[Histonet] RE: Histonet Digest, Vol 74, Issue 12
Moody, Robert
rmoody <@t> ameripath.com
Wed Jan 13 22:08:53 CST 2010
Hi, All we are having problems with chatter in our biopsies their usually on the edge of tissue is the a problem with the cutting or in the handling of the tissue after it is removed from the patient like the biopsies being left out to dry or not put in formalin what are some of your experience with this.
Robert Moody HT ASCP
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of histonet-request <@t> lists.utsouthwestern.edu
Sent: Wednesday, January 13, 2010 10:32 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 74, Issue 12
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Today's Topics:
1. IHC (Marian Powers)
2. RE: Eliminating the edge effect in IHC/IF (C.M. van der Loos)
3. RE: RE: Eliminating the edge effect in IHC/IF
(Sherwood, Margaret )
4. RE: Eliminating the edge effect in IHC/IF (Rene J Buesa)
5. RE: RE: Eliminating the edge effect in IHC/IF
(Della Speranza, Vinnie)
6. RE: RE: Eliminating the edge effect in IHC/IF (Rene J Buesa)
7. RE: RE: Eliminating the edge effect in IHC/IF (Hermina Borgerink)
8. Fw: Waste paraffin and edge effect (Jeffrey Silverman)
9. Re: Sakura iDent (Anne van Binsbergen)
10. Looking for Florida licensed Histologist (Kaye Ryan)
11. RE: Re: Sakura iDent (Cynthia Pyse)
12. AW: [Histonet] Eliminating the edge effect in IHC/IF (Gudrun Lang)
13. Bladder Biopsies (S R)
14. Re: Bladder Biopsies (Lester Raff MD)
15. Re: Eliminating the edge effect in IHC/IF
(histology <@t> medsurgpath.com)
16. Part-time Flex Histologist position (Fallak, Alice)
17. RE: Re: Sakura iDent (Nancy Klemme)
18. Re: RE: [Histonet] Problems with MSB stain (John Kiernan)
19. (Used microtome) - Thanks ... (Massimo)
20. (Used microtome) - Thanks ... (Massimo)
21. AW: [Histonet] Problems with MSB stain (Gudrun Lang)
22. Re: Re: Sakura iDent (Anne van Binsbergen)
----------------------------------------------------------------------
Message: 1
Date: Tue, 12 Jan 2010 14:34:33 -0500
From: Marian Powers <mpowers <@t> dpspa.com>
Subject: [Histonet] IHC
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<5d7de0e61001121134g662d43adr3a7c4cd1b0235854 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Hi: We are looking to start N-ras and B-raf for IHC, is anyone running
these antibodies on paraffin? If so, what clone and vendor are you using?
Thanks in advance!
--
Marian L. Powers, HT(ASCP)
Manager, Technical Operations
Doctors Pathology Services
1253 College Park Drive
Dover, DE 19904
302-677-0000
------------------------------
Message: 2
Date: Tue, 12 Jan 2010 20:54:35 +0100
From: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Message-ID: <9637c5fe5d7dffc7.4b4ce18b <@t> amc.uva.nl>
Content-Type: text/plain; charset=utf-8
Hi all,We also have observed this phenomenon many times. But sorry Colleen and Rene, I don't believe that an fixation issue is the explanation why the edges are sometimes stronger than the rest. To my opinion this is a bit too easy. One of my explanations is that the immuno reagents tend to stick to the edges of the tissue section, especially when no Tween20 is involved. As a result the outer edges might become a little dry during incubation times and cause darker mostly non-specific staining. Always try to cover a section completely, including a rim around the section. However, to be honest, I am sure my explanation is certainly not always appropriate. Anyone else????Cheers,ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone: +31 20 5665631
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>
Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren??? J.
--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:
From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
------------------------------
Message: 3
Date: Tue, 12 Jan 2010 15:28:55 -0500
From: "Sherwood, Margaret " <MSHERWOOD <@t> PARTNERS.ORG>
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>,
<histonet <@t> lists.utsouthwestern.edu>
Cc: kcai <@t> prosci-inc.com
Message-ID:
<073AE2BEA1C2BA4A8837AB6C4B943D9703E23F19 <@t> PHSXMB30.partners.org>
Content-Type: text/plain; charset="us-ascii"
Actually, when I read about the problem, my thought was that if you don't circle
your tissue with an immunopen or wax pencil, then the reagents (esp. primary
antibody)might not cover the tissue evenly as you say. So I tend to agree with
you.
Peggy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van der
Loos
Sent: Tuesday, January 12, 2010 2:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF
Hi all,We also have observed this phenomenon many times. But sorry Colleen and
Rene, I don't believe that an fixation issue is the explanation why the edges
are sometimes stronger than the rest. To my opinion this is a bit too easy. One
of my explanations is that the immuno reagents tend to stick to the edges of the
tissue section, especially when no Tween20 is involved. As a result the outer
edges might become a little dry during incubation times and cause darker mostly
non-specific staining. Always try to cover a section completely, including a rim
around the section. However, to be honest, I am sure my explanation is certainly
not always appropriate. Anyone else????Cheers,ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone: +31 20 5665631
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>
Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren J.
