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The procedure for doing the *measurement* of # decades has been well
described in a few FCSC handouts; even in their "Catalog: Fluorescent
Microbead Standards". It's just a matter of running a mixture of
their calibrated "Quantum" beads and plotting # molecules vs. channel
number on semi-log graph paper. If you extrapolate the (hopefully
straight) line to channels 1 and 256 (or 1024) you can read off the #
decades.
Doing something about it is harder: On the FACStar+ it's not too bad;
there are screwdriver holes on the front edges of the logamp cards
marked "G" for parameters 2, 3, 6 and 7. For the other parameters you
need to guess which of the rectangular blue trimpots to tweak. Just
keep tweaking and measuring until you get 4 decades exactly. It is
helpful to remember that, once you have established that your calibration
points lie on a straight line, you only need to monitor the *distance in
channels between 2 widely spaced peaks* as you continue to tweak; you
don't need to plot the graph each time.
For the FACScan I can't help. For ours we just *measured* the
characteristic without trying to *change* it. I wonder if B_D
service personel would help with this kind of exercise??
Regards, | | < Frank: battye@wehi.edu.au
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
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