Let me know if I can be of further assistance.
Rochelle Diamond
California Institute of Technology
Division of Biology
Flow Cytometry/Cell Sorting Facility
diamond@cco.caltech.edu
On Wed, 9 Aug 1995, Ralph Smialowicz 541-5776 wrote:
> I would appreciate any help with a problem that we have had in preparing
> "clump free" thymocyte suspensions, from mice and rats, for flow
> analysis. We routinely prepare thymocyte suspensions in RPMI followed by
> centrifugation at 200 x g. Cell pellets are resuspended in PBS with 2% FBS,
> 0.1% sodium azide and 0.025% DNase, the latter is added to "prevent"
> clumping. The cells are subsequently centrifuged and resuspended in PBS
> with 2% FBS and 0.1% sodium azide for staining. If we have a clumping
> problem, it usually occurs after the first wash. In general, the clumping
> appears to be associated with cell suspensions with higher than usual cell
> numbers. Any suggestions would be greatly appreciated. Thanks.
>
> Ralph J. Smialowicz
> US EPA
> RTP,NC
>
>
>
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