing the sections in a low pH solution (pH 5) will break the Ag-Ab reaction you obtained with your FITC conjugated Ab. Probably that would be enough. After the treatment (that will affect the quaternary structure of the Ag site) you should place the slide in pH neutral buffer to try to "return" the structure to its natural configuration. If you are targeting another Ag sites, you don't need another HIER treatment, all sites should have become "available" to receive the Abs. Hope this will help you! Ren? J. "Bobrowitz, Carol" wrote: I have already stained FFPE rat kidney's using FITC with a 2 hour heat treatment in citrate buffer ph 6.0. (the staining was great) I am now going to restain the same rat kidney slides with an additional primary using DAB as the chromagen. This primary also requires heat treatment. Am I correct in my thinking that I should repeat the heat treatment again due to the fact that the epitope (binding) sites have already closed after accepting the application of the 1st primary? Any help will be appreciated. Thank you in advance. Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology 414-456-8179 cbobrowi@mcw.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From kemlo.rogerson <@t> waht.swest.nhs.uk Mon Apr 24 09:32:37 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Mon Apr 24 09:31:59 2006 Subject: [Histonet] Bone decalcification Message-ID: Agreed. It is a comment set to humble someone trying to learn and serves only for them to not speak again lest they offend; it is a form of bullying I guess. If I had a dollar for every question I asked that everyone else knew the answer to then I'd be rich; but then those who knew the answer must have asked the question at sometime, mustn't they? Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Deltour, Douglas D. (HM2) [mailto:DDDeltour@mar.med.navy.mil] Sent: Monday, April 24, 2006 3:21 PM To: 'Charles.Embrey'; Harper, Heather A., CIV Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Apr 24 09:33:31 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Apr 24 09:33:38 2006 Subject: [Histonet] Bone decalcification In-Reply-To: <3F500F8B416C554EBB21FF16642F72E959CE55@marxchg03.mar.med.navy.mil> Message-ID: <20060424143331.14949.qmail@web61225.mail.yahoo.com> I seldom intervene in "thread discussions" but this time I want to say that I agree with Douglas (although my agreement has no relevance in itself). Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer. It is my opinion in this subject (that also in itself bears no real weight) that once a question has been answred correctly we all shoul refrain from keep sending the same correct answer, and even more, refrain from making sarcastic or even sometimes insulting comments. And now you all have the opportunity of contradicting what I have just wrote (that also in itself would be absolutely irrelevant!). Ren? J. "Deltour, Douglas D. (HM2)" wrote: "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From Charles.Embrey <@t> carle.com Mon Apr 24 09:47:44 2006 From: Charles.Embrey <@t> carle.com (Charles.Embrey) Date: Mon Apr 24 09:47:52 2006 Subject: [Histonet] Bone decalcification Message-ID: I must apologize to Heather and the rest of the Histonet for my flippant remark. I really didn't mean any harm. I am just having a crappy Monday and my bad mood let the sarcasm slip out. This forum is a great place to share knowledge and I let a moment of un-professionalism get in the way of that. Chuck -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Monday, April 24, 2006 9:34 AM To: Deltour, Douglas D. (HM2); Charles.Embrey; Harper, Heather A., CIV Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I seldom intervene in "thread discussions" but this time I want to say that I agree with Douglas (although my agreement has no relevance in itself). Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer. It is my opinion in this subject (that also in itself bears no real weight) that once a question has been answred correctly we all shoul refrain from keep sending the same correct answer, and even more, refrain from making sarcastic or even sometimes insulting comments. And now you all have the opportunity of contradicting what I have just wrote (that also in itself would be absolutely irrelevant!). Ren? J. "Deltour, Douglas D. (HM2)" wrote: "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From asmith <@t> mail.barry.edu Mon Apr 24 09:53:11 2006 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Mon Apr 24 09:53:16 2006 Subject: [Histonet] Bone decalcification Message-ID: <5D2189E74151CC42BEC02906BA8996322B91CC@exchsrv01.barrynet.barry.edu> Heather's question is not at all unreasonable. Some acids are pretty decent fixatives. While I learned histotechnique back in the Dark Ages, I was taught to fix and decalcify simulataneously. We used 85% saturated aqueous picric acid, 10% formalin, and 5% formic acid. The explosive nature of dry picric acid has made this formula less popular than it used to be. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Charles.Embrey Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 Vickroy.Jim <@t> mhsil.com Mon Apr 24 10:25:08 2006 From: Vickroy.Jim <@t> mhsil.com (Vickroy, Jim) Date: Mon Apr 24 10:25:21 2006 Subject: [Histonet] formalin levels and exhaust systems Message-ID: We are looking into a benchtop exhaust system to absorb and filter formalin fumes at two of our grossing stations. Has anyone ever used the benchtop systems (such as ones by companies such as Mopec or Thermoshandon)? What kind of results did you have with these formalin filtering systems. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. From mprice26 <@t> juno.com Mon Apr 24 10:43:30 2006 From: mprice26 <@t> juno.com (mprice26@juno.com) Date: Mon Apr 24 10:45:20 2006 Subject: [Histonet] RE:Used Shandon Varistain 24-4 Automatic Slide Stainer for sale Message-ID: <20060424.084419.28635.793826@webmail47.nyc.untd.com> We have a 15 year old Shandon Varistain 24-4 automatic Slide stainer for sale for $2000. It is in good working condition. If interested or would like additional information respond to this e-mail message. Thank you. Marsha Price From funderwood <@t> mcohio.org Mon Apr 24 10:47:43 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Mon Apr 24 10:48:05 2006 Subject: [Histonet] Bone decalcification Message-ID: The downside of the world wide web. A phone can be hung up before something regrettable is said. But once the send button is clicked..... >>> "Charles.Embrey" 04/24/06 10:47AM >>> I must apologize to Heather and the rest of the Histonet for my flippant remark. I really didn't mean any harm. I am just having a crappy Monday and my bad mood let the sarcasm slip out. This forum is a great place to share knowledge and I let a moment of un-professionalism get in the way of that. Chuck -----Original Message----- From: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: Monday, April 24, 2006 9:34 AM To: Deltour, Douglas D. (HM2); Charles.Embrey; Harper, Heather A., CIV Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I seldom intervene in "thread discussions" but this time I want to say that I agree with Douglas (although my agreement has no relevance in itself). Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer. It is my opinion in this subject (that also in itself bears no real weight) that once a question has been answred correctly we all shoul refrain from keep sending the same correct answer, and even more, refrain from making sarcastic or even sometimes insulting comments. And now you all have the opportunity of contradicting what I have just wrote (that also in itself would be absolutely irrelevant!). Ren? J. "Deltour, Douglas D. (HM2)" wrote: "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ________________________________ Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Mon Apr 24 11:01:33 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Mon Apr 24 11:01:38 2006 Subject: [Histonet] formalin levels and exhaust systems In-Reply-To: Message-ID: <20060424160133.62176.qmail@web61221.mail.yahoo.com> I bought two large counter-top fume extraction hoods (from Fisher) to be used in the cassetting area. They worked very well and eliminated formalin fumes. We also had a smaller one for coverslipping in the frozen section area. We always found them convenient with the advantage that you can place them were needed. Hope this will help! Ren? J. "Vickroy, Jim" wrote: We are looking into a benchtop exhaust system to absorb and filter formalin fumes at two of our grossing stations. Has anyone ever used the benchtop systems (such as ones by companies such as Mopec or Thermoshandon)? What kind of results did you have with these formalin filtering systems. James R. Vickroy Supervisor - MMC Surgical Pathology 217-788-4046 This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1¢/min. From sbreeden <@t> nmda.nmsu.edu Mon Apr 24 11:33:13 2006 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Mon Apr 24 11:33:19 2006 Subject: [Histonet] Hold on, there! Message-ID: First, the necessary disclaimers: "I do NOT mean to offend; nor do I wish to have my comments misinterpreted in any way, shape, or form; I understand that the English language is difficult to interpret/translate/etc.; this struck me as funny on a Monday; and I just can't help myself..." I just HAVE to ask about the message relating to the woman in Germany who wanted to bury something of her husband's with his remains...please - could it be "head", "heart" or is it "hard"??? I'm chuckling...please join me! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From GauchV <@t> mail.amc.edu Mon Apr 24 11:36:30 2006 From: GauchV <@t> mail.amc.edu (Vicki Gauch) Date: Mon Apr 24 11:36:01 2006 Subject: [Histonet] Giving Specimens to Patients Message-ID: Wanda, We have run into this scenario numerous times and we do not release any specimens that are in formalin. If a patient wishes to have tissue released they have to go through a funeral director . We did have a patient request his arm back and it was decided that he would go through the funeral director who made some type of arrangement so that the arm would be buried with him when the time came. Our hospital has even begun to clamp down on the release of foreign bodies,etc. We offer them photographs of the foreign bodies which satsfies SOME of the patients. It is always an issue though and there doesn't seem to be an easy answer. Vicki AMCH >>> "Smith Wanda" 4/21/2006 1:28:22 PM >>> Happy Friday Histonet, We have aways given gallstones and foreign bodies to patients if they request them, however, I have a patient that wants his rib back that was removed in surgery. It just looks like a rib fracture, no atypia. Our Pathologist don't recommend this, but it is a call from the hospital legal department and I can't get in touch with them today. What is the practice everyone uses for giving/not giving patient's their specimens in formalin? Thanks, Wanda _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ----------------------------------------- CONFIDENTIALITY NOTICE: This email and any attachments may contain confidential information that is protected by law and is for the sole use of the individuals or entities to which it is addressed. If you are not the intended recipient, please notify the sender by replying to this email and destroying all copies of the communication and attachments. Further use, disclosure, copying, distribution of, or reliance upon the contents of this email and attachments is strictly prohibited. To contact Albany Medical Center, or for a copy of our privacy practices, please visit us on the Internet at www.amc.edu. From gcallis <@t> montana.edu Mon Apr 24 11:56:05 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Apr 24 11:56:12 2006 Subject: Another Re: [Histonet] Bone decalcification In-Reply-To: References: Message-ID: <6.0.0.22.1.20060424104158.01b70480@gemini.msu.montana.edu> Acid decalcification BEFORE fixation will macerate your tissues aka protein hydrolysis. It will destroy both soft tissue morphology and nuclear staining, and there is no way you can repair this kind of chemical destruction of tissues. Total fixation is needed to protect the tissues from the effects of acid decalcification. IF the bone is small, i.e. a bone biopsy or cut down into a very small piece, then a formic acid/formalin (combination fixation and decalcification) can be used successfully. This is NOT recommended for really large bones (mfrs even make these recommendations in their instruction sheets). One message kindly provided vendor recommendation for this fix/decal solution. If you do not want to use formic acid/formalin, then fix the bone first in NBF and proceed to a acid decalcifier. Keep in mind, strong acids i.e. HCl (commercial solutions work very well) can damage antigens if bone is left in these solutions too long. For a gentler acid decalcifier, buffered formic acid is available commericially. The recommendation about a textbook is a good one - several devote whole chapters to bone histotechnics, including how to decalcify, etc and what will happen IF you do not perform these procedures carefully. Our personal preference is to totally fix the bone in NBF. To speed up both fixation and decalcification, reduce the size of the bone i.e. cut a representative sample for fixation followed by decalcification. This also helps with the combo fix/decal method. If you suspend the bone, the reagents surround all sides of the bones. The only time I have seen unfixed bone followed by decalcification is with EDTA, and it was a tedious, long drawn out method, used for research projects and NOT the clinical setting where you need a diagnosis asap. The EDTA methods are found in several publicaitons. At 04:40 AM 4/24/2006, you wrote: > I just wanted to know other histo techs opinions on bone decalcification. >I learned in school that you decal the bone than you fix it. I have a >pathologist who claims she learned it the hard way, and that it is better to >fix the bone than decal it. What did you learn on how to do this procedure? >This pathologist claims that fixing prior to decaling keeps the cells more >intact. Any opinion on your procedure or your technique is appreciated. > > > >Heather A. Harper > >Naval Hospital > >Pensacola, FL > >_______________________________________________ >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 406 994-4303 (FAX) From gcallis <@t> montana.edu Mon Apr 24 12:19:42 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Mon Apr 24 12:19:48 2006 Subject: [Histonet] Bone decalcification In-Reply-To: <5D2189E74151CC42BEC02906BA8996322B91CC@exchsrv01.barrynet. barry.edu> References: <5D2189E74151CC42BEC02906BA8996322B91CC@exchsrv01.barrynet.barry.edu> Message-ID: <6.0.0.22.1.20060424111136.01b94eb8@gemini.msu.montana.edu> Allen, You are correct. Zenkers (although mercury is no longer environmentally acceptable) will fix and decalcify, and some have used this in the past for Jamshidi bone biopsy work. Bouins with acetic acid works for tiny decalcifications, and I have seen a Bouins modification using formic acid in place of acetic acid - not too much different than what you suggest here. Neither worked very well on thicker cortical bone, and I would not like to fix a long time in Bouins (72 hours is an upper limit recommended fixation ala histotext books and the bone may not be totally decalcified by then). We store our stock picric acid under a layer of water, check it very frequently and make sure the edges of lids do not seep out picric acid ( sealed with hot paraffin) to exist in powder form on exterior of lid. At 08:53 AM 4/24/2006, you wrote: >Heather's question is not at all unreasonable. Some acids are pretty decent >fixatives. While I learned histotechnique back in the Dark Ages, I was >taught to fix and decalcify simulataneously. We used 85% saturated aqueous >picric acid, 10% formalin, and 5% formic acid. The explosive nature of dry >picric acid has made this formula less popular than it used to be. > >Allen A. Smith, Ph.D. 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 406 994-4303 (FAX) From jcline <@t> wchsys.org Mon Apr 24 12:20:33 2006 From: jcline <@t> wchsys.org (Joyce Cline) Date: Mon Apr 24 12:20:49 2006 Subject: [Histonet] bone marrow Fe stains/Galiotto,Laura In-Reply-To: <270614B321ACB44D8C1D91F4F921FDC3036CBCC2@NCH01EX02.nch.org> Message-ID: <000001c667c3$66ba0af0$1d2a14ac@wchsys.org> It is processed liver. Our path's are acceptable of this type of control. -----Original Message----- From: Galiotto, Laura [mailto:LGaliotto@nch.org] Sent: Thursday, April 20, 2006 3:02 PM To: Joyce Cline Subject: RE: [Histonet] bone marrow Fe stains What type of control is it Aspirate smear or processed liver -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joyce Cline Sent: Thursday, April 20, 2006 12:42 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] bone marrow Fe stains I use only purchased iron controls from H.O.M.E. I use the same control for the aspirate block, the core block and the smears. -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Galiotto, Laura Sent: Wednesday, April 19, 2006 3:58 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] bone marrow Fe stains I have a problem with bone marrow aspirate controls. The problem is that we are limited to the amount of controls we can collect. Even though the tissue control is working if the aspirate control (which are difficult to obtain) does not work we are being asked to repeat the stain until we find a aspirate control that is acceptable. Since I can not guarantee the aspirate control contains an adequate amount of Fe, I will be wasting time and reagents. Can anyone give feedback on how they process stains on bone marrow's, do you only use a tissue control, does anyone know where bone marrow aspirate controls -positive for Iron -can be purchased commercially? Laura Galiotto, HT (ASCP) Histology Facilitator Northwest Community Hospital 800 West Central Road Arlington Heights, Il 60005 847-618-6190 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ***** CONFIDENTIALITY NOTICE ***** This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. From MElliott <@t> mrl.ubc.ca Mon Apr 24 12:29:52 2006 From: MElliott <@t> mrl.ubc.ca (Mark Elliott) Date: Mon Apr 24 12:30:13 2006 Subject: [Histonet] control slides Message-ID: <444CA8A0020000D600005945@mail.mrl.ubc.ca> Does anyone know of a supplier of control slides for influenza and parvo in humans?? Thanks Mark Elliott From DeBrosse_Beatrice <@t> Allergan.com Mon Apr 24 12:53:23 2006 From: DeBrosse_Beatrice <@t> Allergan.com (DeBrosse_Beatrice) Date: Mon Apr 24 12:53:48 2006 Subject: [Histonet] Hold on, there! Message-ID: Sally, I'm glad you have a disclaimer, since I was a bit offended by your remark. Even though Gudrun probably meant "heart" and not "hard", I would imagine you have enough common sense to make out what she meant. At least she communicates, should I say quite well, in English. Since I am Swiss, I can relate a bit, and the difficulty in English is, that a lot of words are pronounced the same, but spelled differently. I've seen a whole lot worse as far as spelling goes on the histonet, and some people were from the USA. Why don't you just give her a break? And would you do as well in German? Beatrice DeBrosse-Serra BS, HT(ASCP)QIHC Allergan, Inc. 2525 Dupont Drive RD-2A Irvine, CA 92612 714-246-5116 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Breeden, Sara Sent: Monday, April 24, 2006 9:33 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Hold on, there! First, the necessary disclaimers: "I do NOT mean to offend; nor do I wish to have my comments misinterpreted in any way, shape, or form; I understand that the English language is difficult to interpret/translate/etc.; this struck me as funny on a Monday; and I just can't help myself..." I just HAVE to ask about the message relating to the woman in Germany who wanted to bury something of her husband's with his remains...please - could it be "head", "heart" or is it "hard"??? I'm chuckling...please join me! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From asmith <@t> mail.barry.edu Mon Apr 24 13:48:42 2006 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Mon Apr 24 13:48:49 2006 Subject: [Histonet] Bone decalcification Message-ID: <5D2189E74151CC42BEC02906BA8996322B91CD@exchsrv01.barrynet.barry.edu> Although every textbook I've seen limits Bouin's fixation to 72 hours or less, I have occasionally gotten away with fixing in Bouin's for a week (168 hours). Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: Gayle Callis [mailto:gcallis@montana.edu] Sent: Monday, April 24, 2006 1:20 PM To: Smith, Allen; Histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Bone decalcification Allen, You are correct. Zenkers (although mercury is no longer environmentally acceptable) will fix and decalcify, and some have used this in the past for Jamshidi bone biopsy work. Bouins with acetic acid works for tiny decalcifications, and I have seen a Bouins modification using formic acid in place of acetic acid - not too much different than what you suggest here. Neither worked very well on thicker cortical bone, and I would not like to fix a long time in Bouins (72 hours is an upper limit recommended fixation ala histotext books and the bone may not be totally decalcified by then). We store our stock picric acid under a layer of water, check it very frequently and make sure the edges of lids do not seep out picric acid ( sealed with hot paraffin) to exist in powder form on exterior of lid. At 08:53 AM 4/24/2006, you wrote: >Heather's question is not at all unreasonable. Some acids are pretty decent >fixatives. While I learned histotechnique back in the Dark Ages, I was >taught to fix and decalcify simulataneously. We used 85% saturated aqueous >picric acid, 10% formalin, and 5% formic acid. The explosive nature of dry >picric acid has made this formula less popular than it used to be. > >Allen A. Smith, Ph.D. 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 406 994-4303 (FAX) 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 sbreeden <@t> nmda.nmsu.edu Mon Apr 24 15:37:25 2006 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Mon Apr 24 15:37:28 2006 Subject: [Histonet] Apology Message-ID: It's a Monday for apology. Apparently my posting about the organ being returned for burial offended a couple of individuals and for that, I'm sorry. I have apologized to the originator as well. My first "flaming" and I'm a little crisp around the edges; but I still like a good chuckle. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From muddymoo <@t> gmail.com Mon Apr 24 16:25:43 2006 From: muddymoo <@t> gmail.com (Alan Bishop) Date: Mon Apr 24 16:25:50 2006 Subject: [Histonet] Giving Specimens to Patients In-Reply-To: References: Message-ID: Both in the UK and NZ you now have legalities in place for patients to request the return of ANY tissues after a visit to the hospital. I was majorly involved in this process in the UK for 3 years after a hospital kept a lot of specimens from children without permission. Due to this consent forms have now changed and the option for return is included, right down to blocks, slides and even trimmings from cutting the blocks if so desired. As Darren says it very similar here in NZ now as well as we have to fit in with the wishes of such a culturally diverse country. One thing though, specimens are never returned in formalin. Always well washed and air dried if the specimen allows for that. Obviously never get rid of all traces of formalin so in the UK we were giving out strict guidlines on what should or should not be done with a specimen and also included the Hazard sheet. Going to far? Probably Cheers Alan From jnocito <@t> satx.rr.com Mon Apr 24 18:57:10 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Mon Apr 24 18:57:18 2006 Subject: [Histonet] Apology References: Message-ID: <000a01c667fa$cdb7d660$e8bd0b43@yourxhtr8hvc4p> Sally, welcome to the "flamed" club. As president of the Histonet Flamed Society, I welcome your new membership. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Breeden, Sara" To: Sent: Monday, April 24, 2006 3:37 PM Subject: [Histonet] Apology It's a Monday for apology. Apparently my posting about the organ being returned for burial offended a couple of individuals and for that, I'm sorry. I have apologized to the originator as well. My first "flaming" and I'm a little crisp around the edges; but I still like a good chuckle. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 From jnocito <@t> satx.rr.com Mon Apr 24 19:02:42 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Mon Apr 24 19:02:49 2006 Subject: [Histonet] repeating AR heat treatment References: <8F78639AC56F4143B267FE5F5A1B92C80E42D3@guyton.phys.mcw.edu> Message-ID: <003c01c667fb$93bc4800$e8bd0b43@yourxhtr8hvc4p> Carol, my experience is that I didn't need to go through the retrieval steps again. I've done more cases than I can count with a pathologist wanting one immuno, reads that one and then orders another one on the same slide. Just my 3 cents worth. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Bobrowitz, Carol" To: "'Histonet (E-mail)" Sent: Friday, April 21, 2006 11:29 AM Subject: [Histonet] repeating AR heat treatment I have already stained FFPE rat kidney's using FITC with a 2 hour heat treatment in citrate buffer ph 6.0. (the staining was great) I am now going to restain the same rat kidney slides with an additional primary using DAB as the chromagen. This primary also requires heat treatment. Am I correct in my thinking that I should repeat the heat treatment again due to the fact that the epitope (binding) sites have already closed after accepting the application of the 1st primary? Any help will be appreciated. Thank you in advance. Carol Ann Bobrowitz Medical College of Wisconsin Department of Physiology 414-456-8179 cbobrowi@mcw.edu _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 From jenbug812 <@t> aol.com Mon Apr 24 21:12:08 2006 From: jenbug812 <@t> aol.com (jenbug812@aol.com) Date: Mon Apr 24 21:12:15 2006 Subject: [Histonet] LOOKINF FOR LUXOL FAST BLUE/PAS Staining Procedure Message-ID: <8C83624CA59E70A-1DAC-2676@mblk-r12.sysops.aol.com> Cannot locate an exact staining procedure for LFB/PASH. Is anyone currently using this stain and if so can you forward me your procedure. Thanks From katri <@t> cogeco.ca Mon Apr 24 22:02:37 2006 From: katri <@t> cogeco.ca (Katri Tuomala) Date: Mon Apr 24 22:02:34 2006 Subject: [Histonet] Hold on, there! References: Message-ID: <00b701c66814$b58e8a10$6a9a9618@Katri> I must admit, I had a chuckle too... Katri ----- Original Message ----- From: "Breeden, Sara" To: Sent: Monday, April 24, 2006 12:33 PM Subject: [Histonet] Hold on, there! First, the necessary disclaimers: "I do NOT mean to offend; nor do I wish to have my comments misinterpreted in any way, shape, or form; I understand that the English language is difficult to interpret/translate/etc.; this struck me as funny on a Monday; and I just can't help myself..." I just HAVE to ask about the message relating to the woman in Germany who wanted to bury something of her husband's with his remains...please - could it be "head", "heart" or is it "hard"??? I'm chuckling...please join me! