From: Dave Coder (dave@nucleus.immunol.washington.edu)
Date: Mon Aug 08 1994 - 18:37:55 EST
There are several issues:
1. do you need to use glutaraldehyde as a fixative?
2. what do you need to measure?
3. if #2, and it's other fluorochromes, then can you use fluorochromes that
fluorescence away from glutaraldehyde fluorescence? That is, dyes the fluoresce
beyond 600nm?
To expand,
1. there are other fixatives that will work just fine for flow cytometry.
Para-formaldehyde has much less autofluorescence; alcohols such as MeOH or EtOH work
well with many cell types.
But if the tissue is always fixed with glutaraldehyde and you have no choice, there
are other strategies.
3. if no other fluorochromes are to be used, then crystal violet may quench the
glutaraldehyde fluoresce. It can be used to quench FITC surface fluorescence while
permitting cytoplasmic FITC fluorescence.
3. if using antibodies, then phycoerythrin (PE) or better, PE-Cy5 conjugates, will
fluoresce beyond the typical glutaraldehyde fluorescence range. In general,
red-emitting dyes are the ones of choice for the problem.
Dave Coder
Univ. of Washington
Begin forwarded message:
Date: Fri, 05 Aug 94 09:01:24 PST
From: "Susan D. DEMAGGIO" <SDDEMAGG@uci.edu>
Subject: Gluteraldehyde autofluorescence quenching
I have been asked by a researcher to see if I can find a way
to quench autofluorescence of gluteraldehyde - or get around
it. Does anyone have any suggestions for us? THANKS
Sue DeMaggio
UC Irvine
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