Staining leukocytes before freezing
Dr M Salmon (salmonm@rheuma.bham.ac.uk)
Tue, 29 Jun 93 11:49:59 BST
In response to Steve Hilliards interesting note about freezing cells after
staining them. We have quite routinely frozen cells after separation from
human blood. Recovery from liquid nitrogen storage is almost invariably
excellent, though we have frequently observed a selective loss of CD16+ NK cells
under these conditions. We have not tried staining the cells before freezing
for two reasons: firstly the freeze-thaw process may dislodge certain antibodies of low affinity, leading to a non-representative result, and secondly because
this limits your options for staining unnecessarily. We usually freeze several
aliquots of 5 million cells. Recovery of one such aliquot is usually sufficient
for a whole range of staining reactions. Staining the cells before freezing
must presumably take up a lot of storage space. The advantage for us is also
that if you make a mess of thawing one aliquot you have severakl[2~[l left which can
still be used for everything. Frozen unlabelled cells from important samples
can also be retained for at least 3 years in case you wish to use them with
currently unavailable antigens.
Best Wishes
Mike Salmon
Department of Rheumatology
Birmingham University
United Kingdom
Email to salmonm@rheuma.bham.ac.uk Voice: 021-414-6781
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