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jkiernan <@t> uwo.ca Mon Apr 24 23:41:13 2006 From: jkiernan <@t> uwo.ca (John Kiernan) Date: Mon Apr 24 23:40:04 2006 Subject: [Histonet] Apology Message-ID: <444DA869.4020703@uwo.ca> You shouldn't feel the need to apologise for amusing all but 2 or 3 of us with a story. John Kiernan London, Canada From djamesnz <@t> orcon.net.nz Mon Apr 24 23:57:30 2006 From: djamesnz <@t> orcon.net.nz (Darren James) Date: Mon Apr 24 23:57:42 2006 Subject: [Histonet] Apology In-Reply-To: <444DA869.4020703@uwo.ca> Message-ID: <001301c66824$c250bba0$0301010a@DAZZA> Here here. Lighten up people :-) -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of John Kiernan Sent: Tuesday, 25 April 2006 4:41 p.m. To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Apology You shouldn't feel the need to apologise for amusing all but 2 or 3 of us with a story. John Kiernan London, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Apr 25 02:25:08 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Apr 25 02:24:31 2006 Subject: [Histonet] Apology Message-ID: Come off it I was the first! If I remember it was a crass remark concerning Sept 11th. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Tuesday, April 25, 2006 12:57 AM To: Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Apology Sally, welcome to the "flamed" club. As president of the Histonet Flamed Society, I welcome your new membership. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Breeden, Sara" To: Sent: Monday, April 24, 2006 3:37 PM Subject: [Histonet] Apology It's a Monday for apology. Apparently my posting about the organ being returned for burial offended a couple of individuals and for that, I'm sorry. I have apologized to the originator as well. My first "flaming" and I'm a little crisp around the edges; but I still like a good chuckle. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Marco.Prunotto <@t> medecine.unige.ch Tue Apr 25 03:33:43 2006 From: Marco.Prunotto <@t> medecine.unige.ch (Marco Prunotto) Date: Tue Apr 25 03:33:53 2006 Subject: [Histonet] pdgf-bb receptor antibodies and inhibitors In-Reply-To: References: Message-ID: <444DDEE7.3000808@medecine.unige.ch> I need help from you as I'm searching specific good antibodies for PDGF-bb receptor (in the phosphorilated and not-phosphorilated forms ) and specific inhibitors for PDGF-bb receptor. thanks a lot marco prunotto University of Geneva College of Medicine From Heather.A.Harper <@t> pcola.med.navy.mil Tue Apr 25 05:54:21 2006 From: Heather.A.Harper <@t> pcola.med.navy.mil (Heather.A.Harper@pcola.med.navy.mil) Date: Tue Apr 25 05:57:53 2006 Subject: [Histonet] Bone decalcification Message-ID: I didn't realize any of the drama about my questioning the proper procedure on decaling bone raised some drama. I work for the Navy hospital, and we do not receive bone that often. I have been here for 6 years and have had 7 pathologists and they have always sawed a section and placed it in decal. I figured that was the way it was done plus that is how I was taught at school. Obviously this is the wrong way of doing it and I want to thank those who told me how it is done properly. As far as everybody who made comments, no biggie. I work for the navy so when it comes to comments, I let it roll right off of me. I am here to do my job and learn and that is why I love this web site, because truthfully, no question is stupid. I have gotten a lot of answers to my questions on this website. Thank you again everybody for your responses. Heather A. Harper Naval Hospital of Florida From Terry.Marshall <@t> rothgen.nhs.uk Tue Apr 25 07:02:29 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Tue Apr 25 07:19:02 2006 Subject: [Histonet] Apology Message-ID: Quite right. It's almost impossible nowadays to say anything without some exquisitely sensitive individual protesting how upset they are. 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: John Kiernan [mailto:jkiernan@uwo.ca] Sent: 25 April 2006 05:41 To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Apology You shouldn't feel the need to apologise for amusing all but 2 or 3 of us with a story. John Kiernan London, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rjbuesa <@t> yahoo.com Tue Apr 25 07:37:20 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Tue Apr 25 07:37:27 2006 Subject: [Histonet] Bone decalcification In-Reply-To: Message-ID: <20060425123720.22465.qmail@web61214.mail.yahoo.com> Dear Heather: Remember this: there are no stupid question, the answers are the ones that can be stupid! Ren? J. Heather.A.Harper@pcola.med.navy.mil wrote: I didn't realize any of the drama about my questioning the proper procedure on decaling bone raised some drama. I work for the Navy hospital, and we do not receive bone that often. I have been here for 6 years and have had 7 pathologists and they have always sawed a section and placed it in decal. I figured that was the way it was done plus that is how I was taught at school. Obviously this is the wrong way of doing it and I want to thank those who told me how it is done properly. As far as everybody who made comments, no biggie. I work for the navy so when it comes to comments, I let it roll right off of me. I am here to do my job and learn and that is why I love this web site, because truthfully, no question is stupid. I have gotten a lot of answers to my questions on this website. Thank you again everybody for your responses. Heather A. Harper Naval Hospital of Florida _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From LGaliotto <@t> nch.org Tue Apr 25 08:33:50 2006 From: LGaliotto <@t> nch.org (Galiotto, Laura) Date: Tue Apr 25 08:33:57 2006 Subject: [Histonet] Giving Specimens to Patients Message-ID: <270614B321ACB44D8C1D91F4F921FDC3036CBCD9@NCH01EX02.nch.org> Look up the EPA and OSHA regulations for handling and disposal of "potentially infective medical waste". -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Alan Bishop Sent: Monday, April 24, 2006 4:26 PM To: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Giving Specimens to Patients Both in the UK and NZ you now have legalities in place for patients to request the return of ANY tissues after a visit to the hospital. I was majorly involved in this process in the UK for 3 years after a hospital kept a lot of specimens from children without permission. Due to this consent forms have now changed and the option for return is included, right down to blocks, slides and even trimmings from cutting the blocks if so desired. As Darren says it very similar here in NZ now as well as we have to fit in with the wishes of such a culturally diverse country. One thing though, specimens are never returned in formalin. Always well washed and air dried if the specimen allows for that. Obviously never get rid of all traces of formalin so in the UK we were giving out strict guidlines on what should or should not be done with a specimen and also included the Hazard sheet. Going to far? Probably Cheers Alan _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Tue Apr 25 09:59:03 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Tue Apr 25 09:59:16 2006 Subject: Question and more on Re: [Histonet] Bone decalcification In-Reply-To: References: Message-ID: <6.0.0.22.1.20060425082350.01b4b110@gemini.msu.montana.edu> Heather A question: Are the bones brought to the lab already immersed in formalin? If your pathologist is used to RECEIVING a bone already in formalin and if he or she (through blind luck rather than knowledge of bone fixation/decalcification/effects of acids on unfixed bone) may have had a partially fixed bone, although more on the exterior rather than interior of sample,and then sawed off a piece and plopped it into acid decalcifier. It is doubtful a huge bone would be fixed throughout the whole or large sample. Your eyes along can help you here. A indication of partially fixed or unfixed bone is the bone interior will appear reddish to pinkish color i.e. bloody - meaning the bone is not fixed, and the sawn piece should be immersed in NBF longer i.e. overnight. Fixed bone takes on the brownish gray look seen with formalin fixed tissues. Take into consideration the dense calcified bone matrix will slow down crosslinking of formalin. If the bone is mostly trabecular bone (mesh of open spaces filled with marrow), then fixation will proceed much faster since there is less bone in the sample. Opening up/bisecting larger bone is always advisable upon receipt, to allow fixative to access interior surfaces - and there are some good saws out there to help you out. MarMed, Cabelas's bench top band saw - both are inexpensive. Yes, you are correct, there are no stupid questions. Repeated questions are not stupid either and I love the diversity of repetitive correct answers on subjects that interest me. These show how people have learned their technics and graciously pass on that information. Gayle Callis HTL, HT, MT(ASCP) Research Histopathology Supervisor Veterinary Molecular Biology Montana State University Bozeman MT 59717 From Janet.Bonner <@t> FLHosp.org Tue Apr 25 10:21:03 2006 From: Janet.Bonner <@t> FLHosp.org (Bonner, Janet) Date: Tue Apr 25 10:22:09 2006 Subject: [Histonet] Apology References: <444DA869.4020703@uwo.ca> Message-ID: Probalbly more than 2 or 3... you forgot the silent majority ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of John Kiernan Sent: Tue 4/25/2006 12:41 AM To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Apology You shouldn't feel the need to apologise for amusing all but 2 or 3 of us with a story. John Kiernan London, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Tue Apr 25 10:22:41 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Tue Apr 25 10:23:28 2006 Subject: [Histonet] Apology References: Message-ID: I feel a Friday Flame-off coming on!!! @) ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kemlo Rogerson Sent: Tue 4/25/2006 3:25 AM To: 'Joe Nocito'; Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Apology Come off it I was the first! If I remember it was a crass remark concerning Sept 11th. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Joe Nocito [mailto:jnocito@satx.rr.com] Sent: Tuesday, April 25, 2006 12:57 AM To: Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Apology Sally, welcome to the "flamed" club. As president of the Histonet Flamed Society, I welcome your new membership. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: "Breeden, Sara" To: Sent: Monday, April 24, 2006 3:37 PM Subject: [Histonet] Apology It's a Monday for apology. Apparently my posting about the organ being returned for burial offended a couple of individuals and for that, I'm sorry. I have apologized to the originator as well. My first "flaming" and I'm a little crisp around the edges; but I still like a good chuckle. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet -- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From kemlo.rogerson <@t> waht.swest.nhs.uk Tue Apr 25 10:40:12 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Tue Apr 25 10:39:41 2006 Subject: [Histonet] Apology Message-ID: Yea just done it! Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Tuesday, April 25, 2006 1:02 PM To: John Kiernan; Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Apology Quite right. It's almost impossible nowadays to say anything without some exquisitely sensitive individual protesting how upset they are. 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: John Kiernan [mailto:jkiernan@uwo.ca] Sent: 25 April 2006 05:41 To: Breeden, Sara Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] Apology You shouldn't feel the need to apologise for amusing all but 2 or 3 of us with a story. John Kiernan London, Canada _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From dsantana <@t> pmaonline.com Tue Apr 25 10:53:58 2006 From: dsantana <@t> pmaonline.com (Santana, Diane) Date: Tue Apr 25 10:57:18 2006 Subject: [Histonet] here is another one Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB0711398B@MAILPMA> ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. From JWEEMS <@t> sjha.org Tue Apr 25 11:00:35 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Tue Apr 25 11:00:40 2006 Subject: [Histonet] here is another one Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD3202637C1D@sjhaexc02.sjha.org> Would a 2000 ml cylinder be tall enough? I've used those in the past. j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Santana, Diane Sent: Tuesday, April 25, 2006 11:54 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] here is another one ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From funderwood <@t> mcohio.org Tue Apr 25 11:20:25 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Tue Apr 25 11:20:53 2006 Subject: [Histonet] here is another one Message-ID: There is a cylindrical vessel used by home brewers to hold a sample of beer so that a hydrometer reading can be taken. If you don't have a homebrew shop locally, there are several on line. Fred >>> "Santana, Diane" 04/25/06 11:53AM >>> ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From POWELL_SA <@t> Mercer.edu Tue Apr 25 11:25:28 2006 From: POWELL_SA <@t> Mercer.edu (Shirley Powell) Date: Tue Apr 25 11:25:59 2006 Subject: [Histonet] here is another one In-Reply-To: <1CD6831EB9B26D45B0A3EAA79F7EBD3202637C1D@sjhaexc02.sjha.org> Message-ID: <01M1OAX42XA28X32PH@Macon2.Mercer.edu> You could go to Wal-Mart and get a plastic container to fit your needs. sp -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Tuesday, April 25, 2006 11:01 AM To: Santana, Diane; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] here is another one Would a 2000 ml cylinder be tall enough? I've used those in the past. j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Santana, Diane Sent: Tuesday, April 25, 2006 11:54 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] here is another one ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From jkiernan <@t> uwo.ca Tue Apr 25 12:15:26 2006 From: jkiernan <@t> uwo.ca (John A. Kiernan) Date: Tue Apr 25 12:15:23 2006 Subject: [Histonet] LOOKINF FOR LUXOL FAST BLUE/PAS Staining Procedure References: <8C83624CA59E70A-1DAC-2676@mblk-r12.sysops.aol.com> Message-ID: <444E592E.BFB9759E@uwo.ca> Nobody seems to have answered this. Assuming that LFB is luxol fast blue for myelin, the following is based on the procedure of Kluver & Barrera (1953) J. Neuropath. Exp. Neurol. 12:400-403. There are other variants; this is the only one I've used myself. Solutions required: Staining solution. Luxol fast blue MBS: 0.25 g 95% ethanol: 250 ml Many workers add glacial acetic acid (2.5 ml) but this does not appear to confer any advantage. The solution keeps for at least 5 years and may be used repeatedly. Differentiating solution (0.05% Li2CO3). Lithium carbonate (Li2CO3): 0.25 g Water: 500 ml This keeps indefinitely and can be used repeatedly, but the solution should be discarded when it is more than faintly blue from extracted dye. Procedure for formaldehyde-fixed tissue: 1. De-wax paraffin sections and take to 100% ethanol. Frozen sections should be dried onto slides (from water) and then equilibrated with 100% ethanol. 2. Stain in luxol fast blue solution in a screw-capped staining jar at 55-60C (in an oven or water bath) for 16-24 hours (overnight). 3. Rinse in 70% ethanol and then in water. 4. Immerse in the differentiating solution until grey and white matter can be distinguished. This commonly takes 2 to 30 seconds for thin paraffin sections, but thick frozen sections may require up to 30 minutes. Move to Step 5 early rather than late, to avoid extracting too much blue dye. 5. Transfer slides to 70% ethanol, two changes, each 1 minute. More dye leaves the sections at this stage. 6. Rinse in water. Check with a hand-lens or low power microscope. Grey matter should be largely unstained; white matter should be a greenish shade of blue. Carefully repeat Steps 4 and 5 if there is still blue colour in neuronal cell bodies in grey matter. 7. Counterstain with an aqueous solution of a contrasting cationic dye at pH 3.5 to 4. (0.5% neutral red, CI 50040, is good) to show Nissl substance of neurons and nuclei of all cells. 8. Wash in water and blot dry. 9. If necessary, differentiate the counterstain in 70% ethanol until only the nuclei and Nissl substance are stained. There should be no "background" colour in the neuropil between neuronal cell bodies in grey matter. The colour of the stained myelin is deepened by counterstaining. 10. Blot dry and dehydrate in three changes of 100% ethanol (each 30 seconds with agitation) or n-butanol (each 3-5 minutes, without agitation). (Use n-butanol if you find that ethanol extracts too much of the counterstain.) 11. Clear in xylene and cover, using a resinous medium. Result: Myelin blue; nuclei and Nissl substance the colour of the counterstain. Comments: 1. Some batches of luxol fast blue MBS do not work very well. For other techniques, see Cook (1974). Churukian (1997) describes a variant in which the staining is accelerated by heating in a microwave oven. Luxol fast blue ARN can be used for myelin staining, with minor variations in technique. 2. Neutral red is an excellent counterstain for luxol fast blue. Another counterstain that is sometimes used is the PAS procedure which provides a pink background in the neuropil, without nuclear or other cellular staining. This seems a rather pointless thing to do. 3. For the mechanism of staining, see R.A.Clasen et al. (1973) J. Neuropath. Exp. Neurol. 32: 271-283. 4. I don't know why this method is still used. Myelin staining with a ferric salt and eriochrome cyanine R (= chromoxane cyanine R, = solochrome cyanine R, = CI Mordant blue 3, CI 43820) has been around since 1965, uses a much less expensive dye whose chemical identity is known, takes only 30 minutes, and gives a stronger blue colour to myelin than luxol fast blue. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ jenbug812@aol.com wrote: > > Cannot locate an exact staining procedure for LFB/PASH. Is anyone currently using this stain and if so can you forward me your procedure. > Thanks > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Janet.Bonner <@t> FLHOSP.ORG Tue Apr 25 13:05:55 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Tue Apr 25 13:08:56 2006 Subject: [Histonet] LOOKINF FOR LUXOL FAST BLUE/PAS Staining Procedure References: <444E592E.BFB9759E@uwo.ca> Message-ID: We do the LFB/PAS by doing the LFB part of the stain and counterstaining with PAS. I'm not sure about the digestion as this could radically change the proceding reactions. from: histonet-bounces@lists.utsouthwestern.edu on behalf of John A. Kiernan Sent: Tue 4/25/2006 1:15 PM To: jenbug812@aol.com Cc: histonet@lists.utsouthwestern.edu Subject: Re: [Histonet] LOOKINF FOR LUXOL FAST BLUE/PAS Staining Procedure Nobody seems to have answered this. Assuming that LFB is luxol fast blue for myelin, the following is based on the procedure of Kluver & Barrera (1953) J. Neuropath. Exp. Neurol. 12:400-403. There are other variants; this is the only one I've used myself. Solutions required: Staining solution. Luxol fast blue MBS: 0.25 g 95% ethanol: 250 ml Many workers add glacial acetic acid (2.5 ml) but this does not appear to confer any advantage. The solution keeps for at least 5 years and may be used repeatedly. Differentiating solution (0.05% Li2CO3). Lithium carbonate (Li2CO3): 0.25 g Water: 500 ml This keeps indefinitely and can be used repeatedly, but the solution should be discarded when it is more than faintly blue from extracted dye. Procedure for formaldehyde-fixed tissue: 1. De-wax paraffin sections and take to 100% ethanol. Frozen sections should be dried onto slides (from water) and then equilibrated with 100% ethanol. 2. Stain in luxol fast blue solution in a screw-capped staining jar at 55-60C (in an oven or water bath) for 16-24 hours (overnight). 3. Rinse in 70% ethanol and then in water. 4. Immerse in the differentiating solution until grey and white matter can be distinguished. This commonly takes 2 to 30 seconds for thin paraffin sections, but thick frozen sections may require up to 30 minutes. Move to Step 5 early rather than late, to avoid extracting too much blue dye. 5. Transfer slides to 70% ethanol, two changes, each 1 minute. More dye leaves the sections at this stage. 6. Rinse in water. Check with a hand-lens or low power microscope. Grey matter should be largely unstained; white matter should be a greenish shade of blue. Carefully repeat Steps 4 and 5 if there is still blue colour in neuronal cell bodies in grey matter. 7. Counterstain with an aqueous solution of a contrasting cationic dye at pH 3.5 to 4. (0.5% neutral red, CI 50040, is good) to show Nissl substance of neurons and nuclei of all cells. 8. Wash in water and blot dry. 9. If necessary, differentiate the counterstain in 70% ethanol until only the nuclei and Nissl substance are stained. There should be no "background" colour in the neuropil between neuronal cell bodies in grey matter. The colour of the stained myelin is deepened by counterstaining. 10. Blot dry and dehydrate in three changes of 100% ethanol (each 30 seconds with agitation) or n-butanol (each 3-5 minutes, without agitation). (Use n-butanol if you find that ethanol extracts too much of the counterstain.) 11. Clear in xylene and cover, using a resinous medium. Result: Myelin blue; nuclei and Nissl substance the colour of the counterstain. Comments: 1. Some batches of luxol fast blue MBS do not work very well. For other techniques, see Cook (1974). Churukian (1997) describes a variant in which the staining is accelerated by heating in a microwave oven. Luxol fast blue ARN can be used for myelin staining, with minor variations in technique. 2. Neutral red is an excellent counterstain for luxol fast blue. Another counterstain that is sometimes used is the PAS procedure which provides a pink background in the neuropil, without nuclear or other cellular staining. This seems a rather pointless thing to do. 3. For the mechanism of staining, see R.A.Clasen et al. (1973) J. Neuropath. Exp. Neurol. 32: 271-283. 4. I don't know why this method is still used. Myelin staining with a ferric salt and eriochrome cyanine R (= chromoxane cyanine R, = solochrome cyanine R, = CI Mordant blue 3, CI 43820) has been around since 1965, uses a much less expensive dye whose chemical identity is known, takes only 30 minutes, and gives a stronger blue colour to myelin than luxol fast blue. -- ------------------------------- John A. Kiernan Department of Anatomy and Cell Biology The University of Western Ontario London, Canada N6A 5C1 kiernan[AT]uwo.ca http://publish.uwo.ca/~jkiernan/ http://instruct.uwo.ca/anatomy/530/index.htm _______________________________ jenbug812@aol.com wrote: > > Cannot locate an exact staining procedure for LFB/PASH. Is anyone currently using this stain and if so can you forward me your procedure. > Thanks > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------- The information contained in this message may be privileged and / or confidential and protected from disclosure. If the reader of this message is not the intended recipient, 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 Michele_Marggi <@t> ssmhc.com Tue Apr 25 13:57:56 2006 From: Michele_Marggi <@t> ssmhc.com (Michele_Marggi@ssmhc.com) Date: Tue Apr 25 13:58:32 2006 Subject: [Histonet] Pneumatic tube Message-ID: Hi everyone, I am wondering who is using a pneumatic tube for sending pathology specimens into the lab. We have a tube system set up throughout the hospital and lab, however we do not use it for the pathology part of the lab due to the risk of formalin spilling and contamination of the tube. Any information, procedures or policies that you have would really be appreciated. Thank you, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. From skouko <@t> po-box.mcgill.ca Tue Apr 25 14:13:38 2006 From: skouko <@t> po-box.mcgill.ca (skouko@po-box.mcgill.ca) Date: Tue Apr 25 14:13:51 2006 Subject: [Histonet] mGluR expression in monkey cerebellum Message-ID: <20060425151338.ocqumw72qs8kg8s0@webmail.mcgill.ca> Dear Histonetters, does anybody happen to know the type of metabotropic glutamate receptor that's most prevalent in the monkey cerebellum? I've ben trying to find that info on pubmed and in neuro textbooks, but can't seem to find the answer. Any help would be much appreciated, Sophia Koukoui, McGill Behavioral Neuroscience. From Janet.Bonner <@t> flhosp.org Tue Apr 25 14:13:21 2006 From: Janet.Bonner <@t> flhosp.org (Bonner, Janet) Date: Tue Apr 25 14:18:24 2006 Subject: [Histonet] Pneumatic tube References: Message-ID: We do not use it at all due to the reasons you listed, as well as the fear of it getting "lost" to another location - ( we're a really big facility ). ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Michele_Marggi@ssmhc.com Sent: Tue 4/25/2006 2:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pneumatic tube Hi everyone, I am wondering who is using a pneumatic tube for sending pathology specimens into the lab. We have a tube system set up throughout the hospital and lab, however we do not use it for the pathology part of the lab due to the risk of formalin spilling and contamination of the tube. Any information, procedures or policies that you have would really be appreciated. Thank you, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From stamptrain <@t> yahoo.com Tue Apr 25 14:25:09 2006 From: stamptrain <@t> yahoo.com (Roger Moretz) Date: Tue Apr 25 14:25:15 2006 Subject: [Histonet] Bouin's vs Davidsons for Testes Message-ID: <20060425192509.88997.qmail@web50305.mail.yahoo.com> We routinely use Bouin's for fixing testes for improved cytoplasmic visualization and evaluation of spermatogenesis. One of the pathologists has asked whether one would get similar results using Davidson's fixative. I did a search of the HistoArchives but didn't come up with results that answered the question (doesn't mean it isn't there and I just plain missed it--just means I didn't find it!). Unfortunately there is no time in which to do the research/evaluation in-house so I am turning to the collective wisdom and knowledge of Histonetters for the answer. Roger Moretz, Ph.D. Dept of Toxicology Boehringer Ingelheim Pharmaceuticals Ridgefield CT __________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com From HornHV <@t> archildrens.org Tue Apr 25 14:25:53 2006 From: HornHV <@t> archildrens.org (Horn, Hazel V) Date: Tue Apr 25 14:26:24 2006 Subject: [Histonet] Pneumatic tube Message-ID: <9AE8AA9E1F644B4AA6C155FB6FD51C63038BE058@EMAIL.archildrens.org> We use the tube for only fresh specimens. No fixed specimens can be tubed. 