[Histonet] llEICA sTAINER

Madary, Joseph MadaryJ <@t> MedImmune.com
Wed Oct 21 12:08:41 CDT 2009


You can just start step one from water as your first step.  Step one Station "wash1" for example.  We use that for frozen sections

Nick Madary, HT/HTL(ASCP)QIHC
Medimmune Histology Mgr, 
OMW, Area 4, Lab 2438
301.398.4745(vm)
301.398.6360(lab)
301.398.9745(fax)
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Today's Topics:

   1. Antigen retrieval question (Connolly, Brett M)
   2. Re: Antigen retrieval question (Ms Janet Tao)
   3. RE: Antigen retrieval question (Connolly, Brett M)
   4. Leica Autostainer (Jennifer Anderson)
   5. Unsubscribe from website (Witt, Dan)
   6. RE: Leica Autostainer (Rathborne, Toni)
   7. Re: Unsubscribe from website (Peter Carroll)
   8. RE: Antigen retrieval question (JR R)
   9. Re: Antigen retrieval question (Rene J Buesa)
  10. RE: Antigen retrieval question (Mejia, Maria)
  11. RE: in search of cd31 for rat tissue (TF) AND novocastra eNOS
      antibody on rat tissues???  (PALMER Jason (SVHM))
  12. Fite control slides (Jennifer Campbell)
  13. Granzyme B and F4/80 immunostaining (Jennifer Campbell)
  14. CD 31 in rats (Melanie Black)
  15. Leica Stainer Question (Breeden, Sara)
  16. Beckstead ZSF   FW: Zn fixative 550523 vs 552658
      (Cormier, Kathleen)
  17. Paraplast (Ian Montgomery)
  18. PRN.POSITION (Stanjeski Paula)


----------------------------------------------------------------------

Message: 1
Date: Tue, 20 Oct 2009 14:21:47 -0400
From: "Connolly, Brett M" <brett_connolly <@t> merck.com>
Subject: [Histonet] Antigen retrieval question
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<63EA0607835FBA4689CEA9EA8B482692026E4C5B <@t> usctmx1141.merck.com>
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Has anyone experienced a "reversal" of citrate HEIR unmasking do to a
prolonged delay in continuing the experiment after the retrieval step? 

Brett M. Connolly, Ph.D.
Research Fellow, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
tel. 215-652-2501 fax. 215-993-6803
brett_connolly <@t> merck.com

  
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Message: 2
Date: Tue, 20 Oct 2009 11:30:55 -0700 (PDT)
From: Ms Janet Tao <tao_janet <@t> yahoo.com>
Subject: Re: [Histonet] Antigen retrieval question
To: histonet <@t> lists.utsouthwestern.edu,	Brett MConnolly
	<brett_connolly <@t> merck.com>
Message-ID: <491199.26795.qm <@t> web57008.mail.re3.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Sounds like the retrieved antigen was lost rather than "reversal"

Janet
QIHC/HTL

--- On Tue, 10/20/09, Connolly, Brett M <brett_connolly <@t> merck.com> wrote:

From: Connolly, Brett M <brett_connolly <@t> merck.com>
Subject: [Histonet] Antigen retrieval question
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, October 20, 2009, 11:21 AM

Has anyone experienced a "reversal" of citrate HEIR unmasking do to a
prolonged delay in continuing the experiment after the retrieval step? 

Brett M. Connolly, Ph.D.
Research Fellow, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
tel. 215-652-2501 fax. 215-993-6803
brett_connolly <@t> merck.com

  
Notice:  This e-mail message, together with any attachments, contains
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------------------------------

Message: 3
Date: Tue, 20 Oct 2009 14:33:04 -0400
From: "Connolly, Brett M" <brett_connolly <@t> merck.com>
Subject: [Histonet] RE: Antigen retrieval question
To: "Morken, Tim" <Timothy.Morken <@t> ucsfmedctr.org>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<63EA0607835FBA4689CEA9EA8B482692026E4C6F <@t> usctmx1141.merck.com>
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Perhaps reversal is not the right word...slides were left overnight in
buffer w/ tween. We got very minimal faint staining compared to previous
runs done in one day.

