[Histonet] Re: Formalin Fixation issues (Bob Richmond)
Amanda Amador
aamador <@t> pcnm.com
Tue Jul 10 12:11:44 CDT 2012
If you have large specimens, it is best to "open" them as soon as possible. If it is a breast, at least do your prelim measurements and slice it open, leaving it still attached, so that the formain can penetrate the tissue. If you go ahead and slice it up, at least wrap it, in order with paper towels, to keep orientation while it fixes.
You can always block it and then let it fix too. You just need to be sure to slice it thin so that it fixes; and make sure you have adequate spacing between cassettes.
You just need to figure out what way is best for your facility.
Amanda Amador, AAS, ASCP(CM)
Sr. Histotechnician
Pathology Consultants of New Mexico
600 N. Richardson
Roswell, NM 88202
575-622-5600, ext 218
________________________________________
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Today's Topics:
1. Re: Formalin Fixation issues (Bob Richmond)
2. RE: Need help with CD40 on frozen sections (Teri Johnson)
3. Re: Formalin Fixation issues (Bob Richmond)
4. Day Shift Histotech Job Near Ithaca, NY (Melissa Phelan)
5. Re: Re: Formalin Fixation issues (Jill Cox)
6. RE: Re: Formalin Fixation issues (Weems, Joyce K.)
7. RE: Re: Formalin Fixation issues (Elizabeth Chlipala)
8. GBS IHC (Richard Cartun)
9. AUTO: Ramona Nelson is out of the office. (returning
07/16/2012) (Ramona_Nelson <@t> bd.com)
10. Subject: Re: [PATHO-L] refreshing perspective on formaldehyde
(White, Lisa M.)
11. Beecher tissue array 1 mm. punch set. (Mohammad Sayeeduddin)
12. CD40 (Reynolds,Donna M)
13. IVD P63 (Cathy Crumpton)
14. human vimentin in mouse tissue (Kim Merriam)
15. Re: IVD P63 (Kim Donadio)
16. RE: IVD P63 (Clare Thornton)
----------------------------------------------------------------------
Message: 1
Date: Mon, 9 Jul 2012 13:25:05 -0400
From: Bob Richmond <rsrichmond <@t> gmail.com>
Subject: [Histonet] Re: Formalin Fixation issues
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<CAOKsRH7dwef0cYYNKuvFjQgxiia_76vHasD-oVwAuk6k=4eMOg <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
Jill Cox asks: >>Is anyone having problems with breast fixation prior
to processing in formalin? For a couple of months now our breast
specimens aren't fixing very well before gross. Our pathologist thinks
they have changed something in the formalin itself. We utilize 15/1
ratio and in some cases let fix over the weekend. This has only been
happening over the last couple of months and can't seem to figure this
out. Any advice or similar problems, would love to hear from you.<<
Breast specimens shouldn't be expected to fix before they're grossed,
or at least before they've been cut into thin slices. Delaying
fixation compromises immunostaining, to say nothing of H & E. They
should be grossed as promptly as possible after they're received, and
should never sit over the weekend without dissection.
I seriously doubt that anything in the formalin has changed. It's your
technique that needs to change. Unfortunately, this is a difficult
task to delegate.
Bob Richmond
Samurai Pathologist
Knoxville TN
------------------------------
Message: 2
Date: Mon, 9 Jul 2012 17:46:25 +0000
From: Teri Johnson <TJohnson <@t> gnf.org>
Subject: [Histonet] RE: Need help with CD40 on frozen sections
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Cc: "Bea DeBrosse-Serra \(BDeBrosse-Serra <@t> isisph.com\)"
<BDeBrosse-Serra <@t> isisph.com>
Message-ID: <9F3CFEE76E51B64991C7485270890B400CDC4A3B <@t> EX4.lj.gnf.org>
Content-Type: text/plain; charset="us-ascii"
Hi Bea!
I just left you a voicemail message. I looked up this antibody data sheet and to say the least the staining on the image is underwhelming!
