Joseph, The ECFP form will not work with your typical flow setup. You'll require a Krypton laser, with optics to provide a 407nm excitation line. Look at J. Dezz Ropp et.al. (Cytometry 21:309-317, 1995; Aequorea green fluorescent protein analysis by flow cytometry) as a good early reference describing these parameters. Keep in mind, though, that this difficulty may be overcome by using the recently introduced DsRed variant (Clontech). While the peak excitation is shown at 558nm, the claim is that it is also nicely excitable at 488nm. MAK. Joseph Webster 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. -- Mark A. KuKuruga, Managing Director University of Michigan Core Flow Cytometry <http://www.cancer.med.umich.edu/flow_cytometry> phone: 734-647-3216 fax: 734-936-7376 kukuru@umich.edu
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