--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:
From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the e-mail
contains patient information, please contact the Partners Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you in error
but does not contain patient information, please contact the sender and properly
dispose of the e-mail.
------------------------------
Message: 4
Date: Tue, 12 Jan 2010 12:40:16 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, "C.M. van der Loos"
<c.m.vanderloos <@t> amc.uva.nl>
Cc: kcai <@t> prosci-inc.com
Message-ID: <661401.32335.qm <@t> web65710.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=utf-8
Hi Dr. van der Loos:
I had also experienced that artifact caused by less than necessary reagents, but that happens??mostly when IHC??is done manually, or when the autostainer delivers less amount than required or programmed.
IF we assume that the colleague with the question did the IHC manually, your explanation can be accepted, otherwise, poor fixation is a valid??cause to the problem.
Ren?? J.
--- On Tue, 1/12/10, C.M. van der Loos <c.m.vanderloos <@t> amc.uva.nl> wrote:
From: C.M. van der Loos <c.m.vanderloos <@t> amc.uva.nl>
Subject: RE: Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com, rjbuesa <@t> yahoo.com, cforster <@t> umn.edu
Date: Tuesday, January 12, 2010, 2:54 PM
Hi all,
We also have observed this phenomenon many times. But sorry Colleen and Rene,??I don't believe??that an??fixation issue??is the explanation why the edges are sometimes stronger than the rest. To my opinion this is??a bit too easy.??One of my explanations is that the immuno reagents tend to stick to the edges of the tissue section, especially when no Tween20 is involved.??As a result??the outer edges might become a little dry during incubation times and cause darker mostly non-specific staining. Always try to cover a section completely,??including a rim around the section.??However, to??be honest,??I am sure my??explanation is certainly not always appropriate. Anyone else????
Cheers,
Chris
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone:?? +31 20 5665631
??
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>
Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren??? J.
--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:
From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
------------------------------
Message: 5
Date: Tue, 12 Jan 2010 15:40:55 -0500
From: "Della Speranza, Vinnie" <dellav <@t> musc.edu>
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: "'C.M. van der Loos'" <c.m.vanderloos <@t> amc.uva.nl>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Cc: "kcai <@t> prosci-inc.com" <kcai <@t> prosci-inc.com>
Message-ID:
<E58D1CD977E29C46A167806513B04577993FF80F08 <@t> EVS5.clinlan.local>
Content-Type: text/plain; charset="utf-8"
I think we'll agree that there are different scenarios so that the solution is not a one size fits all. For example, darkened staining around the periphery of needle core biopsies is not uncommon with even tinctorial stains, often thought to be the result of drying of the tissue during the collection of the sample. Years ago I came across an article maintaining that the dark staining on the periphery of needle cores was in fact due to the "trauma" of the needle cutting into the sampled organ. I've since forgotten the author's name and wish I could get my hands on that reference.
So I agree with Chris that there doesn't appear to be one simple answer to prevent this artifact and while fixation may contribute in some circumstances it's unlikely to be the remedy for all.
Vinnie Della Speranza
Manager for Anatomic Pathology Services
165 Ashley Avenue Suite 309
Charleston, SC 29425
tel. 843-792-6353
fax. 843-792-8974
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van der Loos
Sent: Tuesday, January 12, 2010 2:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF
Hi all,We also have observed this phenomenon many times. But sorry Colleen and Rene, I don't believe that an fixation issue is the explanation why the edges are sometimes stronger than the rest. To my opinion this is a bit too easy. One of my explanations is that the immuno reagents tend to stick to the edges of the tissue section, especially when no Tween20 is involved. As a result the outer edges might become a little dry during incubation times and cause darker mostly non-specific staining. Always try to cover a section completely, including a rim around the section. However, to be honest, I am sure my explanation is certainly not always appropriate. Anyone else????Cheers,ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone: +31 20 5665631
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>
Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren??? J.
--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:
From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 6
Date: Tue, 12 Jan 2010 12:41:24 -0800 (PST)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>,
histonet <@t> lists.utsouthwestern.edu, MargaretSherwood
<MSHERWOOD <@t> PARTNERS.ORG>
Cc: kcai <@t> prosci-inc.com
Message-ID: <842909.92777.qm <@t> web65701.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
Again, provided that you are doing IHC manually.
Ren? J.
--- On Tue, 1/12/10, Sherwood, Margaret <MSHERWOOD <@t> PARTNERS.ORG> wrote:
From: Sherwood, Margaret <MSHERWOOD <@t> PARTNERS.ORG>
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>, histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Date: Tuesday, January 12, 2010, 3:28 PM
Actually, when I read about the problem, my thought was that if you don't circle
your tissue with an immunopen or wax pencil, then the reagents (esp. primary
antibody)might not cover the tissue evenly as you say.? So I tend to agree with
you.
