RE: Minimum number of mouse cells for Flow

From: Snider Denis (sniderd@fhs.csu.mcmaster.ca)
Date: Tue Aug 05 1997 - 13:50:39 EST


Patricia,

We have been analyzing mouse PBMC for a while now.  We take 50-150
microlitre of blood, dropped immediately into EDTA-PBS solution.  Then
it is spun onto Ficoll-H to get rid of RBC and grans.  The resulting
PBMC are very viable.  Usually we have enough cells (2-3 X 10e5 per
tube) for between 2 and 4 tubes.  We often run 4 tube assays on single
isolations.

Good luck.

Denis Snider



 

> -----Original Message-----
> From:	Echeagaray, Patricia L. [SMTP:echeagaray@sri.org]
> Sent:	Friday, July 25, 1997 2:34 PM
> To:	Cytometry Mailing List
> Subject:	Minimum number of mouse cells for Flow
> 
> 
> Greetings! 
> 
> I have a very basic question, and am hoping it is not too off topic
> for
> this group. I am being asked to analyze by flow, mouse lymphocytes.
> They
> will first require a density gradient procedure to isolate the
> lymphoctes.  
> 
> I will have very little volume of blood to use, and would like to have
> an idea as to the minimum volume of whole blood I would need in order
> to
> analyze for CD4, CD8 and CD19 (Ts and total Bs).  I sense I can do
> this
> in 2 tubes (experimental and control), so I will need enough for both
> tubes.
> 
> Does any of you help me?
> 
> Please e-mail me privately at echeagaray@sri.org
> 
> Thanks in advance!
> 
> P. Echeagaray



This archive was generated by hypermail 2b29 : Wed Apr 03 2002 - 11:49:59 EST