Luciferase detection

schupp at nauvax.ucc.nau.edu schupp at nauvax.ucc.nau.edu
Mon Nov 28 08:48:29 EST 1994


Enrico,
        In situ detection depends on the level of luciferase production and
your detection device.  Very high levels are needed for naked eye detection.
You might try using direct exposure onto polaroid type 612 film.  This film is
very sensitive, with a ASA of 20,000.  Your culture dish must be placed directlyon top of the film, assuming it has a clear bottom.  The most sensitive in situ
detection device available is called a Charge Coupled Device (CCD) camera.
They are quite expensive.  The luciferin is usually added as a spray or wash
at 1mM in an appropriate buffer.  This should not kill the cells.
        I have not worked with mammalian cells, just bacterial cells.  I have
used the film for detection, though.  A quick lit search should be enough to
at least get you started in the right direction.  Hope this helps.

Jim Schupp



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