Peggy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van der
Loos
Sent: Tuesday, January 12, 2010 2:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF
Hi all,We also have observed this phenomenon many times. But sorry Colleen and
Rene, I don't believe that an fixation issue is the explanation why the edges
are sometimes stronger than the rest. To my opinion this is a bit too easy. One
of my explanations is that the immuno reagents tend to stick to the edges of the
tissue section, especially when no Tween20 is involved. As a result the outer
edges might become a little dry during incubation times and cause darker mostly
non-specific staining. Always try to cover a section completely, including a rim
around the section. However, to be honest, I am sure my explanation is certainly
not always appropriate. Anyone else????Cheers,ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone:? +31 20 5665631
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>
Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren? J.
--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:
From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the e-mail
contains patient information, please contact the Partners Compliance HelpLine at
http://www.partners.org/complianceline . If the e-mail was sent to you in error
but does not contain patient information, please contact the sender and properly
dispose of the e-mail.
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 7
Date: Tue, 12 Jan 2010 16:03:13 -0500
From: "Hermina Borgerink" <hborgeri <@t> wfubmc.edu>
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: "Rene J Buesa" <rjbuesa <@t> yahoo.com>, "C.M. van der Loos"
<c.m.vanderloos <@t> amc.uva.nl>, <histonet <@t> lists.utsouthwestern.edu>,
"MargaretSherwood" <MSHERWOOD <@t> PARTNERS.ORG>
Cc: kcai <@t> prosci-inc.com
Message-ID:
<9AEEF1FB6254224AA355ED285F8491653EE04680 <@t> EXCHVS2.medctr.ad.wfubmc.edu>
Content-Type: text/plain; charset="iso-8859-1"
I do my immuno staining manually using ProbeOn Plus slides which employ the capillary action principle. I always monitor the taking up/whicking off of my solutions for each pair of slides throughout the entire staining process and know that my sections never dry out (I incubate using moist chambers with plenty of fluid). I work at a research/diagnostic medical school facility where our primary focus is on experimental tissues, using standardized and rigid guidelines for fixation: 24 hours at 4 degrees overnight in 4% PF, followed by post fixation in 70% ethanol for a few days. Never see the "edge effect" with these sections. However, I do occasionally see the effect using archival diagnostic samples that were fixed in 10% NBF for a minimum of 48 hours, and probably longer. I have therefore always perceived this effect to be caused by extended, rather than incomplete fixation.
Hermina
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Rene J Buesa
Sent: Tuesday, January 12, 2010 3:41 PM
To: C.M. van der Loos; histonet <@t> lists.utsouthwestern.edu; MargaretSherwood
Cc: kcai <@t> prosci-inc.com
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
Again, provided that you are doing IHC manually.
Ren? J.
--- On Tue, 1/12/10, Sherwood, Margaret <MSHERWOOD <@t> PARTNERS.ORG> wrote:
From: Sherwood, Margaret <MSHERWOOD <@t> PARTNERS.ORG>
Subject: RE: [Histonet] RE: Eliminating the edge effect in IHC/IF
To: "C.M. van der Loos" <c.m.vanderloos <@t> amc.uva.nl>, histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Date: Tuesday, January 12, 2010, 3:28 PM
Actually, when I read about the problem, my thought was that if you don't circle
your tissue with an immunopen or wax pencil, then the reagents (esp. primary
antibody)might not cover the tissue evenly as you say.? So I tend to agree with
you.
Peggy
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of C.M. van der
Loos
Sent: Tuesday, January 12, 2010 2:55 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: kcai <@t> prosci-inc.com
Subject: [Histonet] RE: Eliminating the edge effect in IHC/IF
Hi all,We also have observed this phenomenon many times. But sorry Colleen and
Rene, I don't believe that an fixation issue is the explanation why the edges
are sometimes stronger than the rest. To my opinion this is a bit too easy. One
of my explanations is that the immuno reagents tend to stick to the edges of the
tissue section, especially when no Tween20 is involved. As a result the outer
edges might become a little dry during incubation times and cause darker mostly
non-specific staining. Always try to cover a section completely, including a rim
around the section. However, to be honest, I am sure my explanation is certainly
not always appropriate. Anyone else????Cheers,ChrisChris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterdam
The Netherlands
phone:? +31 20 5665631
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu, Karen Cai <kcai <@t> prosci-inc.com>
Usually that is the result of incomplete fixation. Check your fixation protocol.
Ren? J.
--- On Mon, 1/11/10, Karen Cai <kcai <@t> prosci-inc.com> wrote:
From: Karen Cai <kcai <@t> prosci-inc.com>
Subject: [Histonet] Eliminating the edge effect in IHC/IF
To: histonet <@t> lists.utsouthwestern.edu
Date: Monday, January 11, 2010, 2:00 PM
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
The information in this e-mail is intended only for the person to whom it is
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------------------------------
Message: 8
Date: Tue, 12 Jan 2010 14:30:46 -0800 (PST)
From: Jeffrey Silverman <pathmaster <@t> yahoo.com>
Subject: [Histonet] Fw: Waste paraffin and edge effect
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <13966.65872.qm <@t> web111106.mail.gq1.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1
---
Hello everyone,
?