I would try fixing the cryosections with Beckstead's zinc prior to staining, and you can even consider fixing the spleens with Zinc and then cryoprotecting with sucrose prior to freezing. It appears they recommend using a biotinylated secondary and then streptavidin-HRP for staining. You might see if that helps as well.
Teri
Teri Johnson, HT(ASCP)QIHC
GNF Histology Lab Manager
Genomics Institute of the Novartis Research Foundation
858-332-4752
------------------------------
Message: 3
Date: Mon, 9 Jul 2012 13:51:10 -0400
From: Bob Richmond <rsrichmond <@t> gmail.com>
Subject: [Histonet] Re: Formalin Fixation issues
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<CAOKsRH4+MW_cyH=TgpZQE=BGrBRd8AVuay+Cs-yJ-gSD=kkcGw <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1
At this point I'm not sure I understand what Jill Cox's problem is.
Forgive me if I keep blaming the pathologist - since I am one! - but
in my experience most of these breast problems originate at the gross
desk.
Is your pathologist cutting the fatty breast tissue thin enough? This
is always a challenge, even after you've done it as long as I have. If
your pathologist is cramming the cassettes full of fat, then that's
where the problem lies.
Do you prepare your own neutral buffered formalin, or buy it? I
prepared a lot of it years ago, and supervised others who did, and I
learned the hard way that mistakes are easy to make when you brew your
own. If you buy your NBF ready made, as most people do nowadays, then
read the label carefully. Have you changed brands recently?
Bob Richmond
Samurai Pathologist
Knoxville TN
***************************************************
On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox <jcox90 <@t> yahoo.com> wrote:
> I think you misunderstood what I meant. I know it's not supposed to be fixed
> before grossing, it's also not getting fixed after processing. But thanks
> for your two cents..
>
> From: Bob Richmond <rsrichmond <@t> gmail.com>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Monday, July 9, 2012 10:25 AM
> Subject: [Histonet] Re: Formalin Fixation issues
>
> Jill Cox asks: >>Is anyone having problems with breast fixation prior
> to processing in formalin? For a couple of months now our breast
> specimens aren't fixing very well before gross. Our pathologist thinks
> they have changed something in the formalin itself. We utilize 15/1
> ratio and in some cases let fix over the weekend. This has only been
> happening over the last couple of months and can't seem to figure this
> out. Any advice or similar problems, would love to hear from you.<<
>
> Breast specimens shouldn't be expected to fix before they're grossed,
> or at least before they've been cut into thin slices. Delaying
> fixation compromises immunostaining, to say nothing of H & E. They
> should be grossed as promptly as possible after they're received, and
> should never sit over the weekend without dissection.
>
> I seriously doubt that anything in the formalin has changed. It's your
> technique that needs to change. Unfortunately, this is a difficult
> task to delegate.
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
------------------------------
Message: 4
Date: Mon, 09 Jul 2012 13:54:26 -0400
From: Melissa Phelan <melissa <@t> alliedsearchpartners.com>
Subject: [Histonet] Day Shift Histotech Job Near Ithaca, NY
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <CC209312.1E445%melissa <@t> alliedsearchpartners.com>
Content-Type: text/plain; charset="US-ASCII"
Hi Everyone,
I have a Day Shift Monday-Friday Histotech job opening about 45 miles
Southeast of Ithaca, NY (About an hour North of Scranton, PA). This position
requires a NY license and ASCP certification. Please message me for details
if you are interested!
To view a complete list of Allied Search Partners current openings go to:
http://www.alliedsearchpartners.com/careers.php
Melissa Phelan
LinkedIn: http://www.linkedin.com/in/melissaphelan
President, Laboratory Staffing
Allied Search Partners
P: 888.388.7571
F: 888.388.7572
M: 407.697.1175
www.alliedsearchpartners.com <http://www.alliedsearchpartners.com/>
------------------------------
Message: 5
Date: Mon, 9 Jul 2012 11:00:41 -0700 (PDT)
From: Jill Cox <jcox90 <@t> yahoo.com>
Subject: Re: [Histonet] Re: Formalin Fixation issues
To: Bob Richmond <rsrichmond <@t> gmail.com>, "Histonet <@t> Lists. Edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<1341856841.88220.YahooMailNeo <@t> web161606.mail.bf1.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Yes Bob, the sections are good and thin. We use Scientific Products, 10% NBF. Just seems like the Formalin is watered down. Going to try a different brand. I have worked for Pathologist's that cram cassettes but this is not the case here.
