Joe,
The 'low' power HeNe does a pretty good job. Note the comparison of 200mW of 647nm light from a krypton laser and 35mW of 633nm light from a HeNe in terms of the resultant brightness of CD4-APC. (Signal to noise with the HeNe is clearly superior despite the fact that the thymus cells sat for 7hours before being run on the FACStar+.) APC/Cy7 is about as good, but there is an edge if can throw more photons at the dye. Needless to say, optimal filters and more efficient detectors will give you the most bang for the buck.
(From the web page I did while at Univ. of Washington:
http://nucleus.immunol.washington.edu/Research_facilities/instrume.htm
the plots are about in the middle of the page.)
Dave
----------------
David M. Coder, Ph.D.
email: dcoder1@qwest.net
tel./messages: 206-499-3446
----- Original Message -----
From: "Joseph Webster" <J.Webster@centenary.usyd.edu.AU>
To: cyto-inbox
Sent: Thursday, August 09, 2001 4:37 PM
Subject: Phar Red supplementary question
>
> Hi Nicole Et Al
> Can you please clarify "high" vs "low" laser power in this context?
>
> I have one of those OLD BD/Spectra Physics HeNe lasers that you
> can't kill with a big stick, still giving about 45mW after all these
> years...
> I was hoping to use that for APC & APC-Cy7 soon; will it do, or do
> we need 100mW that won't last?
>
> Joseph.
>
> At 11:30 9/8/2001, Nicole Baumgarth wrote:
> {in relation to another question...}
>
> >As for the general "disappointing" staining with Cy7APC.....can you
> >increase laser power? All of the tandem-dyes that I have tried seem to be
> >doing increadibly much better at higher laser power. The more the better it
> >seems. This is really a limitation for use of the dyes when you try to use
> >them with low-power lasers.
>
>
>
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