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First, after 4-color upgrade the FL3 channel has a 670LP rather than 650LP
filter. This will probably be BETTER for PE/Cy5 in that PE emmission will be
slightly reduced in FL3. There are no other issues in 3-color applications.
Second, the system HAS inter-beam compensation. The compensation value for PE/
Cy5 conjugates is quite high because of the direct red excitation of Cy5. I
have used comp values like 60% on only one instrument - also only two PE/Cy5
conjugates from only one vendor - "your mileage may vary". Very bright PE/Cy5
conjugates could thus be a problem but dimmer ones seem OK. A strategy where
the PE/Cy5 reagent subsets the APC-positives would be more robust than vice
versa.
Conclusion, PE/Cy5 can be used for 3-color as always - PE/Cy5 is usable, but
PerCP has compensation advantages, in 4-color applications.
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web