Brett 

-----Original Message-----
From: Morken, Tim [mailto:Timothy.Morken <@t> ucsfmedctr.org] 
Sent: Tuesday, October 20, 2009 2:29 PM
To: Connolly, Brett M; histonet <@t> lists.utsouthwestern.edu
Subject: RE: Antigen retrieval question

Brett, how long? I've left them overnight before proceeding without any
problems. 

It would not be "re-fixation" as with formalin. However, if it is in a
detergent buffer it might cause some other kind of denaturation.


Tim Morken
Supervisor, Histology / IPOX
UCSF Medical Center
San Francisco, CA  
 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Connolly, Brett M
Sent: Tuesday, October 20, 2009 11:22 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Antigen retrieval question

Has anyone experienced a "reversal" of citrate HEIR unmasking do to a
prolonged delay in continuing the experiment after the retrieval step? 

Brett M. Connolly, Ph.D.
Research Fellow, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
tel. 215-652-2501 fax. 215-993-6803
brett_connolly <@t> merck.com

  
Notice:  This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station,
New Jersey, USA 08889), and/or its affiliates (which may be known
outside the United States as Merck Frosst, Merck Sharp & Dohme or
MSD and in Japan, as Banyu - direct contact information for affiliates
is
available at http://www.merck.com/contact/contacts.html) 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
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------------------------------

Message: 4
Date: Tue, 20 Oct 2009 11:39:45 -0700
From: Jennifer Anderson <janderson <@t> halozyme.com>
Subject: [Histonet] Leica Autostainer
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<5F7CC9B788911848A79BC83453A3D6530360B5D7D4 <@t> Tomlinson.hti.com>
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Good Afternoon Histonet.

Our lab just purchased a Leica XL for routine H&E and hopefully Trichrome.  I have never used this instrument before, and was hoping for some direction on where to optimally place reagents in the machine for the best (and easiest) staining runs.

Thanks so much for your help.

Jennifer M. Anderson, Scientist
Halozyme Therapeutics, Inc.
11388 Sorrento Valley Road
San Diego, CA 92121
858-704-8333
janderson <@t> halozyme.com<mailto:janderson <@t> halozyme.com>



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Message: 5
Date: Tue, 20 Oct 2009 13:42:29 -0500
From: "Witt, Dan" <dwitt <@t> sebh.org>
Subject: [Histonet] Unsubscribe from website
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
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	<FB8FCBE5DD96E34087D2574186FFBCCBE21546D0 <@t> EXCHANGERG1.sebdom1.ad.sebh.org>
	
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I would appreciate you removing me from the Histonet website


Dan Witt PA, HT (ASCP)

Pathologists Assistant / Histology Coordinator

St. Elizabeth's Hospital

211 S. 3 rd Street

Belleville, IL 62220

234-2120 Ext. 1369 or 1192



------------------------------

Message: 6
Date: Tue, 20 Oct 2009 14:48:44 -0400
From: "Rathborne, Toni" <trathborne <@t> somerset-healthcare.com>
Subject: RE: [Histonet] Leica Autostainer
To: "Jennifer Anderson" <janderson <@t> halozyme.com>,
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<E78340C766A5284D999F5F5891DDF8900BAF8EF0 <@t> smcmail.somerset-healthcare.com>
	
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Contact Carolyn Doan at Leica, she's an expert at setting up staining protocols. 

Carolyn.Doan <@t> leica-microsystems.com

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Jennifer
Anderson
Sent: Tuesday, October 20, 2009 2:40 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Leica Autostainer


Good Afternoon Histonet.

Our lab just purchased a Leica XL for routine H&E and hopefully Trichrome.  I have never used this instrument before, and was hoping for some direction on where to optimally place reagents in the machine for the best (and easiest) staining runs.

Thanks so much for your help.

