[Histonet] cytospins

[Melissa Mazan]

Hi all,
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
Tel:508-839-5395
Email: melissa.mazan <@t> tufts.edu
Fax:508-839-7922