Eukolight kit
JoAnne Thomas (jthomas@path.som.sunysb.edu)
Tue, 30 Jul 1996 11:02:36 EST5EDT
Hi All!
I have a user who has been assessing viability of neural cells using
Molecular Probes Eukolight kit which uses Calcein AM for live cell
measurement and Ethidium homodimer for dead cell measurement. My
question is this: Since ethidium staining is related to leakage into
the cell as a result of altered membrane integrity and calcein is
related to cytoplasmic esterase activity, what are the kinetics with
respect to the sequence of events? Is it possible for cells to stain
with ethidium while still maintaining esterase activity; hence stain
with both dyes, i.e. damaged but not dead? Since these samples were
run on a FACscan which cannot compensateFL1 and FL3 and there is
significant spectral overlap between these dyes it was not possible
to look at the dual color staining by flow. However using
fluorescence microscopy we definitely saw dual staining. Could it be
that the cells were damaged but still had some esterase activity? I
would appreciate any responses from people who have worked with this
kit before. Thanks-Joanne Thomas
CD ROM Vol 2 was produced by 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:(317) 494-0757; FAX (317) 494-0517; Web http://www.cyto.purdue.edu
EMAIL robinson@flowcyt.cyto.purdue.edu
