glutaraldehyde fixation

Jan Keij keij at telomere.lanl.gov
Fri Nov 8 10:03:39 EST 1996




I am in search of a low molecular weight molecule that will 'instantly'
inhibit glutaraldehyde fixation.

The experiments involve a series of 1 ml glutaraldehyde dilutions ranging
from 0.0001 to 1 % in which 1E+06 cells are fixed for 30 minutes on ice.
After 2 washes with PBS, the cells are run on the FACSCalibur and the
changes in autofluorescence are anaylzed. So far we've noticed a non-linear
relation between the % glutaraldehyde and the autofluorescence intensity.
Some of these results could be caused by the unintended incubations with
lower % glutaraldehyde during the washes, which take as much time as the
incubation.

Rather than run the whole series through the Calibur without washing, I'd
like to block the fixation, with a small molecule that will not influence
the results.

Jan.



Jan F. Keij
Los Alamos National Laboratory
Life Sciences Division
LS-5, Cytometry, MS M888
Los Alamos, NM 87544
USA

tel     : (505)-667-3526
fax     : (505)-665-6894
e-mail  : keij at telomere.lanl.gov





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