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> Has anyone seen PE overlap into the Texas Red channel on a 2-laser sorter?
Yep. And vice-versa.
> Does anyone know what's happening?
> How do I fix it?
Unfortunately we haven't kept much hard data but, anecdotally... we
seem to see it to different extents from different PE reagents from
different sources. These phycobili's are from bacteria and algae so
it would be my (unsubstantiated) suspicion that there could be more
molecular species than one in any "pure" PE sample (APC is a member of
the same family). Of course I'm just guessing. It could also be that
your dye laser beam is of short enough wavelength to excite a bit of
the PE; the PE absorption curves tail out to about 600nm or more (ref
Molecular Probes Handbook of Fluorescent Probes...1992-1994 pg 77).
If that is the case you might wind the wavelength up a bit (we use
605-607nm), being careful not to impinge on the bottom edge of your
Texas red optical filter.
Regards,
\ / < Flow Systems Laboratory
Frank Battye \__/ <<<<< The Walter & Eliza Hall Institute
Robyn Muir ----------!!<<<<<<<< Voice: 61_3_9345 2540
Dora Constantinou /!!\ <<<<< Fax: 61_3_9347 0852
o !! \ < IN:: "facs_copy@wehi.edu.au"
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