CONFOCALMICROSCOPY Archives

March 2000

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Subject:
From:
"Dr. Sudipta Maiti" <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Fri, 3 Mar 2000 21:44:13 +0530
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Well, you have to know the Forster radius for the GFP-dye pair (can be
calculated from the spectra), but usually for dyes with good spectral
overlap, something <5nm should be good. A quick back of the envelope
calculation would mean (in solution) 1 rhodamine molecule (Rhodamine6G
should have good overlap with EGFP) per 50 cubic nm, or 2X10^22
molecules/liter, i.e. 30mM. The concentration of acceptor molecules thus
have to be very high. Self quenching also occurs by Forster transfer and
will happen, but you should see EGFP-Rhodamine transfer dominate at
appropriate concentrations.
Sudipta

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