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One-Photon and Two-Photon Excitation of Fluorescent Proteins

  • R. NifosìEmail author
  • V. Tozzini
Chapter
Part of the Springer Series on Fluorescence book series (SS FLUOR, volume 11)

Abstract

Fluorescent proteins (FPs) offer a wide palette of colors for imaging applications. One purpose of this chapter is to review the variety of FP spectral properties, with a focus on their structural basis. Fluorescence in FPs originates from the autocatalytically formed chromophore. Several studies exist on synthetic chromophore analogs in gas phase and in solution. Together with the X-ray structures of many FPs, these studies help to understand how excitation and emission energies are tuned by chromophore structure, protonation state, and interactions with the surrounding environment, either solvent molecules or amino acids residues. The increasing use of FPs in two-photon microscopy also prompted detailed investigations of their two-photon excitation properties. The comparison with one-photon excitation reveals nontrivial features, which are relevant both for their implications in understanding multiphoton properties of fluorophores and for application purposes.

Keywords

Fluorescent proteins Chromophore structures Computational studies Isolated chromophores Multiphoton spectroscopy Structure-property relationship Spectral tuning 

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© Springer-Verlag Berlin Heidelberg 2011

Authors and Affiliations

  1. 1.NEST CNR-NANOPisaItaly

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