Fluorochromes choice

David Dombkowski dombkowski at helix.mgh.harvard.edu
Thu Oct 31 14:54:23 EST 2002


Michal,

  It is certainly possible to do 6-color on a BD LSR with 488nm,633nm
and 325nm lasers. With a little thought and minor work the instrument
can be configured to see 4 fluorochromes on 488nm laser 1
fluorochrome on 633nm and 1 fluorochrome on 325nm laser. Software
compensation as with FlowJo will work well to do final compensation.
Please be as creative with the instrumentation and good things will
come from it!

David



Hi Michal,

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
NIH/NIAMS
Building 10, Room 6D57
9000 Rockville Pike
Bethesda, MD 20892
(301) 594-3537
fischer1 at mail.nih.gov

>  ----------
>  From:		Michal Abel
>  Sent:		Wednesday, October 30, 2002 2:21 PM
>  To:	Cytometry Mailing List
>  Subject:	Fluorochromes choice
>
>
>  Dear experts in flow cytometry,
>
>  I have a possibility to use 6 colors FACS. The trouble is that I am not
>  sure it is possible with the filters I have.
>  I would very much appreciate any help.
>  The instrument has three lasers: 325, 488 and 633 nm and I can use
>  following filters:
>  380/LP
>  400/40
>  424/44
>  500/11
>  510/20
>  530/28
>  575/15
>  610/20
>  660/13
>  670/LP
>  682/13
>  Is it possible to find 6 different flourochromes, which would be
>  efficiently differentiated with these filters/lasers ?
>
>  Thank you very much for your help.
>
>  Michal
>
>  --
>  Michal Abel, MD, PhD
>  Laboratory of Clinical Immunology
>  INSERM U25
>  Necker Hospital, 161 Rue de Sèvres. Paris
>  tel: 33 1 42 19 28 87
>  fax: 33 1 44 49 53 74
--
David M. Dombkowski
dombkowski at helix.mgh.harvard.edu
Flow Cytometry-Pathology-CNY rm7017
Massachusetts General Hospital-East
149 13th Street
Charlestown, MA 02129
Tel (617)-726-1683
Fax (617)-724-3164



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