[Histonet] cytospin slide staining

Terri Braud tbraud <@t> holyredeemer.com
Thu Oct 19 07:32:04 CDT 2006

I'm presuming your cytospins are fixed in 95% Alcohol?  Sometimes antibodies needing antigen retrieval for formalin fixed paraffin embedded slides will not need any antigen retrieval, or will need an entirely different protocol. Its often hit and miss trials. I've found that exposure to methanol is the kiss of death for many antibodies that will work with ethanol fixed protocols. If you are working in human tissues, you can make a decent control by making a cell suspension from minced skin, but of course, best of all would be cervical cancer cells, suspended in RPMI, and make your cytospin controls. 
From a proud LSU alumnus
Terri L. Braud, HT(ASCP)
Anatomic Pathology Supv.
Laboratory, Holy Redeemer Hospital
1648 Huntingdon Pike
Meadowbrook, PA 19046
(215) 938-3689

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of Dahui You
Sent: Wednesday, October 18, 2006 8:08 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] cytospin slide staining

Hello, histonetters:
Has anyone done cytospin slide staining before? What antigen retrieval method you used?
I am staining bronchoalveolar lavage fluid slides with anti-active caspase3 and none of my experimental and control slides showed positive.
I am wondering if that is real or I did it wrong since I don't have any positive control slides for this antibody and I never did this kind of staining before.
Any suggestion will be greatly appreciated!
Have a nice day.

Dahui You

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