novice on FRET

Janos Szollosi szollo at JAGUAR.DOTE.HU
Fri Oct 1 10:37:58 EST 1999

Dear Oliver,

I have performed a lot of FRET measurements for mapping various antigens on
the cell surface. In the old days I used mostly the FITC/TRITC pair because
in a flow cytometer, where I have most of the measurements, these dyes fits
the available laser lights.

I have not heard about the "ideal" acceptor you would like to use.  In a
more sophisticated, lifetime or time-gated instrument there is a way to
come close to this ideal situation:

You can use a TBP-EU3+ donor which has a long lifetime and a very selective
emission spectra (composed of lines) and an ACP acceptor.  Using
time-gating you can detect fluorescence on the acceptor side only if the
long lifetime donor is present.  In addition you will get rid of the prompt
autofluorescence as well. This pair was described in: Clin. Chem.
39:1953-1959, 1993.

You can find more donor-acceptor pairs in my recent review paper and book
Cytometry, 34:159-179, (1998)
Current Protocols in Cytometry (managing editor Paul Robinson) 1998,
Supplement 9,  (1999) Chapter 1.12  Principles of Resonance Energy Transfer.

If you need reprints please send me your address and I will mail them to you!



At 03:00 PM 10/1/99 +0200, you wrote:
>has someone anytime done Fluorescence Energy Transfer (FRET)? I've read that
>mostly you can observe reduction in donor emission but what I need is a
>strong increase in acceptor emission. It would be best if the acceptor
>wouldn't emit anything without the donor. Can somoeone give me a tip for a
>suitable pair of dyes?
>Oliver Schaefer
>Heinrich-Heine-Universitaet Duesseldorf
Janos Szollosi
Department of Biophysics and Cell Biology
University Medical School of Debrecen
Nagyerdei krt. 98, P.O.Box 39,
H-4012 Debrecen,
Fax/Phone: (36) (52) 412-623
e-mail: szollo at jaguar.dote.hu

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