99% of our specimens are hand carried to our lab. Hazel Horn -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Bonner, Janet Sent: Tuesday, April 25, 2006 2:13 PM To: Michele_Marggi@ssmhc.com; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Pneumatic tube We do not use it at all due to the reasons you listed, as well as the fear of it getting "lost" to another location - ( we're a really big facility ). ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Michele_Marggi@ssmhc.com Sent: Tue 4/25/2006 2:57 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Pneumatic tube Hi everyone, I am wondering who is using a pneumatic tube for sending pathology specimens into the lab. We have a tube system set up throughout the hospital and lab, however we do not use it for the pathology part of the lab due to the risk of formalin spilling and contamination of the tube. Any information, procedures or policies that you have would really be appreciated. Thank you, Michele Marggi Surgical Pathology Supervisor St. Marys Hospital Medical Center 707 S Mills Street Madison WI 53715 Telephone: 608.258.6930 Fax: 608.258.6268 ----------------------------------------- Confidentiality Notice: This email message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original 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 ------------------------------------------------------------------------------ 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 Janet.Bonner <@t> FLHOSP.ORG Tue Apr 25 14:27:03 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Tue Apr 25 14:30:29 2006 Subject: [Histonet] here is another one References: <4C96AA62BADFD81180CB0002A5AD67FB0711398B@MAILPMA> Message-ID: Can you just pour some out of the container into a 250ml graduated cylinder and then use the hydrometer? How big is your hydrometer (I may be missing something here). What do you mean the containers aren't deep enough? ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Santana, Diane Sent: Tue 4/25/2006 11:53 AM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] here is another one ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gu.lang <@t> gmx.at Tue Apr 25 14:41:01 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Tue Apr 25 14:41:09 2006 Subject: [Histonet] Cd34 in trephine bone marrow Message-ID: <001301c668a0$2fcce110$eeeea8c0@SERVER01> Hi, Can anyone share a protocol for CD34 on decalcified FFPE human bone marrow biopsies? We have a benchmark XT and decalcify with formic acid. Are there any hints especially refering to CD34 in acid-decalcified tissue? Thank you Gudrun Lang From failm <@t> musc.edu Tue Apr 25 14:58:11 2006 From: failm <@t> musc.edu (Mildred Fail) Date: Tue Apr 25 15:37:52 2006 Subject: [Histonet] Cd34 in trephine bone marrow Message-ID: We use formic We use CD34 purchased from Becton-Dickinson. 1:200, citrate buffer for 20 minutes in the steamer. We also use formic acid to decalcify bone marrows. Rena Fail >>> "Gudrun Lang" 04/25/06 03:41PM >>> Hi, Can anyone share a protocol for CD34 on decalcified FFPE human bone marrow biopsies? We have a benchmark XT and decalcify with formic acid. Are there any hints especially refering to CD34 in acid-decalcified tissue? Thank you Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From failm <@t> musc.edu Tue Apr 25 15:01:57 2006 From: failm <@t> musc.edu (Mildred Fail) Date: Tue Apr 25 15:38:00 2006 Subject: [Histonet] Cd34 in trephine bone marrow Message-ID: We also use formic acid to decalcify our bone marrows. We do IHC on 2-3 bm cases a day with good results. Our CD34 is purchased from Becton-dickinson and used at a 1:200 dilution. Hier using citrate buffer for 20 minutes in a steamer, cool down for 10 minutes. Rena Fail >>> "Gudrun Lang" 04/25/06 03:41PM >>> Hi, Can anyone share a protocol for CD34 on decalcified FFPE human bone marrow biopsies? We have a benchmark XT and decalcify with formic acid. Are there any hints especially refering to CD34 in acid-decalcified tissue? Thank you Gudrun Lang _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From BSylinda <@t> aol.com Tue Apr 25 18:02:47 2006 From: BSylinda <@t> aol.com (BSylinda@aol.com) Date: Tue Apr 25 18:03:00 2006 Subject: [Histonet] Immuno Stainers Message-ID: <378.1da89e5.31800497@aol.com> Hello Histoland, Can any of you give me any personal experience with the Biogenix 6000 immunostainer. We have this instrument and certain places the slides are placed we are getting uneven staining. Have tried pap pen, barrier slides, and nothing seems to be working. Anyone with knowledge of this instrument would you please email me personally. Thanks in advance!! Sylinda Battle, HT (ASCP) From marjoh3 <@t> telus.net Tue Apr 25 20:21:10 2006 From: marjoh3 <@t> telus.net (Marilyn Johnson) Date: Tue Apr 25 20:21:12 2006 Subject: [Histonet] Duffy's Stain for Eosinophils/HOPS Stain Message-ID: <001601c668cf$b3a56dc0$6401a8c0@VALUED20606295> Hi Histonetters, Does anyone know if Duffy's stain for eosinophils or HOPS stain are commercially available and if so, how stable are these solutions and how are the results compared to the home-made stains? Please include company names and order numbers. Any replies would be greatly appreciated. Thank you. Marilyn Johnson Edmonton, Alberta, Canada. From eric <@t> ategra.com Tue Apr 25 21:09:38 2006 From: eric <@t> ategra.com (Eric Dye) Date: Tue Apr 25 20:57:38 2006 Subject: [Histonet] Histology Managers, Supervisors, and Bench Techs needed-Hiring now Message-ID: <20060426015729.TOIT20587.rrcs-fep-12.hrndva.rr.com@Pamsgx110> I'm presently on a search for my best client companies seeking HistoTech Supervisors and Histo Bench Techs. These are mostly fulltime permanent positions. Most of my Histo Techs positions are clinical, although I do have a few Temporary and Research Histo Tech positions as well. Most Histo jobs are dayshift (Except where indicated below) Here are some of my Hottest Histology Jobs: 1. Ohio (Southwestern Ohio) (Full-time, Perm, Histo Manager) 2. Northern New Jersey (Full-time, Perm and Temp (2 openings), Bench Histo Tech, 1st shift) 3. Rhode Island - Providence (Full-time, Perm, Bench Histo Tech, 2nd Shift) 4. Northern New Jersey (Full-time, Perm, Bench Histo Tech) 5. Ohio (Southwestern Ohio) (Full-time, Perm and Temp (2 openings), Bench Histo Tech, 1st shift) 6. New York (Long Island) (Full-time, Perm, SUPERVISOR Histo Tech) 7. Virginia(South - Coastal) (Full-time, Perm, Bench Histo Tech) 8. Virginia - Metro DC (Full-time, Perm, Lead Histo Tech) 9. South Florida (Full-time, Perm, Supervisor Histo Tech) 10. South Florida (Full-time, Perm, Bench Histo Tech) 11. Mass (Boston) (Part-time, Perm, Bench Histo Tech) 12. West Virginia (Full-time, Perm, Bench Histo Tech) 13. Texas (Austin area) (Full-time, Perm, Supervisor and Bench Histo Tech) 14. Georgia (Atlanta area) (Full-time, Perm, Bench Histotech) 15. New Hampshire (Mohs and Routine Bench Histotech,Perm and Temp) 16. Florida. (South East Coast, Perm, Bench Histotech) 17. Florida. (Central East Coast, Temp, Bench Histo Tech) 18. Columbus (Central) (Temp, Bench Histotech) The clients are currently interviewing Histology candidates - so if you are interested, please call me today at 1-800-466-9919 ext 223 Thank you, Eric Dye-Sr Allied Healthcare Recruiter 1-800-466-9919 ext 223 --------------------------------------------------------------- Ategra Systems Inc Specialists in Permanent & Contract Staffing 800 466 9919 ext 223 - voice 407 671 6075 - fax eric@ategra.com To Learn More About Ategra: http://www.ategra.com ---------------------------------------------------------------- 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 edgg07 <@t> hotmail.com Tue Apr 25 22:43:07 2006 From: edgg07 <@t> hotmail.com (bill walker) Date: Tue Apr 25 22:43:14 2006 Subject: [Histonet] DAKO Message-ID: I spoke with a colleague on the East Coast and she informed me that DAKO has another recall for Envision. Has anyone else heard? Thanks Bill _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ From vanann702 <@t> skmc.gov.ae Wed Apr 26 01:31:52 2006 From: vanann702 <@t> skmc.gov.ae (Anne Van Binsbergen) Date: Wed Apr 26 01:30:49 2006 Subject: [Histonet] bcl-6 Message-ID: Hi Histonetters This was a request from a Pathologist 'regarding bcl 6 which as you know is a notoriously weak / difficult marker: Cellmarque (www.cellmarque.com; also www.bcl-6.com) is promoting a new clone (GI191E/A8) which - according to the company - yields better staining' Any comments / advice / experiences to share? Annieinarabia Anne S van Binsbegen Anatomical Pathology Laboratory Shaikh Khalifa Medical City PO BOX 51900 Abu Dhabu, U.A.E. ph: +971 2 6102695 mob:+971 50 3162804 fax +971 2 6104989 dance like no one is watching Sheikh Khalifa Medical City (SKMC) Disclaimer Notice: This mail message (including any attachments) is intended only for the designated recipient only and may contain privileged, proprietary, or otherwise private information. If you are not the addressee you must not copy, distribute, disclose or use any of the information in it. If you have received it in error, please notify the sender immediately and delete the original. Real women don't have hot flushes, they have power surges From ian.montgomery <@t> bio.gla.ac.uk Wed Apr 26 04:03:32 2006 From: ian.montgomery <@t> bio.gla.ac.uk (Ian Montgomery) Date: Wed Apr 26 04:03:42 2006 Subject: Fw: [Histonet] Apology Message-ID: <004301c66910$4bd23b60$532ed182@ibls.gla.ac.uk> Interesting use of language, US you are "flamed" while on this green and beautiful island you are "roasted." The position on the body where the "roasting" takes place determines the degree. For example, on the ears is a light "roasting," while on the bum is considered major. Over the years I've been "roasted" many times, first by my mother, then my wife, now even my daughter practices on me. At work, well, the skilled technician knows how to massage errors and avoid the necessary pain. Ian. Dr. Ian Montgomery, Histotechnology, IBLS Support Unit, Graham Kerr Building, Tel: 4652, 6644. ian.montgomery@bio.gla.ac.uk ----- Original Message ----- From: "Joe Nocito" To: "Breeden, Sara" ; Sent: Tuesday, April 25, 2006 12:57 AM Subject: Re: [Histonet] Apology > Sally, > welcome to the "flamed" club. As president of the Histonet Flamed Society, I > welcome your new membership. > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > ----- Original Message ----- > From: "Breeden, Sara" > To: > Sent: Monday, April 24, 2006 3:37 PM > Subject: [Histonet] Apology > > > It's a Monday for apology. Apparently my posting about the organ being > returned for burial offended a couple of individuals and for that, I'm > sorry. I have apologized to the originator as well. My first "flaming" > and I'm a little crisp around the edges; but I still like a good > chuckle. > > Sally Breeden, HT(ASCP) > New Mexico Department of Agriculture > Veterinary Diagnostic Services > POBox 4700 > Albuquerque, NM 87196 > (505)841-2576 > (505)841-2518 FAX > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > No virus found in this incoming message. > Checked by AVG Free Edition. > Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From mverdu <@t> histopat.es Wed Apr 26 05:11:47 2006 From: mverdu <@t> histopat.es (Montse verdu) Date: Wed Apr 26 05:12:12 2006 Subject: [Histonet] p27 clone DSC72 -human prostate Message-ID: <000201c66919$d9a3fe20$3800a8c0@Histopat.com> Dear histonetters, Anybody has experience with p27 clone DSC72 different than Calbiochem antibody? We use these antibody on formalin-fixed, paraffin-embedded human prostate carcinoma stained, using avidin-biotin peroxidase complex and DAB chromogen, and we are satisfied with the results, but we have problems with the distributor of these product (frequently we received products with an expiration date from before at the delivery date) and for these reason we needs to change of antibody. Thanks for your help. Montse Verd? histopat LABORATORIS Tel. 932033000 Fax 932802160 mverdu@histopat.es From cormier <@t> MIT.EDU Wed Apr 26 05:32:28 2006 From: cormier <@t> MIT.EDU (Kathy Cormier) Date: Wed Apr 26 05:32:42 2006 Subject: [Histonet] Immuno Stainers References: <378.1da89e5.31800497@aol.com> Message-ID: <001e01c6691c$b8114c60$92003712@mit.edu> Hi Sylinda, We have the i6000 as well. We have had similiar problems. We use it on large tissue sections, such as whole mouse colons. We do not use the optimax buffer as it was a bit too pricey. We have found that using PBS w/ Tween 20 as a diluent helps to mitigate the uneven staining. The tween 20 helps to "spread" the reagent evenly. We did check to ensure that the machine was completely level and that the black removable racks inside the machine were correctly "seated" inside the machine. Also ensure that your antigen retrival methods are effective, as that can cause uneven staining too. Hope that this helps! Kathy Cormier DCM MIT ----- Original Message ----- From: To: Sent: Tuesday, April 25, 2006 7:02 PM Subject: [Histonet] Immuno Stainers > Hello Histoland, > > Can any of you give me any personal experience with the Biogenix 6000 > immunostainer. We have this instrument and certain places the slides are > placed we > are getting uneven staining. Have tried pap pen, barrier slides, and > nothing seems to be working. Anyone with knowledge of this instrument > would you > please email me personally. Thanks in advance!! > > Sylinda Battle, HT (ASCP) > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From Arfa.R.Maqsood <@t> manchester.ac.uk Wed Apr 26 07:14:19 2006 From: Arfa.R.Maqsood <@t> manchester.ac.uk (Arfa R Maqsood) Date: Wed Apr 26 07:14:55 2006 Subject: [Histonet] fixing rat tibia Message-ID: Hi, Could anyone advise me regarding the time required to fix a rat tibia. Can it be fixed whole or is it best to slice in half longitudinally. Also is 70% ethanol appropriate as a fixative for bones that have been labelled with Oxytetracycline. Another dilema I have is that I dont have access to a specialist tools, should I need to bisect the tibia in half. What tools could be used to cut the tibia in half longitudinally without cracking the bone. I am new to all this so any any help / suggestions will be appreciated. Thanks Ash ARFA.R.MAQSOOD ENDOCRINE SCIENCE RESEARCH GROUP UNIVERSITY OF MANCHESTER OXFORD ROAD MANCHESTER M13 9PT TEL:+44 (0)161 275 5177 FAX:+44 (0)161 275 5958 From la.sebree <@t> hosp.wisc.edu Wed Apr 26 07:56:46 2006 From: la.sebree <@t> hosp.wisc.edu (Sebree Linda A.) Date: Wed Apr 26 07:56:56 2006 Subject: [Histonet] bcl-6 Message-ID: Anne, We use this antibody on our Benchmark and XT immunostainers with standard CC1 (EDTA) HIER and it works well. We found it too weak to use on our NexES with off-line retrieval. Feel free to contact us with any questions you might have. Linda Sebree, HT(ASCP) University of Wisconsin Hospital & Clinics IHC/ISH Laboratory A4/204-3224 600 Highland Ave. Madison, WI 53792 (608)265-6596 FAX: (608)262-7174 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Anne Van Binsbergen Sent: Wednesday, April 26, 2006 1:32 AM To: histonet@lists.utsouthwestern.edu; histonet-request@lists.utsouthwestern.edu Cc: Ridhwaan Arries Subject: [Histonet] bcl-6 Hi Histonetters This was a request from a Pathologist 'regarding bcl 6 which as you know is a notoriously weak / difficult marker: Cellmarque (www.cellmarque.com; also www.bcl-6.com) is promoting a new clone (GI191E/A8) which - according to the company - yields better staining' Any comments / advice / experiences to share? Annieinarabia Anne S van Binsbegen Anatomical Pathology Laboratory Shaikh Khalifa Medical City PO BOX 51900 Abu Dhabu, U.A.E. ph: +971 2 6102695 mob:+971 50 3162804 fax +971 2 6104989 dance like no one is watching Sheikh Khalifa Medical City (SKMC) Disclaimer Notice: This mail message (including any attachments) is intended only for the designated recipient only and may contain privileged, proprietary, or otherwise private information. If you are not the addressee you must not copy, distribute, disclose or use any of the information in it. If you have received it in error, please notify the sender immediately and delete the original. Real women don't have hot flushes, they have power surges _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Terry.Marshall <@t> rothgen.nhs.uk Wed Apr 26 07:26:48 2006 From: Terry.Marshall <@t> rothgen.nhs.uk (Marshall Terry Dr, Consultant Histopathologist) Date: Wed Apr 26 08:09:46 2006 Subject: [Histonet] Apology Message-ID: " Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer." That comment assumes that all receive e-mails in a logical, time ordered manner. This, because of the multitude of hops messages take through various servers to get to the final destination, is simply incorrect. Therefore, one may think that one is the first to reply because no other reply is, as yet, seen. Indeed, I frequently see replies before I see the question. 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: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: 24 April 2006 15:34 To: Deltour, Douglas D. (HM2); 'Charles.Embrey'; Harper, Heather A., CIV Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I seldom intervene in "thread discussions" but this time I want to say that I agree with Douglas (although my agreement has no relevance in itself). Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer. It is my opinion in this subject (that also in itself bears no real weight) that once a question has been answred correctly we all shoul refrain from keep sending the same correct answer, and even more, refrain from making sarcastic or even sometimes insulting comments. And now you all have the opportunity of contradicting what I have just wrote (that also in itself would be absolutely irrelevant!). Ren? J. "Deltour, Douglas D. (HM2)" wrote: "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From STapper <@t> slhduluth.com Wed Apr 26 08:09:51 2006 From: STapper <@t> slhduluth.com (Tapper, Sheila) Date: Wed Apr 26 08:09:58 2006 Subject: [Histonet] CD38 antibody Message-ID: I am looking for a CD38 antibody that is IVD. What are people using? My source for IVD antibody has discontinued the antibody. Any help would be appreciated. Thanks!! Sheila From kemlo.rogerson <@t> waht.swest.nhs.uk Wed Apr 26 08:16:21 2006 From: kemlo.rogerson <@t> waht.swest.nhs.uk (Kemlo Rogerson) Date: Wed Apr 26 08:15:48 2006 Subject: [Histonet] Apology Message-ID: Aren't you watering that stuff down again Terry? Are the replies in pink by any chance? Oops better say something meaningful before I get flamed. I agree.... totally. PS sometimes you see the question answer it, scroll down and there is what you just said, but in American. Question? Does it matter? Geez. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Wednesday, April 26, 2006 1:27 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Apology " Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer." That comment assumes that all receive e-mails in a logical, time ordered manner. This, because of the multitude of hops messages take through various servers to get to the final destination, is simply incorrect. Therefore, one may think that one is the first to reply because no other reply is, as yet, seen. Indeed, I frequently see replies before I see the question. 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: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: 24 April 2006 15:34 To: Deltour, Douglas D. (HM2); 'Charles.Embrey'; Harper, Heather A., CIV Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I seldom intervene in "thread discussions" but this time I want to say that I agree with Douglas (although my agreement has no relevance in itself). Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer. It is my opinion in this subject (that also in itself bears no real weight) that once a question has been answred correctly we all shoul refrain from keep sending the same correct answer, and even more, refrain from making sarcastic or even sometimes insulting comments. And now you all have the opportunity of contradicting what I have just wrote (that also in itself would be absolutely irrelevant!). Ren? J. "Deltour, Douglas D. (HM2)" wrote: "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From Rcartun <@t> harthosp.org Wed Apr 26 09:06:28 2006 From: Rcartun <@t> harthosp.org (Richard Cartun) Date: Wed Apr 26 09:07:03 2006 Subject: [Histonet] DAKO In-Reply-To: References: Message-ID: <444F462402000009000AD7F5@hcnwgwds01.hh.chs> Yes, I just saw the "Urgent IVD Recall" letter yesterday. The recall is for two lots (065022 & 085032) of EnVision+/HRP anti-mouse. Richard Richard W. Cartun, Ph.D. Director, Immunopathology & Histology Assistant Director, Anatomic Pathology Hartford Hospital 80 Seymour Street Hartford, CT 06102 (860) 545-1596 (860) 545-0174 Fax >>> "bill walker" 04/25/06 11:43 PM >>> I spoke with a colleague on the East Coast and she informed me that DAKO has another recall for Envision. Has anyone else heard? Thanks Bill _________________________________________________________________ Express yourself instantly with MSN Messenger! Download today - it's FREE! http://messenger.msn.click-url.com/go/onm00200471ave/direct/01/ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From rachael_emerson <@t> urmc.rochester.edu Wed Apr 26 09:34:12 2006 From: rachael_emerson <@t> urmc.rochester.edu (Rachael Emerson) Date: Wed Apr 26 09:34:37 2006 Subject: [Histonet] ICAM-1 Message-ID: Hello. Has anyone successfully used ICAM-1 antibody, either rat anti-mouse from eBioscience (#14-0541) or goat anti-mouse from Santa Cruz (#sc-1511)? I have tried using both on frozen sections and on paraffin sections, but the staining is inconsistent. Thanks! Rachael -- Rachael L. Emerson Center for Pediatric Biomedical Research University of Rochester Medical Center 575 Elmwood Avenue MRBX 1.11301 Rochester, NY 14642 From Janet.Bonner <@t> FLHosp.org Wed Apr 26 09:35:35 2006 From: Janet.Bonner <@t> FLHosp.org (Bonner, Janet) Date: Wed Apr 26 09:36:19 2006 Subject: [Histonet] Apology References: Message-ID: Thank you....I was starting to get confused! ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Kemlo Rogerson Sent: Wed 4/26/2006 9:16 AM To: 'Marshall Terry Dr, Consultant Histopathologist'; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Apology Aren't you watering that stuff down again Terry? Are the replies in pink by any chance? Oops better say something meaningful before I get flamed. I agree.... totally. PS sometimes you see the question answer it, scroll down and there is what you just said, but in American. Question? Does it matter? Geez. Kemlo Rogerson Pathology Manager Ext 3311 DD 01934 647057 Mob 07749 754194 Infinite patience yields immediate results. -- Marianne Williamson This e-mail is confidential and privileged. If you are not the intended recipient please accept my apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform me that this message has gone astray before deleting it. Thank you for your co-operation -----Original Message----- From: Marshall Terry Dr, Consultant Histopathologist [mailto:Terry.Marshall@rothgen.nhs.uk] Sent: Wednesday, April 26, 2006 1:27 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Apology " Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer." That comment assumes that all receive e-mails in a logical, time ordered manner. This, because of the multitude of hops messages take through various servers to get to the final destination, is simply incorrect. Therefore, one may think that one is the first to reply because no other reply is, as yet, seen. Indeed, I frequently see replies before I see the question. 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: Rene J Buesa [mailto:rjbuesa@yahoo.com] Sent: 24 April 2006 15:34 To: Deltour, Douglas D. (HM2); 'Charles.Embrey'; Harper, Heather A., CIV Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I seldom intervene in "thread discussions" but this time I want to say that I agree with Douglas (although my agreement has no relevance in itself). Time and again I see that after a question it follows a correct answer and, in spite of that, people keep repeating the same thing, like if they don't want to waste the opportunity of appearing knowledgeable by repeating a correct answer. It is my opinion in this subject (that also in itself bears no real weight) that once a question has been answred correctly we all shoul refrain from keep sending the same correct answer, and even more, refrain from making sarcastic or even sometimes insulting comments. And now you all have the opportunity of contradicting what I have just wrote (that also in itself would be absolutely irrelevant!). Ren? J. "Deltour, Douglas D. (HM2)" wrote: "I don't know what school you went to but I would want my money back." Comments like this may tend to keep people from asking questions. This is a resource for even the simplest questions. I consider this forum a learning tool and when people can not ask these questions because they are made to feel foolish by others then something is wrong. There are no stupid questions..... If you can't say anything nice..... Douglas Deltour HT(ASCP) -----Original Message----- From: Charles.Embrey [mailto:Charles.Embrey@carle.com] Sent: Monday, April 24, 2006 9:24 AM To: Heather.A.Harper@pcola.med.navy.mil Cc: Histonet@pathology.swmed.edu Subject: RE: [Histonet] Bone decalcification I don't know what school you went to but I would want my money back. The last thing you want is to put unfixed, fragile cells into acid and allow them to rot while the bone decals. What would be left to fix later? Chuck -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Heather.A.Harper@pcola.med.navy.mil Sent: Monday, April 24, 2006 5:40 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Bone decalcification I just wanted to know other histo techs opinions on bone decalcification. I learned in school that you decal the bone than you fix it. I have a pathologist who claims she learned it the hard way, and that it is better to fix the bone than decal it. What did you learn on how to do this procedure? This pathologist claims that fixing prior to decaling keeps the cells more intact. Any opinion on your procedure or your technique is appreciated. Heather A. Harper Naval Hospital Pensacola, FL _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@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 sbreeden <@t> nmda.nmsu.edu Wed Apr 26 10:08:00 2006 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Wed Apr 26 10:08:04 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: Looking for source for GRAM Stain control slides or a known positive tissue to make control block. Vendor sources seem to be quite pricey. Thanks in advance. