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@telomere.lanl.gov