3 questions

From: Robinson, Howard (Howard.Robinson@joslin.harvard.edu)
Date: Mon Jul 31 2000 - 15:12:50 EST


1.)Attempting to use all the laser lines simultaneously has not been easy.
I try to resolve 8 peak rainbow beads (Spherotech)for the UV using a 450/65
BP, the peaks are all scrunched together.  I attempted to use a 488 blocker
with the 450/65 BP, because I thought the 488 was causing the problem, but
ended up loosing all my signal.  I'm wondering if a 488 shortpass would work
in front of the UV pathway to keep out 488 from bleeding through.  Does
anyone have any ideas?

2.)I'm also interested in finding a bead that I can run after setting the
PMT voltages and parameter gains with unstained cells.  I'd like to have a
fluorescent particle control that I run at the beginning and end of each set
of samples, utilizing all the settings I run the samples under, to ensure my
customers the machine is not causing any anomalies that may arise from time
to time in the experiments.  I would like to have a negative bead and a
positive bead that is excited by a UV line, 488 line, and 633 line.  Can
anyone offer me any advice?

3.) Lastly, I'm looking for an alignment bead for when I set-up to run
APC-Cy7.  Does anyone know of a bead that emits at 767 nm and excited by a
633nm laser line?



Thanks,

Howard


Howard Robinson
Flow Cytometry Core Facility
Joslin Diabetes Center
Room 491
One Joslin Place
Boston, MA 02215

Howard.Robinson@Joslin.Harvard.Edu
(617)-732-2400 (Ext. 4516)



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