Colleagues -
Has anyone used Luciferin/Luciferase in flow cytometry? The
chemiluminescence is at 560nm so it seems we should be able to detect it.
Suggestions?
Julie
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Julie A. Auger
Director, Immunology Applications Core Facility
University of Chicago
5841 S. Maryland Ave. MC-1089
Chicago, IL 60637
e-mail: jauger@flowcity.bsd.uchicago.edu
telephone: 773-702-9212 lab 773-702-9261 office
fax: 773-702-3701
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Luciferase is certainly a very (most) sensitive reporter gene using the standard fluorometer benchtop assay.
Possible methodology
Would you switch off the laser or reduce its power?
Would you need a dark room as well ?
Is the transit time in the orifice too short to collect enough photons?
Could you increase the voltage on the PMTs to detect chemiluminescence?
What signal would you threshold on?
Could you use scanning laser cytometry?
Signal/noise without laser excitation? Could you get better sensitiviI'm also interested. Is this a stupid question since I haven't seen any
replies yet. The cytometry mailing list is always so eager to give advice!
ty in comparison to other methods?
I must appologise for so many questions!
Many thanks for your interest
Robert Nordon
MB BS, BMedSci, PhD
Research Fellow
Graduate School of Biomedical Engineering
University of New South Wales
Sydney, 2052.
Australia
tel: 61-2-9385-3906
fax: 61-2-9663-2108
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