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We recently needed to run samples stained with FITC, Tricolor and
Texas Red, a choice forced on us by availability of reagents. We
avoided the problem of 2nd laser excitation of Tricolor and its spill
into the Texas Red channel by treating the Tricolor like Cy5, exciting
it with the 2nd laser, which then allowed us to compensate against
Texas Red. Used this way the signal/background looked about as good
as Tricolor excited with laser1. Yet we see very few people reporting
results using Cy5 itself. Which brings us to the question: In
*actual* laboratory bench terms, how does Cy5 compare to APC (when
excited sub-optimally by dye laser at 605nm) for:
a: Ease and stability of conjugation.
b: Intensity of fluorescence.
And further, how does Cy3 compare to PE in the same terms.
It would seem there may sometimes be advantages in Cy3 and Cy5 being
so much smaller molecules than the phycobilis
\ / < Flow Systems Laboratory
Frank Battye \__/ <<<<< The Walter & Eliza Hall Institute
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o !! \ < IN:: "facs_copy@wehi.edu.au"
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