Re: 8 day old embryonic cells

Dear Joan,

I am worry that your client would need many, many embryos to run flow. Wouldn't be easier
to do apoptosis assay in whole embryos by immunohisto, i.e. TUNEL in situ?

My two cents

Corrado



Corrado M. Cilio, M.D., Ph.D. 
Assistant Professor 
Cellular Autoimmunity Unit 
Dept. of Clinical Sciences/Paediatrics 
Malmo University Hospital 
Lund University 
205 02 Malmo, Sweden 
Tel: office +46-40332395; mobil: +46-70-4330338 
"Judge a men by his questions rather than his answers" - Voltaire


----- Original Message -----
From: Joan Kalnitsky <jkalnits@vt.edu>
Date: Thursday, November 10, 2005 7:41 pm
Subject: 8 day old embryonic cells

>	  I have a client who would like to use Flow to determine 
> apoptosis 
> in her embryonic cells.  Her window of experimental time is day 8 
> and day 
> 9.  Her experience is that something as gentle as PBS lyses her 
> cells 
> almost immediately.  She is also concerned that multiple washings 
> for a 
> staining protocol would destroy any cell population she might be 
> able to 
> collect.
>	  My questions are 1) does any one have a method for 
> dissociating 
> embryonic cells for use in flow cytometry, and 2) does anyone have 
> a 
> suggestion for which "assay for apoptosis" would be best suited 
> for these 
> cells.
>	  As always, Thanks in advance for any and all offerings of 
> assistance.	      Joan K.
> 
> 
> 
> 
> 
> Joan Kalnitsky
> Flow Cytometry Lab Supervisor
> Virginia Maryland Regional College of Veterinary Medicine
> Duckpond Drive
> Blacksburg, Va. 24061
> Phone (540) 231-4115
> 
> 
>	  "It is better to serve than to receive."  B. Borg 
> 
>