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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
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web