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>I read about freezing cells for controls in texts of all sorts, but none
>describe a procedure for doing so. We want to freeze not only peripheral
>blood leukocytes, but also BM and tissues (mostly lymph nodes).
> If someone would be so kind as to send a reference to a procedure, or
attach
>
>a description of a procedure, I would be most appreciative.
We have had good luck freezing PBLs in 10% DMSO and complete medium (RPMI
1640 with 10-15% serum). Freezing rate is important however. Surround the
freezing vials with ~1cm of styrofoam (we use styrofoam trays from Corning or
Falcon 12x75 mm conical centrifuge tubes) and place them in a -80 C freezer
overnight, then transfer them to liquid nitrogen or a -135 C freezer. Thaw
rapidly in a 37 C water bath. With this procedure we typically achieve
2.5-3% C.V. for PI staining of hypotonically lysed cells.
Thomas L. Morgan, Ph.D.
Regional Research Lab
Kaiser Permanente Medical Center
1515 N. Vernont Ave
Los Angeles, CA 90027
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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
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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