RE: Fluorochromes choice

From: Derek Davies (daviesd2@cancer.org.uk)
Date: Fri Nov 01 2002 - 11:25:11 EST


Hi Michal and Randy,

I assume Michal that you have a BD LSR? I also assume, Randy, that you
have some sort of issue with it. To describe the laser configuration as
'totally useless' is not only provocative but also implies that 6 colour
work is impossible on the LSR. Yes, the 325nm excitation isnt optimal
for a lot of the dyes available but there is sufficient excitation to
allow the use of Hoechst, DAPI, Indo, Alexa350 ands AMCA. I have
certainly managed to do 6 colour combinations eg Indo (blue and green
emission), FITC, PE, PE/Cy5 and APC or TP3.

The 325nm laser may not be ideal and will presumably be replaced by a
newer solid state 350ish one soon but I - and probably more importantly
my users - have found it to be useful especially in doing analytical
experiments that were once occupying my sorters.

Derek


On Thu, 31 Oct 2002, Fischer, Randy (NIH/NIAMS) wrote:
> You are just another in the line of researchers who has an instrument which
> has used a misleading terminology in its description.  It does have three
> lasers, and two are fairly useful although there is no compensation
> circuitry between the 488 and 633 lasers (problem for APC and PE-Cy5
> channels).  The third laser as mentioned above is a "UV", but the problem is
> that the "UV" laser is too far down in the "UV" spectrum to be beneficial
> for the "UV" excitable fluorochromes. So, unfortunately, this laser
> configuration is totally useless when trying to use the currently available
> fluorochromes to do 6-color staining.
> Randy T. Fischer

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Derek Davies                       Voice: (44) 020 7269 3394
FACS Laboratory,                   FAX: (44) 020 7269 3100
Cancer Research UK,                e_mail:derek.davies@cancer.org.uk
London Research Institute,	   mobile: 07790 604112
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Web Page: http://sci.cancerresearchuk.org/axp/facs/davies/index.html

In tenebris lux
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