FW: [Histonet] Immuno Controls

Fri May 30 10:46:33 CDT 2008

-----Original Message-----
From: Thomas Jasper 
Sent: Friday, May 30, 2008 8:46 AM
To: 'Charles, Roger'
Subject: RE: [Histonet] Immuno Controls

Hey Roger,

Have been following this thread and your post has convinced me to
finally weigh-in.  I'm a little unclear when you say you put both a
positive and a negative on one slide.  When you say per run or per
antibody, this assumes the per run to be one antibody only?  The
negative should be from your target (patient), I say target as with
animal work I'm not sure if it's research or individual vet diagnostic
cases.

Also a slide designated as a negative should get all reagents sans the
antibody/antibodies that are being run.  If you've got both on one slide
this seems impossible to me.  The positive control must receive the
antibody!?  My understanding of running a negative is to demonstrate
that the reagents being run are not contaminated or are going to cause
any sort of false positive.  Therefore, what is seen as positive
staining on the target (patient) slides is a true positive.

I also don't have a problem with folks putting a positive control on
each slide, however this can be cumbersome and/or not an option with
large sections of tissue.  I believe; as long as a known positive
control, is run per antibody, per run, whether it's on the same slide as
the target (patient) tissue or on a separate slide, and that known
positive lights up, it's all good.

I do not worry too much about machine malfunction.  If for some reason
you are seeing less than desirable staining or unexpected results,
machine malfunction is only one path of investigation for
troubleshooting.  Prior to IHC being automated there were a lot more
problems.  What automation has done has given us greater consistency and
helped to eliminate variables, thus improving the staining overall.
This also narrows down any necessary troubleshooting with problematic
staining.

I'm sure there are folks out there that will disagree with me.  That's
ok, if what people are doing works for them, their scope of service,
application etc., so be it.  I know you are not in the clinical world of
human histology, but for us this is the most efficient and successful
approach to meeting our needs for IHC.  Hope this helps.

Tom J.

Thomas Jasper HT (ASCP) BAS
Histology Supervisor
Central Oregon Regional Pathology Services Bend, Oregon 97701
541/693-2677
tjasper <@t> copc.net  

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Charles,
Roger
Sent: Friday, May 30, 2008 7:54 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: FW: [Histonet] Immuno Controls

Hello all,
I've read many posts on this subject of people putting controls on each
slide because machines do make mistakes.  I do not do this; I use one
slide as a control (which contains both negative and positive tissue)
for each run or each different antibody.  What I would like to know from
those putting controls on each slide is if they ever seen one or more
slides where the control did not work but yet controls on other slides
from the same run and same antibody did.  Convince me to change my ways.
Thanks 

Roger Charles
Microbiologist
Pennsylvania Veterinary Laboratory
2305 N Cameron St
Harrisburg, PA 17110
717-787-8808

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mickie
Johnson
Sent: Friday, May 30, 2008 10:34 AM
To: 'Cheri Miller'; 'Happel, James F.';
Histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Immuno Controls

Hi Histonetters,

I would like to add my 2 cents worth. When I started IHC it was by hand
and the tech was sure they had put primary Ab, etc on each slide. Then
we went automated with a Ventana ES. We learned the hard way that
dispensers may or may not dispense their contents and that this could
vary from slide to slide on the same run! This is when we began to put a
control slide on each positive patient slide for each antibody run on
that patient and block.
Machines make mistakes too.

Thanks.

Best Regards,

Mickie

Mickie Johnson, B.S., HTL(ASCP)
Mohs Histology Consulting Services, LLC
  & Mohs Lab Staffing
2507 S. Manito Blvd.
Spokane, WA 99203
509-954-7134
FAX   509-624-3926
Web: www.mohshistogyconsulting.com & www.mohslabstaffing.com
Email: mickie25 <@t> netzero.net

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Cheri
Miller
Sent: Friday, May 30, 2008 4:08 AM
To: 'Happel, James F.'; Histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Immuno Controls

I put control tissue on each slide I stain except the neg controls.

Cheryl Miller HT (ASCP)
Histology Supervisor
Physicians Laboratory,P.C.
Omaha, Ne. 
402 738 5052
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Happel,
James F.
Sent: Thursday, May 29, 2008 2:11 PM
To: Histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Immuno Controls

Good Day Histonetters!  For immuno cases, how are you handling the
control tissue question?  Are you putting a piece of control tissue on
each slide or are you using a control slide for each ab performed on a
given day.  We are using Ventana'e Benchmark for the lion's share of our
immunos.

Thanks!

James Happel
Massachusetts General Hospital

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