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From rjbuesa <@t> yahoo.com Wed Apr 26 10:42:14 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Wed Apr 26 10:42:18 2006 Subject: [Histonet] GRAM Stain Control Slides In-Reply-To: Message-ID: <20060426154214.388.qmail@web61223.mail.yahoo.com> Sara: I used for years sections from cases of acute appendicities. They contain a host of both Gram + and - bacteria. I hope this will help you! Ren? J. "Breeden, Sara" wrote: Looking for source for GRAM Stain control slides or a known positive tissue to make control block. Vendor sources seem to be quite pricey. Thanks in advance. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1¢/min. From ROrr <@t> enh.org Wed Apr 26 11:22:56 2006 From: ROrr <@t> enh.org (Orr, Rebecca) Date: Wed Apr 26 11:23:00 2006 Subject: [Histonet] Hydrometer Message-ID: Heather, I have a small one that does fit in the container...it did come from Fisher, but I've had it awhile...there's no part number on it. I was also thinking that Wine makers would use them, try surfing a wine making site to see if they have a size that would fit into the container Becky Orr Evanston Hospital Message: 24 Date: Tue, 25 Apr 2006 11:53:58 -0400 From: "Santana, Diane" Subject: [Histonet] here is another one To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB0711398B@MAILPMA> Content-Type: text/plain; charset="iso-8859-1" ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. From JWEEMS <@t> sjha.org Wed Apr 26 11:34:39 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Apr 26 11:34:43 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD3202637C5D@sjhaexc02.sjha.org> A section of processed Slim Jim works well. J:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, Sara Sent: Wednesday, April 26, 2006 11:08 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GRAM Stain Control Slides Looking for source for GRAM Stain control slides or a known positive tissue to make control block. Vendor sources seem to be quite pricey. Thanks in advance. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 settembr <@t> umdnj.edu Wed Apr 26 11:51:40 2006 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Wed Apr 26 11:52:17 2006 Subject: [Histonet] p27 clone DSC72 -human prostate Message-ID: I do use p27, but not your clone. I use clone SX53G8 from Dako. They are a great company to deal with; very user friendly. This p27 is IVD cat.3 M7203; I use it at 1:80 with their Target Retrieval Solution and incubate for 15 min. room temp. Dana Settembre University Hospital - UMDNJ Newark, NJ USA >>> Montse verdu 04/26/06 6:11 AM >>> Dear histonetters, Anybody has experience with p27 clone DSC72 different than Calbiochem antibody? We use these antibody on formalin-fixed, paraffin-embedded human prostate carcinoma stained, using avidin-biotin peroxidase complex and DAB chromogen, and we are satisfied with the results, but we have problems with the distributor of these product (frequently we received products with an expiration date from before at the delivery date) and for these reason we needs to change of antibody. Thanks for your help. Montse Verd? histopat LABORATORIS Tel. 932033000 Fax 932802160 mverdu@histopat.es _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jqb7 <@t> cdc.gov Wed Apr 26 11:46:50 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/NCID/VR)) Date: Wed Apr 26 11:59:41 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: That doesn't demonstrate both Gram-positives and negatives, does it? Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Wednesday, April 26, 2006 12:35 PM To: Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] GRAM Stain Control Slides A section of processed Slim Jim works well. J:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, Sara Sent: Wednesday, April 26, 2006 11:08 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GRAM Stain Control Slides Looking for source for GRAM Stain control slides or a known positive tissue to make control block. Vendor sources seem to be quite pricey. Thanks in advance. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 gcallis <@t> montana.edu Wed Apr 26 12:03:49 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Apr 26 12:04:02 2006 Subject: [Histonet] fixing rat tibia In-Reply-To: <200604261218.k3QCIeZD013189@mail2.msu.montana.edu> References: <200604261218.k3QCIeZD013189@mail2.msu.montana.edu> Message-ID: <6.0.0.22.1.20060426093626.01b5b150@gemini.msu.montana.edu> 70% alcohol fixation to preserve the tetracycline - bones can sit in this for several days, and I would opt for longer time due to bone density. People also use alcoholic formalin which does not remove the fluorescent label. After fixation is complete, you should transfer bones from this fixative into 70% fixative. The alcoholic formalin improves morphology for other staining from the same block. If you use this, Anatech has a superb commercial formulation, and to test how long it takes to fix bone, work with several tibias (unlabeled) and open one sample every day for 1,2,3,etc days. If the bone is still bloody inside, then you need to fix longer, it should appear grayish, as formalin fixed tissues would appear when well fixed. You should not have to bisect the rat tibia. That is difficult to do and to avoid shattering bone, you could purchase a little hobby saw, like a Dremel rotating tool/hand saw, but you need to find one that has a tiny little saw blade for cutting. Unfortunately, the small saw blade for Dremel was discontinued, probably for safety reasons, but look for a similar blade at a hobby shop or a similar handtool setup. You may find a blade that fits the Dremel tool. Surprisingly, hobby shops for woodworking, etc often have minitools suitable for bone work, just wear eye safety wear, things tend to fly around and spray blood and bone dust, very messy. Good luck At 06:14 AM 4/26/2006, you wrote: >Hi, > Could anyone advise me regarding the time required to fix a rat >tibia. Can it be fixed whole or is it best to slice in half longitudinally. >Also is 70% ethanol appropriate as a fixative for bones that have >been labelled with Oxytetracycline. Another dilema I have is that I >dont have access to a specialist tools, should I need to bisect the >tibia in half. What tools could be used to cut the tibia in half >longitudinally without cracking the bone. I am new to all this so any >any help / suggestions will be appreciated. Thanks > Ash >ARFA.R.MAQSOOD >ENDOCRINE SCIENCE RESEARCH GROUP >UNIVERSITY OF MANCHESTER >OXFORD ROAD >MANCHESTER >M13 9PT > >TEL:+44 (0)161 275 5177 >FAX:+44 (0)161 275 5958 > >_______________________________________________ >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 406 994-4303 (FAX) From jhaviland <@t> mdanderson.org Wed Apr 26 12:06:02 2006 From: jhaviland <@t> mdanderson.org (jhaviland@mdanderson.org) Date: Wed Apr 26 12:06:13 2006 Subject: [Histonet] scanners Message-ID: Hello: My boss just informed me that he wants to barcode all of our tissue samples in our tumor bank. I was wondering if anyone had any recommendations of a good scanner. Thanks Joie Haviland, HT (ASCP) Senior Research Assistant MD Anderson Cancer Research Center From DDDeltour <@t> mar.med.navy.mil Wed Apr 26 12:09:56 2006 From: DDDeltour <@t> mar.med.navy.mil (Deltour, Douglas D. (HM2)) Date: Wed Apr 26 12:10:34 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE5D@marxchg03.mar.med.navy.mil> I have only seen Gram-positive in the Slim Jim. Let me know if you find negative and I will give it another try. Douglas Deltour HT(ASCP) -----Original Message----- From: Bartlett, Jeanine (CDC/NCID/VR) [mailto:jqb7@cdc.gov] Sent: Wednesday, April 26, 2006 12:47 PM To: Weems, Joyce; Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] GRAM Stain Control Slides That doesn't demonstrate both Gram-positives and negatives, does it? Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, Joyce Sent: Wednesday, April 26, 2006 12:35 PM To: Breeden, Sara; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] GRAM Stain Control Slides A section of processed Slim Jim works well. J:>) Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, Sara Sent: Wednesday, April 26, 2006 11:08 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] GRAM Stain Control Slides Looking for source for GRAM Stain control slides or a known positive tissue to make control block. Vendor sources seem to be quite pricey. Thanks in advance. Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Wed Apr 26 12:12:25 2006 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Wed Apr 26 12:12:33 2006 Subject: [Histonet] Hydrometer In-Reply-To: Message-ID: <4.3.2.7.2.20060426100638.021e0808@algranth.inbox.email.arizona.edu> Heather, I measure the concentration of the alcohols on my stain line with a hydrometer. I just pour alcohol into a 250 ml cylinder (usually a little over the 250 line) and drop in the hydrometer and give it a twirl. The hydrometer came from Fisher, catalog # 11-590. Andi At 11:22 AM 4/26/2006 -0500, Orr, Rebecca wrote: >Heather, >I have a small one that does fit in the container...it did come from >Fisher, but I've had it awhile...there's no part number on it. > >I was also thinking that Wine makers would use them, try surfing a wine >making site to see if they have a size that would fit into the container > >Becky Orr >Evanston Hospital > >Message: 24 >Date: Tue, 25 Apr 2006 11:53:58 -0400 >From: "Santana, Diane" >Subject: [Histonet] here is another one >To: "'histonet@lists.utsouthwestern.edu'" > >Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB0711398B@MAILPMA> >Content-Type: text/plain; charset="iso-8859-1" > >ok I have tried to find this out on my own with no luck.... and I DO >feel >stupid in asking, but here goes. >I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we >are >concerned that there is some X contamination. So my pathologist want me >to >test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The >problem is this, if I try to measure the solutions inside the container >they >aren't deep enough... and either is a 2 qt or 4000ml container. Is there >a >smaller one, does anyone else have one they use. Sorry the one I order >is >catalog number 11590. >Please any help is welcome... don't try to figure out the X >contamination >thing. I know I just am one of those unlucky ones who got a lemon, I >just >have to prove it. >Have a great Tuesday, >Diane Santana >PMA >Haverhill, Mass. > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From vazquezr <@t> ohsu.edu Wed Apr 26 12:12:55 2006 From: vazquezr <@t> ohsu.edu (Robyn Vazquez) Date: Wed Apr 26 12:13:26 2006 Subject: [Histonet] Hydrometer Message-ID: Heather I have one for alcohol from fisher scientific part number is 11-590 Robyn OHSU >>> "Orr, Rebecca" 4/26/2006 9:22 AM >>> Heather, I have a small one that does fit in the container...it did come from Fisher, but I've had it awhile...there's no part number on it. I was also thinking that Wine makers would use them, try surfing a wine making site to see if they have a size that would fit into the container Becky Orr Evanston Hospital Message: 24 Date: Tue, 25 Apr 2006 11:53:58 -0400 From: "Santana, Diane" Subject: [Histonet] here is another one To: "'histonet@lists.utsouthwestern.edu'" Message-ID: <4C96AA62BADFD81180CB0002A5AD67FB0711398B@MAILPMA> Content-Type: text/plain; charset="iso-8859-1" ok I have tried to find this out on my own with no luck.... and I DO feel stupid in asking, but here goes. I need to measure my alcohol% on my tissue processor ( Sakura VIP 5) we are concerned that there is some X contamination. So my pathologist want me to test them daily. So I ordered a alcohol hydrometer 0-100 by fisher. The problem is this, if I try to measure the solutions inside the container they aren't deep enough... and either is a 2 qt or 4000ml container. Is there a smaller one, does anyone else have one they use. Sorry the one I order is catalog number 11590. Please any help is welcome... don't try to figure out the X contamination thing. I know I just am one of those unlucky ones who got a lemon, I just have to prove it. Have a great Tuesday, Diane Santana PMA Haverhill, Mass. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From algranth <@t> u.arizona.edu Wed Apr 26 12:14:02 2006 From: algranth <@t> u.arizona.edu (Andrea Grantham) Date: Wed Apr 26 12:14:09 2006 Subject: [Histonet] GRAM Stain Control Slides In-Reply-To: Message-ID: <4.3.2.7.2.20060426101253.021e21f0@algranth.inbox.email.arizona.edu> I had my students try this and they got gram positives - didn't see any gram negatives. Hot dogs worked the same. Andi At 12:46 PM 4/26/2006 -0400, Bartlett, Jeanine (CDC/NCID/VR) wrote: >That doesn't demonstrate both Gram-positives and negatives, does it? > > >Jeanine Bartlett, BS, HT(ASCP) >Centers for Disease Control and Prevention >1600 Clifton Road, MS/G-32 >18/SB-114 >Atlanta, GA 30333 >(404) 639-3590 >jeanine.bartlett@cdc.hhs.gov > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, >Joyce >Sent: Wednesday, April 26, 2006 12:35 PM >To: Breeden, Sara; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] GRAM Stain Control Slides > >A section of processed Slim Jim works well. J:>) > > >Joyce Weems >Pathology Manager >Saint Joseph's Hospital of Atlanta >404-851-7376 >404-851-7831 - fax > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, >Sara >Sent: Wednesday, April 26, 2006 11:08 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] GRAM Stain Control Slides > >Looking for source for GRAM Stain control slides or a known positive >tissue to make control block. Vendor sources seem to be quite pricey. >Thanks in advance. > >Sally Breeden, HT(ASCP) >New Mexico Department of Agriculture >Veterinary Diagnostic Services >POBox 4700 >Albuquerque, NM 87196 >(505)841-2576 >(505)841-2518 FAX > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message may >be privileged and is confidential information intended for the use of >the addressee listed above. If you are neither the intended recipient >nor the employee or agent responsible for delivering this message to the >intended recipient, you are hereby notified that any disclosure, >copying, distribution or the taking of any action in reliance on the >contents of this information is strictly prohibited. If you have >received this communication in error, please notify us immediately by >replying to the message and deleting it from your computer. Thank you. >Saint Joseph's Health System, Inc. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html From arunams <@t> interchange.ubc.ca Wed Apr 26 12:41:22 2006 From: arunams <@t> interchange.ubc.ca (arunams) Date: Wed Apr 26 12:41:29 2006 Subject: [Histonet] Job opening in Vancouver BC Message-ID: <12652023.1146073282825.JavaMail.myubc2@portal9.itservices.ubc.ca> Position: Histotechnologist Company: Wax-it Histology Service Inc. Wax-it Histology Services is a fast growing Vancouver based contract research organization that is providing histology related solutions to academic institutes, hospitals, biotechnology and pharmaceutical industries. We are specialized in both animal and plant tissue related solutions. Wax-it Histology Services has an immediate opening for a full time histotechnologist . Major Responsibilities The successful incumbent will be responsible for routine histology methods such as grossing tissue, processing, sectioning, cryo processing and sectioning, immunohistochemistry and special histological stains. The position also involves the evaluation and development of new technologies and includes routine QC testing. Emphasis will be placed on the maintenance of neat, accurate and precise documentation of all work performed. Other research and general lab duties will be performed as required. Requirements The successful candidate will be a qualified histotechnologist with at least a BSc and 2-4 years relevant work experience, with basic knowledge of standard laboratory procedures including lab safety. He or She should have strong organizational, multi-tasking and time management abilities. Further to that the candidate will have good problem solving skills and posses well-developed communication skills. The candidate must also be competent and self motivated and be able to integrate into a fast growing and dynamic company with the ability to work independently or collaboratively in a team environment. Your resume must clearly demonstrate that you meet all of the above qualifications and preferences. We thank all applicants for their interest, but only those candidates short-listed will be contacted for an interview. Please send your resume, cover letter and three referees with salary expectations to: Address: Wax-it Histology Services 310-2386 East Mall Vancouver, BC, V6T 1Z3 E-mail: jobs@waxitinc.com From Sandra.Harrison3 <@t> va.gov Wed Apr 26 12:46:31 2006 From: Sandra.Harrison3 <@t> va.gov (Harrison, Sandra C.) Date: Wed Apr 26 12:46:38 2006 Subject: [Histonet] Histology Position in Minneapolis, MN. Message-ID: <736E8889E98B8F4FBBD29FDFEC8074BA49C051@VHAV23MSGA2.v23.med.va.gov> Dear Histonetters, The VA hospital in Minneapolis, MN will be posting a Histology position, in the next week or so. This position requires someone with their HT registry and at least a few years of experience working in general Histology. This employee will rotate through all the various benches including embedding, microtomy, special stains and assisting with gross. No immuno experience is required, although you will have the opportunity of training into our busy immunohistochemistry lab, at some point. Please call 612-467-2449 for more information about the position, the benefits and how to apply. If you prefer to send your contact information and resume electronically, my e-mail address is Sandra.Harrison3@va.gov. Sincerely, Sandra C. Harrison, HTL Supervisor of Anatomical and Surgical Pathology From Charlene.Henry <@t> STJUDE.ORG Wed Apr 26 12:53:06 2006 From: Charlene.Henry <@t> STJUDE.ORG (Henry, Charlene) Date: Wed Apr 26 12:53:11 2006 Subject: [Histonet] Position available in Memphis TN Message-ID: <5CB39BCA5724F349BCB748675C6CA1A2099A1C21@SJMEMXMB02.stjude.sjcrh.local> We currently have a HT 2 position available at St Jude Children's Research Hospital. If anyone is interested in the position, go to www.stjude.org and clink on Job Openings. Thanks, Charlene Henry HT (ASCP), QIHC Histology/Immunohistochemistry Section Head Department of Pathology St. Jude Children's Research Hospital 901-495-3191 fax 901-495-3100 From PMonfils <@t> Lifespan.org Wed Apr 26 13:06:20 2006 From: PMonfils <@t> Lifespan.org (Monfils, Paul) Date: Wed Apr 26 13:06:30 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: <09C945920A6B654199F7A58A1D7D1FDE017176E4@lsexch.lsmaster.lifespan.org> Excellent gram controls are pretty easy to make. I order cultures of gram positive and gram negative organisms from a biological supply company. I get them in liquid culture (broth), not on agar plates. They cost less than $10.00 per culture. I spin down the cultures to concentrate the organisms, remove most of the supernatant, resuspend the organisms by vortexing, then inject the concentrated culture into a small block of fresh lung. I push the needle almost all the way through, then withdraw the needle as I inject, in order to distribute the organisms through the tissue. Lung works best because there are a lot of small spaces into which the culture can flow. Gently knead the tissue a bit with a fingertip (gloved of course) to further distribute the organisms. Drop the lung into fixative, let it fix, then process and embed as usual. I inject gram positive organisms into one piece of tissue, gram negatives into another, then after processing, cut the tissue into smaller square pieces, and embed a gram positive piece and a gram negative piece side by side in one paraffin block. An easier way is to purchase cultures on agar plates, fix them as they are, then process and embed small pieces of the agar with the organisms on it. But even though it is a little more involved, I like the more "natural" look of the organisms in tissue. Also, in using normal lung you don't have to deal with all the cellular infiltrate, necrosis, exudates, etc. that would be present in an actual massive bacterial lung infection. For gram positive I favor Bacillus subtilis and Streptococcus epidermidis. For gram negative I use Escherichia coli or Enterobacter aerogenes, and Neisseria subflava. I like to have both a bacillus (rod) and a coccus (sphere) in both the gram positive and the gram negative tissue. I get my cultures from: http://www.ctvalleybio.com From Kathy.Johnston <@t> CLS.ab.ca Wed Apr 26 13:18:18 2006 From: Kathy.Johnston <@t> CLS.ab.ca (Kathy.Johnston@CLS.ab.ca) Date: Wed Apr 26 13:18:25 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE0136531F@mail1.calgary.com> We make our own as well. I ask our Micro department to innoculate a broth with E. Coli, and Staph or Strep, and when it's ready, throw in some fresh cubed placenta. I'll let it sit over night at RT and then fix and process it. Works beautifully, and is very inexpensive. Kathy Johnston Tech II Special Stains Anatomic Pathology Calgary Laboratory Services #9 - 3535 Research Road NW Calgary, AB. 403-770-3572 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Monfils, Paul Sent: April 26, 2006 12:06 PM To: 'histonet@lists.utsouthwestern.edu' Subject: RE: [Histonet] GRAM Stain Control Slides Excellent gram controls are pretty easy to make. I order cultures of gram positive and gram negative organisms from a biological supply company. I get them in liquid culture (broth), not on agar plates. They cost less than $10.00 per culture. I spin down the cultures to concentrate the organisms, remove most of the supernatant, resuspend the organisms by vortexing, then inject the concentrated culture into a small block of fresh lung. I push the needle almost all the way through, then withdraw the needle as I inject, in order to distribute the organisms through the tissue. Lung works best because there are a lot of small spaces into which the culture can flow. Gently knead the tissue a bit with a fingertip (gloved of course) to further distribute the organisms. Drop the lung into fixative, let it fix, then process and embed as usual. I inject gram positive organisms into one piece of tissue, gram negatives into another, then after processing, cut the tissue into smaller square pieces, and embed a gram positive piece and a gram negative piece side by side in one paraffin block. An easier way is to purchase cultures on agar plates, fix them as they are, then process and embed small pieces of the agar with the organisms on it. But even though it is a little more involved, I like the more "natural" look of the organisms in tissue. Also, in using normal lung you don't have to deal with all the cellular infiltrate, necrosis, exudates, etc. that would be present in an actual massive bacterial lung infection. For gram positive I favor Bacillus subtilis and Streptococcus epidermidis. For gram negative I use Escherichia coli or Enterobacter aerogenes, and Neisseria subflava. I like to have both a bacillus (rod) and a coccus (sphere) in both the gram positive and the gram negative tissue. I get my cultures from: http://www.ctvalleybio.com _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From jqb7 <@t> cdc.gov Wed Apr 26 13:15:12 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/NCID/VR)) Date: Wed Apr 26 13:30:25 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: That's what I heard.....I'll have to try it myself. Jeanine Bartlett, BS, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Wednesday, April 26, 2006 1:14 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] GRAM Stain Control Slides I had my students try this and they got gram positives - didn't see any gram negatives. Hot dogs worked the same. Andi At 12:46 PM 4/26/2006 -0400, Bartlett, Jeanine (CDC/NCID/VR) wrote: >That doesn't demonstrate both Gram-positives and negatives, does it? > > >Jeanine Bartlett, BS, HT(ASCP) >Centers for Disease Control and Prevention >1600 Clifton Road, MS/G-32 >18/SB-114 >Atlanta, GA 30333 >(404) 639-3590 >jeanine.bartlett@cdc.hhs.gov > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, >Joyce >Sent: Wednesday, April 26, 2006 12:35 PM >To: Breeden, Sara; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] GRAM Stain Control Slides > >A section of processed Slim Jim works well. J:>) > > >Joyce Weems >Pathology Manager >Saint Joseph's Hospital of Atlanta >404-851-7376 >404-851-7831 - fax > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, >Sara >Sent: Wednesday, April 26, 2006 11:08 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] GRAM Stain Control Slides > >Looking for source for GRAM Stain control slides or a known positive >tissue to make control block. Vendor sources seem to be quite pricey. >Thanks in advance. > >Sally Breeden, HT(ASCP) >New Mexico Department of Agriculture >Veterinary Diagnostic Services >POBox 4700 >Albuquerque, NM 87196 >(505)841-2576 >(505)841-2518 FAX > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message may >be privileged and is confidential information intended for the use of >the addressee listed above. If you are neither the intended recipient >nor the employee or agent responsible for delivering this message to the >intended recipient, you are hereby notified that any disclosure, >copying, distribution or the taking of any action in reliance on the >contents of this information is strictly prohibited. If you have >received this communication in error, please notify us immediately by >replying to the message and deleting it from your computer. Thank you. >Saint Joseph's Health System, Inc. