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
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