From: Bob Richmond <rsrichmond <@t> gmail.com>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Monday, July 9, 2012 10:51 AM
Subject: [Histonet] Re: Formalin Fixation issues
At this point I'm not sure I understand what Jill Cox's problem is.
Forgive me if I keep blaming the pathologist - since I am one! - but
in my experience most of these breast problems originate at the gross
desk.
Is your pathologist cutting the fatty breast tissue thin enough? This
is always a challenge, even after you've done it as long as I have. If
your pathologist is cramming the cassettes full of fat, then that's
where the problem lies.
Do you prepare your own neutral buffered formalin, or buy it? I
prepared a lot of it years ago, and supervised others who did, and I
learned the hard way that mistakes are easy to make when you brew your
own. If you buy your NBF ready made, as most people do nowadays, then
read the label carefully. Have you changed brands recently?
Bob Richmond
Samurai Pathologist
Knoxville TN
***************************************************
On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox <jcox90 <@t> yahoo.com> wrote:
> I think you misunderstood what I meant. I know it's not supposed to be fixed
> before grossing, it's also not getting fixed after processing. But thanks
> for your two cents..
>
> From: Bob Richmond <rsrichmond <@t> gmail.com>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Monday, July 9, 2012 10:25 AM
> Subject: [Histonet] Re: Formalin Fixation issues
>
> Jill Cox asks: >>Is anyone having problems with breast fixation prior
> to processing in formalin? For a couple of months now our breast
> specimens aren't fixing very well before gross. Our pathologist thinks
> they have changed something in the formalin itself. We utilize 15/1
> ratio and in some cases let fix over the weekend. This has only been
> happening over the last couple of months and can't seem to figure this
> out. Any advice or similar problems, would love to hear from you.<<
>
> Breast specimens shouldn't be expected to fix before they're grossed,
> or at least before they've been cut into thin slices. Delaying
> fixation compromises immunostaining, to say nothing of H & E. They
> should be grossed as promptly as possible after they're received, and
> should never sit over the weekend without dissection.
>
> I seriously doubt that anything in the formalin has changed. It's your
> technique that needs to change. Unfortunately, this is a difficult
> task to delegate.
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 6
Date: Mon, 9 Jul 2012 18:33:56 +0000
From: "Weems, Joyce K." <Joyce.Weems <@t> emoryhealthcare.org>
Subject: RE: [Histonet] Re: Formalin Fixation issues
To: "'Jill Cox'" <jcox90 <@t> yahoo.com>, Bob Richmond
<rsrichmond <@t> gmail.com>, "Histonet <@t> Lists. Edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<E3A4EBD57A691646BCCED4AA5911A0301A96E9BF <@t> e14mbx12n.Enterprise.emory.net>
Content-Type: text/plain; charset="us-ascii"
I would also call the company to see if they have had others with problems. Has it all been the same lot number?
Joyce Weems
Pathology Manager
678-843-7376 Phone
678-843-7831 Fax
joyce.weems <@t> emoryhealthcare.org
www.saintjosephsatlanta.org
5665 Peachtree Dunwoody Road
Atlanta, GA 30342
This e-mail, including any attachments is the property of Saint Joseph's Hospital and is intended for the sole use of the intended recipient(s). It may contain information that is privileged and confidential. Any unauthorized review, use, disclosure, or distribution is prohibited. If you are not the intended recipient, please delete this message, and reply to the sender regarding the error in a separate email.
