Re: Sorting by Chemiluminescence

Tim Kute wrote:

>Has anyone ever tried to sort cells using chemiluminescence as the indicator?
>
>An investigator has some cells that have been transfected with 
>luceriferase and could mix the the cells with the substrate prior to 
>putting on the machine.  Is there a known protocol for seperating positive 
>cells from the negative transfected cells?
>
>If yes,  Please send me a article or protocol.

Lindqvist C, Karp M, Åkerman K, Oker-Blom C: Flow cytometric analysis of 
bioluminescence emitted by recombinant baculovirus-infected insect cells. 
Cytometry 15:207-212, 1994

These authors apparently detected low levels of bioluminescence in a flow 
cytometer, although the cells to which luciferin was added were measured in 
a 488 nm laser beam, complicating the issue. In general, bioluminescence 
lifetimes are very long (seconds to minutes), and one would not expect to 
collect many bioluminescent photons in the few microseconds it takes for a 
cell to pass through the observation region of the cytometer. You'd be 
better off imaging a large number of cells for a much longer time and 
selecting the bright ones with laser capture or old-fashioned 
microdissection, because, if you slowed the flow cytometer down enough to 
get good bioluminescence signals as a basis for sorting, your analysis rate 
would be one cell every few seconds.

-Howard