Jennifer M. Anderson, Scientist
Halozyme Therapeutics, Inc.
11388 Sorrento Valley Road
San Diego, CA 92121
858-704-8333
janderson <@t> halozyme.com<mailto:janderson <@t> halozyme.com>



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The information transmitted in this email is confidential and is intended only for the person(s) or entity to which it is addressed. Delivery of this message to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or any applicable privilege. Any review, retransmission, dissemination or other use of, or taking of any action in reliance upon, this information by individuals or entities other than the intended recipient is prohibited by Halozyme and may be in violation of applicable laws. If you received this in error, please contact the sender and delete/destroy this email.
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Message: 7
Date: Tue, 20 Oct 2009 15:02:10 -0400
From: Peter Carroll <carrolpb <@t> umdnj.edu>
Subject: Re: [Histonet] Unsubscribe from website
To: "Witt, Dan" <dwitt <@t> sebh.org>
Cc: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
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There is a link at the bottom of this and every message on this list.
Click it.
Scroll to the bottom.
Unsubscribe yourself.
Easy, right?

Witt, Dan wrote:
> I would appreciate you removing me from the Histonet website
>
>
> Dan Witt PA, HT (ASCP)
>
> Pathologists Assistant / Histology Coordinator
>
> St. Elizabeth's Hospital
>
> 211 S. 3 rd Street
>
> Belleville, IL 62220
>
> 234-2120 Ext. 1369 or 1192
>
> _______________________________________________
> Histonet mailing list
> Histonet <@t> lists.utsouthwestern.edu
> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>
>
>   




------------------------------

Message: 8
Date: Tue, 20 Oct 2009 12:43:22 -0700
From: JR R <rosenfeldtek <@t> hotmail.com>
Subject: [Histonet] RE: Antigen retrieval question
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <BAY135-W1F475B7969F14ABC6D89FDBC00 <@t> phx.gbl>
Content-Type: text/plain; charset="iso-8859-1"






Perhaps the antigen is soluble and diffused out of the
tissue into the buffer?

 

Jerry Ricks

Research Scientist

University
 of Washington

Department of Pathology

 

histonet <@t> lists.utsouthwestern.edu

 

 

Perhaps reversal is not the right word...slides were left overnight in

buffer w/ tween. We got very minimal faint staining compared to previous

runs done in one day.

 

Brett 

 

-----Original Message-----

From: Morken, Tim [mailto:Timothy.Morken <@t> ucsfmedctr.org] 

Sent: Tuesday, October 20, 2009 2:29 PM

To: Connolly, Brett M; histonet <@t> lists.utsouthwestern.edu

Subject: RE: Antigen retrieval question

 

Brett, how long? I've left them overnight before proceeding without any

problems. 

 

It would not be "re-fixation" as with formalin. However, if it is in a

detergent buffer it might cause some other kind of denaturation.

 

 

Tim Morken

Supervisor, Histology / IPOX

UCSF Medical Center

San Francisco, CA  

 

 

-----Original Message-----

From: histonet-bounces <@t> lists.utsouthwestern.edu

[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of

Connolly, Brett M

Sent: Tuesday, October 20, 2009 11:22 AM

To: histonet <@t> lists.utsouthwestern.edu

Subject: [Histonet] Antigen retrieval question

 

Has anyone experienced a "reversal" of citrate HEIR unmasking do to a

prolonged delay in continuing the experiment after the retrieval step? 

 

Brett M. Connolly, Ph.D.

Research Fellow, Imaging Dept.

Merck & Co., Inc.

