Luciferase detection

Hi everyone,

 

I've been asked to post this question about luciferase detection:

 

« Luciferase reporter gene expression can be detected and quantified with very high sensitivity in bioluminescence tests from whole wells containing cells or organs. However, it is not useful when evaluating events at the single cell level, due principally to the low emission intensity of the substrate luciferin. I read the thread of Nan Jiang dec 2002 but was wondering if things had evolved since then. In other words, I would like to know if someone has found a way of differentiating luciferase positive and luciferase negative cells by cytometry: either in microscopy or flow. It could be by the direct detection of activated luciferin substrate, but also by using antibodies against luciferase. >From my own searches, there are few antibodies (so if someone already knows of a nice working one for FACS) and we found one protocol suggesting a 10 min acquisition under the microscope to detect the activated substrate. Any help would be appreciated, 

 

Thank you!

 

 

Danièle

Danièle Gagné

Conseillère Technique Cytométrie

IRIC, Université de Montréal

Bureau 1404, Pavillon Marcelle-Coutu

Tél.: (514) 343-6111 x1-8094

Fax: (514) 343-7780