Re: Dead time

From: Dave (dcoder@u.washington.edu)
Date: Fri Apr 30 1999 - 11:01:11 EST


An anecdotal report with APC/Cy7 detection is that a 710 SP DC with a 780BP40
did not give any signal when APC/Cy7 was excited by 35mW of 633nm light, whereas
it does work well with a 700LPDC and 780BP40 when excited by 647nm light.  One
recommended filter combination (though not tested by me) is 710  LPDC and a
750LP filter in front of the PMT.  You may also want a PMT with enhanced
red-sensitivity. A Hamamatsu R3896 has better performance at the red end than
the commonly used R928.

Dave Coder
dcoder@u.washington.edu


----- Original Message -----
From: Joseph Trotter <trotter@scripps.edu>
To: cyto-inbox
Sent: Wednesday, April 28, 1999 6:35 PM
Subject: Re: Dead time


>
>
> Dear Sasha,
>
> I forgot an important point - we have 4 PMT detectors on laser 1, and
> have measured PE-Cy7 off the 1st laser as well as APC-Cy7 off the
> 2nd laser. We use the same 780/60 BP filter for both tandem conjugates
> involving Cy7 and the 710 LP dichroic as well. I have not tested a 710 SP
> dichroic for Cy7 reagents. You ought to use a BP in addition to your
> mirror to minimize leakage from other areas of the spectrum. We have left the
> standard 610 SP in place to reflect signal to the 710LP dichroic.
>
> Sorry for the omission,
>
> Joe



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