PO Box 4, WP-44K

West Point, PA 19486

tel. 215-652-2501 fax. 215-993-6803

brett_connolly <@t> merck.com

 

 		 	   		  
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Message: 9
Date: Tue, 20 Oct 2009 12:47:41 -0700 (PDT)
From: Rene J Buesa <rjbuesa <@t> yahoo.com>
Subject: Re: [Histonet] Antigen retrieval question
To: histonet <@t> lists.utsouthwestern.edu,	Brett MConnolly
	<brett_connolly <@t> merck.com>
Message-ID: <936839.34911.qm <@t> web65701.mail.ac4.yahoo.com>
Content-Type: text/plain; charset=iso-8859-1

Consider the following sequence:
1- the NBF corsslinked the antigens making them "more" stable, "insolubable".
2- you did HIER and that crosslinkage disappeared, making the antigen "vulnerable" or able to be dissolved
3- you left them in buffer while "vulnerable". Does not sound to you as if the antigens were "diluted", "dissolved" in the buffer causing a weaker reaction than usual?
That would be my explanation, but to your original question ("Have you experienced...") my answer is no, I always proceeded with the IHC test immediately after HIER. Maybe that is why it has not happened to me.
Just a thought!
René J.

--- On Tue, 10/20/09, Connolly, Brett M <brett_connolly <@t> merck.com> wrote:


From: Connolly, Brett M <brett_connolly <@t> merck.com>
Subject: [Histonet] Antigen retrieval question
To: histonet <@t> lists.utsouthwestern.edu
Date: Tuesday, October 20, 2009, 2:21 PM


Has anyone experienced a "reversal" of citrate HEIR unmasking do to a
prolonged delay in continuing the experiment after the retrieval step? 

Brett M. Connolly, Ph.D.
Research Fellow, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
tel. 215-652-2501 fax. 215-993-6803
brett_connolly <@t> merck.com

  
Notice:  This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station,
New Jersey, USA 08889), and/or its affiliates (which may be known
outside the United States as Merck Frosst, Merck Sharp & Dohme or
MSD and in Japan, as Banyu - direct contact information for affiliates is
available at http://www.merck.com/contact/contacts.html) 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
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message in error, please notify us immediately by reply e-mail and
then delete it from your system.
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Message: 10
Date: Tue, 20 Oct 2009 15:09:51 -0700
From: "Mejia, Maria" <Maria.Mejia <@t> ucsf.edu>
Subject: [Histonet] RE: Antigen retrieval question
To: "Connolly, Brett M" <brett_connolly <@t> merck.com>, "Morken, Tim"
	<Timothy.Morken <@t> ucsfmedctr.org>, "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<BD7845710F83B94796F4534A6D6EC71C0FF5022BC0 <@t> EX02.net.ucsf.edu>
Content-Type: text/plain; charset="us-ascii"

Brett,

We mostly work & do IHC on 40um free-floating brain sections.  These sections were cut from fixed brain blocks.  Sometimes, not on
a regular basis, but sometimes we have had to place our sections in (plain) PBS solution & stored at 4C - overnight before continuing the IHC
protocol with no effect to the staining quality.  Keep in mind, that this was done before the primary antibody. We have also placed our cut
and unstained sections in plate wells (5ml of PBS) - again at 4C for 2 days before starting our IHC protocol.  We use quite a number of 
different antibodies & stain using polymer.

I hope this helps.

Maria Mejia
Histology Manager
Department of Neurosurgery
UCSF
Sf, CA 94103
 
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Connolly, Brett M [brett_connolly <@t> merck.com]
Sent: Tuesday, October 20, 2009 11:33 AM
To: Morken, Tim; histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] RE: Antigen retrieval question

Perhaps reversal is not the right word...slides were left overnight in
buffer w/ tween. We got very minimal faint staining compared to previous
runs done in one day.

Brett

-----Original Message-----
From: Morken, Tim [mailto:Timothy.Morken <@t> ucsfmedctr.org]
Sent: Tuesday, October 20, 2009 2:29 PM
To: Connolly, Brett M; histonet <@t> lists.utsouthwestern.edu
Subject: RE: Antigen retrieval question

Brett, how long? I've left them overnight before proceeding without any
problems.

It would not be "re-fixation" as with formalin. However, if it is in a
detergent buffer it might cause some other kind of denaturation.