Our?system's ?hazardous chemical waste consultants changed our method of disposal- we had been solidifying the paraffin in a hood and then red bagging it, ie treating it as regulated medical (biohazard) waste.
?
But, due to it's contamination with xylene, the very reason for our disposing of it, the paraffin is really hazardous chemical waste. Believe it or not, EPA regs prohibit most hospitals from treating their chemical waste and the consultants worried that solidifying it under the hood was considered "treating" the waste.? Yeah right!!? ?Anyway, the blocks of solidified paraffin are now dumped in?it's own specially labelled drum and has become a part of our hazardous waste stream, manifested like the xylene and dye waste. I'd be careful about putting hazardous chemicals into?regulated medical waste.
?
Interestingly, our system recently inquired if formalin in discarded surgicals needs to be decanted before disposal- many hospitals are doing just that, but most of those, but not all, recycle formalin (yuk!).?Our disposal facility informed us that as long as the containers are plastic, ie are flammamble, they can be incinerated with the formalin and there is no need to decant. ?Glory be!!
?
In IHC slides, edge effect, or more intense, sometimes?nonspecific, staining at the periphery of the tissue, can be caused? by more intense fixation of a block at the periphery,?drying out of the edges of a block before fiation, ?and/or by some degree of drying of antibodies and detection chemistry reagents during?staining- this cause is more common in manually stained slides rather than those stained on automated stainers. ?
?
Also electrocautery?used during the excision can also cause intense staining at the edges of specimens.
?
Jeffrey S. Silverman HT HTL QIHC (ASCP)
Southside Hospital- NSLIJ Health System
Pathologists' Assistant and Laboratory Safety Officer
------------------------------
Message: 9
Date: Wed, 13 Jan 2010 13:15:36 +0400
From: Anne van Binsbergen <annigyg <@t> gmail.com>
Subject: [Histonet] Re: Sakura iDent
To: Denise Van Eaton <dvaneaton <@t> littonlab.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<f8332fbe1001130115l1020ce1do6680da466ee09de4 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Hi Denise
sadly no resolution
also sadly very little response from the histonet - except for Rene - he and
i did some troubleshooting to try to see if i had missed anything - we had
the same thoughts
so ... i am still sitting with the iDent in its box, unused, until someone,
somewhere can convince me that the print does not wipe off with xylene -
which will be hard to do seeing that i wiped it off myself and saw with my
own eyes that xylene removes the ink!!
Sakura - if you are reading this - i am not a happy camper
Annie
2010/1/12 Denise Van Eaton <dvaneaton <@t> littonlab.com>
> Hi Anne,
>
> We have been checking into cassette printers. Obviously, if we can bring
> one in for a demonstration we will but I thought the iDent looked like a
> good place to start. I noticed your problem (on the Histonet) with the ink
> rubbing off after the xylene. I never saw any responses from the rest of the
> Histonetters... did you resolve the problem? Assuming you (or Sakura) did,
> what was the trick?
>
> Thank you in advance for sharing your "fix" for a potential problem.
>
> Denise Van Eaton HT(ASCP)
> Litton Pathology Associates
>
--
Anne van Binsbergen (Hope)
Abu Dhabi
UAE
------------------------------
Message: 10
Date: Wed, 13 Jan 2010 08:58:54 -0500
From: "Kaye Ryan" <kryan <@t> nfderm.com>
Subject: [Histonet] Looking for Florida licensed Histologist
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<DCB07C3EF84D92439E5A453D8D5B5EB004ADD2 <@t> exchserver.NFDA.com>
Content-Type: text/plain; charset="us-ascii"
North Florida Dermatology Associates located in Jacksonville, Florida is
looking for a full time histologist. Must be eligible for Florida State
Technologist license. The lab is a small private lab with good benefits
and minimal stress. Responsibilities include routine histology and some
IHC techniques. If interested, please contact me or our Personnel
Director at 904-398-0547, ext. 1106.
Kaye Ryan
North Florida Dermatology Associates
Histology Lab Supervisor
(904) 398-0547 Ext. 2004
------------------------------
Message: 11
Date: Wed, 13 Jan 2010 09:03:34 -0500
From: "Cynthia Pyse" <cpyse <@t> x-celllab.com>
Subject: RE: [Histonet] Re: Sakura iDent
To: "'Anne van Binsbergen'" <annigyg <@t> gmail.com>, "'Denise Van Eaton'"
<dvaneaton <@t> littonlab.com>
Cc: "'histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID: <000001ca9459$32066300$96132900$@com>
Content-Type: text/plain; charset="us-ascii"
Anne
You might want to try going up the Sakura chain of command. I had trouble
with a coverslipper, after contacting the regional manager it was amazing
the service it received. Good luck.
Cindy Pyse, CLT, HT (ASCP)
Histology Supervisor
X-Cell Laboratories
e-mail cpyse <@t> x-celllab.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anne van
Binsbergen
Sent: Wednesday, January 13, 2010 4:16 AM
To: Denise Van Eaton
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Sakura iDent
Hi Denise
sadly no resolution
also sadly very little response from the histonet - except for Rene - he and
i did some troubleshooting to try to see if i had missed anything - we had
the same thoughts
so ... i am still sitting with the iDent in its box, unused, until someone,
somewhere can convince me that the print does not wipe off with xylene -
which will be hard to do seeing that i wiped it off myself and saw with my
own eyes that xylene removes the ink!!
