I recommend against freezing at all (presumably you are talking about using DMSO or other method similar to freezing tissue culture cells). Even storing cells on ice too long causes loss of subpopulations and modulations of antigen expression, you would have to validate that what you measure on sorted cells that were frozen is the same as fresh cells. If you can validate this, then you will have to thaw and use immediately, and take great care to exclude dead cells using propidium iodide ======================================================= Rachel M. Gerstein, Ph.D. Department of Molecular Genetics and Microbiology Graduate Program in Immunology/Virology University of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655-0002 (508) 856-1044 (508) 856-5920 (FAX) > ---------- > From: Capocasale, Renold [CNTUS] > Sent: Tuesday, December 3, 2002 12:00 PM > To: Cytometry Mailing List > Subject: sorting fozen murine B cells > > Hello to all, > I would like to sort primary splenic B cells that will have been frozen prior to sorting. The cells will be frozen for less than a week prior to sorting. In particular, should the cells be thawed and immediately sorted or rather should a recovery period be implemented and for how long? > > Also, has anyone tried surface staining immediately before freezing followed by sorting at a later date? > > Any recommendations on the nuances of this type of sorting would be much appreciated. > Thanks > > Renold Capocasale > Senior Associate Scientist > Experimental Pathology/ Cell Sorter Core Facility > Centocor, Inc > a Division of Johnson & Johnson > phone # 610 - 651- 6421 > fax # 610 - 651- 7363 > >
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