Tim Morken
Supervisor, Histology / IPOX
UCSF Medical Center
San Francisco, CA


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Connolly, Brett M
Sent: Tuesday, October 20, 2009 11:22 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Antigen retrieval question

Has anyone experienced a "reversal" of citrate HEIR unmasking do to a
prolonged delay in continuing the experiment after the retrieval step?

Brett M. Connolly, Ph.D.
Research Fellow, Imaging Dept.
Merck & Co., Inc.
PO Box 4, WP-44K
West Point, PA 19486
tel. 215-652-2501 fax. 215-993-6803
brett_connolly <@t> merck.com


Notice:  This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station,
New Jersey, USA 08889), and/or its affiliates (which may be known
outside the United States as Merck Frosst, Merck Sharp & Dohme or
MSD and in Japan, as Banyu - direct contact information for affiliates
is
available at http://www.merck.com/contact/contacts.html) 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.
_______________________________________________
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http://lists.utsouthwestern.edu/mailman/listinfo/histonet

Notice:  This e-mail message, together with any attachments, contains
information of Merck & Co., Inc. (One Merck Drive, Whitehouse Station,
New Jersey, USA 08889), and/or its affiliates (which may be known
outside the United States as Merck Frosst, Merck Sharp & Dohme or
MSD and in Japan, as Banyu - direct contact information for affiliates is
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------------------------------

Message: 11
Date: Wed, 21 Oct 2009 10:08:54 +1100
From: "PALMER Jason (SVHM)" <Jason.PALMER <@t> svhm.org.au>
Subject: [Histonet] RE: in search of cd31 for rat tissue (TF) AND
	novocastra eNOS antibody on rat tissues??? 
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<EEE122D54674BF4CA53C0D05F9BB939E0A24CBDA <@t> SVHM-EXCHCCR0.svhm.schs.org.au>
	
Content-Type: text/plain; charset="us-ascii"

To possibly kill (or at least wound a little?) two birds with the one stone... 

We have used BD Transduction anti-eNOS, # 610296 successfully on FFPE rat tissue, if that is any help.  It seems to label the endothelium in all vessels in the tissues we have looked at so far, as far as we can tell (eNOS being a constitutive form of NOS) and therefore may be considered a vascular marker.  

Cheers,
Jason

Jason Palmer
Histology Laboratory Coordinator
O'Brien Institute
42 Fitzroy St, Fitzroy Victoria 3065
Australia
tel +61 3 9288 4045
fax +61 3 9416 0926
email: jason.palmer <@t> svhm.org.au

?????? [Histonet] in search of cd31 for rat tissue

Has anyone used RECA-1 (from Novus) to stain endothelial cells in rat
tissue.  I also heard about RDI-RTCD31-3A12 from Fitzgerald but it requires
Zinc fixative and frozen sections, I would really like to use ffpe rat
tissue already archived???
Regards,
Patsy
------------------------------
From: "Nejat Yilmaz" <nyilmaz <@t> mersin.edu.tr>
Subject: [Histonet] novocastra eNOS antibody on rat tissues???
Dear All,
We're planning to study FFPE rat aorta specimens with anti endothelial
nitric oxide synthase (NOS-3) antibody. We want to perform that with
Novocastra brand anti-eNOS antibody (Product Code: NCL-NOS-3). According to
datasheet the antibody works on human tissues only, other species not tested
yet. Does anybody have any experince with this antibody on different species
other than human?
Thanks in advance.

Necat Yilmaz MD PhD
University of Mersin School of Medicine

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------------------------------

Message: 12
Date: Tue, 20 Oct 2009 19:14:50 -0400
From: "Jennifer Campbell" <rcampbe <@t> frontiernet.net>
Subject: [Histonet] Fite control slides
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <DJEHLBGJPFDJAHIABGPJOEIPCIAA.rcampbe <@t> frontiernet.net>
Content-Type: text/plain;	charset="iso-8859-1"

I am looking for a reputable vendor for Fite control slides. My pathologist
would prefer they be "leprosy containing skin slides".

I also need the vendor to provide a positvely stained slide with the
procedure as well.

