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Summary: NEST Scientific Report 2007-200987
P
rogress in fluorescence imaging is enabling the study of biological events at
unprecedented detail, thanks to novel microscopy techniques that provide imaging
with nanoscale resolution. 3D imaging of living cells within intact tissues, organs,
and whole animals, is accessible through multiphoton excitation. The remarkably
rich photophysics of Fluorescent Proteins (FPs) makes them flexible enough to be used
in conjunction with these advanced spectroscopic approaches. Here we report on our
progress in understanding FPs multiphoton excitation spectra and in unraveling the
photophysics behind photoactivation/deactivation of some FPs, a property enabling their
use as fluorophores for nano-resolution imaging. This analysis allows us to develop new
reversibly switchable FP mutants.
Photoswitching and cis-trans photoisomer-
ization
Recently, a novel approach to in
vivo imaging has emerged with the
development of new FPs that can be
reversibly or irreversibly photoconverted
between two optical states, adding a
new temporal dimension to imaging
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