Hi Joseph,
you could look at this paper- :
Beavis AJ, Kalejta RF.
Simultaneous analysis of the cyan, yellow and green
fluorescent proteins by flow
cytometry using single-laser excitation at 458 nm.
Cytometry. 1999 Sep 1;37(1):68-73.
PMID: 10451508; UI: 99382298
CFP- EX: major peak 433nm, minor peak 453nm-- EM: major peak 475nm,
minor peak 501nm. I run ECFP and EYFP together using dual laser 457nm and
488nm. The best emission filter I found for CFP is a 505/10 BP. The
emission signal is not as good as the YFP but easily resolved from the neg
population. You can, as you'll see if you read the above paper, also excite
GFP at 457nm.
regards
Ann
At 14:58 08.05.00 +1000, you wrote:
>
>I have a user who wishes to run Clontech EGFP and ECFP together; we
>can run EGFP O/K but we have not yet tried ECFP & I'm dubious....
>
>Has anyone used ECFP (cyan) in a flow cytometer?
>If so, what laser & wavelength excitation?
>
>The Clontech website says excitation is 433 with a secondary
>absorption peak at 453.
>No spectra shown, so I don't know how broad the absorption curve
>might be.
>We don't have any 433 line available, would the argon 457 line work?
>
>Any ideas &/or experience?
>
>Thanks, Joseph.
>--
>Joseph Webster, Flow Cytometry Facility
>Centenary Institute, Sydney AUSTRALIA.
>
>
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