Re: Power requirements for Alexa350 excitation

From: David Coder (dcoder@u.washington.edu)
Date: Thu May 10 2001 - 18:28:01 EST


To come into this late, with the LSR I too, have seen only disappointingly dim
fluorescence of Alexa 350-labeled HLA-DR. It was about 2-4 times brighter than
background, but the same cells with the same antibody conjugated to other
fluorochromes (FITC, PE, Alexa 633, or Alexa 680) were 100 to 500 times brighter
than background.

If anyone has seen better results, what were the details?

Dave
===========
David M. Coder, Ph.D.
Director, Cell Analysis Facility
Univ. of Washington School of Medicine
1959 NE Pacific Street
H474A Box 357650
Seattle WA 98195

tel. 206-685-3014
email: dcoder@u.washington.edu
----- Original Message -----
From: Joseph Webster <J.Webster@centenary.usyd.edu.AU>
To: cyto-inbox
Sent: Wednesday, May 09, 2001 3:21 PM
Subject: Re: Power requirements for Alexa350 excitation


>
> At 21:47 9/5/2001, David_C_McFarland@sbphrd.com wrote:
> >Wayne,
> >What wavelength does your UV laser operate at?  I'm about to try Alexa 350
> >on an LSR which has a 325nm UV laser and it has been intimated to me that
> >it may not work very well.  And I won't have the option of adjusting
> >output.  I for one would be interested in your results.
> >David McFarland
>
> We did try Alexa 350 on an LSR, the best I can say is
> "disappointing..."
> There was a detectable signal, but not enough separation
> to do anything useful; if I remember correctly we used
> something like CD4.
>
> I plan to try it on the sorter next time I run an argon
> laser in multi-line UV, that may not be soon though.
>
> Joseph.
>
> --
> Joseph Webster, Flow Cytometry Facility
> Centenary Institute, Sydney AUSTRALIA.
>
>



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