side population wtih LSR

David_Matsuyama@BDIS.Com David_Matsuyama at BDIS.Com
Tue Mar 5 14:58:35 EST 2002


BD will soon release a filter set and modifications to detect the SP population.
As you can see from the plots the data compares favorably to the FACSVantage SE.
The HeCd presents no problems or issues in detecting the Side Population, the
statistics compare to the FACSVantage SE running at 30mW for the UV beam.  The
data is courtesy of Teresa Hawley (American Red Cross).


(Embedded image moved to file: pic06194.pcx)

David Matsuyama
BD Biosciences
Assoc Product Manager
San Jose, CA






Derek Davies <daviesd2 at cancer.org.uk> on 02/28/2002 02:08:57 AM

Please respond to derek.davies at cancer.org.uk

To: cyto-inbox
cc:    (bcc: David Matsuyama/SANJ/BDX)
Subject:  Re: side population wtih LSR




Hi Kimmo,

I think your problems are stemming (no pun intended, honest!) from the
filter set-up of the LSR. If you look at the laser paths, the UV signals
are directed to the FL4/5 PMTs via a 510LP filter so only fluorescence
below 510nm will get to the detectors. As you say, this is a bit of a
problem when looking for signals in the far red in the SP population.
The problem will be worse if you alter the filter in front of FL4 to say
a 650LP; keeping a 510/20 there will give you some signal but probably
not far enough out in the red for the 'classical' SP population.

There is no off the shelf solution as far as I can see other than
modifying the LSR considerably - I believe BD may be working on this. It
would be an adavnatge to be able to do SP analysis on a benchtop and I
belive that the Cytomation CyAn may be better equipped to do this -
currently we do all stem cell analysis on the MoFlo.

Derek

On Wed, 27 Feb 2002, Porkka Kimmo wrote:
> Has anyone have experience on running a side population analysis of human
> bone marrow with the BD LSR? I'm using the Hoecst 33342 dye and a FITC cell
> surface marker and following the published protocol to the letter, but still
> cannot see any Hoechst fluorescence on the red channel. Filter setup?
> Software compensation? Any help would be much appreciated.

************************************************************************
Derek Davies                       Voice: (44) 020 7269 3394
FACS Laboratory,                   FAX: (44) 020 7269 3100
Cancer Research UK,                e_mail:derek.davies at cancer.org.uk
London Research Institute,       mobile: 07790 604112
44 Lincolns Inn Fields,
London, UK.

Web Page: http://sci.cancerresearchuk.org/axp/facs/davies/index.html

In tenebris lux
*************************************************************************






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