Thank you/APC-PE

Pizzo,Eugene Pizzo at nso1.uchc.edu
Tue Dec 2 10:44:45 EST 1997



Colleagues,

Thanks to all who answered my follow up question to the need
to compensate APC from Pe-Cy5. The initial replys that suggested
there might be a persistence of signals generated by the blue beam
that then entered the FL4 channel although debunked by Dr. Shapiro,
fluorescence signals lasting on the order of nanoseconds and the
travel time between the two beams lasting on the order of microseconds,
nonetheless gave me food for thought and were much appreciated.

Tom Frey's explanation is quite clear. I think I was misled by the graphic
used in class which illustrated signals from both beams being overlayed
during the Time-Delay calibration on the Calibur. If I now understand Tom
correctly the signals generated from the two beams are not electronically
processed simultaneously - such would in fact prevent the instrument
from distinguishing red excitation from blue excitation - they are compared
simultaneously to permit analog subtraction for interbeam compensation.

Gene Pizzo/UCONN Health




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