single cell preparation

From: Eckstein, Volker (Volker.Eckstein@med.uni-heidelberg.de)
Date: Mon Sep 02 2002 - 04:46:09 EST


Hi everybody,

I know a lot of people are still on holidays but nervertheless I am posting
my question again. I've got no replies yet.

Dear experts,

is there anybody who has experience or a good working protocol in preparing
single cells from stomach carcinoma tissue or kidney carcinoma tissue for
further flow cytometric anaylsis?
I have a client who started to do this but the first attempts looked -
let's say not as what we assume and expect is a nice single cell suspension
for flow analysis. A lot of debris and a lot of aggregates also seen during
counting the cells on the microscope. Even after filtering the 'suspension'
I can't identify a certain cell population. And there also is no difference
between permeabilized (for cytoplasmic antigen staining) and
nonpermeabilized 'cells'.

I appreciate any help

Many thanks in advance

Regards

Volker (a little bit stressed about this isssue)

Volker Eckstein PhD
Dept. of Internal Medicine V
Medical school of the university
University of Heidelberg
GERMANY
volker_eckstein@med.uni-heidelberg.de



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