[Histonet] mouse IHC background

Madary, Joseph MadaryJ <@t> MedImmune.com
Mon Sep 17 12:09:32 CDT 2007

Maybe a mouse adsorbed secondary is all you need.  Vector makes them.

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Subject: Histonet Digest, Vol 46, Issue 30

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Today's Topics:

   1. RE: Surgical specimens (Debbie Hogg)
   2. Background on FFPE mouse tissue (Gale, Nadia)
   3. RE: Background on FFPE mouse tissue (Monfils, Paul)
   4. Re: Re: negative controls (Amos Brooks)
   5. Formalin & FNA (Satterfield, Marirose)
   6. RE: I think I will give it an acronym (Amos Brooks)


Message: 1
Date: Mon, 17 Sep 2007 10:45:34 -0400
From: "Debbie Hogg" <debbie_hogg <@t> hotmail.com>
Subject: RE: [Histonet] Surgical specimens
To: histonet <@t> lists.utsouthwestern.edu
Message-ID: <BAY144-F401909C01348CED4F4D1999ABF0 <@t> phx.gbl>
Content-Type: text/plain; format="flowed"

   I  am  hospital  based  here  in  Canada  and  Surgical  specimens
   delivered to our pathology lab by circulator's.  Our assistant does
   to the Ambulatory Care Unit to pick up the small specimens.

   The  last hospital that I worked at also had designated times that
   surgicals  would  be  delivered to the Path. lab.  Technologist's
   more  than enough work to keep them busy without running to the OR

   Debbie Hogg

   Put Your Face In Your Space [1]with Windows Live Spaces!


   1. http://g.msn.com/8HMBENCA/2728??PS=47575


Message: 2
Date: Mon, 17 Sep 2007 08:47:33 -0700
From: "Gale, Nadia" <Ngale <@t> bccancer.bc.ca>
Subject: [Histonet] Background on FFPE mouse tissue
To: <histonet <@t> lists.utsouthwestern.edu>
<E76535B27E4D5A428863F402EC2E0C291ADFFC <@t> srvex03.phsabc.ehcnet.ca>
Content-Type: text/plain;	charset="us-ascii"

Hello Histonetters,
I am relatively new to the mouse world of IHC having come from a
clinical background into research and have what might be a simple
problem... I am experiencing heavy background with the majority of mouse
tissues I am staining.  I am using the Ventana Discovery system with
their universal DAB kit and "titrate" 100uL of antibody diluent onto my
negative control slides.  I've tried a biotin blocking kit as well (even
though it doesn't look like biotin, thought I'd give it a try) and
observed no decrease in the amount of background staining.
Any ideas for me?
Thanks in advance!
Nadia Gale
Lead Histotechnologist
Centre for Translational and Applied Genomics (CTAG)
3427-600 West 10th Avenue
Vancouver, BC V5Z 4E6
tel 604-877-6000 ext 2426
fax 604-877-6089
ngale <@t> bccancer.bc.ca


Message: 3
Date: Mon, 17 Sep 2007 12:31:58 -0400
From: "Monfils, Paul" <PMonfils <@t> Lifespan.org>
Subject: RE: [Histonet] Background on FFPE mouse tissue
To: <histonet <@t> lists.utsouthwestern.edu>
<4EBFF65383B74D49995298C4976D1D5E273CE3 <@t> LSRIEXCH1.lsmaster.lifespan.org>
Content-Type: text/plain;	charset="iso-8859-1"

Are you using mouse monoclonal antibodies?  Heavy background staining is
a common problem when your antibody was produced in the same species
that your tissue sample came from.  The anti-mouse secondary antibody
can't distinguish between the applied mouse primary and the endogenous
immunoglobulins naturally present in the mouse tissue.  You can avoid
this either by using a mouse-on-mouse kit, available from several
vendors, or by using primariy antibodies made in other species.


Message: 4
Date: Mon, 17 Sep 2007 12:40:24 -0400
From: "Amos Brooks" <amosbrooks <@t> gmail.com>
Subject: Re: [Histonet] Re: negative controls
To: "godsgalnow <@t> aol.com" <godsgalnow <@t> aol.com>
Cc: histonet <@t> lists.utsouthwestern.edu
	<582736990709170940y19d73ff4re2eab837bd7a2277 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1


   Is there any possibility that the multiple cores are from different
of the prostate that may have metastasises from other places like kidney
   Ultimately you can do this how you like. It is just important to know
limitations of your testing and be aware that this can bite you in the
later on.

