Re: Pyronin Y into APC?

From: facs_copy (facs_copy@wehi.edu.au)
Date: Thu Feb 21 2002 - 21:59:10 EST


Ron,
Your problem sounds like one we had with our LSR whereby the
laser1-excited PI of one cell would leak into the APC detector at the
red laser time point of the previous cell.  This random occurrence was
cell rate dependent; the faster the cell rate, the greater the chance of
a coincidence and the higher the percentage of spurious APC "positives".
 The (completely effective) fix was a modified APC channel optical
filter holder, supplied by BD, which had a slightly lower pinhole and
better blocked any laser1 spillover (from *any* cell) just by virtue of
the geometry.  A quick workaround may be to tape over the top half of
your APC pinhole.

Frank Battye
WEHI Cytometry



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