Formation
The
FRET (Fluorescence Resonance Energy Transfer) is a phenomenon of the
quantum mechanics which occurs between a fluorescent donor (green)
and a fluorescent acceptor (red) which are very close (distance
<100Å).
When
the emission spectrum donor overlaps the spectrum of excitation of
the acceptor. (Förster, 1948). Under ideal conditions of FRET,
the excitation of D causes a transfer of energy (not photons) towards
A, who emits in his wavelengths. We can use this phenomenon to study
with the confocal microscope the interactions between two molecules.
We couple the molecules which interest us with fluorochromes
carefully selected. If they are close, the acceptor "ignites",
if not it remains "extinct".
Formation
3D/4D
FLIM in the undercellulary compartments of the alive cells.
Images
FLIM will be obtained with a space resolution in space 3D resolution
or 4D (3D+time), by using systems of confocal microscopy (confocal
laser scanning) for alive cells.
By
visualizing the undercellular compartments with molecules of
GFP-combined with the proteines specific of these compartments, the
cinetiques ones of the molecular interactions detectées by
FLIM can be assigned with certain undercellular compartments.
