Joseph, We have been able to detect expression of eCFP in cells using a FACS Vantage. We tuned the argon laser to 458 nm and used a 480/30 bandpass filter to collect the eCFP fluorescence. We also found that eGFP and eYFP could be excited at 458 nm which allows the simultaneous analysis of these three fluorescent proteins using a single excitation wavelength. This was published recently in Cytometry: Beavis AJ and Kalejta RF. Simultaneous analysis of the cyan, yellow and green fluorescent proteins by flow cytometry using single-laser excitation at 458 nm. Cytometry 37(1), pp 68-73. 1999. Details can also be found on my web page at: http://www.molbio.princeton.edu/facility/flowcyt/3FP.html Please do not hesitate to call me if you have any further questions. regards Andy Beavis ******************************************************************* Andrew J. Beavis Manager, Flow Cytometry Core Facility Princeton University Department of Molecular Biology Lewis Thomas Laboratory Washington Road Princeton, NJ 08544 U.S.A. Phone: (609) 258 1695 Fax: (609) 258 5323 E-mail. Abeavis@molbio.princeton.edu Web: http://www.molbio.princeton.edu/facility/flowcyt/index.html ******************************************************************* -----Original Message----- From: Joseph Webster [mailto:J.Webster@centenary.usyd.edu.AU] Sent: Monday, May 08, 2000 12:59 AM To: cyto-inbox Subject: ECFP Anyone? 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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