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jill Cox
Sent: Monday, July 09, 2012 2:01 PM
To: Bob Richmond; Histonet <@t> Lists. Edu
Subject: Re: [Histonet] Re: Formalin Fixation issues
Yes Bob, the sections are good and thin. We use Scientific Products, 10% NBF. Just seems like the Formalin is watered down. Going to try a different brand. I have worked for Pathologist's that cram cassettes but this is not the case here.
From: Bob Richmond <rsrichmond <@t> gmail.com>
To: histonet <@t> lists.utsouthwestern.edu
Sent: Monday, July 9, 2012 10:51 AM
Subject: [Histonet] Re: Formalin Fixation issues
At this point I'm not sure I understand what Jill Cox's problem is.
Forgive me if I keep blaming the pathologist - since I am one! - but in my experience most of these breast problems originate at the gross desk.
Is your pathologist cutting the fatty breast tissue thin enough? This is always a challenge, even after you've done it as long as I have. If your pathologist is cramming the cassettes full of fat, then that's where the problem lies.
Do you prepare your own neutral buffered formalin, or buy it? I prepared a lot of it years ago, and supervised others who did, and I learned the hard way that mistakes are easy to make when you brew your own. If you buy your NBF ready made, as most people do nowadays, then read the label carefully. Have you changed brands recently?
Bob Richmond
Samurai Pathologist
Knoxville TN
***************************************************
On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox <jcox90 <@t> yahoo.com> wrote:
> I think you misunderstood what I meant. I know it's not supposed to be
> fixed before grossing, it's also not getting fixed after processing.
> But thanks for your two cents..
>
> From: Bob Richmond <rsrichmond <@t> gmail.com>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Monday, July 9, 2012 10:25 AM
> Subject: [Histonet] Re: Formalin Fixation issues
>
> Jill Cox asks: >>Is anyone having problems with breast fixation prior
> to processing in formalin? For a couple of months now our breast
> specimens aren't fixing very well before gross. Our pathologist thinks
> they have changed something in the formalin itself. We utilize 15/1
> ratio and in some cases let fix over the weekend. This has only been
> happening over the last couple of months and can't seem to figure this
> out. Any advice or similar problems, would love to hear from you.<<
>
> Breast specimens shouldn't be expected to fix before they're grossed,
> or at least before they've been cut into thin slices. Delaying
> fixation compromises immunostaining, to say nothing of H & E. They
> should be grossed as promptly as possible after they're received, and
> should never sit over the weekend without dissection.
>
> I seriously doubt that anything in the formalin has changed. It's your
> technique that needs to change. Unfortunately, this is a difficult
> task to delegate.
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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Message: 7
Date: Mon, 9 Jul 2012 12:54:10 -0600
From: Elizabeth Chlipala <liz <@t> premierlab.com>
Subject: RE: [Histonet] Re: Formalin Fixation issues
To: 'Bob Richmond' <rsrichmond <@t> gmail.com>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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There is a real nice article on fixation and handling of breast lumpectomy samples in the Journal of Histotechnology by Dr. Stephen Ruby. It goes over a nice technique we used to use on those samples, worked great. It did use alcoholic formalin which I suspect can't be used currently if you have not validated for that fixative for ER/PR etc. But the overall technique can be used with 10% NBF. The thickness of samples is key as Bob pointed out. We had a new pathologist on gross and the next day we had about 20 or 30 recuts on their breast samples. That number of reacts was uncommon. I needed to figure out what was the issue and we found out it was the size of tissue that was placed into the cassettes. I explained that they needed to place smaller and thinner pieces of tissue in the cassette. Once I did that we did not have anymore problems with their samples. The article information is below.
Paper Towel "Sandwich" - Alcoholic Formalin Fixation for Breast Biopsies
Author: Ruby, Stephen G.
Source: Journal of Histotechnology, Number 1, March 1999 , pp. 49-51(3)
Publisher: Maney Publishing
Liz
Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308-1592
(303) 682-3949 office
(303) 682-9060 fax
(303) 881-0763 cell
www.premierlab.com
Ship to address:
1567 Skyway Drive, Unit E
Longmont, CO 80504
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Bob Richmond
Sent: Monday, July 09, 2012 11:51 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Re: Formalin Fixation issues
At this point I'm not sure I understand what Jill Cox's problem is.