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JWEEMS <@t> sjha.org Wed Apr 26 13:35:20 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Wed Apr 26 13:35:31 2006 Subject: [Histonet] GRAM Stain Control Slides Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD3202637C74@sjhaexc02.sjha.org> It has been some years since I used them - but I swear both bugs were there then. Perhaps there has been a change. The placenta idea sounded good - unless you're where you don't have placentas - like here! j Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Bartlett, Jeanine (CDC/NCID/VR) Sent: Wednesday, April 26, 2006 2:15 PM To: Andrea Grantham; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] GRAM Stain Control Slides That's what I heard.....I'll have to try it myself. Jeanine Bartlett, BS, HT(ASCP) (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Andrea Grantham Sent: Wednesday, April 26, 2006 1:14 PM To: histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] GRAM Stain Control Slides I had my students try this and they got gram positives - didn't see any gram negatives. Hot dogs worked the same. Andi At 12:46 PM 4/26/2006 -0400, Bartlett, Jeanine (CDC/NCID/VR) wrote: >That doesn't demonstrate both Gram-positives and negatives, does it? > > >Jeanine Bartlett, BS, HT(ASCP) >Centers for Disease Control and Prevention >1600 Clifton Road, MS/G-32 >18/SB-114 >Atlanta, GA 30333 >(404) 639-3590 >jeanine.bartlett@cdc.hhs.gov > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Weems, >Joyce >Sent: Wednesday, April 26, 2006 12:35 PM >To: Breeden, Sara; histonet@lists.utsouthwestern.edu >Subject: RE: [Histonet] GRAM Stain Control Slides > >A section of processed Slim Jim works well. J:>) > > >Joyce Weems >Pathology Manager >Saint Joseph's Hospital of Atlanta >404-851-7376 >404-851-7831 - fax > >-----Original Message----- >From: histonet-bounces@lists.utsouthwestern.edu >[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Breeden, >Sara >Sent: Wednesday, April 26, 2006 11:08 AM >To: histonet@lists.utsouthwestern.edu >Subject: [Histonet] GRAM Stain Control Slides > >Looking for source for GRAM Stain control slides or a known positive >tissue to make control block. Vendor sources seem to be quite pricey. >Thanks in advance. > >Sally Breeden, HT(ASCP) >New Mexico Department of Agriculture >Veterinary Diagnostic Services >POBox 4700 >Albuquerque, NM 87196 >(505)841-2576 >(505)841-2518 FAX > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >Confidentiality Notice ** The information contained in this message may >be privileged and is confidential information intended for the use of >the addressee listed above. If you are neither the intended recipient >nor the employee or agent responsible for delivering this message to the >intended recipient, you are hereby notified that any disclosure, >copying, distribution or the taking of any action in reliance on the >contents of this information is strictly prohibited. If you have >received this communication in error, please notify us immediately by >replying to the message and deleting it from your computer. Thank you. >Saint Joseph's Health System, Inc. > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet ..................................................................... : Andrea Grantham, HT(ASCP) Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212) P.O. Box 245044 : : (voice: 520-626-4415) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: algranth@u.arizona.edu) : :...................................................................: http://www.cba.arizona.edu/histology-lab.html _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet 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 gcallis <@t> montana.edu Wed Apr 26 13:52:35 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Wed Apr 26 13:52:45 2006 Subject: Appendix for Re: [Histonet] GRAM Stain Control Slides In-Reply-To: <20060426154214.388.qmail@web61223.mail.yahoo.com> References: <20060426154214.388.qmail@web61223.mail.yahoo.com> Message-ID: <6.0.0.22.1.20060426124552.01b5ba18@gemini.msu.montana.edu> Appendix works very well, but a veterinary laboratory situation, getting appendix may be difficult. For vet diagnostic work, we used a section of bowel or try small intestine, do not rinse it excessively before fixation in NBF. Cow, dog, pig, sheep, any animal should work - I would go for the species that is easiest to section. There are several ways to make controls from pure cultures too and you can have your microbiology people help with this. I think Histonet archives has some of these methods, then you guarantee Gram pos and negative being there, and do a large number of blocks at one time. At 09:42 AM 4/26/2006, you wrote: >Sara: > I used for years sections from cases of acute appendicities. They > contain a host of both Gram + and - bacteria. > I hope this will help you! > Ren? J. 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 406 994-4303 (FAX) From amoklebu <@t> seattlecca.org Wed Apr 26 17:02:56 2006 From: amoklebu <@t> seattlecca.org (Moklebust, Amanda C) Date: Wed Apr 26 17:03:10 2006 Subject: [Histonet] Identifying epithelial cells in urine cell block Message-ID: <3B294E8385BB664B82F756C009D15A2F115C30@storm.seattlecca.org> Dear Histonetters, I am looking for a way to determine if the epithelial cells in a voided urine cell block are from the bladder or collecting ducts of the kidneys. Do any of you know of an antibody that will stain epithelial cells from the kidney but not the bladder, or vice versa? Thank you in advance for your suggestions. Amanda Moklebust 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 ja.mitchell <@t> hosp.wisc.edu Wed Apr 26 17:32:34 2006 From: ja.mitchell <@t> hosp.wisc.edu (Mitchell Jean A.) Date: Wed Apr 26 17:32:40 2006 Subject: [Histonet] NSH Awards Deadline Message-ID: <583D3E9A1E843445BD54E461D1A2F6F3192C1ABF@UWHIS-XCHNG2.uwhis.hosp.wisc.edu> As a reminder: May 1st is the deadline for submitting nominations/applications for the 2006 NSH Awards and Scholarship program. Submissions must be postmarked by May 1, 2006 for this years consideration. Applications and criteria are listed at www.nsh.org. Feel free to contact me with any questions. Jean Mitchell Chair, NSH Awards Committee From mbmphoto <@t> gmail.com Wed Apr 26 22:00:16 2006 From: mbmphoto <@t> gmail.com (Maria Mejia) Date: Wed Apr 26 21:59:40 2006 Subject: [Histonet] need to section -80C frozen tissue on sliding microtome Message-ID: My question is a bit odd, but here goes. Over the years, I've gained a lot of experience sectioning fixed brain on the sliding microtome. These specimens were always frozen from a 30% sucrose/PBS solution & sectioned on the microtome. Now, for the past 2 months I've been working in a new lab that has a very large tissue bank. All of the primate tissue is fixed in zamboni fixative and they go through the usual 30% sucrose solution before they are snap frozen and stored in a -80 C sub zero freezer. If I need to cryostat section this type of frozen tissue, I know that once I remove the tissue from the -80C storage area I have to leave it in the cryostat at least 1 hour before sectioning it. Now, my question is - if I need to use the sliding microtome to section these -80C frozen tissues- how do I proceed with them so I can section them?????????? They are already frozen, but I still need to attach them to the sliding tissue stage - how? And, they maybe too cold to section?? Please any suggestions, and tips you can provide will be greatly appreciated. Yours Maria Bartola Mejia Department of Neurosurgery University of California San Francisco San Francisco, CA 94103 From louise.renton <@t> gmail.com Thu Apr 27 03:11:58 2006 From: louise.renton <@t> gmail.com (louise renton) Date: Thu Apr 27 03:12:02 2006 Subject: [Histonet] secondary antibody Message-ID: Dear immuno experts, Please help my soggy xylene soaked brain work out what is probably a simple question. If I am using a polyclonal antibody on mouse tissue (reactivity is established) , and detecting with Vector's ABC kit, what biotinylated seconday do I use? goat or swine anti rabbit? something else??? ? PS I have a mouse on mouse kit already -are there regaents in there that i could sucessfully use?? PPS. This is the first time i have done immuno in other species, I was used to work on human tissue alone hence my confusion Thanks in advance -- Louise Renton Bone Research Unit University of the Witwatersrand Johannesburg South Africa "Does a conceited cowboy, only ride on pompous grass?. From Jacqueline.Malam <@t> rli.mbht.nhs.uk Thu Apr 27 03:44:08 2006 From: Jacqueline.Malam <@t> rli.mbht.nhs.uk (Malam Jacqueline) Date: Thu Apr 27 03:45:09 2006 Subject: [Histonet] Gram stain control tissue Message-ID: Have heard about injecting fresh placenta with mixed cultures from Micro and incubating it overnight, or using an inflamed appendix; but the best we have tried is some nice, ripe blue cheese - process it (wrap in speciwrap or similar as it can be a bit crumbly), block it out and section it as normal - works a treat and you don't need its permission to use it, and if you have any left over and you left your lunch at home, it goes down well with a nice wedge of brown bread, plum chutney and a cold glass of cider! Jacqui DISCLAIMER: This e-mail is confidential and privileged. If you are not the intended recipient please accept our apologies; please do not disclose, copy or distribute information in this e-mail or take any action in reliance on its contents: to do so is strictly prohibited and may be unlawful. Please inform postmaster@rli.mbht.nhs.uk that this message has gone astray before deleting it. Comments or opinions expressed in this email are those of their respective contributors only. The views expressed do not represent the views of the Trust, its management or employees. Morecambe Bay Hospitals NHS Trust is not responsible and disclaims any and all liability for the content of comments written within.Thank you for your co-operation. From DDDeltour <@t> mar.med.navy.mil Thu Apr 27 07:28:03 2006 From: DDDeltour <@t> mar.med.navy.mil (Deltour, Douglas D. (HM2)) Date: Thu Apr 27 07:28:29 2006 Subject: [Histonet] Sakura: Tissue-Tek Xpress Rapid Tissue Processor Message-ID: <3F500F8B416C554EBB21FF16642F72E959CE60@marxchg03.mar.med.navy.mil> Can I get some realistic feedback from the Tissue-Tek Xpress users? What are your actual processing times for biopsies and surgical? What are the negatives? How did you adjust your staffing schedules? Any feedback would be appreciated. Thanks DOUGLAS D. DELTOUR HT(ASCP) NMC PORTSMOUTH VA (757)953-1525 From abright <@t> brightinstruments.com Thu Apr 27 09:32:21 2006 From: abright <@t> brightinstruments.com (Alan Bright) Date: Thu Apr 27 09:22:15 2006 Subject: [Histonet] need to section -80C frozen tissue on sliding microtome Message-ID: Dear Maria, Although I have a broad knowledge of cryostats & microtomes and some useful sectioning techniques plus the fact I have not seen any answers yet to your problem. Here are my two penny's worth followed after your text with _ _ _ _ Now, my question is - if I need to use the sliding microtome to section these -80C frozen tissues- how do I proceed with them so I can section them?????????? They are already frozen, but I still need to attach them to the sliding tissue stage - how? _ _ _ _As the specimen is frozen and the stage should be too, you will be unable to use embedding medium to adhere the specimen to the tissue stage of the microtome. Therefore you will have to use water from a syringe or pipette and be very quick in placing the specimen in place prior to the water freezing. And, they maybe too cold to section??_ _ _ _ Yes they will be too cold, so you need to measure the temperature of the specimen with a thermometer or electronic probe until it rises to the desired temperature, this is the only way you will know accurately, as it will always be a different timing unless the specimens are all the same size, the tissue stage is always at the same temperature and the ambient too. 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 Skype User ID: dazzle0 -----Original Message----- From: Maria Mejia [mailto:mbmphoto@gmail.com] Sent: 27 April 2006 04:00 To: histonet@lists.utsouthwestern.edu Subject: [Histonet] need to section -80C frozen tissue on sliding microtome My question is a bit odd, but here goes. Over the years, I've gained a lot of experience sectioning fixed brain on the sliding microtome. These specimens were always frozen from a 30% sucrose/PBS solution & sectioned on the microtome. Now, for the past 2 months I've been working in a new lab that has a very large tissue bank. All of the primate tissue is fixed in zamboni fixative and they go through the usual 30% sucrose solution before they are snap frozen and stored in a -80 C sub zero freezer. If I need to cryostat section this type of frozen tissue, I know that once I remove the tissue from the -80C storage area I have to leave it in the cryostat at least 1 hour before sectioning it. Now, my question is - if I need to use the sliding microtome to section these -80C frozen tissues- how do I proceed with them so I can section them?????????? They are already frozen, but I still need to attach them to the sliding tissue stage - how? And, they maybe too cold to section?? Please any suggestions, and tips you can provide will be greatly appreciated. Yours Maria Bartola Mejia Department of Neurosurgery University of California San Francisco San Francisco, CA 94103 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gpbnas <@t> yahoo.es Thu Apr 27 11:00:27 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Thu Apr 27 11:00:32 2006 Subject: [Histonet] Good TUNEL staining but very few Active Caspase-3 positive cells!!! Message-ID: <20060427160027.99051.qmail@web26206.mail.ukl.yahoo.com> Hello histoneters, I am studying apoptosis in mouse kidney samples, and while TUNEL staining looks pretty good (positive cells checked by morphology with DAPI), staining with Anti-Active-Caspase-3 Ab from Cell Signaling shows very low number of positive cells. I have tried frozen sections fixed with Acetone or Paraformaldehyde as well as parafin embeded sections boiled in citrate buffer for 10 min. Positive cells look very good, with nice cytoplasmic staining, but I would have expected a higher number of positive cells (See J Pathol 2003; 199: 221?228 for a comparison of TUNEL and Active Caspase-3 staining). I am quite sure that TUNEL staining is specific as I titrated TdT concentration and nuclei look apoptotic with DAPI. How could I improve Active Caspase-3 staining? Any ideas are most welcome! Guillermo --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From Ebreisch <@t> chsd.org Thu Apr 27 11:28:34 2006 From: Ebreisch <@t> chsd.org (Breisch, Eric) Date: Thu Apr 27 11:28:47 2006 Subject: [Histonet] double blades for rotary paraffin microtome Message-ID: <00D7298B3825D511BF540008C75F8A3C0CAD82E8@excluster.chsd.org> Hi Histologists, This is a generalised question regarding microtomes and blades. Has anyone ever heard of a rotary microtome that uses or used double edged blades (not unlike the razor blades currently being marketed for shaving) to cut paraffin blocks? More specifically have double edged blades ever been used to cut paraffin blocks. Your help and input is greatly appreciated. Regards, Eric A. Breisch, Ph.D. Clinical Anatomist Department of Pathology Associate Clinical Professor of Anatomy, UCSD-SOM Children's Hospital-San Diego 3020 Children's Way, MC5007 San Diego, CA 92123 Phone: 858-966-5944 From asmith <@t> mail.barry.edu Thu Apr 27 11:40:16 2006 From: asmith <@t> mail.barry.edu (Smith, Allen) Date: Thu Apr 27 11:40:21 2006 Subject: [Histonet] double blades for rotary paraffin microtome Message-ID: <5D2189E74151CC42BEC02906BA8996322B91D1@exchsrv01.barrynet.barry.edu> Many (40?) years ago, I saw a blade holder designed to allow the use of double-edged razor blades for cutting paraffin sections. My recollection is that the professor told me that he used it only for teaching students how to use amicrotome. Allen A. Smith, Ph.D. Professor of Anatomy Barry University School of Graduate Medical Sciences Podiatric Medicine and Surgery Miami Shores, Florida 33161 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Breisch, Eric Sent: Thursday, April 27, 2006 12:29 PM To: 'histonet@lists.utsouthwestern.edu' Subject: [Histonet] double blades for rotary paraffin microtome Hi Histologists, This is a generalised question regarding microtomes and blades. Has anyone ever heard of a rotary microtome that uses or used double edged blades (not unlike the razor blades currently being marketed for shaving) to cut paraffin blocks? More specifically have double edged blades ever been used to cut paraffin blocks. Your help and input is greatly appreciated. Regards, Eric A. Breisch, Ph.D. Clinical Anatomist Department of Pathology Associate Clinical Professor of Anatomy, UCSD-SOM Children's Hospital-San Diego 3020 Children's Way, MC5007 San Diego, CA 92123 Phone: 858-966-5944 _______________________________________________ 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 rjbuesa <@t> yahoo.com Thu Apr 27 12:02:55 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Apr 27 12:03:01 2006 Subject: [Histonet] double blades for rotary paraffin microtome In-Reply-To: <00D7298B3825D511BF540008C75F8A3C0CAD82E8@excluster.chsd.org> Message-ID: <20060427170255.18589.qmail@web61215.mail.yahoo.com> Eric: American Optical Co., in the early 1950s, manufactured a blade holder for the regular 2 edge razor (shaving) blade to be used in the Spencer-AO 860 horizontal microtome. The holder was able to have a very good rgip on the blades than although very thin, could be used for routine sectioning without vibrations. I have heard of no other type of 2 edges microtome blade, though. On a separate e-mail I am sending you a photo of this type of holder for rotary microtomes. Hope this will help you! Ren? J. "Breisch, Eric" wrote: Hi Histologists, This is a generalised question regarding microtomes and blades. Has anyone ever heard of a rotary microtome that uses or used double edged blades (not unlike the razor blades currently being marketed for shaving) to cut paraffin blocks? More specifically have double edged blades ever been used to cut paraffin blocks. Your help and input is greatly appreciated. Regards, Eric A. Breisch, Ph.D. Clinical Anatomist Department of Pathology Associate Clinical Professor of Anatomy, UCSD-SOM Children's Hospital-San Diego 3020 Children's Way, MC5007 San Diego, CA 92123 Phone: 858-966-5944 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- New Yahoo! Messenger with Voice. Call regular phones from your PC and save big. From TJJ <@t> Stowers-Institute.org Thu Apr 27 12:04:29 2006 From: TJJ <@t> Stowers-Institute.org (Johnson, Teri) Date: Thu Apr 27 12:04:46 2006 Subject: [Histonet] Re: Good TUNEL staining but very few Active Caspase-3 positive cells Message-ID: Guillermo, Good question! I'll take a stab at it and will read with interest any additional commentary on this. We found to get good staining with Cleaved Caspase-3 in paraffin section we needed to add a brief pepsin digestion in addition to the HIER to achieve staining. Without it we found we needed to use tyramide enhancement to see it in fluorescence. I think the antibody really does exhibit sensitivity issues. Otherwise, caspase-3 is activated early in the apoptotic process. It could be that it is no longer in its activated state in later stages of apoptosis, whereas the TUNEL will show positive across a wide range of apoptosis stages. I'm an amateur on the mechanism of this event, and welcome Dr. Cartun or anyone with more advanced knowledge of this to confirm or disagree with my conjecture. Best wishes, Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 From livieira <@t> ualg.pt Thu Apr 27 12:10:43 2006 From: livieira <@t> ualg.pt (Lina Vieira) Date: Thu Apr 27 12:12:41 2006 Subject: [Histonet] Sudan Black for lipids Message-ID: <002801c66a1d$84d671c0$2914100a@labhistologia> Hi, We have samples of fish gonads imersed in 70% etanol and fixed in Bouin?s fixative, it is possible to get acceptable results for lipids localization with Sudan Black in that samples? Any protocols, ideas, and/or suggestions will be greatly appreciated! Thank in advance, Lina Vieira University of Algarve From rjbuesa <@t> yahoo.com Thu Apr 27 12:21:42 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Thu Apr 27 12:21:53 2006 Subject: [Histonet] Sudan Black for lipids In-Reply-To: <002801c66a1d$84d671c0$2914100a@labhistologia> Message-ID: <20060427172142.52250.qmail@web61221.mail.yahoo.com> Lina: Sudan Black is used to detect complex/insoluble fats in processed tissues. If the fish gonads contain them, they will stain. All depends in the type of fat the gonads contain. With Sudan Black lipofuscin, lipoproteins and any insoluble fat will appear from violet-black to black. You can find a good protocol in Sheehan & Hrapchak (1973): 127-8, or in any histotechnology book. Hope this will help you! Ren? J. Lina Vieira wrote: Hi, We have samples of fish gonads imersed in 70% etanol and fixed in Bouin?s fixative, it is possible to get acceptable results for lipids localization with Sudan Black in that samples? Any protocols, ideas, and/or suggestions will be greatly appreciated! Thank in advance, Lina Vieira University of Algarve _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Yahoo! Messenger with Voice. Make PC-to-Phone Calls to the US (and 30+ countries) for 2?/min or less. From gu.lang <@t> gmx.at Thu Apr 27 12:23:52 2006 From: gu.lang <@t> gmx.at (Gudrun Lang) Date: Thu Apr 27 12:23:56 2006 Subject: [Histonet] CD34 in trephine bone marrow Message-ID: <000401c66a1f$5b1bb230$eeeea8c0@SERVER01> Hi, Short time ago I've asked for CD34 protocols. Thank you for the answers, but I still have troubles. Our treatment: Trephine bone marrow biopsies - ca. 3-4 mm in diameter, 15-25 mm long Fixation - 12 h 8% neutral buffered formaldehyd Decal - 6-8 h "Cal-rite", 5-10% formic acid and formaldehyd, roomtemp. Dehydration and infiltration in paraffin - VIP over night Immunostainer - benchmark XT CD34, Dako QBEND10 1:50 I've tried several modification with the protocols. In fact the tonsil on the same slide shows positiv cells with every method, also in the bone marrow biopsy there are positiv endothels (no retrieval, 24 min), but the lymphoblasts are all negative. I think the antibody is allright, but something has happend to the lymphoblasts due to the pretreatment. Any suggestions are appreciated. Thank you Gudrun Lang Akh Linz Austria From Joyce.Rush <@t> sjmcmn.org Thu Apr 27 12:24:22 2006 From: Joyce.Rush <@t> sjmcmn.org (Rush, Joyce) Date: Thu Apr 27 12:24:27 2006 Subject: [Histonet] One more system about returning tissue Message-ID: <2337362E8548BE4C85EE5DD5D526B085744393@sjw3smail2.SJMCMN.ORG> We are a Catholic hospital so some of our tissue handling is "Church" related. We save all POC's and, when we have enough, take them to a local funeral director who cremates them. The remains are placed on a gravesite which was purchased and maintained by the "Empty Arms Society." If a family wants to make private arrangements for a POC they may do so but, must go through a licensed, funeral director. Other requests are handled using CAP rules. We offer pictures. This has worked quite well and considerably cut down on handling difficult requests. When push comes to shove our Medical director makes the decision. Joyce Joyce A. Rush, BS, MT(ASCP) Laboratory Manager St Joseph's Medical Center 523 North Third Street Brainerd, MN 56401 Office:218-828-7500 Fax: 218-828-7510 DISCLAIMER: This email and any files transmitted with it are confidential and intended solely for the use of histonet@lists.utsouthwestern.edu. If you have received this email in error please notify Rush, Joyce and destroy/delete the message. Please note that any views or opinions presented in this email are solely those of the Author and do not necessarily represent those of the company. Finally, the recipient should check this email and any attachments for the presence of viruses. The Medical Center accepts no liability for any damage caused by any virus transmitted by this email. St. Joseph's Medical Center, 523 N. 3rd street, Brainerd, MN 56401 From sbreeden <@t> nmda.nmsu.edu Thu Apr 27 13:02:41 2006 From: sbreeden <@t> nmda.nmsu.edu (Breeden, Sara) Date: Thu Apr 27 13:02:48 2006 Subject: [Histonet] GRAM Thanks Message-ID: Thanks to all of you that answered my cry for help with finding GRAM control. I have stocked up on Slim Jims, hot dogs and we're going to have a cookout for Fuming Friday. I'm thinking that beer brats would make a good test subject, too.. Happy GRAMming! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX From funderwood <@t> mcohio.org Thu Apr 27 13:29:30 2006 From: funderwood <@t> mcohio.org (Fred Underwood) Date: Thu Apr 27 13:29:50 2006 Subject: [Histonet] GRAM Thanks Message-ID: Mmmmm.. How long of a drive is it from Dayton Ohio to Albuquerque? p.s. Go Isotopes! >>> "Breeden, Sara" 04/27/06 02:02PM >>> Thanks to all of you that answered my cry for help with finding GRAM control. I have stocked up on Slim Jims, hot dogs and we're going to have a cookout for Fuming Friday. I'm thinking that beer brats would make a good test subject, too.. Happy GRAMming! Sally Breeden, HT(ASCP) New Mexico Department of Agriculture Veterinary Diagnostic Services POBox 4700 Albuquerque, NM 87196 (505)841-2576 (505)841-2518 FAX _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From laurie.colbert <@t> huntingtonhospital.com Thu Apr 27 13:34:42 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Thu Apr 27 13:34:52 2006 Subject: [Histonet] POC for Genetic Studies Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653B0D@EXCHANGE1.huntingtonhospital.com> This question is for those of you who work in hospitals and send out POC tissue for genetic studies/chromosome analysis. Does the tissue get sent out directly from Labor and Delivery, or does the tissue go through Pathology and Pathology sends it out? From JWEEMS <@t> sjha.org Thu Apr 27 13:57:46 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Thu Apr 27 13:57:52 2006 Subject: [Histonet] POC for Genetic Studies Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD3202637CAF@sjhaexc02.sjha.org> We send through Pathology. Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Laurie Colbert Sent: Thursday, April 27, 2006 2:35 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] POC for Genetic Studies This question is for those of you who work in hospitals and send out POC tissue for genetic studies/chromosome analysis. Does the tissue get sent out directly from Labor and Delivery, or does the tissue go through Pathology and Pathology sends it out? _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From Kristopher.Kalleberg <@t> unilever.com Thu Apr 27 14:01:06 2006 From: Kristopher.Kalleberg <@t> unilever.com (Kalleberg, Kristopher) Date: Thu Apr 27 14:01:11 2006 Subject: [Histonet] TUNEL staining Message-ID: <0E6BC087F70F9C47ACFF2C203D6E329CA2C341@NTRSEVS30002.s3.ms.unilever.com> Guillermo, I am currently performing TUNEL on human skin post UV irradiation and have had inconclusive results. I was using the Promega TUNEL assay kit and also tried a formamide induced TUNEL assay with differing results. I also have read in the literature that the kit and most TUNEL assays are nonspecific (necrotic cells also show positive staining). The formamide induced protocol that I have only tried twice has shown no positive staining yet. I was just wondering which protocol you use for TUNEL and how certain are you that it is specific for apoptotic cells? Maybe we can help each other. Kris Kalleberg Research Associate Unilever R&D 40 Merritt Blvd. Trumbull, CT 06457 203 381 5765 From Bauer.Karen <@t> mayo.edu Thu Apr 27 14:01:22 2006 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Thu Apr 27 14:01:32 2006 Subject: [Histonet] POC for Genetic Studies References: <0BE6ADFAE4E7E04496BF21ABD346628005653B0D@EXCHANGE1.huntingtonhospital.com> Message-ID: Hi Laurie, We are the ones responsible for sending out specimens for chromosome analysis. We have Hank's Solution sent to us by Mayo Clinic, and when we are notified that a specimen is coming for chromosomal analysis, we gross in the (fresh/unfixed) specimen and put recognizable chorionic villi into the solution. Paperwork for Mayo is filled out by our secretaries and we have it sent to them by courier. Good Luck, Karen Bauer HT(ASCP) Histology Supervisor Luther Hospital Eau Claire, WI ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie Colbert Sent: Thu 4/27/2006 1:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] POC for Genetic Studies This question is for those of you who work in hospitals and send out POC tissue for genetic studies/chromosome analysis. Does the tissue get sent out directly from Labor and Delivery, or does the tissue go through Pathology and Pathology sends it out? _______________________________________________ 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 NDhatt <@t> qltinc.com Thu Apr 27 14:02:53 2006 From: NDhatt <@t> qltinc.com (NDhatt@qltinc.com) Date: Thu Apr 27 14:03:01 2006 Subject: [Histonet] CD3 antibody for FFPE rabbit tissue Message-ID: Hi All, I am looking for a CD3 antibody to use on FFPE rabbit tissue. Most antibodies I have looked into are tested in human. Thanks Narinder Dhatt Preclinical - Histology QLT Inc. Vancouver, BC Canada This electronic transmission (including any and all attachments) is intended solely for the use of the individual or entity to whom it is addressed and may contain information that is privileged and/or confidential. If you are not the intended recipient of this electronic transmission, you are hereby notified that any disclosure, copying or distribution, or the taking of any action in reliance upon the contents of this electronic transmission, is strictly prohibited, and you are further requested to purge this electronic transmission and all copies thereof from your computer system. From laurie.colbert <@t> huntingtonhospital.com Thu Apr 27 14:15:04 2006 From: laurie.colbert <@t> huntingtonhospital.com (Laurie Colbert) Date: Thu Apr 27 14:15:10 2006 Subject: [Histonet] Additional question on POC for Genetic Studies Message-ID: <0BE6ADFAE4E7E04496BF21ABD346628005653B11@EXCHANGE1.huntingtonhospital.com> All of the responses I've received regarding what department sends out tissue for genetic studies state that Pathology sends the tissue out. We are told that the tissue needs to be sent out as soon as possible. What do you do if the tissue comes in at night or on the weekend? From gpbnas <@t> yahoo.es Thu Apr 27 14:16:37 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Thu Apr 27 14:16:42 2006 Subject: [Histonet] Re: Good TUNEL staining but very few Active Caspase-3 positive cells In-Reply-To: Message-ID: <20060427191637.96769.qmail@web26202.mail.ukl.yahoo.com> Teri, What you say seems to make perfect sense in terms of kinetics of apoptosis, but actually in the paper I found comparing both methods the result was the opposite (ie. Greater number of Active Casp-3+ than TUNEL+ cells). However the authors did not postulate any possible mechanism to explain this... Could you send more details on the pepsin digestion you used to improve sensitivity of Ab? Did you try Proteinase K treatment of samples? Thanks for your input, Guillermo "Johnson, Teri" escribi?: Guillermo, Good question! I'll take a stab at it and will read with interest any additional commentary on this. We found to get good staining with Cleaved Caspase-3 in paraffin section we needed to add a brief pepsin digestion in addition to the HIER to achieve staining. Without it we found we needed to use tyramide enhancement to see it in fluorescence. I think the antibody really does exhibit sensitivity issues. Otherwise, caspase-3 is activated early in the apoptotic process. It could be that it is no longer in its activated state in later stages of apoptosis, whereas the TUNEL will show positive across a wide range of apoptosis stages. I'm an amateur on the mechanism of this event, and welcome Dr. Cartun or anyone with more advanced knowledge of this to confirm or disagree with my conjecture. Best wishes, Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64133 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From Janet.Bonner <@t> FLHOSP.ORG Thu Apr 27 14:18:46 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Thu Apr 27 14:20:18 2006 Subject: [Histonet] POC for Genetic Studies References: <0BE6ADFAE4E7E04496BF21ABD346628005653B0D@EXCHANGE1.huntingtonhospital.com> Message-ID: All of our specimens go through the Pathology Lab Sent-Out Department after being described by the PA. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie Colbert Sent: Thu 4/27/2006 2:34 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] POC for Genetic Studies This question is for those of you who work in hospitals and send out POC tissue for genetic studies/chromosome analysis. Does the tissue get sent out directly from Labor and Delivery, or does the tissue go through Pathology and Pathology sends it out? _______________________________________________ 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, 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 KMB1904 <@t> aol.com Thu Apr 27 14:23:34 2006 From: KMB1904 <@t> aol.com (KMB1904@aol.com) Date: Thu Apr 27 14:23:50 2006 Subject: [Histonet] Expired antibodies...for immunos.. Message-ID: <400.4dfa38.31827436@aol.com> How does JCAHO handle expired antibodies if you have them...and are using them... Is it ok ....or not?? Kat From Janet.Bonner <@t> FLHOSP.ORG Thu Apr 27 14:26:46 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Thu Apr 27 14:27:34 2006 Subject: [Histonet] Additional question on POC for Genetic Studies References: <0BE6ADFAE4E7E04496BF21ABD346628005653B11@EXCHANGE1.huntingtonhospital.com> Message-ID: We refrigerate it. ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie Colbert Sent: Thu 4/27/2006 3:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Additional question on POC for Genetic Studies All of the responses I've received regarding what department sends out tissue for genetic studies state that Pathology sends the tissue out. We are told that the tissue needs to be sent out as soon as possible. What do you do if the tissue comes in at night or on the weekend? _______________________________________________ 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, 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 Bauer.Karen <@t> mayo.edu Thu Apr 27 14:41:28 2006 From: Bauer.Karen <@t> mayo.edu (Bauer, Karen) Date: Thu Apr 27 14:41:34 2006 Subject: [Histonet] Additional question on POC for Genetic Studies References: <0BE6ADFAE4E7E04496BF21ABD346628005653B11@EXCHANGE1.huntingtonhospital.com> Message-ID: We are called in. Since our histology dept. shuts down at 5:30 pm, the histologist on call would have to come in and gross the tissue and select the appropriate sample for chromosome analysis. If the specimen came to the lab at, let's say 2 am., we would instruct the night personnel to place the specimen in the histology refrigerator and we would gross it in when we arrived in the lab at 4:30 am. There are times when the OB physician demands the specimen get immediate attention, so we can be called in at any time to do them. Karen ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie Colbert Sent: Thu 4/27/2006 2:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Additional question on POC for Genetic Studies All of the responses I've received regarding what department sends out tissue for genetic studies state that Pathology sends the tissue out. We are told that the tissue needs to be sent out as soon as possible. What do you do if the tissue comes in at night or on the weekend? _______________________________________________ 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 gpbnas <@t> yahoo.es Thu Apr 27 14:44:09 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Thu Apr 27 14:44:14 2006 Subject: [Histonet] TUNEL staining In-Reply-To: <0E6BC087F70F9C47ACFF2C203D6E329CA2C341@NTRSEVS30002.s3.ms.unilever.com> Message-ID: <20060427194409.85079.qmail@web26204.mail.ukl.yahoo.com> Hi Kristofer, I used Roche In situ cell death detection kit Cat No 11 684 817 910. I modified protocol by permeabilizing paraformaldehyde-fixed frozen sections with Triton 0.5% for 10 min at RT, and reducing TdT concentration 1:10 from manufacturers recommendation. With that I was able to obtain 100% positive cells in DNase I treated samples, thus ensuring efficient permeabilization and activity of enzime. I might still be artificially detecting an excessive number of cells with these conditions, although numbers I found in mouse kidney are in agreement with what is described in literature (an indeed very low number of positive cells). As a way to increase especificity I always counterstain with DAPI, and most green TUNEL+ cells have a more whitish nucleus when seen with DAPI filter. Still you can have cells with DNA damage unrelated to apoptosis and they will be TUNEL+. Because of all this issues I wanted to confirm results with an additional technique such as Active Caspase-3, but before assuming unspecific TUNEL staining in all Casp3 negative but TUNEL+ cells, I wanted to make sure I got optimal working conditions for Anti-active Caspase-3 Ab. After optimizing fixation and permeabilization conditions to get good positive controls with DNase treated samples you can always titrate TdT concentration to obtain maximum signal to noise ratio as shown in the paper I mentioned before. If you need any further details of have any suggestions please tell me. Best regards, Guillermo "Kalleberg, Kristopher" escribi?: Guillermo, I am currently performing TUNEL on human skin post UV irradiation and have had inconclusive results. I was using the Promega TUNEL assay kit and also tried a formamide induced TUNEL assay with differing results. I also have read in the literature that the kit and most TUNEL assays are nonspecific (necrotic cells also show positive staining). The formamide induced protocol that I have only tried twice has shown no positive staining yet. I was just wondering which protocol you use for TUNEL and how certain are you that it is specific for apoptotic cells? Maybe we can help each other. Kris Kalleberg Research Associate Unilever R&D 40 Merritt Blvd. Trumbull, CT 06457 203 381 5765 _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From Don.Birgerson <@t> leica-microsystems.com Thu Apr 27 14:59:22 2006 From: Don.Birgerson <@t> leica-microsystems.com (Don.Birgerson@leica-microsystems.com) Date: Thu Apr 27 14:59:32 2006 Subject: [Histonet] double blades for rotary paraffin microtome In-Reply-To: <00D7298B3825D511BF540008C75F8A3C0CAD82E8@excluster.chsd.org> Message-ID: Hi Dr Breisch, What you are asking about was prevalent in the days before the "disposable" blade as we know it. The old A O microtomes offered a "razor blade holder" . These holders were seen around the college labs due to the cost of steel blades and the danger in handling the reusable knives. The current disposable blades are 3 or 4 times sharper than a razor blade and all of our rotary microtomes offer a holder designed and machine to fit the current microtomes. Don Birgerson Leica Microsystems Technical Assistance Center Don.Birgerson@Leica-Microsystems.Com 1-800-248-0123 ext 5918 "Breisch, Eric" To Sent by: "'histonet@lists.utsouthwestern.edu histonet-bounces@ '" lists.utsouthwest ern.edu cc Subject 04/27/2006 11:28 [Histonet] double blades for rotary AM paraffin microtome Hi Histologists, This is a generalised question regarding microtomes and blades. Has anyone ever heard of a rotary microtome that uses or used double edged blades (not unlike the razor blades currently being marketed for shaving) to cut paraffin blocks? More specifically have double edged blades ever been used to cut paraffin blocks. Your help and input is greatly appreciated. Regards, Eric A. Breisch, Ph.D. Clinical Anatomist Department of Pathology Associate Clinical Professor of Anatomy, UCSD-SOM Children's Hospital-San Diego 3020 Children's Way, MC5007 San Diego, CA 92123 Phone: 858-966-5944 _______________________________________________ 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 settembr <@t> umdnj.edu Thu Apr 27 16:14:28 2006 From: settembr <@t> umdnj.edu (Dana Settembre) Date: Thu Apr 27 16:15:23 2006 Subject: [Histonet] Expired antibodies...for immunos.. Message-ID: Hello Kat, JCAHO does not allow anything expired to be used for patients Antibodies cannot be used either, if they are expired. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> 04/27/06 3:23 PM >>> How does JCAHO handle expired antibodies if you have them...and are using them... Is it ok ....or not?? Kat _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From jnocito <@t> satx.rr.com Thu Apr 27 17:27:11 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Thu Apr 27 17:27:18 2006 Subject: [Histonet] Expired antibodies...for immunos.. References: <400.4dfa38.31827436@aol.com> Message-ID: <002a01c66a49$ba8541d0$1bb30b43@yourxhtr8hvc4p> Oh boy, I get to get flamed again. JCAHO and CAP do not let you use expired reagents, including antibodies. therefore, everyone gets to through money down the drain time and time again. I called CAP and talked to a person who got very defensive when I asked him to give the rule that says we can not use expired antibodies. He referred me to the U.S. National Register. I'm sorry, I don't have that information with right now but I'm sure someone will let me know. My beef is, if you run a known positive control with every run, you are, in essence, revalidating that antibody. Once that antibody doesn't work, replace it with one that has not expired. I know this causes a repeat and maybe a delay in reporting the results, but ask your pathologists if they would want a one day delay, or throw thousands of dollars down the sink. When I was supervising the immuno lab at the AFIP, we froze concentrated antibodies at -70 C and they still worked up to 10 years later. Now, you can't even do that. A big waste, that what it is, a big waste. Let Flaming Friday begin. Joe Nocito BS, HT(ASCP)QIHC Histology Manager Pathology Reference Lab San Antonio, TX ----- Original Message ----- From: To: Sent: Thursday, April 27, 2006 2:23 PM Subject: [Histonet] Expired antibodies...for immunos.. > How does JCAHO handle expired antibodies if you have them...and are using > them... > Is it ok ....or not?? > Kat > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > Internal Virus Database is out-of-date. > Checked by AVG Free Edition. > Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 > > From tahseen <@t> brain.net.pk Thu Apr 27 17:32:03 2006 From: tahseen <@t> brain.net.pk (Muhammad Tahseen) Date: Thu Apr 27 17:32:38 2006 Subject: [Histonet] Reference for ER,PR,HER-2/neu,p53,& EGFR Message-ID: <006801c66a4a$694e4860$2fa2fea9@p> Hello histoneters, I was wondering if any one has reference(s) of ER,PR,HER-2/neu,p53 & EGFR for metaplastic breast cancer study. Thanks in advance. Muhammad Tahseen Histology Supervisor, SKMCH&RC Lahore,Pakistan. From LuckG <@t> empirehealth.org Thu Apr 27 18:10:32 2006 From: LuckG <@t> empirehealth.org (Luck, Greg D.) Date: Thu Apr 27 18:10:42 2006 Subject: [Histonet] Additional question on POC for Genetic Studies Message-ID: Oh why not jump in, That's why every hospital based anatomic pathology dept must have an on-call pathologist and tech (e.g. the pathologist to select the appropriate tissue sample and a tech to follow-up with coordinating the transport and /or the optimal preservation of the sample) until it can be dealt with during regular hours of operation. You already should have a pathologist on call for frozens (e.g. organ harvesting) and a tech (e.g. if the tissue processor goes down). Seems straight forward to me. That's one the costs of doing business for our patients and clients in an acute care setting of health care. Greg Luck Anatomic Path Spvr Deaconess Med Cntr 800 W. 5th Ave Spokane, WA 99204 Offc 509.473.7077 Fax 509.473.7133 luckg@empirehealth.org www.deaconessmedicalcenter.org -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] Sent: Thursday, April 27, 2006 12:41 PM To: Laurie Colbert; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] Additional question on POC for Genetic Studies We are called in. Since our histology dept. shuts down at 5:30 pm, the histologist on call would have to come in and gross the tissue and select the appropriate sample for chromosome analysis. If the specimen came to the lab at, let's say 2 am., we would instruct the night personnel to place the specimen in the histology refrigerator and we would gross it in when we arrived in the lab at 4:30 am. There are times when the OB physician demands the specimen get immediate attention, so we can be called in at any time to do them. Karen ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Laurie Colbert Sent: Thu 4/27/2006 2:15 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] Additional question on POC for Genetic Studies All of the responses I've received regarding what department sends out tissue for genetic studies state that Pathology sends the tissue out. We are told that the tissue needs to be sent out as soon as possible. What do you do if the tissue comes in at night or on the weekend? _______________________________________________ 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. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From bhewlett <@t> cogeco.ca Thu Apr 27 20:15:20 2006 From: bhewlett <@t> cogeco.ca (Bryan Hewlett) Date: Thu Apr 27 20:15:29 2006 Subject: [Histonet] Expired antibodies...for immunos.. References: <400.4dfa38.31827436@aol.com> <002a01c66a49$ba8541d0$1bb30b43@yourxhtr8hvc4p> Message-ID: <002401c66a61$38564e30$6500a8c0@mainbox> Hey Joe, I'm with you! The disposal of a expensive, validated, working reagent, just because of some mindless arbitrary date rule makes absolutely no sense! I thought that we were in the era of evidence-based medicine, whatever happened? Bryan Hewlett Consultant technologist Anatomic Pathology Quality Management Program - Laboratory Services Ontario, Canada. ----- Original Message ----- From: "Joe Nocito" To: ; Sent: Thursday, April 27, 2006 6:27 PM Subject: Re: [Histonet] Expired antibodies...for immunos.. > Oh boy, > I get to get flamed again. > JCAHO and CAP do not let you use expired reagents, including antibodies. > therefore, everyone gets to through money down the drain time and time > again. > I called CAP and talked to a person who got very defensive when I asked > him to give the rule that says we can not use expired antibodies. He > referred me to the U.S. National Register. I'm sorry, I don't have that > information with right now but I'm sure someone will let me know. > My beef is, if you run a known positive control with every run, you > are, in essence, revalidating that antibody. Once that antibody doesn't > work, replace it with one that has not expired. I know this causes a > repeat and maybe a delay in reporting the results, but ask your > pathologists if they would want a one day delay, or throw thousands of > dollars down the sink. > When I was supervising the immuno lab at the AFIP, we froze > concentrated antibodies at -70 C and they still worked up to 10 years > later. Now, you can't even do that. A big waste, that what it is, a big > waste. > Let Flaming Friday begin. > > Joe Nocito BS, HT(ASCP)QIHC > Histology Manager > Pathology Reference Lab > San Antonio, TX > > ----- Original Message ----- > From: > To: > Sent: Thursday, April 27, 2006 2:23 PM > Subject: [Histonet] Expired antibodies...for immunos.. > > >> How does JCAHO handle expired antibodies if you have them...and are using >> them... >> Is it ok ....or not?? >> Kat >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> -- >> Internal Virus Database is out-of-date. >> Checked by AVG Free Edition. >> Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 >> >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > From sohail_e <@t> yahoo.com Thu Apr 27 22:41:07 2006 From: sohail_e <@t> yahoo.com (sohail ejaz) Date: Thu Apr 27 22:41:13 2006 Subject: [Histonet] Protocol for MOM kit ( whole mount staining) Message-ID: <20060428034107.84674.qmail@web30606.mail.mud.yahoo.com> I am trying whole mount staining of mouse Retina, Trachea and 20 micrometer tissues with vWF (Rabbit anti human von Willibrand factor polyclonal antibody, Chemicon international) SMA (Monoclonal anti-actin, alpha smooth muscle antibody produced in mouse) and i need to use MOM kit (Vector) for this purpose. They recommend to incubate the samples for just 30 min with primary antibody. I wonder if this time is enough..................?? Can anyone guide me a good protocol regarding this issue. Thanks Dr. Sohail --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From c.m.vanderloos <@t> amc.uva.nl Fri Apr 28 01:47:25 2006 From: c.m.vanderloos <@t> amc.uva.nl (C.M. van der Loos) Date: Fri Apr 28 01:47:42 2006 Subject: [Histonet] CD3 antibody for FFPE rabbit tissue Message-ID: <33f60f33f96f.33f96f33f60f@amc.uva.nl> Dear Narinder, It's a wild guess and haven't tried it, but perhaps worthwhile to test (....and better than nothing): The LabVision CD3 rabbit monoclonal antibody (clone SP7) reacts very well with a lot of different species (human, rat, mouse), so to my opinion there is a good chance it reacts with rabbit CD3 as well. Just test it on some lymphoid rabbit tissue and look through heavy background staining (we have a rabbit-on-rabbit situation here and you will observe a lot of plasmacells and staining near vessels!) whether or not T-cells are positive. If this is the case you need the Zenon (Molecular Probes/Invitrogen) rabbit-on-rabbit kit for proper visualization. Please let us know if this approach works for you. Good luck! Chris van der Loos, PhD Dept. of Pathology Academic Medical Center M2-230 Meibergdreef 9 NL-1105 AZ Amsterdam The Netherlands Date: Thu, 27 Apr 2006 12:02:53 -0700 From: NDhatt@qltinc.com Subject: [Histonet] CD3 antibody for FFPE rabbit tissue To: histonetlistsutsouthwesternedu Hi All, I am looking for a CD3 antibody to use on FFPE rabbit tissue. Most antibodies I have looked into are tested in human. Thanks Narinder Dhatt Preclinical - Histology QLT Inc. Vancouver, BC Canada From lpwenk <@t> sbcglobal.net Fri Apr 28 04:48:24 2006 From: lpwenk <@t> sbcglobal.net (Lee & Peggy Wenk) Date: Fri Apr 28 04:50:29 2006 Subject: [Histonet] Expired antibodies...for immunos.. In-Reply-To: <400.4dfa38.31827436@aol.com> Message-ID: <005d01c66aa8$e4cfa020$ac022643@HPPav2> JCAHO, CAP, etc. all refer labs back to the CMS (Center for Medicare and Medicaid Services) and HHS (Health and Human Services), which go back to the Federal government regulations, as laid out in 42CFR 493 (Code of Federal Regulations - Laboratory Requirements): Specifically, 42CFR 493.1252(d) (1252 = Standard on Tests systems, Equipment, Instruments, Reagents, Materials, and Supplies) (d) = "Reagents, solutions, culture media, control materials, calibration materials, and other supplies must not be used when they have exceeded their expiration date, have deteriorated, or are of substandard quality." http://www.access.gpo.gov/nara/cfr/waisidx_04/42cfr493_04.html (Hope the link works.) Sorry, there's no exception for costly IHC or ISH solutions that cost a lot of money and that we know we can "prove" are still good. The only thing I can see us histotechs doing is persuading the manufacturers to sell the reagents in smaller quantities. (Which they already have, over the years.) Peggy A. Wenk, HTL(ASCP)SLS William Beaumont Hospital Royal Oak, MI 48073 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of KMB1904@aol.com Sent: Thursday, April 27, 2006 3:24 PM To: histonet@pathology.swmed.edu Subject: [Histonet] Expired antibodies...for immunos.. How does JCAHO handle expired antibodies if you have them...and are using them... Is it ok ....or not?? Kat _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From JCarpenter764 <@t> aol.com Fri Apr 28 06:45:14 2006 From: JCarpenter764 <@t> aol.com (JCarpenter764@aol.com) Date: Fri Apr 28 06:45:30 2006 Subject: [Histonet] NC NCSHT meeting.... Message-ID: <2f9.4187ff5.31835a4a@aol.com> Goodmorning, I was wondering if anyone could tell me the company who manufactures the IHC machine that can run three different runs at one time. This machine was absolutely wonderful. It had three different slots for slides that held 10 a piece and I can not find the brochure I picked up..of course. It also had little slide covers that fit over the slide in the machine. If anyone can help I would greatly appreciate it. Thanks a bunch Jennell From mikael.niku <@t> helsinki.fi Fri Apr 28 07:14:28 2006 From: mikael.niku <@t> helsinki.fi (Mikael Niku) Date: Fri Apr 28 07:14:37 2006 Subject: [Histonet] Fixing cells on coverslips for immunos Message-ID: <44520724.8030506@helsinki.fi> Dear Histonetters, a basic question: how to make cultured cells stay on coverslips for immufluorescence staining? We are culturing cells on lysine-coated glass coverslips and trying to immunostain them. We know the antibody works with acetone fixation and probably not with PFA (at least not in the case of tissue sections). The problem is, most of cells drop off during the staining. What can we do? I have done mainly tissue sections, so I'm in trouble with this one. Currently we have tried like this: cells were washed with PBS, transferred to acetone for 10 min at +4C, back to PBS, followed with a very basic indirect immunofluorescence staining (with 30-minute antibody incubations, whole procedure within one day), and Faramount embedding. This works well for cytospin preps (the difference is that the cytospins, of course, are dried before the fixation; and we use Superfrost slides for them). Is it possible (or even common) that an antibody might work with cell culture samples using a quick PFA (paraformaldehyde) fixation, although it only works with non-crosslinking fixatives for tissue sections? With best regards, Mikael -- //////////////////////////////////////////////////////////// Mikael Niku URL: www.helsinki.fi/~mniku/ University of Helsinki Dept. Basic Veterinary Sciences - Mit?k? mielt? olen l?nsimaisesta sivistyksest?? Minusta se olisi erinomainen ajatus! - Gandhi //////////////////////////////////////////////////////////// From shive003 <@t> umn.edu Fri Apr 28 09:39:33 2006 From: shive003 <@t> umn.edu (Jan Shivers) Date: Fri Apr 28 09:39:47 2006 Subject: [Histonet] Expired antibodies...for immunos.. References: <005d01c66aa8$e4cfa020$ac022643@HPPav2> Message-ID: <002e01c66ad1$90f0d810$a1065486@auxs.umn.edu> And let us not forget that it's the antibody manufacturers who set these expiration dates. Is there an actual