I'm doing immunofluorescence on cytospins - and have had several problems.
First, we're looking at a rare cell from a FACS sort - about 1000-1500
cells per ml. We're getting a big loss when we cytocentrifuge - about
60% loss. Does anyone have a recommendation for a cytocentrifuge that
will conserve cells better than this?
Second, how important is it to fix cells immediately? Our FACS sorts are
done in Boston, then we travel about an hour to our lab. It is really
tough to fix cells in Boston - our procedure is Cytofix at 4C for 20
minutes, then acetone at 4C for 10 minutes, followed by PBS wash, block,
and overnight primaries. Will we experience serious cell deterioration
if we don't fix them for an hour to two hours after sorting?
I really appreciate any input. Melissa
Melissa R. Mazan, DVM, Diplomate ACVIM
Associate Professor and Director of Equine Sports Medicine
Tufts Cummings School of Veterinary Medicine
200 Westboro Road
North Grafton, MA 01536
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