Re: reagent and cell durability questions

From: Simon Monard (smonard@trudeauinstitute.org)
Date: Wed Apr 10 2002 - 08:14:30 EST


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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