Just my $0.02

On 9/17/07, godsgalnow <@t> aol.com <godsgalnow <@t> aol.com> wrote:
> But what if all of the blocks are prostate core biopsies?
> Roxanne
> -----Original Message-----
> From: Amos Brooks <amosbrooks <@t> gmail.com>
> To: histonet <@t> lists.utsouthwestern.edu
> Sent: Sat, 15 Sep 2007 11:16 pm
> Subject: [Histonet] Re: negative controls
> Hypothetically:
>      Let's say an endometriosis case has several parts as they
commonly do.
> A) Uterus bx (obviously), B) Ovary, C) Lg Colon bx, D) Kidney E) Liver
> So an IHC is ordered on the case and randomly part A is chosen as a
> representative negative control. Have you ever noticed how much
> biotin is in kidney & liver? Since there is not a negative control on
> there is no real way of knowing that the labeling that one would see
> is not real.
>      Honestly having a negative on each block is best, if not for eash
> being run (a tall order for sure!). Before polymer detection became
> I once used kidney as a positive control for CD10. It labels the brush
> boarder of the proximal convuluded tubule really nicely. Problem is
> this is also a great place to find endogenous biotin! This really sold
me on
> the use of negative controls. Thanks Mary!
> Have a nice day,
> Amos
> Message: 1
> Date: Fri, 14 Sep 2007 15:35:48 -0400
> From: godsgalnow <@t> aol.com
> Subject: [Histonet] negative controls
> To: histonet <@t> lists.utsouthwestern.edu
> Message-ID: <8C9C51D962FF31B-284-1FE4 <@t> webmail-md17.sysops.aol.com
> >
> Content-Type: text/plain; charset="us-ascii"
> How many non-CAP labs out there are doing negative controls on every
> that you do an antibody on?
> If you have a case that goes A-J and they are all 1 block each and you
do an
> AE1/AE3 on blocks B,C,F,& J, will you do a negative on every block or
> one for the entire case?
> This has been an ongoing debate with us.? We typincally only do 1
> control for the case and given the above example, it might be on block
> We ususally cut an extra on every block we cut and when the IHC is
> we just go and pull it and for the negative, we just grab whatever
slide is
> left to run the negative on.? What is everyone else doing??
> Roxanne
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> Histonet <@t> lists.utsouthwestern.edu
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>  ------------------------------
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> !


Message: 5
Date: Mon, 17 Sep 2007 12:37:45 -0400
From: "Satterfield, Marirose" <Marirose.Satterfield <@t> MercyMemorial.org>
Subject: [Histonet] Formalin & FNA
To: <histonet <@t> lists.utsouthwestern.edu>
<BBD46D81DE497443B2FAAF7DB7782FD824D2B6 <@t> MMHEXCHVS1.mercymemorial.org>
Content-Type: text/plain;	charset="us-ascii"

Actually I have two different questions. 


#1 Who performs the FNA procedures at your facility?


#2 How many people have their microtomy, staining, tissue processing and
grossing all in one room?


I am doing a little bit of research and any answers will help.



Mari S.



Message: 6
Date: Mon, 17 Sep 2007 12:47:35 -0400
From: "Amos Brooks" <amosbrooks <@t> gmail.com>
Subject: RE: [Histonet] I think I will give it an acronym
To: histonet <@t> lists.utsouthwestern.edu
	<582736990709170947p15ceb7fcg1327ed43ae033966 <@t> mail.gmail.com>
Content-Type: text/plain; charset=ISO-8859-1

Or better yet :

(let's see how many geeks we have on the histonet)

Message: 20
Date: Mon, 17 Sep 2007 15:40:39 +0100
From: "Edwards, R.E." <ree3 <@t> leicester.ac.uk>
Subject: RE: [Histonet] I think I will give it an acronym
To: "Bernice Frederick" <b-frederick <@t> northwestern.edu>, "Rene J Buesa"
       <rjbuesa <@t> yahoo.com>, "Gayle Callis" <gcallis <@t> montana.edu>,
       <curtis.king <@t> thermofisher.com>,
<Histonet <@t> lists.utsouthwestern.edu>
       <DC88BEDFD1FC3F468D0376A7C75465F7101C7143 <@t> Saffron.cfs.le.ac.uk>
Content-Type: text/plain;       charset="iso-8859-1"

  How  about "HISTOSTUFFRUS"


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