SP with LSR

David Dombkowski dombkows at helix.mgh.harvard.edu
Wed Mar 6 07:47:31 EST 2002



Jim,

  I suppose that if you co-layer the UV and HeNe you will have a working
solution but there are other considerations. I and others do not want to
sacrifice the HeNe APC signal along with Hoechst. Your solution would not
allow this as Red fluorescence of Hoechst would interfere with Red
fluorescence of APC. I think that another pinhole in front of PMT to allow
for UV signal would be sufficient. This is the kind of problem we need to
address with B.D. to improve on the standard configuration. I would also
like to use LSR for four dyes off of Argon and two off of HeNe for 6-color.

David



If you co-layer the UV and the HeNe in the 3rd position you can detect the
red signal from the HO dye in FL3.  My LSR is configured with a change in
laser alignment and pinholes to allow UV/HeNe laser to be seen in FL3.  Not
so hard to do.

Contact  BD if unable to re-align yourself.

Jim Houston
> ----------
> From:		David Dombkowski
> Sent:		Monday, March 4, 2002 7:32 AM
> To:	Cytometry Mailing List
> Subject:	SP with LSR
>
>
> Kimmo,
>
>     You will not see red fluorescence with Hoechst 33342 with the
> configuration of 2 PMT UV, 3 or 4 PMT Argon and 1 PMT He-Ne. The red
> fluorescence is too high for dichroic to reflect to PMT for UV. You need
> to
> be able to collect this signal in PMT usually dedicated to Argon or He-Ne
> laser. There is no pinhole for UV fluorescence in these channels. You need
> a custon configuration which may be achieved if you cantact Becton
> Dickinson.
>
> David

David M. Dombkowski
dombkowski at helix.mgh.harvard.edu
Flow Cytometry-Pathology-CNY rm7017
149 13th Street
Massachusetts General Hospital-East
Charlestown, MA 02129
Tel. (617)726-1683
Fax.(617)724-3164






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