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_/ _/_/ * _/_/_/ * _ /*
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* Dennis J. Young Voice : (619) 822-0407 *
* Flow Cytometry Core Facility FAX : (619) 822-0412 *
* University of California, San Diego USA e-mail: djyoung@ucsd.edu *
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>>>>>
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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>Date: Wed, 9 Aug 1995 09:10:10 EST
>From: Ralph Smialowicz 541-5776 <SMIALOWICZ@am.herl.epa.gov>
>Subject: Rodent thymocyte suspension
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web