Sakura - if you are reading this - i am not a happy camper
Annie
2010/1/12 Denise Van Eaton <dvaneaton <@t> littonlab.com>
> Hi Anne,
>
> We have been checking into cassette printers. Obviously, if we can bring
> one in for a demonstration we will but I thought the iDent looked like a
> good place to start. I noticed your problem (on the Histonet) with the ink
> rubbing off after the xylene. I never saw any responses from the rest of
the
> Histonetters... did you resolve the problem? Assuming you (or Sakura) did,
> what was the trick?
>
> Thank you in advance for sharing your "fix" for a potential problem.
>
> Denise Van Eaton HT(ASCP)
> Litton Pathology Associates
>
--
Anne van Binsbergen (Hope)
Abu Dhabi
UAE
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 12
Date: Wed, 13 Jan 2010 15:32:19 +0100
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: AW: [Histonet] Eliminating the edge effect in IHC/IF
To: "'Karen Cai'" <kcai <@t> prosci-inc.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <21FEAA12A3C44D2FBE8EC603949E9CB3 <@t> dielangs.at>
Content-Type: text/plain; charset="iso-8859-1"
My favourite for intense specific staining at the edges vs. faint specific
staining in the center is poor fixation.
I think drying artefacts or crush artefacts would lead rather to unspecific
background staining.
Gudrun Lang
Histolab, Akh Linz, Austria
-----Urspr?ngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Karen Cai
Gesendet: Montag, 11. J?nner 2010 20:01
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Eliminating the edge effect in IHC/IF
Hi,
I have a question for the generous input. When I do the IHC or IF, it
seems very common that the intensity of the edge area of the tissue is
always stronger than the central tissue part. Is it possible to
eliminate this and make the staining evenly distributed around the whole
tissue section?
Your kind help is greatly appreciated,
Thanks in advance,
Best,
Karen
Karen Cai
Research Scientist
Prosci Incorporated
(858) 513-2638 x 204
(858) 513-2692 Fax
<http://www.prosci-inc.com> www.prosci-inc.com
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 13
Date: Wed, 13 Jan 2010 09:44:03 -0500
From: S R <sjkitten <@t> live.com>
Subject: [Histonet] Bladder Biopsies
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BAY133-W22498DA6FFEA03DC0245BCB36B0 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"
Hello Everyone!
I have a couple of questions. I work for a urology group. As of right now i only work with prostate bx's. They want to add Bladder bx's into the mix. As of right now i microwave process them. I was wondering if anyone eles does this and if you use a different procedure than you would use for processing the prostate biopsies, and would anyone be willing to share?
Thanks in Advanced
Sammy
_________________________________________________________________
Your E-mail and More On-the-Go. Get Windows Live Hotmail Free.
http://clk.atdmt.com/GBL/go/196390709/direct/01/
------------------------------
Message: 14
Date: Wed, 13 Jan 2010 09:09:54 -0600
From: "Lester Raff MD" <LRaff <@t> uropartners.com>
Subject: [Histonet] Re: Bladder Biopsies
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<FE0368FE5AA6684DB5990BBEED742C5306EE5D <@t> UPHQMSX01.uropartners.local>
Content-Type: text/plain; charset="us-ascii"
We microwave process our bladder biopsies on the same cycle as prostate
biopsies.
Lester J. Raff, MD
Medical Director
UroPartners Laboratory
2225 Enterprise Dr. Suite 2511
Westchester, Il 60154
Tel 708.486.0076
Fax 708.486.0080
------------------------------
Message: 15
Date: Wed, 13 Jan 2010 07:50:50 -0800 (PST)
From: histology <@t> medsurgpath.com
Subject: Re: [Histonet] Eliminating the edge effect in IHC/IF
To: "Karen Cai" <kcai <@t> prosci-inc.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<61410.68.178.72.125.1263397850.squirrel <@t> webmail.integra.net>
Content-Type: text/plain;charset=iso-8859-1
Hi Karen/Histonet,
This is a problem we have been working on for some time with our IHC. We
have ruled out fixation as the source of the problem, and we are in Oregon
so we don't believe we are having a drying out issue. I have also tried
different detection kits with both having this artifact. Recently we were
wondering if it could be a problem with our blower. Is it possible that it
is blowing too hard, or that the rinses are too hard? We have tried hand
staining twice, once with perfect results, once with the same artifact.
Any additional advice you can give will be greatly appreciated.
Thank you, once again,
Katelin Lester, HTL(ASCP)
MedSurg Pathology Associates, Inc.
(formerly Cutting Edge Histology)
(503)443-2157
> Hi,
> I have a question for the generous input. When I do the IHC or IF, it
> seems very common that the intensity of the edge area of the tissue is
> always stronger than the central tissue part. Is it possible to
> eliminate this and make the staining evenly distributed around the whole
> tissue section?
