Single cell luciferase bioluminescence detection. Is it possible? - (Sep/11/2007 )

Hello!

I have just transfected (electroporation) my Jurkat cells with a plasmid construct for luciferase expression (NFAT consensus sequence Luc expression).

After maintaining the cells in hygromycin for selection, I have got my positive signal (10.000 RLU) in the bioluminometer after PMA stimulation of the cells. So far, so good!

I would like now, for future experiments, to measure the % of luc+ cells since the luminescence gives me only some light signal but not information on % Luc+ cells. Is it possible by microscopy (I mean, Bioluminescence measurement, not immunohistochemistry for luciferase)? Does anyone have any protocol for doing it? I know it is not possible by FACS, but I am not sure if for microscopy it is also true.

Thank you!

Yes, if you have a very expensive CCD device and you will be willing to work under total darkness. Regular microscope is not good enough.