From: MORGANTL@aol.com
Date: Tue Dec 19 1995 - 01:07:31 EST
In a message dated 95-12-08 15:22:47 EST, you write: >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
This archive was generated by hypermail 2.1.6 : Thu Jan 01 2004 - 17:30:40 EST
