[Histonet] RE: ICC on cytology preps

From:Malam Jacqueline

We occasionally get asked for ICC on cyt preps and, as these don't need ag.
retrieval, I felt the positive controls should also be a cyt prep to be
valid. We have found that if you receive cellular fluids (eg malig pleural
or ascitic) in cyt fix, after they have been diagnosed you can test their
positivity for the antigen in question and, instead of disposing of them,
they can be kept to use as positive control material. The dilution of the
primary antisera may need adjusting so it's worth doing a titration. Also,
we suspend the fluid in saline, spin down, discard the supernatant and
resuspend x3 to get rid of the protein in the fluid which can cause
background staining and interfere with the picture. The downside is if you
can't get hold of a fluid which is positive for the antigen you need, or if
your test arrives ready prepared on the slide, or is a fluid which only has
scanty cells - you may lose some in washing with saline. You could always
make a cell block with agar (if you have enough cells) and fix in formalin
etc then you can do your routine protocols. Good luck!

-----Original Message-----
From: histonet-request@lists.utsouthwestern.edu
Sent: 19 July 2005 18:02
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 20, Issue 23

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Today's Topics:

   1. Re: Cytology Immuno's (Richard Cartun)
   2. Re: Bodies in Morgue (chiggerson@memhosp.com)


Message: 1
Date: Tue, 19 Jul 2005 11:20:03 -0400
From: "Richard Cartun" 
Subject: Re: [Histonet] Cytology Immuno's
To: ,
Content-Type: text/plain; charset=US-ASCII

I use paraffin sections for positive controls and, yes, I consider it
adequate for confirming that the test worked.  Obviously, an internal
positive control is best, but is not always present.  Each laboratory needs
to validate the individual antibodies on cytologic specimens using known
cases.  In our experience, some antibodies don't work very well on
alcohol-fixed cytology specimens and, therefore, should be used with
caution.  I have not been able to confirm this, but I don't think "IVD"
labels for antibodies used on formalin-fixed, paraffin-embedded tissue
sections apply when used on cytology specimens.


Richard W. Cartun, Ph.D.
Director, Immunopathology & Histology
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

>>> Jean Gillson  07/19/05 10:40AM >>>
Can I ask you all, what do you use as controls when doing ICC on cytology
cytospin preps or smears?  Since there isn't much sample, do you use
paraffin sections as positive controls and is that adequate control
measures?  We currently only use paraffin cytoblocks but considering ICC on
thin layer cytology preps and wondering what control material to use.
Thanks for your help.

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Message: 2
Date: Tue, 19 Jul 2005 11:35:58 -0500
From: chiggerson@memhosp.com
Subject: [Histonet] Re: Bodies in Morgue
To: histonet@lists.utsouthwestern.edu
Content-Type: text/plain; charset="US-ASCII"

Bodies are checked in and released by Security.


----- Forwarded by Cindy Higgerson/6550/Pmmc on 07/19/2005 11:33 AM -----

Date: Mon, 18 Jul 2005 16:37:34 +0000
From: "Paula Wilder" 
Subject: [Histonet] Bodies in morgue - who does the checking?
To: histonet@lists.utsouthwestern.edu
Content-Type: text/plain; format=flowed

Hi everyone!

Any feedback on whose responsibility it is to check bodies in the morgue 
your respective instituition would be greatly appreciated.  So far, by 
phoning neighboring hospitals, I have found that Security, a diener 
or the Pathology Assistants are the ones responsible.  Any help in this 
would truly be greatly appreciated!  Thanks so much!

Paula Wilder
St.Joseph Medical Center
Towson, MD 21204


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