Hey Maciej Are you using decent quality fresh formaldehyde? If you use EM grade and make it up fresh there shouldn't be much change over a few days. Polyscience and Tousimis make EM grade formaldehyde in handy glass vials.You can also fix for an hour or two then wash out and suspend your cells in PBS. Freezing sounds like a lot of trouble. I don't know about re-dissolving antibodies after they have dried up, I suspect they will still work if you add the appropriate amount of water as long as you can get the antibodies to dissolve. Keep us posted I'd be interested in your findings. Best Simon Simon Monard FACS Lab Manager Trudeau Institute Saranac Lake NY12983 Ph 518 891 3080 X352 >>> maciej simm <flowlab@myflow.net> - 4/8/02 10:32 AM >>> This is a two parter: 1 - can stained PBMC's be frozen for later analysis? I have people coming in over the weekend who have to stain their experiments. When I acquire these in the morning on Monday, they don't look very good (typically 48-72 hrs after fixing). Is there a way to preserve fluorescence by freezing the cells? 2 - someone in the lab distributed mucho dollars worth of antibodies for 'future stainings' to 'save time'; of course the antibodies dried up into colorful little pellets on the bottoms of the staining tubes. Will these be still usable after mixing them with blood/cell cultures ? or are there irreversible confirmation changes? Thanks for any insights. Maciej -- Maciej S. Simm Senior Technologist Weill Cornell Medical College Pediatrics Hematology/Oncology Immunology Laboratory/Flow Cytometry Unit 525 East 68th Street N-805 New York, NY 10021 voice (212)746 3428 fax (212)746 8573 --
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