>
> Your kind help is greatly appreciated,
>
>
> Thanks in advance,
>
> Best,
> Karen
>
> Karen Cai
> Research Scientist
> Prosci Incorporated
> (858) 513-2638 x 204
> (858) 513-2692 Fax
> <http://www.prosci-inc.com> www.prosci-inc.com
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
------------------------------
Message: 16
Date: Wed, 13 Jan 2010 10:14:18 -0600
From: "Fallak, Alice" <Alice.Fallak <@t> uhsi.org>
Subject: [Histonet] Part-time Flex Histologist position
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<C97AC55513965A41BC143DA68E0B7D405D6103 <@t> KMCEXCH.uhsi.local>
Content-Type: text/plain; charset="us-ascii"
We have a part-time/Flex Histologist position open at United Hospital
and Medical Center in Kenosha,Wi.
Anyone interested can contact our Human Resource Dept. at (262)656-2116.
Alice Fallak
Histology coordinator
United Hosp. & Med. Center
------------------------------
Message: 17
Date: Wed, 13 Jan 2010 08:21:12 -0800
From: Nancy Klemme <NKlemme <@t> sakuraus.com>
Subject: RE: [Histonet] Re: Sakura iDent
To: Anne van Binsbergen <annigyg <@t> gmail.com>, Denise Van Eaton
<dvaneaton <@t> littonlab.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<782E3A02C2EB2347BEA6DEA69DC7AB8669D9A4399E <@t> sfamail.SAKURAUS.LOCAL>
Content-Type: text/plain; charset="us-ascii"
Dear Annie,
I am writing you from California where our Wednesday workday is beginning and you are preparing for your Wednesday to draw to a close.
Although the majority of Sakura Finetek products are carried by all Sakura Finetek entities, the iDent is not one that is available from Sakura Finetek USA in North America. If most Histonet subscribers are from the U.S., that may explain the "very little response from the histonet" that you noted.
I am sorry about your cassette labeler issues and have forwarded your posting to the gentleman who is in charge of Sakura Finetek Middle East. His name is Mr. Ghaleb Alkhaseb and I am sure he will be in contact with you very soon after reading it. I do not know what Sakura contact information you have been using, but I know that Mr. Alkhaseb will make sure that you will have the most current information for a more direct company contact. I do expect your iDent situation will be corrected and resolved to your satisfaction very soon.
>From 12 hours behind you, I offer my kindest regards.
Nancy Klemme, HT(ASCP)
Edu. Svcs. Dir. - Sakura Finetek USA, Inc. - Torrance, CA
800-725-8723 x7879
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anne van Binsbergen
Sent: Wednesday, January 13, 2010 1:16 AM
To: Denise Van Eaton
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Sakura iDent
Hi Denise
sadly no resolution
also sadly very little response from the histonet - except for Rene - he and
i did some troubleshooting to try to see if i had missed anything - we had
the same thoughts
so ... i am still sitting with the iDent in its box, unused, until someone,
somewhere can convince me that the print does not wipe off with xylene -
which will be hard to do seeing that i wiped it off myself and saw with my
own eyes that xylene removes the ink!!
Sakura - if you are reading this - i am not a happy camper
Annie
2010/1/12 Denise Van Eaton <dvaneaton <@t> littonlab.com>
> Hi Anne,
>
> We have been checking into cassette printers. Obviously, if we can bring
> one in for a demonstration we will but I thought the iDent looked like a
> good place to start. I noticed your problem (on the Histonet) with the ink
> rubbing off after the xylene. I never saw any responses from the rest of the
> Histonetters... did you resolve the problem? Assuming you (or Sakura) did,
> what was the trick?
>
> Thank you in advance for sharing your "fix" for a potential problem.
>
> Denise Van Eaton HT(ASCP)
> Litton Pathology Associates
>
--
Anne van Binsbergen (Hope)
Abu Dhabi
UAE
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 18
Date: Wed, 13 Jan 2010 11:24:19 -0500
From: John Kiernan <jkiernan <@t> uwo.ca>
Subject: Re: RE: [Histonet] Problems with MSB stain
To: Stephen.Eyres <@t> sanofi-aventis.com
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <fc2bbb6c1666f.4b4dad63 <@t> uwo.ca>
Content-Type: text/plain; CHARSET=US-ASCII
The unduly blue sections are all from Animals 12, 13 and 14. Anything different about these specimens? The blue and red sections surely must look different under the microscope.
John Kiernan
Anatomy, UWO
London, Canada
= = =
----- Original Message -----
From: Stephen.Eyres <@t> sanofi-aventis.com
Date: Wednesday, January 13, 2010 10:12
Subject: RE: [Histonet] Problems with MSB stain
To: jkiernan <@t> uwo.ca
> Hi John,
>
> Many thanks for the rapid response. The slides look fine down the microscope except for a few that macroscopically look blue. However because the MSBs will be sent away for evaluation, we want to send a consistent set of slides. I attach a picture to show the problem. I can also scan a slide if you prefer, but the image size is large. The paper was very helpful.
