Sam As a general rule I always try to reflect the dimmest signal and pass the brighter signal. The loss is most significant when passing thru a filter. Larry At 07:50 PM 4/18/00 -0400, you wrote: > >Hi Group > I'm trying to collect DAPI and APC signals on the second (and third) >laser lines of a FACStar Plus in a manner very similar to that described by >Moore et al in: > Cytometry (32) 1: 57-65 , May 1998. The method describes the >collection of DAPI and APC fluorescence from cells excited by collinear >beams of a mixed gas laser for UV (350-356 nm) and a HeNe laser (633 nm). > >In this technical note, the filters adjacent to the relevant PMT's are >described (660DF20 for APC and 450DF20 for DAPI), but the splitter between >the two PMT's is not mentioned. > I have looked around and cannot find a reference as to the "proper >thing to do" in this case. > > The DAPI signal is very bright... should I reflect it and filter the >APC fluorescence or should I filter the DAPI and reflect the APC? > >Is there a general rule that I could apply based on wavelength regardless of >"brightness"? > > Thanks in Advance, > > I look forward to your input. > > Cheers, > Sam >Sam Witherspoon >sw11527@glaxowellcome.com > >Dept. of Receptor Biochemistry Tel. 919-483-3078 >Glaxo Wellcome R&D Page 919-857-7768 >5 Moore Dr. Fax 919-483-0585 >RTP, NC 27709 > Larry W. Arnold, Ph.D. Res. Assoc. Prof. Director, Flow Cytometry Facility Department of Microbiology and Immunology CB# 7290 University of North Carolina Chapel Hill, NC 27599 Phone: 919-966-1530 FAX: 919-962-8103
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