luciferin-esters and flow cytometry?

Garry P. Nolan gnolan at cmgm.stanford.edu
Sun Sep 17 14:44:50 EST 1995


In article <1995Sep13.140818.6455 at wisipc.weizmann.ac.il>, David de Graaf
<bmgraaf at dapsas1.weizmann.ac.il> wrote:

> Dear Netters, I have heard that it is possible to assay luciferase
> activity by
> ordinary flow cytometry in individual cells using luciferin-esters.
> 
> Although I have found one reference, which I do not have handy, for
> analyzing
> luciferase expression by FACS, these good people perform the assay on sf9
> insect cells. Does anyone have experience with such an assay system on
> mammalian cells? Thanks in advance,


I had not thought the light output high enough for FACS.  I found the
reference that maybe you were referring to (In Cytometry?).  They use the
polyhedron promoter, which is pretty strong).  Do you know if anyone has
used weaker promoters?  I will look up the paper since it seems
interesting, so thanks for noting this interesting finding.  I wonder if
the light output they see can be "enhanced" by a fluorescence aspect,
since in many luciferase reactions there is a transient intermediate that
is fluorescent.

I will try this in mammalian cells with a CMV promoter soon, once I find
that cytometry paper.

-- 
Garry P. Nolan, Ph.D.
Assistant Professor
Dept. of Molecular Pharmacology
Dept. of Microbiology and Immunology
Stanford Universoty School of Medicine
Stanford, CA 94305
gnolan at cmgm.stanford.edu



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