Forgive me if I keep blaming the pathologist - since I am one! - but
in my experience most of these breast problems originate at the gross
desk.
Is your pathologist cutting the fatty breast tissue thin enough? This
is always a challenge, even after you've done it as long as I have. If
your pathologist is cramming the cassettes full of fat, then that's
where the problem lies.
Do you prepare your own neutral buffered formalin, or buy it? I
prepared a lot of it years ago, and supervised others who did, and I
learned the hard way that mistakes are easy to make when you brew your
own. If you buy your NBF ready made, as most people do nowadays, then
read the label carefully. Have you changed brands recently?
Bob Richmond
Samurai Pathologist
Knoxville TN
***************************************************
On Mon, Jul 9, 2012 at 1:38 PM, Jill Cox <jcox90 <@t> yahoo.com> wrote:
> I think you misunderstood what I meant. I know it's not supposed to be fixed
> before grossing, it's also not getting fixed after processing. But thanks
> for your two cents..
>
> From: Bob Richmond <rsrichmond <@t> gmail.com>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Monday, July 9, 2012 10:25 AM
> Subject: [Histonet] Re: Formalin Fixation issues
>
> Jill Cox asks: >>Is anyone having problems with breast fixation prior
> to processing in formalin? For a couple of months now our breast
> specimens aren't fixing very well before gross. Our pathologist thinks
> they have changed something in the formalin itself. We utilize 15/1
> ratio and in some cases let fix over the weekend. This has only been
> happening over the last couple of months and can't seem to figure this
> out. Any advice or similar problems, would love to hear from you.<<
>
> Breast specimens shouldn't be expected to fix before they're grossed,
> or at least before they've been cut into thin slices. Delaying
> fixation compromises immunostaining, to say nothing of H & E. They
> should be grossed as promptly as possible after they're received, and
> should never sit over the weekend without dissection.
>
> I seriously doubt that anything in the formalin has changed. It's your
> technique that needs to change. Unfortunately, this is a difficult
> task to delegate.
>
> Bob Richmond
> Samurai Pathologist
> Knoxville TN
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
------------------------------
Message: 8
Date: Mon, 09 Jul 2012 16:32:58 -0400
From: "Richard Cartun" <Rcartun <@t> harthosp.org>
Subject: [Histonet] GBS IHC
To: "Histonet" <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <4FFB07BA.7400.0077.1 <@t> harthosp.org>
Content-Type: text/plain; charset=US-ASCII
Does anyone know of a good antibody to Group B Streptococci (GBS) that works on formalin-fixed, paraffin embedded tissue? I had a wonderful mAb from a company in Canada, but they went out of business. Thank you.
Richard
Richard W. Cartun, MS, PhD
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT 06102
(860) 545-1596 Office
(860) 545-2204 Fax
------------------------------
Message: 9
Date: Tue, 10 Jul 2012 07:50:59 -0400
From: Ramona_Nelson <@t> bd.com
Subject: [Histonet] AUTO: Ramona Nelson is out of the office.
(returning 07/16/2012)
To: histonet <@t> lists.utsouthwestern.edu
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<OF74F07663.41232527-ON85257A37.004117E7-85257A37.004117E7 <@t> bd.com>
Content-Type: text/plain; charset="US-ASCII"
I am out of the office until 07/16/20
I
Note: This is an automated response to your message &quo t;Histonet Digest, Vol 104, Issue 10" sent on 7/9/2012 1:00:01 PM.
This is the only notification you will receive whi person is away.
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Message: 10
Date: Tue, 10 Jul 2012 09:45:31 -0400
From: "White, Lisa M." <Lisa.White3 <@t> va.gov>
Subject: [Histonet] Subject: Re: [PATHO-L] refreshing perspective on
formaldehyde
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<2B2ECF33934F5D4996D8BE03EFDF397609E9E660 <@t> VHAV09MSGA3.v09.med.va.gov>
Content-Type: text/plain; charset="us-ascii"
For the paranoid out there: imagine what a lawyer will do to a hospital
in court when Formalin (or other proper fixative) was not allowed to be
utilized and the specimen is ruined. The lawsuit settlement will make a
fine from OSHA look like buying a candy bar. More than likely it will
be a member of or government or high society type who is "wronged"
before changes are made.
