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