Jim,
Most procedures for TUNEL ("Tdt-Utp Nick End Labeling" - but I'm told that's
a misnomer anyway) are also designed to counterstain the DNA and have
ethanol as part of the fixation method, which can cause a loss of many
cell-surface antigens. Have them look at:
Sgonc R, Boeck G, Dietrich H, Gruber J, Recheis H, and Wick G: Simultaneous
determination of cell surface antigens and apoptosis. Trends in Genetics,
10:41-42, 1994.
The above uses a brief paraformaldehyde fixation followed by triton to
permeabilize and should work better to retain the surface. We have also had
success using the commercial product FACSLYSE from BD as a
fixation/peabilization reagent. See:
Syrjala MT, Tiirkainen M, Jansson SE, and Krusius T: Flow cytometric
analysis of terminal deoxynucleotidyl transferase. A simplified method.
American Journal of Clincial Pathology, 99:298-303, 1993.
Good luck,
Neal
-- Neal A. Benson Biotechnology Program, University of Florida E-mail: nbenson@biotech.ufl.edu
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