It is hard to believe that a hospital would allow the fixative to be
removed from all areas. Yes everyone who is utilizing the fixative
needs to be educated in spill clean-up and have spill kits maintained in
their area. It is a small price to pay to protect the integrity of the
specimen. What if it was your loved ones specimen? I have heard of
others leaving the taps open. The remedy was to buy bottles with screw
caps with the bonus of the fixative not being as heavy.
Lisa White, HT(ASCP)
Supervisory HT
James H. Quillen VAMC
PO Box 4000
Corner of Veterans Way and Lamont
PLMS 113
Mountain Home, TN 37684
423-979-3567
423-979-3401 fax
------------------------------
Message: 11
Date: Tue, 10 Jul 2012 07:02:17 -0700 (PDT)
From: Mohammad Sayeeduddin <sayeeduddin <@t> sbcglobal.net>
Subject: [Histonet] Beecher tissue array 1 mm. punch set.
To: histonet <@t> lists.utsouthwestern.edu
Message-ID:
<1341928937.86834.YahooMailClassic <@t> web81506.mail.mud.yahoo.com>
Content-Type: text/plain; charset=us-ascii
Hi Histonet members, I am in trouble. I need 1 mm. tissue array punch set ( Beecher manual instrument ). Either you can sell us one or exchange for a box of ten sets of 0.6mm size punch sets. Please contact me at sayeeduddin <@t> sbcglobal.net
Thanks in advance for your response.
Sayeed.
------------------------------
Message: 12
Date: Tue, 10 Jul 2012 09:32:51 -0500
From: "Reynolds,Donna M" <dreynold <@t> mdanderson.org>
Subject: [Histonet] CD40
To: "'histonet <@t> lists.utsouthwestern.edu'"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<785BBF0C5F49CE41BA74460A43A08F0232223A8783 <@t> DCPWVMBXC0VS3.mdanderson.edu>
Content-Type: text/plain; charset="us-ascii"
What are you using for Endogenous blocking? If you are using methanol with H2O2 that may be your problem. Many CD antibodies are very sensitive to the methanol. Try PBS or whatever buffer you are using to dilute the H202. I would also try going straight form the -20 into cold acetone without air drying and fix for10 min.
Donna Reynolds HT (ASCP)
U. T. M.D. Anderson Cancer Center
Chief histology Tech, Department Cancer Biology
713-792-8106
------------------------------
Message: 3
Date: Mon, 9 Jul 2012 07:55:16 -0700
From: Bea DeBrosse-Serra <BDeBrosse-Serra <@t> isisph.com>
Subject: [Histonet] Need help with CD40 on frozen sections
To: "'Histonet <@t> lists.utsouthwestern.edu'"
<Histonet <@t> lists.utsouthwestern.edu>
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<493CAA64F203E14E8823737B9EE0E25F092BA8FBFB <@t> EXCHMB01.isis.local>
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Histonetters,
I need some help with CD40 on frozen sections (spleen, non-treated). I cut my frozens, let them briefly air dry and keep them at -20?C. When I use them for staining, I air dry them, fix them in cold acetone for 5 minutes and start the staining process. I use the primary AB from BD Pharmingen, # 550285 at various dilution, incubate them overnight at 4?C, use the secondary HRP conjugated antibody, DAB, counterstain and don't get any staining. Does anyone have any pointers, suggestions?
