>Hi, someone here is trying to image a double label
>with Cy3 and FITC and is experiencing bleed through
>from Cy3 into FITC channel. I haven't seen the samples
>but I think that is to be expected. Does anyone know
>or have experience with this? I suspect going to Cy5
>would be a better choice but they wanted to use Cy3
>because of results of tests with various secondaries.
>Any experiences appreciated thanks dave smith
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Try using Lissamine-Rhodamine in conjunction with FITC for double label. If
you have a BioRAD, use the K1 and K2 filter sets with an Argon-Krypton
laser. Lissamine-Rhodamine is shifted to a longer wavelength than ordinary
Rhodamine, and produces very little bleedthrough. If you only have an Argon
laser, however, you may have problems.
                                H.Karten
 
 
 Harvey J. Karten, M.D.
 Dept. of Neurosciences
 Univ.California San Diego
 La Jolla, CA 92093-0608
 EMail: [log in to unmask]
 Phone: (619)-534-4938
 FAX: (619)-534-6602