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
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:16 EST