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@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@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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