Thank you in advance

Jen Campbell




------------------------------

Message: 13
Date: Tue, 20 Oct 2009 18:22:26 -0700
From: "Jennifer Campbell" <jcampbell <@t> vdxpathology.com>
Subject: [Histonet] Granzyme B and F4/80 immunostaining
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<5658CBDB9EAE6545ABE50D2563D81BF8181FB8 <@t> VDXSERVER01.vdxpathology.local>
	
Content-Type: text/plain;	charset="iso-8859-1"

Hi All,
 
  I'm currently trying to work up Granzyme B and F4/80 on FFPE mouse tissue.  I seemed to get a bit of non-specific staining for both and was wondering if there is any way I can eliminate some of this.  For Granzyme B I am using a rabbit polyclonal from Abcam.  My protocol goes like this: H202 block, citrate buffer HIER, Powerblock (casein-based, non-serum protein block), primary antibody, secondary antibody (rabbit-on rodent HRP polymer, from biocare), DAB, counterstain.  My F4/80 is a rat anti-mouse primary from Serotec and the protocol is as follows: H202 block, citrate buffer HIER, Powerblock, avidin/biotin blocks, primary antibody, secondary antibody (Goat anti-rat, biotinylated, mouse-absorbed, from Biocare), HRP Label, DAB, counterstain.  Is it true that if I switched to a serum block from the species in which the secondary was raised, instead of my non-serum protein block, it may help?  Thank you in advance!
 
Jennifer


------------------------------

Message: 14
Date: Wed, 21 Oct 2009 09:13:19 +0200
From: Melanie Black <Melanie.Black <@t> uct.ac.za>
Subject: [Histonet] CD 31 in rats
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <639A6331-F3C7-4823-8D7C-38825F051AB8 <@t> uct.ac.za>
Content-Type: text/plain;	charset=US-ASCII;	delsp=yes;	format=flowed

Hi Patsy

I have tried RECA-1 and not had great success. The Fitzgerald one is  
the best and we have researched this plenty.

One option for PFA fixed rat tissue is the Abcam VWF ( ab6994). I use  
it 1:400 overnight incubation of primary with 10 minutes proteinase K  
before hand, followed by Donkey anti-Rabbit Cy 3 (fluorescent stain).  
Works well.

Melanie.


Melanie Black
082 469 3352

Cardiovascular Research Unit
3rd Floor; Chris Barnard Building
Medical School;
Observatory. 7925.
University of Cape Town.
South Africa.





------------------------------

Message: 15
Date: Wed, 21 Oct 2009 06:48:57 -0600
From: "Breeden, Sara" <sbreeden <@t> nmda.nmsu.edu>
Subject: [Histonet] Leica Stainer Question
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<4D14F0FC9316DD41972D5F03C070908B02E46BAC <@t> nmdamailsvr.nmda.ad.nmsu.edu>
	
Content-Type: text/plain;	charset="us-ascii"

I use the Leica AutoStainer XL (and love it).  I need to program it for
a run FROM water through the H&E part through to XYL.  I have a program
TO water for special stains, IHC, etc., but have not programmed it for a
run when I mess up and forget to reset for the routine H&E run after
I've done a run to water run.  If you've already figured this out, I'd
appreciate the steps - I'm not lazy, I'm just not a programmer.  Can
anyone help a technologically-challenged person such as myself?  Thanks.

 

Sally Breeden, HT(ASCP)

NM Dept. of Agriculture

Veterinary Diagnostic Services

PO Box 4700

Albuquerque, NM  87106

505-841-2576

 



------------------------------

Message: 16
Date: Wed, 21 Oct 2009 11:11:16 -0400
From: "Cormier, Kathleen" <Kathleen.Cormier <@t> crl.com>
Subject: [Histonet] Beckstead ZSF   FW: Zn fixative 550523 vs 552658
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<7862ACBA8359E04FA9B0C5375A2E6F466A7191 <@t> shr-sv0014.na01.crl.com>
Content-Type: text/plain;	charset="us-ascii"

Follow up to the Beckstead Zinc IHC fixative saga.....BD Pharmingen
states that their IHC Zinc fix does contain the proper ratio/ingredients
to make it identical to the Beckstead fix. See below...Thanks to
everyone!!