>
> Cheers
>
> Steve
>
> From: John Kiernan [mailto:jkiernan <@t> uwo.ca]
> Sent: Wednesday, January 13, 2010 6:23 AM
> To: Eyres, Stephen R&D/GB
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: Re: [Histonet] Problems with MSB stain
>
> What do the slides look like under the microscope? In a normal liver there shouldn't be much collagen (blue) and there should be plenty of cytoplasm (red). If the preceding nuclear stain was too strong, that could put unwanted bluish shades in the cytoplasm.
>
> Trichrome methods aren't at their best after formaldehyde fixation; the colours can be improved by soaking the hydrated sections in saturated aqueous picric acid for 30 min or so at 55-60C before staining. Bouin's solution is also used for this purpose, and it is said that the same effect can be achieved with citrate buffer pH4 or with 1% iodine in 2% aqueous KI. See Yu Y, Chapman CM (2003) Masson trichrome stain: postfixation substitutes. J. Histotechnol. 26: 131-134. I've not yet tried these alternatives to picric acid; has anyone else?
>
> Lendrum's 1962 paper, which includes colour photos, includes many technical tips. More than 40 years ago a med lab technician in Birmingham, UK, suggested this method to me as a way to impart different colours to gliosis (red) and collagenous scarring (blue) in sections of the brains of frogs and other animals in which I'd made surgical lesions dorsal to the optic chiasma. The MSB method showed these changes very well. I recall that we had to buy about 500 grams of brilliant crystal scarlet 6R because it was then cheaper as an an article of commerce than as a bioogical stain.
>
> I have appended a PDF of Lendrum et al 1962. This won't gwt through to all Histonetters, but it might make it to Stephen.Eyres <@t> sanofi-aventis.com.
>
>
> John Kiernan
> Anatomy, UWO
> London, Canada
> = = =
> ----- Original Message -----
> From: Stephen.Eyres <@t> sanofi-aventis.com
> Date: Tuesday, January 12, 2010 11:06
> Subject: [Histonet] Problems with MSB stain
> To: histonet <@t> lists.utsouthwestern.edu
>
> > Hi,
> >
> > I have just encountered a problem I have never seen before and would
> > welcome opinions as to the cause. I work in pharma R&D and had a
> > requestto perform MSBs on some liver slides. The results varied
> > within the run,
> > with a distinct difference in the overall colour of the
> > slides; some
> > appearing blue and others red. The person performing the
> > staining is
> > experienced, and our neqas scores are good. When we investigated the
> > problem the only difference we could identify was that the bluer
> > slideswere stained after 40 minutes drying after being
> > cut, whereas the
> > redder slides were cut the day before. This suggests that maybe slides
> > were incompletely dried and that this affected the staining. Comments
> > please.
> >
> > Cheers
> >
> > Steve
> > _______________________________________________
> > Histonet mailing list
> > Histonet <@t> lists.utsouthwestern.edu
> > http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 19
Date: Wed, 13 Jan 2010 08:50:18 -0800 (PST)
From: Massimo <max_histo_00 <@t> yahoo.it>
Subject: [Histonet] (Used microtome) - Thanks ...
To: histonet <@t> lists.utsouthwestern.edu
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <939066.56789.qm <@t> web23307.mail.ird.yahoo.com>
Content-Type: text/plain; charset=utf-8
... to all those who kindly responded at my email.
Best Regards,
Massimo Tosi
=================================================
In ricordo di "Nice":
"E Argo, che aveva visto Odisseo dopo vent???anni,
fu preso dal Fato della nera morte."
Omero, Odissea, XVII, 290-327
------------------------------
Message: 20
Date: Wed, 13 Jan 2010 08:50:18 -0800 (PST)
From: Massimo <max_histo_00 <@t> yahoo.it>
Subject: [Histonet] (Used microtome) - Thanks ...
To: histonet <@t> lists.utsouthwestern.edu
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <939066.56789.qm <@t> web23307.mail.ird.yahoo.com>
Content-Type: text/plain; charset=utf-8
... to all those who kindly responded at my email.
Best Regards,
Massimo Tosi
=================================================
In ricordo di "Nice":
"E Argo, che aveva visto Odisseo dopo vent???anni,
fu preso dal Fato della nera morte."
Omero, Odissea, XVII, 290-327
------------------------------
Message: 21
Date: Wed, 13 Jan 2010 17:58:18 +0100
From: "Gudrun Lang" <gu.lang <@t> gmx.at>
Subject: AW: [Histonet] Problems with MSB stain
To: <Stephen.Eyres <@t> sanofi-aventis.com>
Cc: histonet <@t> lists.utsouthwestern.edu
Message-ID: <76730F9EC40D4109BDEB62A3B9762C60 <@t> dielangs.at>
Content-Type: text/plain; charset="iso-8859-1"
Hi Steve,
I have noticed that the CAB-stain, which is also a trichrome-stain (like
Trichrome Gomori), shows a blue overall appearence after NBF-fixation alone.