science behind determining an antibody's 'lifespan', or is it more of a business profit decision? It's certainly cost-inefficient for a lab to pay upwards of $500 for an antibody which you only use a few times before its expiration date hits, yet the doctors/clients want these tests available. If we can't change the wording on the rules and regulations in regards to using antibodies past their expiration date, despite proof of their continued reactivity... then we should encourage the manufacturers to set more reasonable expiration dates.... or sell in smaller quantities, as Peggy stated below. Jan Shivers ----- Original Message ----- From: "Lee & Peggy Wenk" To: ; Sent: Friday, April 28, 2006 4:48 AM Subject: RE: [Histonet] Expired antibodies...for immunos.. > JCAHO, CAP, etc. all refer labs back to the CMS (Center for Medicare and > Medicaid Services) and HHS (Health and Human Services), which go back to > the > Federal government regulations, as laid out in 42CFR 493 (Code of Federal > Regulations - Laboratory Requirements): > > Specifically, 42CFR 493.1252(d) > (1252 = Standard on Tests systems, Equipment, Instruments, Reagents, > Materials, and Supplies) > > (d) = "Reagents, solutions, culture media, control materials, calibration > materials, and other supplies must not be used when they have exceeded > their > expiration date, have deteriorated, or are of substandard quality." > > http://www.access.gpo.gov/nara/cfr/waisidx_04/42cfr493_04.html > > (Hope the link works.) > > Sorry, there's no exception for costly IHC or ISH solutions that cost a > lot > of money and that we know we can "prove" are still good. > > The only thing I can see us histotechs doing is persuading the > manufacturers > to sell the reagents in smaller quantities. (Which they already have, over > the years.) > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > KMB1904@aol.com > Sent: Thursday, April 27, 2006 3:24 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Expired antibodies...for immunos.. > > How does JCAHO handle expired antibodies if you have them...and are using > them... > Is it ok ....or not?? > Kat > _______________________________________________ > Histonet mailing list > Histonet@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 gayle.brosnanwatters <@t> sru.edu Fri Apr 28 09:43:52 2006 From: gayle.brosnanwatters <@t> sru.edu (gayle brosnanwatters) Date: Fri Apr 28 09:44:09 2006 Subject: [Histonet] where can I send my student? Message-ID: <8172F14C39FCAF4E99E266F6756A368307412CD4@msfexch01.srunet.sruad.edu> Hi, folks, I'm back after a long absence, and at a new address. Because I am only a psychologist, I have no idea where to send my student who really wants to spend her life in a research laboratory doing histology! She is currently graduating from Slippery Rock University with a degree in psychology, but I have sent her to the chemistry department and she has taken chemistry and is now taking two organic chemistry classes as a post bac in order to get herself somewhat more ready to do what she has discovered she loves to do. She has spent the last four years here with me doing work with mice, and is currently sectioning mouse brains on my ancient ultramicrotome and using a simple Richardson stain. She loves working at the scope counting injured cells! So, where can she go for additional training? Is this something she needs a masters to do, or are there training programs available that will teach her immuno and fluorescent and all those other things that I've heard of but have no idea how to do? She's a fantastic young woman who washed mouse cages for me for over a year when we had no cage washer, and is very dedicated and willing to do the "grunt work" that others disdain. Any suggestions? Gayle Gayle L. Brosnan-Watters, PhD Assistant Professor Dept of Psychology Treasurer, Faculty for Undergraduate Neuroscience 226 Vincent Science Hall Slippery Rock University Slippery Rock, PA 16057 gayle.brosnanwatters@sru.edu 724-738-2529 - office 724-738-4807 - fax From JWEEMS <@t> sjha.org Fri Apr 28 09:45:19 2006 From: JWEEMS <@t> sjha.org (Weems, Joyce) Date: Fri Apr 28 09:45:26 2006 Subject: [Histonet] Expired antibodies...for immunos.. Message-ID: <1CD6831EB9B26D45B0A3EAA79F7EBD3202637CD7@sjhaexc02.sjha.org> (d) = "Reagents, solutions, culture media, control materials, calibration materials, and other supplies must not be used when they have exceeded their expiration date, have deteriorated, or are of substandard quality." I'm wondering if this phrase could be interpreted to be ONE of the three reasons listed because the conjunction is OR. This could be proven and documented by reasons given earlier. Joyce Joyce Weems Pathology Manager Saint Joseph's Hospital of Atlanta 404-851-7376 404-851-7831 - fax -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Jan Shivers Sent: Friday, April 28, 2006 10:40 AM To: histonet Subject: Re: [Histonet] Expired antibodies...for immunos.. And let us not forget that it's the antibody manufacturers who set these expiration dates. Is there an actual science behind determining an antibody's 'lifespan', or is it more of a business profit decision? It's certainly cost-inefficient for a lab to pay upwards of $500 for an antibody which you only use a few times before its expiration date hits, yet the doctors/clients want these tests available. If we can't change the wording on the rules and regulations in regards to using antibodies past their expiration date, despite proof of their continued reactivity... then we should encourage the manufacturers to set more reasonable expiration dates.... or sell in smaller quantities, as Peggy stated below. Jan Shivers ----- Original Message ----- From: "Lee & Peggy Wenk" To: ; Sent: Friday, April 28, 2006 4:48 AM Subject: RE: [Histonet] Expired antibodies...for immunos.. > JCAHO, CAP, etc. all refer labs back to the CMS (Center for Medicare and > Medicaid Services) and HHS (Health and Human Services), which go back to > the > Federal government regulations, as laid out in 42CFR 493 (Code of Federal > Regulations - Laboratory Requirements): > > Specifically, 42CFR 493.1252(d) > (1252 = Standard on Tests systems, Equipment, Instruments, Reagents, > Materials, and Supplies) > > (d) = "Reagents, solutions, culture media, control materials, calibration > materials, and other supplies must not be used when they have exceeded > their > expiration date, have deteriorated, or are of substandard quality." > > http://www.access.gpo.gov/nara/cfr/waisidx_04/42cfr493_04.html > > (Hope the link works.) > > Sorry, there's no exception for costly IHC or ISH solutions that cost a > lot > of money and that we know we can "prove" are still good. > > The only thing I can see us histotechs doing is persuading the > manufacturers > to sell the reagents in smaller quantities. (Which they already have, over > the years.) > > Peggy A. Wenk, HTL(ASCP)SLS > William Beaumont Hospital > Royal Oak, MI 48073 > > > > -----Original Message----- > From: histonet-bounces@lists.utsouthwestern.edu > [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of > KMB1904@aol.com > Sent: Thursday, April 27, 2006 3:24 PM > To: histonet@pathology.swmed.edu > Subject: [Histonet] Expired antibodies...for immunos.. > > How does JCAHO handle expired antibodies if you have them...and are using > them... > Is it ok ....or not?? > Kat > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet Confidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you. Saint Joseph's Health System, Inc. From gpbnas <@t> yahoo.es Fri Apr 28 10:02:30 2006 From: gpbnas <@t> yahoo.es (Guillermo Palao) Date: Fri Apr 28 10:02:36 2006 Subject: [Histonet] Protocol for MOM kit ( whole mount staining) In-Reply-To: <20060428034107.84674.qmail@web30606.mail.mud.yahoo.com> Message-ID: <20060428150230.33266.qmail@web26202.mail.ukl.yahoo.com> In my experience trying to block deposited endogenous mouse IgG in mouse kidney from being detected with secondary anti-mouse Abs MOM kit did not work. I tried increasing concentration of blocking reagent up to 6 times what they recommended together with increased blocking times, without result. Primary Ab incubation was not important because the Ab was quite good and it worked in 1 h RT or O/N 4 ?C. Of course your level of background will depend on the amount of endogenous IgG in your mouse samples. I have now changed to rabbit mAbs with Jackson?s secondary Abs pre-adsorbed with mouse Ags. Best regards, Guillermo sohail ejaz escribi?: I am trying whole mount staining of mouse Retina, Trachea and 20 micrometer tissues with vWF (Rabbit anti human von Willibrand factor polyclonal antibody, Chemicon international) SMA (Monoclonal anti-actin, alpha smooth muscle antibody produced in mouse) and i need to use MOM kit (Vector) for this purpose. They recommend to incubate the samples for just 30 min with primary antibody. I wonder if this time is enough..................?? Can anyone guide me a good protocol regarding this issue. Thanks Dr. Sohail --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- LLama Gratis a cualquier PC del Mundo. Llamadas a fijos y m?viles desde 1 c?ntimo por minuto. http://es.voice.yahoo.com From Janet.Bonner <@t> FLHOSP.ORG Fri Apr 28 10:30:49 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Apr 28 10:31:05 2006 Subject: [Histonet] where can I send my student? References: <8172F14C39FCAF4E99E266F6756A368307412CD4@msfexch01.srunet.sruad.edu> Message-ID: Well, she initially majored in the correct field of study to enter Histology. A BS or MS is desirable with majors in Biology/Chemistry. There are assorted schools/courses of study for Histology scattered about the US. One is in Jacksonville, FL and we have one here at Florida Hospital although we only take five students per year (and this coming year was just selected). Indiana University also has a good program, which is the program we have connected to to teach our students. A search online should help as well. Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of gayle brosnanwatters Sent: Fri 4/28/2006 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] where can I send my student? Hi, folks, I'm back after a long absence, and at a new address. Because I am only a psychologist, I have no idea where to send my student who really wants to spend her life in a research laboratory doing histology! She is currently graduating from Slippery Rock University with a degree in psychology, but I have sent her to the chemistry department and she has taken chemistry and is now taking two organic chemistry classes as a post bac in order to get herself somewhat more ready to do what she has discovered she loves to do. She has spent the last four years here with me doing work with mice, and is currently sectioning mouse brains on my ancient ultramicrotome and using a simple Richardson stain. She loves working at the scope counting injured cells! So, where can she go for additional training? Is this something she needs a masters to do, or are there training programs available that will teach her immuno and fluorescent and all those other things that I've heard of but have no idea how to do? She's a fantastic young woman who washed mouse cages for me for over a year when we had no cage washer, and is very dedicated and willing to do the "grunt work" that others disdain. Any suggestions? Gayle Gayle L. Brosnan-Watters, PhD Assistant Professor Dept of Psychology Treasurer, Faculty for Undergraduate Neuroscience 226 Vincent Science Hall Slippery Rock University Slippery Rock, PA 16057 gayle.brosnanwatters@sru.edu 724-738-2529 - office 724-738-4807 - fax _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From gcallis <@t> montana.edu Fri Apr 28 11:14:51 2006 From: gcallis <@t> montana.edu (Gayle Callis) Date: Fri Apr 28 11:15:02 2006 Subject: [Histonet] CD3 antibody for FFPE rabbit tissue In-Reply-To: <33f60f33f96f.33f96f33f60f@amc.uva.nl> References: <33f60f33f96f.33f96f33f60f@amc.uva.nl> Message-ID: <6.0.0.22.1.20060428101055.01b4ec88@gemini.msu.montana.edu> Can this antibody be purchased biotinylated? If so, then you can work with it in the same way as mouse on mouse kits with rabbit serum instead of mouse serum added, not so much fun. However, there was a publication in Lab Vision, a now defunct journal and this method is also in Gu's Analytical Morphology book, from Biotechniques for doing rabbit on rabbit IHC. The Xenon technology sounds much easier and readily available. At 12:47 AM 4/28/2006, you wrote: > Dear Narinder, > > It's a wild guess and haven't tried it, but perhaps worthwhile to test > (....and better than nothing): > > The LabVision CD3 rabbit monoclonal antibody (clone SP7) reacts very > well with a lot of different species (human, rat, mouse), so to my > opinion there is a good chance it reacts with rabbit CD3 as well. > Just test it on some lymphoid rabbit tissue and look through heavy > background staining (we have a rabbit-on-rabbit situation here and you > will observe a lot of plasmacells and staining near vessels!) whether > or not T-cells are positive. If this is the case you need the Zenon > (Molecular Probes/Invitrogen) rabbit-on-rabbit kit for proper > visualization. Please let us know if this approach works for you. > > Good luck! > > Chris van der Loos, PhD > Dept. of Pathology > Academic Medical Center M2-230 > Meibergdreef 9 > NL-1105 AZ Amsterdam > The Netherlands > > > Date: Thu, 27 Apr 2006 12:02:53 -0700 > From: NDhatt@qltinc.com > Subject: [Histonet] CD3 antibody for FFPE rabbit tissue > To: histonetlistsutsouthwesternedu > Hi All, > I am looking for a CD3 antibody to use on FFPE rabbit tissue. Most > antibodies I have looked into are tested in human. > Thanks > Narinder Dhatt > Preclinical - Histology > QLT Inc. > Vancouver, BC > Canada >_______________________________________________ >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 406 994-4303 (FAX) From rjbuesa <@t> yahoo.com Fri Apr 28 11:23:53 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Apr 28 11:23:59 2006 Subject: [Histonet] Expired antibodies...for immunos.. In-Reply-To: Message-ID: <20060428162353.25906.qmail@web61225.mail.yahoo.com> Hi all: I want to jump into this thread that has already been a subject of discussion on November of last year. 1- I had never this expriration problem because our IHC volume was large, but is is something a great concern to small laboratories, some of which have opted not to do IHC and send it to a reference lab. 2- The experiration date is set by the manufacturer, but no manufacturer says which studies are behing their dating system. 3- CAP has now stablished a rigid rule about not allowing "expired" Abs to be used BUT the same CAP in thei Laboratory Improvement Programs published in 1998 the following: Tubbs et al. (1998): "Extension of Useful Reagent Shelf Life Beyond Manufacturer's Recommendations" Arch Pathol Lab Med. 122: 1051-52 and concluded: "The data suggest review of the Health Care Financing Administration's ruling on extending the useful reagent shelf life beyond manufacturers recommendations. Similar studies using more inherently quantitative methodology are suggested" (sic.) 4- The next year Balaton et al. published a paper under the title of: "Satisfactory performance of primary antibodies beyond manufacturers' recommended expiration dates" Applied Immunohistochemistry & Molecular Morphology 7(3):221-25 They tested 65 antibodies that had expired 6 to 134 months previously (average of 33 months) and concluded that"....These results indicate that primary antibodies have a shelf life significantly longer than the one specified by the manufacturers...."(sic.). 5- It is worth noting that both papers stress the fact that those expiration dates are manufacturers' recommendations, they are not written on stone. The one now doing the writing on stone is CAP after sponsoring a contradictory study. So, when you decide to flame Joe Nocito, please leave a space for me. Joe's suggestion seems to be very reasonable. Another thing, please use cajun sauce for my roasting. Best wishes Ren? J. Dana Settembre wrote: Hello Kat, JCAHO does not allow anything expired to be used for patients Antibodies cannot be used either, if they are expired. Dana Settembre University Hospital-UMDNJ Newark, NJ >>> 04/27/06 3:23 PM >>> How does JCAHO handle expired antibodies if you have them...and are using them... Is it ok ....or not?? Kat _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Get amazing travel prices for air and hotel in one click on Yahoo! FareChase From weneng2004 <@t> yahoo.com Fri Apr 28 11:24:42 2006 From: weneng2004 <@t> yahoo.com (wen eng) Date: Fri Apr 28 11:24:47 2006 Subject: [Histonet] slides shipping Message-ID: <20060428162442.86148.qmail@web53406.mail.yahoo.com> Hello histonetter, How do you ship the paraffin slides or frozen section slides? Dry ice, white ice or room temperature? Thanks, Wen --------------------------------- Blab-away for as little as 1?/min. Make PC-to-Phone Calls using Yahoo! Messenger with Voice. From rjbuesa <@t> yahoo.com Fri Apr 28 11:26:47 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Fri Apr 28 11:26:50 2006 Subject: [Histonet] where can I send my student? In-Reply-To: Message-ID: <20060428162647.31482.qmail@web61213.mail.yahoo.com> Janet: Barry University and Miami Dade College, both in Miami, have reputable histology courses. Ren? J. "Bonner, Janet" wrote: Well, she initially majored in the correct field of study to enter Histology. A BS or MS is desirable with majors in Biology/Chemistry. There are assorted schools/courses of study for Histology scattered about the US. One is in Jacksonville, FL and we have one here at Florida Hospital although we only take five students per year (and this coming year was just selected). Indiana University also has a good program, which is the program we have connected to to teach our students. A search online should help as well. Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of gayle brosnanwatters Sent: Fri 4/28/2006 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] where can I send my student? Hi, folks, I'm back after a long absence, and at a new address. Because I am only a psychologist, I have no idea where to send my student who really wants to spend her life in a research laboratory doing histology! She is currently graduating from Slippery Rock University with a degree in psychology, but I have sent her to the chemistry department and she has taken chemistry and is now taking two organic chemistry classes as a post bac in order to get herself somewhat more ready to do what she has discovered she loves to do. She has spent the last four years here with me doing work with mice, and is currently sectioning mouse brains on my ancient ultramicrotome and using a simple Richardson stain. She loves working at the scope counting injured cells! So, where can she go for additional training? Is this something she needs a masters to do, or are there training programs available that will teach her immuno and fluorescent and all those other things that I've heard of but have no idea how to do? She's a fantastic young woman who washed mouse cages for me for over a year when we had no cage washer, and is very dedicated and willing to do the "grunt work" that others disdain. Any suggestions? Gayle Gayle L. Brosnan-Watters, PhD Assistant Professor Dept of Psychology Treasurer, Faculty for Undergraduate Neuroscience 226 Vincent Science Hall Slippery Rock University Slippery Rock, PA 16057 gayle.brosnanwatters@sru.edu 724-738-2529 - office 724-738-4807 - 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 --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. From jqb7 <@t> cdc.gov Fri Apr 28 11:40:02 2006 From: jqb7 <@t> cdc.gov (Bartlett, Jeanine (CDC/NCID/VR)) Date: Fri Apr 28 11:43:19 2006 Subject: [Histonet] where can I send my student? Message-ID: www.nsh.org lists approved programs for schools of histotechnology. Jeanine Bartlett, BS, HT(ASCP) Centers for Disease Control and Prevention 1600 Clifton Road, MS/G-32 18/SB-114 Atlanta, GA 30333 (404) 639-3590 jeanine.bartlett@cdc.hhs.gov -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Rene J Buesa Sent: Friday, April 28, 2006 12:27 PM To: Bonner, Janet; gayle brosnanwatters; histonet@lists.utsouthwestern.edu Subject: RE: [Histonet] where can I send my student? Janet: Barry University and Miami Dade College, both in Miami, have reputable histology courses. Ren? J. "Bonner, Janet" wrote: Well, she initially majored in the correct field of study to enter Histology. A BS or MS is desirable with majors in Biology/Chemistry. There are assorted schools/courses of study for Histology scattered about the US. One is in Jacksonville, FL and we have one here at Florida Hospital although we only take five students per year (and this coming year was just selected). Indiana University also has a good program, which is the program we have connected to to teach our students. A search online should help as well. Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of gayle brosnanwatters Sent: Fri 4/28/2006 10:43 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] where can I send my student? Hi, folks, I'm back after a long absence, and at a new address. Because I am only a psychologist, I have no idea where to send my student who really wants to spend her life in a research laboratory doing histology! She is currently graduating from Slippery Rock University with a degree in psychology, but I have sent her to the chemistry department and she has taken chemistry and is now taking two organic chemistry classes as a post bac in order to get herself somewhat more ready to do what she has discovered she loves to do. She has spent the last four years here with me doing work with mice, and is currently sectioning mouse brains on my ancient ultramicrotome and using a simple Richardson stain. She loves working at the scope counting injured cells! So, where can she go for additional training? Is this something she needs a masters to do, or are there training programs available that will teach her immuno and fluorescent and all those other things that I've heard of but have no idea how to do? She's a fantastic young woman who washed mouse cages for me for over a year when we had no cage washer, and is very dedicated and willing to do the "grunt work" that others disdain. Any suggestions? Gayle Gayle L. Brosnan-Watters, PhD Assistant Professor Dept of Psychology Treasurer, Faculty for Undergraduate Neuroscience 226 Vincent Science Hall Slippery Rock University Slippery Rock, PA 16057 gayle.brosnanwatters@sru.edu 724-738-2529 - office 724-738-4807 - 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 --------------------------------- Yahoo! Messenger with Voice. PC-to-Phone calls for ridiculously low rates. _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From micro <@t> superlink.net Fri Apr 28 11:45:42 2006 From: micro <@t> superlink.net (Markus F. Meyenhofer) Date: Fri Apr 28 11:46:05 2006 Subject: [Histonet] Expired antibodies...for immunos.. References: <005d01c66aa8$e4cfa020$ac022643@HPPav2> <002e01c66ad1$90f0d810$a1065486@auxs.umn.edu> Message-ID: <00f101c66ae3$31841ce0$bc893cd1@DJ4VDH31> The same goes for lab chemicals! Like Sodium Chloride, after millions of years in the ground, it will expire in a few years! That's when bureaucrats with no technical ideas and common sense make rules! Markus F. Meyenhofer ----- Original Message ----- From: "Jan Shivers" To: "histonet" Sent: Friday, April 28, 2006 10:39 AM Subject: Re: [Histonet] Expired antibodies...for immunos.. > And let us not forget that it's the antibody manufacturers who set these > expiration dates. Is there an actual science behind determining an > antibody's 'lifespan', or is it more of a business profit decision? It's > certainly cost-inefficient for a lab to pay upwards of $500 for an > antibody which you only use a few times before its expiration date hits, > yet the doctors/clients want these tests available. > > If we can't change the wording on the rules and regulations in regards to > using antibodies past their expiration date, despite proof of their > continued reactivity... then we should encourage the manufacturers to set > more reasonable expiration dates.... or sell in smaller quantities, as > Peggy stated below. > > Jan Shivers > > > ----- Original Message ----- > From: "Lee & Peggy Wenk" > To: ; > Sent: Friday, April 28, 2006 4:48 AM > Subject: RE: [Histonet] Expired antibodies...for immunos.. > > >> JCAHO, CAP, etc. all refer labs back to the CMS (Center for Medicare and >> Medicaid Services) and HHS (Health and Human Services), which go back to >> the >> Federal government regulations, as laid out in 42CFR 493 (Code of Federal >> Regulations - Laboratory Requirements): >> >> Specifically, 42CFR 493.1252(d) >> (1252 = Standard on Tests systems, Equipment, Instruments, Reagents, >> Materials, and Supplies) >> >> (d) = "Reagents, solutions, culture media, control materials, calibration >> materials, and other supplies must not be used when they have exceeded >> their >> expiration date, have deteriorated, or are of substandard quality." >> >> http://www.access.gpo.gov/nara/cfr/waisidx_04/42cfr493_04.html >> >> (Hope the link works.) >> >> Sorry, there's no exception for costly IHC or ISH solutions that cost a >> lot >> of money and that we know we can "prove" are still good. >> >> The only thing I can see us histotechs doing is persuading the >> manufacturers >> to sell the reagents in smaller quantities. (Which they already have, >> over >> the years.) >> >> Peggy A. Wenk, HTL(ASCP)SLS >> William Beaumont Hospital >> Royal Oak, MI 48073 >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> KMB1904@aol.com >> Sent: Thursday, April 27, 2006 3:24 PM >> To: histonet@pathology.swmed.edu >> Subject: [Histonet] Expired antibodies...for immunos.. >> >> How does JCAHO handle expired antibodies if you have them...and are using >> them... >> Is it ok ....or not?? >> Kat >> _______________________________________________ >> Histonet mailing list >> Histonet@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 michael.owen <@t> fda.hhs.gov Fri Apr 28 11:50:24 2006 From: michael.owen <@t> fda.hhs.gov (Owen, Michael P) Date: Fri Apr 28 11:51:13 2006 Subject: [Histonet] Cleaning Methods for Dye Solutions Message-ID: Dear List Members, Is there a current list of methods to use for cleaning dye solutions from glassware and on countertops? Thank you in advance. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov From ploykasek <@t> phenopath.com Fri Apr 28 12:19:12 2006 From: ploykasek <@t> phenopath.com (Patti Loykasek) Date: Fri Apr 28 12:19:39 2006 Subject: [Histonet] where can I send my student? In-Reply-To: <8172F14C39FCAF4E99E266F6756A368307412CD4@msfexch01.srunet.sruad.edu> Message-ID: There is a new program starting up in Sept. near Seattle, WA. The program will be at Bates Technical College just south of Tacoma, WA. By the way, they are accepting applications for an instructor. Patti Loykasek BS, HTL, QIHC PhenoPath Laboratories Seattle, WA > Hi, folks, > > I'm back after a long absence, and at a new address. > Because I am only a psychologist, I have no idea where to send my > student who really wants to spend her life in a research laboratory > doing histology! She is currently graduating from Slippery Rock > University with a degree in psychology, but I have sent her to the > chemistry department and she has taken chemistry and is now taking two > organic chemistry classes as a post bac in order to get herself > somewhat more ready to do what she has discovered she loves to do. She > has spent the last four years here with me doing work with mice, and is > currently sectioning mouse brains on my ancient ultramicrotome and using > a simple Richardson stain. She loves working at the scope counting > injured cells! > > So, where can she go for additional training? Is this > something she needs a masters to do, or are there training programs > available that will teach her immuno and fluorescent and all those other > things that I've heard of but have no idea how to do? She's a fantastic > young woman who washed mouse cages for me for over a year when we had no > cage washer, and is very dedicated and willing to do the "grunt work" > that others disdain. Any suggestions? > > Gayle > > > > Gayle L. Brosnan-Watters, PhD > > Assistant Professor > > Dept of Psychology > > Treasurer, Faculty for Undergraduate Neuroscience > > 226 Vincent Science Hall > > Slippery Rock University > > Slippery Rock, PA 16057 > > gayle.brosnanwatters@sru.edu > > 724-738-2529 - office > > 724-738-4807 - fax > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet ------------------------------------------------------------------------- This e-mail message, including any attachments, is for the sole use of the intended recipients and may contain privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by e-mail and destroy all copies of the original message, or you may call PhenoPath Laboratories, Seattle, WA U.S.A. at (206) 374-9000. From Janet.Bonner <@t> FLHOSP.ORG Fri Apr 28 12:28:03 2006 From: Janet.Bonner <@t> FLHOSP.ORG (Bonner, Janet) Date: Fri Apr 28 12:28:28 2006 Subject: [Histonet] Cleaning Methods for Dye Solutions References: Message-ID: Alconox, bleach, eradosol, are good general cleaners. Tarn-ex (found at your local Walgreens or CVS) removes silver stains (silver nitrate-GMS). Micro stains (carbol fuchsin AFB), especially, are removed for the most part by 1% Acid Alcohol(1% HCl in 70% EtOH). Some dishwashing liquids also are good cleaners. Janet ________________________________ From: histonet-bounces@lists.utsouthwestern.edu on behalf of Owen, Michael P Sent: Fri 4/28/2006 12:50 PM To: 'Histonet' Subject: [Histonet] Cleaning Methods for Dye Solutions Dear List Members, Is there a current list of methods to use for cleaning dye solutions from glassware and on countertops? Thank you in advance. Michael P. Owen, Regulatory Microbiologist U.S. FDA Pacific Regional Lab Northwest 22201 23rd Drive SE Bothell, WA 98021-4421 Phone: 425-483-4865 E-Mail: michael.owen@fda.hhs.gov _______________________________________________ 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, 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 Michael.Rice <@t> holy-cross.com Fri Apr 28 12:39:59 2006 From: Michael.Rice <@t> holy-cross.com (Rice, Michael) Date: Fri Apr 28 12:40:22 2006 Subject: [Histonet] meditech vs copath advise Message-ID: <3BC92F29BE821745AB15E04C98EE028D6D392B@HCH2KMAIL.holy-cross.com> Hi all Many thanks to all of you out there who supplied me with the information to help keep Copath in my laboratory. A decision will be made next week and I am very hopeful that administration will take your input to heart. The best comment that I received back was: "run as fast as you can" Thanks again Mike Michael Rice CT.HT(ASCP) Supervisor Of Pathology Holy Cross Hospital Ft Lauderdale, Fl 33308 954.776.3070 ====================== Confidentiality Notice: Prepared in Anticipation of Litigation. Attorney-Client Privileged. This e-mail message, including any attachments, is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply e-mail and destroy all copies of the original. ====================== From JMacDonald <@t> mtsac.edu Fri Apr 28 14:22:52 2006 From: JMacDonald <@t> mtsac.edu (Jennifer MacDonald) Date: Fri Apr 28 14:23:16 2006 Subject: [Histonet] where to send students Message-ID: Harford Community College has an online program. Here for the information. [1]http://www.harford.edu/Department/CBS/histotech/onlineprogram.as Indiana University has a program w online. Here is the URL for that prog [2]http://bulletin.iupui.edu/200 References 1. 3D"http://www.harford.ed=/ 2. 3D"http://bulletin.iupui.edu/2004-html/medicine/his From jnocito <@t> pathreflab.com Fri Apr 28 14:27:12 2006 From: jnocito <@t> pathreflab.com (Joe Nocito) Date: Fri Apr 28 14:24:54 2006 Subject: [Histonet] Expired antibodies...for immunos.. In-Reply-To: <00f101c66ae3$31841ce0$bc893cd1@DJ4VDH31> Message-ID: Hear, hear Markus -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Markus F. Meyenhofer Sent: Friday, April 28, 2006 11:46 AM To: Jan Shivers; histonet Subject: Re: [Histonet] Expired antibodies...for immunos.. The same goes for lab chemicals! Like Sodium Chloride, after millions of years in the ground, it will expire in a few years! That's when bureaucrats with no technical ideas and common sense make rules! Markus F. Meyenhofer ----- Original Message ----- From: "Jan Shivers" To: "histonet" Sent: Friday, April 28, 2006 10:39 AM Subject: Re: [Histonet] Expired antibodies...for immunos.. > And let us not forget that it's the antibody manufacturers who set these > expiration dates. Is there an actual science behind determining an > antibody's 'lifespan', or is it more of a business profit decision? It's > certainly cost-inefficient for a lab to pay upwards of $500 for an > antibody which you only use a few times before its expiration date hits, > yet the doctors/clients want these tests available. > > If we can't change the wording on the rules and regulations in regards to > using antibodies past their expiration date, despite proof of their > continued reactivity... then we should encourage the manufacturers to set > more reasonable expiration dates.... or sell in smaller quantities, as > Peggy stated below. > > Jan Shivers > > > ----- Original Message ----- > From: "Lee & Peggy Wenk" > To: ; > Sent: Friday, April 28, 2006 4:48 AM > Subject: RE: [Histonet] Expired antibodies...for immunos.. > > >> JCAHO, CAP, etc. all refer labs back to the CMS (Center for Medicare and >> Medicaid Services) and HHS (Health and Human Services), which go back to >> the >> Federal government regulations, as laid out in 42CFR 493 (Code of Federal >> Regulations - Laboratory Requirements): >> >> Specifically, 42CFR 493.1252(d) >> (1252 = Standard on Tests systems, Equipment, Instruments, Reagents, >> Materials, and Supplies) >> >> (d) = "Reagents, solutions, culture media, control materials, calibration >> materials, and other supplies must not be used when they have exceeded >> their >> expiration date, have deteriorated, or are of substandard quality." >> >> http://www.access.gpo.gov/nara/cfr/waisidx_04/42cfr493_04.html >> >> (Hope the link works.) >> >> Sorry, there's no exception for costly IHC or ISH solutions that cost a >> lot >> of money and that we know we can "prove" are still good. >> >> The only thing I can see us histotechs doing is persuading the >> manufacturers >> to sell the reagents in smaller quantities. (Which they already have, >> over >> the years.) >> >> Peggy A. Wenk, HTL(ASCP)SLS >> William Beaumont Hospital >> Royal Oak, MI 48073 >> >> >> >> -----Original Message----- >> From: histonet-bounces@lists.utsouthwestern.edu >> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of >> KMB1904@aol.com >> Sent: Thursday, April 27, 2006 3:24 PM >> To: histonet@pathology.swmed.edu >> Subject: [Histonet] Expired antibodies...for immunos.. >> >> How does JCAHO handle expired antibodies if you have them...and are using >> them... >> Is it ok ....or not?? >> Kat >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From vd38 <@t> georgetown.edu Fri Apr 28 15:23:37 2006 From: vd38 <@t> georgetown.edu (Vernon Dailey) Date: Fri Apr 28 15:24:30 2006 Subject: [Histonet] Brain IHC Message-ID: <445279C9.6000108@georgetown.edu> Hi Histonet, Is it necessary to use antigen retreival techniques on brain tissue fixed in 4% paraform, cryoprotected in 30% sucrose and sectioned on a cryostat for free floating IHC. I am interested in staining for Mineralocorticoid receptor and DA D2 receptor. Thanks in advance, Vernon From meint002 <@t> umn.edu Fri Apr 28 15:47:30 2006 From: meint002 <@t> umn.edu (meint002) Date: Fri Apr 28 15:47:47 2006 Subject: [Histonet] Y-Chromosome DNA probes for FISH studies Message-ID: <200604282047.k3SKlUG1027130@sarge.software.umn.edu> Dear Histos, We are going to be doing FISH studies of tissue from female subjects who received cord blood transplants from male donors. I am beginning to research y-chromosome probes which will be used to detect the amount of the donor cells which have engrafted into the female receipient tissue. I am looking for recommendations as to a good Y-chromosome DNA probe and the vendor that manufactures it. If you have a great procedure for FISH concerning the same, I would be very grateful to hear from you! Many thanks for any advice you can give concerning this topic. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center From Tracey.Lenek <@t> CLS.ab.ca Fri Apr 28 16:49:37 2006 From: Tracey.Lenek <@t> CLS.ab.ca (Tracey.Lenek@CLS.ab.ca) Date: Fri Apr 28 16:49:42 2006 Subject: [Histonet] Waterbaths and hot plates Message-ID: <4BC300747AF87A48BCDF8E48BC2885CE02C1D6B1@mail1.calgary.com> We are looking to replace some tissue floatation waterbaths and slide hot plates in our department. Can anyone recommend a supplier for a rectangular waterbath preferably without a glass insert as well as some larger hot plates. Thanks in advance. Tracey CONFIDENTIALITY NOTICE: This e-mail and any files/attachments may contain confidential, personal and/or privileged information intended for a specific purpose and recipient. If you are not the intended recipient do not disclose, copy, retain, distribute, use or modify any of the contents of this transmission. If you received this transmission in error please notify me immediately by return e-mail or telephone and destroy the entire transmission and any copies produced. Thank you. From tpmorken <@t> labvision.com Fri Apr 28 18:16:32 2006 From: tpmorken <@t> labvision.com (Morken, Tim - Labvision) Date: Fri Apr 28 18:16:45 2006 Subject: [Histonet] test Message-ID: <0556BE8AC5551E4E8AF6BB9E42509BA20328D895@usca0082k08.labvision.apogent.com> test From Linresearch <@t> aol.com Fri Apr 28 18:16:44 2006 From: Linresearch <@t> aol.com (Linresearch@aol.com) Date: Fri Apr 28 18:17:03 2006 Subject: [Histonet] CD Abs Message-ID: <393.253becc.3183fc5c@aol.com> Hi, I need to develop several CD Abs on FFPE rodent tissues. Would anyone be willing to share their protocol and/or Ab source for: CD3 CD4 CD8 CD21 CD45 Thanks, Linda From hbeard27 <@t> hotmail.com Sat Apr 29 06:09:56 2006 From: hbeard27 <@t> hotmail.com (Helen Beard) Date: Sat Apr 29 06:10:07 2006 Subject: [Histonet] microglial marker In-Reply-To: Message-ID: I am looking for a microglial marker to use on 4% PFA perfused, frozen &/or vibratome mouse brain sections. I have tried MAC-1 from Serotec with pretty disappointing results. Any suggestions would be greatly appreciated. Helen ______________________________________________________________ From bjessee <@t> omnisonics.com Sat Apr 29 06:41:31 2006 From: bjessee <@t> omnisonics.com (Bret Jessee) Date: Sat Apr 29 06:41:47 2006 Subject: [Histonet] Evan's Blue, then H&E? Message-ID: I'm using in vivo perfusion of Evan's Blue to assess damage of vascular endothelium by endovascular devices. Once I have done a gross exam of tissues explanted en bloc, can standard H&E procedures (10% NBF fixation, paraffin embedding, H&E staining) be used without interference from the Evan's Blue? Thanks, BRET C. Bret Jessee, Ph.D. Director Preclinical Affairs OmniSonics Medical Technologies, Inc. 66 Concord Street, Suite A Wilmington, MA 01887 TEL: 978-657-9980 x332 CELL: 978-394-3175 Notice: This e-mail message and any attachments to it may contain legally privileged confidential information. If you are not the intended recipient, you must not review, retransmit, convert to hard copy, copy, use or disseminate this e-mail or any attachments. If you have received this e-mail in error, please immediately notify us by return e-mail or by telephone at 978-657-9980 and delete this message. From rjbuesa <@t> yahoo.com Sat Apr 29 07:33:59 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Apr 29 07:34:04 2006 Subject: [Histonet] Brain IHC In-Reply-To: <445279C9.6000108@georgetown.edu> Message-ID: <20060429123359.11307.qmail@web61220.mail.yahoo.com> Vernon: You will need to do antigen retrieval depending on the nature of your target substance and the possibility it was crosslinked with the paraform. If such a crosslinkage took place you will need to do antigen retrieval to restore the targeted substance to its natural condition and full reactive potential. The tricky thing would be to do it as a "free floating" technique. I would try to do your staining without the antigen retrieval first and if your results are below what you should expect (comparing with other published results) then I would attempt the antigen retrieval, that I would do at the lowest temperature possible. Hope this will help you! Ren? J. Vernon Dailey wrote: Hi Histonet, Is it necessary to use antigen retreival techniques on brain tissue fixed in 4% paraform, cryoprotected in 30% sucrose and sectioned on a cryostat for free floating IHC. I am interested in staining for Mineralocorticoid receptor and DA D2 receptor. Thanks in advance, Vernon _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Talk is cheap. Use Yahoo! Messenger to make PC-to-Phone calls. Great rates starting at 1¢/min. From rjbuesa <@t> yahoo.com Sat Apr 29 07:35:33 2006 From: rjbuesa <@t> yahoo.com (Rene J Buesa) Date: Sat Apr 29 07:35:37 2006 Subject: [Histonet] Y-Chromosome DNA probes for FISH studies In-Reply-To: <200604282047.k3SKlUG1027130@sarge.software.umn.edu> Message-ID: <20060429123533.79512.qmail@web61211.mail.yahoo.com> I believe Abbot labs have such a probe. Ren? J. meint002 wrote: Dear Histos, We are going to be doing FISH studies of tissue from female subjects who received cord blood transplants from male donors. I am beginning to research y-chromosome probes which will be used to detect the amount of the donor cells which have engrafted into the female receipient tissue. I am looking for recommendations as to a good Y-chromosome DNA probe and the vendor that manufactures it. If you have a great procedure for FISH concerning the same, I would be very grateful to hear from you! Many thanks for any advice you can give concerning this topic. Joyce Meints Histologist University of Minnesota Paul and Sheila Wellstone Muscular Dystrophy Center _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet --------------------------------- Love cheap thrills? Enjoy PC-to-Phone calls to 30+ countries for just 2?/min with Yahoo! Messenger with Voice. From histosci <@t> shentel.net Sat Apr 29 07:42:12 2006 From: histosci <@t> shentel.net (HSRL) Date: Sat Apr 29 07:42:27 2006 Subject: [Histonet] Sakura DRS-601 stainer Message-ID: <000e01c66b8a$59889520$0200a8c0@HSRLMAIN> Dear Netters, We have a Sakura DRS-601 slide stainer we would like to sell. It is about 9 years old and has been maintained in a GLP laboratory since day 1. It is in great shape (even the plastic top is in perfect condition). It can be programmed for multiple staining protocols. We are selling it because we purchased a linear stainer. If you are interested, please call or e-mail me directly. Have a good day. -Tom Tom Galati Laboratory Director HSRL, Inc.- A GLP Compliant Contract Laboratory/Archive 5930 Main Street Mount Jackson, Virginia 22842 540.477.4440 540.477.4448 tomgalati@hsrl.org www.hsrl.org From jnocito <@t> satx.rr.com Sat Apr 29 18:05:53 2006 From: jnocito <@t> satx.rr.com (Joe Nocito) Date: Sat Apr 29 18:06:06 2006 Subject: [Histonet] Expired antibodies...for immunos.. References: <400.4dfa38.31827436@aol.com> <002a01c66a49$ba8541d0$1bb30b43@yourxhtr8hvc4p> <002401c66a61$38564e30$6500a8c0@mainbox> Message-ID: <004601c66be1$774d3500$1bb30b43@yourxhtr8hvc4p> only when it's convenient for the other person. Joe ----- Original Message ----- From: "Bryan Hewlett" To: "Joe Nocito" ; ; Sent: Thursday, April 27, 2006 8:15 PM Subject: Re: [Histonet] Expired antibodies...for immunos.. > Hey Joe, > > I'm with you! > The disposal of a expensive, validated, working reagent, just because of > some mindless arbitrary date rule makes absolutely no sense! > I thought that we were in the era of evidence-based medicine, whatever > happened? > > Bryan Hewlett > Consultant technologist > Anatomic Pathology > Quality Management Program - Laboratory Services > Ontario, Canada. > > ----- Original Message ----- > From: "Joe Nocito" > To: ; > Sent: Thursday, April 27, 2006 6:27 PM > Subject: Re: [Histonet] Expired antibodies...for immunos.. > > >> Oh boy, >> I get to get flamed again. >> JCAHO and CAP do not let you use expired reagents, including antibodies. >> therefore, everyone gets to through money down the drain time and time >> again. >> I called CAP and talked to a person who got very defensive when I >> asked him to give the rule that says we can not use expired antibodies. >> He referred me to the U.S. National Register. I'm sorry, I don't have >> that information with right now but I'm sure someone will let me know. >> My beef is, if you run a known positive control with every run, you >> are, in essence, revalidating that antibody. Once that antibody doesn't >> work, replace it with one that has not expired. I know this causes a >> repeat and maybe a delay in reporting the results, but ask your >> pathologists if they would want a one day delay, or throw thousands of >> dollars down the sink. >> When I was supervising the immuno lab at the AFIP, we froze >> concentrated antibodies at -70 C and they still worked up to 10 years >> later. Now, you can't even do that. A big waste, that what it is, a big >> waste. >> Let Flaming Friday begin. >> >> Joe Nocito BS, HT(ASCP)QIHC >> Histology Manager >> Pathology Reference Lab >> San Antonio, TX >> >> ----- Original Message ----- >> From: >> To: >> Sent: Thursday, April 27, 2006 2:23 PM >> Subject: [Histonet] Expired antibodies...for immunos.. >> >> >>> How does JCAHO handle expired antibodies if you have them...and are >>> using >>> them... >>> Is it ok ....or not?? >>> Kat >>> _______________________________________________ >>> Histonet mailing list >>> Histonet@lists.utsouthwestern.edu >>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >>> >>> >>> -- >>> Internal Virus Database is out-of-date. >>> Checked by AVG Free Edition. >>> Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 >>> >>> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > > _______________________________________________ > Histonet mailing list > Histonet@lists.utsouthwestern.edu > http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > -- > Internal Virus Database is out-of-date. > Checked by AVG Free Edition. > Version: 7.1.385 / Virus Database: 268.4.3/317 - Release Date: 4/18/2006 > From jcox90 <@t> yahoo.com Sat Apr 29 18:15:21 2006 From: jcox90 <@t> yahoo.com (Jill Cox) Date: Sat Apr 29 18:15:24 2006 Subject: [Histonet] Histology Tech position in Phoenix Arizona Message-ID: <20060429231521.58554.qmail@web52114.mail.yahoo.com> We are a new private lab in Tempe and have an opening for a Histologist. We offer excellent salary and benefits along with relocation assistance. Please call or email with any questions. Jill Jill Cox HT (ASCP) Clin-Path Diagnostics LLC 2109 S 48th St. Suite 102 Tempe Arizona 85282 602-319-6471 Lab Manager From anh2006 <@t> med.cornell.edu Sat Apr 29 19:57:37 2006 From: anh2006 <@t> med.cornell.edu (Andrea T. Hooper) Date: Sat Apr 29 19:56:58 2006 Subject: [Histonet] Re: lymphatics In-Reply-To: <4344A583.842FBFA@uwo.ca> References: <8D6F233E2A5D574B929F3944F3316FD006310469@EXCH2K3.utmb.edu> <4344A583.842FBFA@uwo.ca> Message-ID: Hi Histonet, Has anyone performed the 5-nucleotidase stains John referred to below. I am really interested in this method of distinguising BV from LV. If so does someone have a working protocol I can work from or words of wisdom?? Thanks in advance, Andrea >Dear Hal Hawkins, > >The endothelium of lymphatic vessels can be stained by >enzyme activity histochemistry. The enzyme is a >5-nucleotidase (belongs to the phosphatase family). >Ordinarily this type of method is carried out with frozen >sections of tissue that has been minimally fixed in >formaldehyde or glutaraldehyde, or with unfixed cryostat >sections that have been briefly fixed in cold acetone. The >enzyme in lymphatics may be fairly robust because its >activity can survive embedding in glycol methacrylate. > >The references below may help. If you choose one of these >methods it will be important to consult the original >publication because there's more than one sort of >5-nucleotidase. Each ref is followed by brief notes that I >took when reading the papers. [An R in the acquisition >number means I've got a Reprint of the whole paper; an A >usually means I have a copy of the paper's Abstract. No >letter with the acquisition number usually means I've seen >the whole paper and took notes in the library.] > >8194R. Kato,S; Yasunaga,A; Uchida,U (1991): >Enzyme-histochemical method for identification of lymphatic >capillaries. Lymphology 24, 125-129. > Glycol methacrylate-embedded sections stained for >5'-nucleotidase & alkaline phosphatase. 5-N-ase in lymph >capills; AlkP-ase in blood capills. > >9239R. Nishida,S; Ohkuma,M (1993): Enzyme-histochemical >staining of dermal lymphatic capillaries by guanylate >cyclase. Lymphology 26, 195-199. > Enzyme histochemical staining distinguishes lymph from >blood capillaries. Says 5-nucleotidase & adenylate cyclase >do so too. (Used unfixed cryostat sections.) > >9665R. Okada,E (1994): An improved enzyme-histochemical >method for identification of lymphatic capillaries on >paraffin sections. Lymphology 27, Suppl, 732-735. > Staining of blood & lymph capillaries. Enzyme >histochemistry for 5-nucleotidase in presence of levamisole >for lymph capills; alkaline phosphatase for blood capills. > >11114A. Miura,M; Kato,S; von Ludinghausen,M (1998): >Lymphatic drainage of the cerebrospinal fluid from monkey >spinal meninges with special reference to the distribution >of the epidural lymphatics. Arch. Histol. Cytol. 61(3, Aug), >277-286. > 5'-nucleotidase staining for lymphatics; alk phosphatase >for blood capillaries (Kato et al '91,'93). Also traced >carbon particles from cisterna magna. Lymphatics and carbon >found on surfaces of cervical & thoracic (most at brachial >plexus levels) roots; not lumbosacral. Epidural lymphatics >most developed on dorsal surface of lower cervical dura. > >I hope this helps. >-- >------------------------------- John A. Kiernan -- From kelseymichele <@t> hotmail.com Sun Apr 30 14:00:45 2006 From: kelseymichele <@t> hotmail.com (Kelsey Peek) Date: Sun Apr 30 14:00:53 2006 Subject: [Histonet] Histo Question Message-ID: What is the most difficult stain to work with and what tissue is it most often used? _________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/ From cfavara <@t> niaid.nih.gov Sun Apr 30 18:07:24 2006 From: cfavara <@t> niaid.nih.gov (Favara, Cynthia (NIH/NIAID) [E]) Date: Sun Apr 30 18:07:32 2006 Subject: [Histonet] microglial marker In-Reply-To: Message-ID: WAKO - Iba c Cynthia Favara NIAID/NIH/RML/LPVD 903 South 4th Street Hamilton, MT 59840 406-363-9317 Disclaimer: The information in this e-mail and any of its attachments is confidential and may contain sensitive information. It should not be used by anyone who is not the original intended recipient. If you have received this e-mail in error please inform the sender and delete it from your mailbox or any other storage devices. National Institute of Allergy and Infectious Diseases shall not accept liability for any statements made that are sender's own and not expressly made on behalf of the NIAID by one of its representatives -----Original Message----- From: Helen Beard [mailto:hbeard27@hotmail.com] Sent: Saturday, April 29, 2006 4:10 AM To: histonet@pathology.swmed.edu Subject: [Histonet] microglial marker I am looking for a microglial marker to use on 4% PFA perfused, frozen &/or vibratome mouse brain sections. I have tried MAC-1 from Serotec with pretty disappointing results. Any suggestions would be greatly appreciated. Helen ______________________________________________________________ _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet From AnthonyH <@t> chw.edu.au Sun Apr 30 18:28:29 2006 From: AnthonyH <@t> chw.edu.au (Tony Henwood) Date: Sun Apr 30 18:28:46 2006 Subject: [Histonet] NC NCSHT meeting.... Message-ID: It's the Vision Biosystems Bond: http://www.vision-bio.com/product_bondmax.html Regards Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC) Laboratory Manager & Senior Scientist The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: 612 9845 3306 Fax: 612 9845 3318 -----Original Message----- From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of JCarpenter764@aol.com Sent: Friday, 28 April 2006 9:45 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] NC NCSHT meeting.... Goodmorning, I was wondering if anyone could tell me the company who manufactures the IHC machine that can run three different runs at one time. This machine was absolutely wonderful. It had three different slots for slides that held 10 a piece and I can not find the brochure I picked up..of course. It also had little slide covers that fit over the slide in the machine. If anyone can help I would greatly appreciate it. Thanks a bunch Jennell _______________________________________________ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you are not the intended recipient, please delete it and notify the sender. Views expressed in this message and any attachments are those of the individual sender, and are not necessarily the views of The Children's Hospital at Westmead This note also confirms that this email message has been virus scanned and although no computer viruses were detected, the Childrens Hospital at Westmead accepts no liability for any consequential damage resulting from email containing computer viruses. **********************************************************************