Thank you in advance,
Bea
Beatrice DeBrosse-Serra HT(ASCP)QIHC
Isis Pharmaceuticals
Antisense Drug Discovery
2855 Gazelle Ct.
Carlsbad, CA 92010
760-603-2371
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Message: 13
Date: Tue, 10 Jul 2012 15:51:00 +0000
From: Cathy Crumpton <cathy.crumpton <@t> tuality.org>
Subject: [Histonet] IVD P63
To: "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
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<AC889E5706400B438D42F8A07AE3AF8B01787246 <@t> EXMBXSRV01.corp.tuality.net>
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I was just told by my Biocare rep that today was the last day they are selling P63 antibody. I would like to purchase IVD P63 from another vendor, who else sells it? I have having a few problems finding good info just using Google. What is your lab using for diagnostic purposes?
Cathy Crumpton HT(ASCP), Lead Histotechnician
Tuality Community Hospital
503-681-1292
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Message: 14
Date: Tue, 10 Jul 2012 08:48:54 -0700 (PDT)
From: Kim Merriam <kmerriam2003 <@t> yahoo.com>
Subject: [Histonet] human vimentin in mouse tissue
To: Histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<1341935334.82981.YahooMailNeo <@t> web130104.mail.mud.yahoo.com>
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Greetings!
?
I want to stain for human vimentin in mouse tissues (human fibroblasts).? I many years ago, I used?a biotinylated V9 clone for just this purpose and it worked great.??I tried the?biotiylated V9 clone from abcam, but I am not getting any staining at all.? Any suggestions?
?
Kim
?
Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA?
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Message: 15
Date: Tue, 10 Jul 2012 09:02:38 -0700 (PDT)
From: Kim Donadio <one_angel_secret <@t> yahoo.com>
Subject: Re: [Histonet] IVD P63
To: Cathy Crumpton <cathy.crumpton <@t> tuality.org>,
"histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<1341936158.34785.YahooMailNeo <@t> web112318.mail.gq1.yahoo.com>
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Ventana has gotten the sole patton for that antibody now I beleive, or will have soon.
________________________________
From: Cathy Crumpton <cathy.crumpton <@t> tuality.org>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, July 10, 2012 11:51 AM
Subject: [Histonet] IVD P63
I was just told by my Biocare rep that today was the last day they are selling P63 antibody.? I would like to purchase IVD P63 from another vendor, who else sells it?? I have having a few problems finding good info just using Google.? What is your lab using for diagnostic purposes?
Cathy Crumpton HT(ASCP), Lead Histotechnician
Tuality Community Hospital
503-681-1292
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Histonet <@t> lists.utsouthwestern.edu
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------------------------------
Message: 16
Date: Tue, 10 Jul 2012 12:48:20 -0400
From: Clare Thornton <CThornton <@t> dahlchase.com>
Subject: RE: [Histonet] IVD P63
To: 'Kim Donadio' <one_angel_secret <@t> yahoo.com>, Cathy Crumpton
<cathy.crumpton <@t> tuality.org>, "histonet <@t> lists.utsouthwestern.edu"
<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
<C9D78FFC9D668B4CBEA4405F846975040109B4F73615 <@t> iris.dahlchase.net>
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I just spoke with my Biocare rep. He said it is untrue at this time, and they are still selling p63.
Clare J. Thornton, HTL(ASCP),QIHC
Assistant Histology Supervisor
Dahl-Chase Diagnostic Services
417 State Street, Suite 540
Bangor, ME 04401
cthornton <@t> dahlchase.com
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Kim Donadio
Sent: Tuesday, July 10, 2012 12:03 PM
To: Cathy Crumpton; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] IVD P63
Ventana has gotten the sole patton for that antibody now I beleive, or will have soon.
________________________________
From: Cathy Crumpton <cathy.crumpton <@t> tuality.org>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Sent: Tuesday, July 10, 2012 11:51 AM
Subject: [Histonet] IVD P63
I was just told by my Biocare rep that today was the last day they are selling P63 antibody.? I would like to purchase IVD P63 from another vendor, who else sells it?? I have having a few problems finding good info just using Google.? What is your lab using for diagnostic purposes?
Cathy Crumpton HT(ASCP), Lead Histotechnician
Tuality Community Hospital
503-681-1292
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet
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End of Histonet Digest, Vol 104, Issue 11
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