Washing Cell Clusters

Robert Goodman robgood at bestweb.net
Mon Feb 16 00:52:47 EST 2004


"Dan B." <dburgess at bcm.tmc.edu> wrote in message
news:b1538149.0402151540.5102ae9f at posting.google.com...

> Can anyone suggest a method to wash relatively fragile cell
> clusters (to remove Paraformaldehyde and replace with PBS)
> without centrifugation?  I don't want to disturb the morphology
> before microscopy and photographing.  These are round clusters
> of about 200-300 cells held together by somewhat weak adhesion
> proteins.

And they're floating in liquid?  How about dialysis?  1,000 vols. PBS twice
should probably be more than adequate.  Or use non-centrifugating
ultrafiltration apparatus.  I'm assuming you can get the clusters into
either a dialysis bag or filtration apparatus.

What I don't understand is your need to remove paraformaldehyde before
microscopy.  Is that related to the in situ enzyme activity business someone
asked about in another thread?

Robert





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