After re-fixation in Bouin the staining shows normal red staining of
liverslides. Re-fixation is like "antigen-retrieval" and opens the right
binding sites for the dyes.
Did you re-fix your slides in Bouin? Is there a difference in fixation of
the specimen?
Gudrun Lang
-----Urspr?ngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von
Stephen.Eyres <@t> sanofi-aventis.com
Gesendet: Dienstag, 12. J?nner 2010 17:05
An: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] Problems with MSB stain
Hi,
I have just encountered a problem I have never seen before and would
welcome opinions as to the cause. I work in pharma R&D and had a request
to perform MSBs on some liver slides. The results varied within the run,
with a distinct difference in the overall colour of the slides; some
appearing blue and others red. The person performing the staining is
experienced, and our neqas scores are good. When we investigated the
problem the only difference we could identify was that the bluer slides
were stained after 40 minutes drying after being cut, whereas the
redder slides were cut the day before. This suggests that maybe slides
were incompletely dried and that this affected the staining. Comments
please.
Cheers
Steve
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 22
Date: Wed, 13 Jan 2010 21:25:28 +0400
From: Anne van Binsbergen <annigyg <@t> gmail.com>
Subject: Re: [Histonet] Re: Sakura iDent
To: Nancy Klemme <NKlemme <@t> sakuraus.com>
Cc: "histonet <@t> lists.utsouthwestern.edu"
<Histonet <@t> lists.utsouthwestern.edu>, Denise Van Eaton
<dvaneaton <@t> littonlab.com>
Message-ID:
<f8332fbe1001130925s7fcb6db8xdff1e632ad45a3f1 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Dear Nancy
thank you for your email - yes, I know Ghaleb well and have had a working
relationship with him for many years here in the middle east. He is indeed
trying to extract some sort of resolution from the Sakura European Head
office in Zoeterwoude - Ralph, Wim, Chris and Kees-Jan ....my local vendor
here in the UAE is excellent and he has witnessed my print rubbing off the
cassette face when gently brushed with xylene on gloved finger or a cotton
swab.
I must just add that it is very strange for me to be battling with a company
I have avidly supported and promoted for so many years.
I hope that we will resolve this issue and that i will add yet another
Sakura workhorse to my already large Sakura stable.
kind regards
Annie
2010/1/13 Nancy Klemme <NKlemme <@t> sakuraus.com>
> Dear Annie,
>
> I am writing you from California where our Wednesday workday is beginning
> and you are preparing for your Wednesday to draw to a close.
>
> Although the majority of Sakura Finetek products are carried by all Sakura
> Finetek entities, the iDent is not one that is available from Sakura Finetek
> USA in North America. If most Histonet subscribers are from the U.S., that
> may explain the "very little response from the histonet" that you noted.
>
> I am sorry about your cassette labeler issues and have forwarded your
> posting to the gentleman who is in charge of Sakura Finetek Middle East.
> His name is Mr. Ghaleb Alkhaseb and I am sure he will be in contact with
> you very soon after reading it. I do not know what Sakura contact
> information you have been using, but I know that Mr. Alkhaseb will make sure
> that you will have the most current information for a more direct company
> contact. I do expect your iDent situation will be corrected and resolved to
> your satisfaction very soon.
>
> From 12 hours behind you, I offer my kindest regards.
>
> Nancy Klemme, HT(ASCP)
> Edu. Svcs. Dir. - Sakura Finetek USA, Inc. - Torrance, CA
> 800-725-8723 x7879
>
> -----Original Message-----
> From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:
> histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Anne van
> Binsbergen
> Sent: Wednesday, January 13, 2010 1:16 AM
> To: Denise Van Eaton
> Cc: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] Re: Sakura iDent
>
> Hi Denise
> sadly no resolution
> also sadly very little response from the histonet - except for Rene - he
> and
> i did some troubleshooting to try to see if i had missed anything - we had
> the same thoughts
> so ... i am still sitting with the iDent in its box, unused, until someone,
> somewhere can convince me that the print does not wipe off with xylene -
> which will be hard to do seeing that i wiped it off myself and saw with my
> own eyes that xylene removes the ink!!
>
> Sakura - if you are reading this - i am not a happy camper
>
> Annie
> 2010/1/12 Denise Van Eaton <dvaneaton <@t> littonlab.com>
>
> > Hi Anne,
> >
> > We have been checking into cassette printers. Obviously, if we can bring
> > one in for a demonstration we will but I thought the iDent looked like a
> > good place to start. I noticed your problem (on the Histonet) with the
> ink
> > rubbing off after the xylene. I never saw any responses from the rest of
> the
> > Histonetters... did you resolve the problem? Assuming you (or Sakura)
> did,
> > what was the trick?
> >
> > Thank you in advance for sharing your "fix" for a potential problem.
> >
> > Denise Van Eaton HT(ASCP)
> > Litton Pathology Associates
> >
>
>
>
> --
> Anne van Binsbergen (Hope)
> Abu Dhabi
> UAE
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
--
Anne van Binsbergen (Hope)
Abu Dhabi
UAE
------------------------------
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End of Histonet Digest, Vol 74, Issue 12
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