 

Kathy Cormier

Histology Manager

Charles River Laboratories

251 Ballardvale Street 

Wilmington, MA 01887

Ph: 781-222-6803

Fax: 978-988-8793

kathleen.cormier <@t> crl.com <BLOCKED::mailto:kathleen.cormier <@t> crl.com> 

 

Experience the new www.criver.com <http://www.criver.com/>  

 

Accelerating Drug Development. Exactly.

Notice - This email and any files transmitted with it are confidential
and may contain privileged and/or proprietary information. You must not
disclose this message to another party without Charles River's express
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message in error, please notify Charles River immediately, and delete it
from your system.

________________________________

From: Sun_Min_Lee <@t> bd.com [mailto:Sun_Min_Lee <@t> bd.com] 
Sent: Tuesday, October 20, 2009 12:13 PM
To: Cormier, Kathleen
Subject: Zn fixative 550523 vs 552658

 


Hi Kathleen, 

Thank you for your interest in BD Biosciences Pharmingen product.
Further to your inquiry, the products cat. no. 550523 (1x Zn Fixative)
and cat. no. 552658 (10X Zn Fixative) are very similar (almost the same)
in terms of ingredients. Both products contain the same quantity of
Calcium Acetate, Zinc Acetate, and Zinc Chloride per 1x  Liter. 

Hope this would be helpful and please let us know if you have any
further question. 

Best Regards,

Sun Min Lee, PhD 
Scientist 
Technical Service, Research Reagents and Applications 
BD Biosciences 
Tel: 877 232 8995, option 3, option 2; Fax: 408-954-2722 
Email:  Sun_Min_Lee <@t> bd.com, ResearchApplications <@t> bd.com
<mailto:ResearchApplications <@t> bd.com>  
  
Please complete a brief survey <https://forms.bd.com/bdbtac/>  to
provide feedback on your experience with technical service.

________________________________

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------------------------------

Message: 17
Date: Wed, 21 Oct 2009 16:15:27 +0100
From: "Ian Montgomery" <ian.montgomery <@t> bio.gla.ac.uk>
Subject: [Histonet] Paraplast
To: <histonet <@t> lists.utsouthwestern.edu>
Message-ID: <10A1FA34C5234A85A9792C3AD5C19D30 <@t> IBLS.GLA.AC.UK>
Content-Type: text/plain;	charset="us-ascii"

            Has anyone noticed changes in tissue fluorescence if using
Paraplast, Paraplast plus or Paraplast extra? E.g. increased
autofluorescence, increased or decreased signal after staining, anything
really. 

Ian.   

 

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

 



------------------------------

Message: 18
Date: Wed, 21 Oct 2009 11:17:32 -0500
From: Stanjeski Paula <Paula.Stanjeski <@t> HCAhealthcare.com>
Subject: [Histonet] PRN.POSITION
To: "histonet <@t> lists.utsouthwestern.edu"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID:
	<14D454E82AC4804CA378B2ABDC7A3B5D872BD7F57E <@t> FWDCWPMSGCMS10.hca.corpad.net>
	
Content-Type: text/plain; charset="us-ascii"

Hello,

We are seeking a PRN histotech for Tuesday and Thursday of each week,6:00 am to 2:30 pm.

Must be a team player and be flexible to fill in other shifts when needed,other shifts would be 7:00 am to 3:30
And 8:30 am to 5:00pm  Monday thru Friday only. (when other employees need time off)


Possible after 5pm call for occasional late surgery cases with frozen sections.. Hospital based Pathology Lab,routine procedures. Located in Hudson, Florida approximately 35 miles North of Tampa,Florida.

Interested applicants fax Resume to 727-819-2943 /attention Paula



------------------------------

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End of Histonet Digest, Vol 71, Issue 21
****************************************



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