1. Two-photon excitation action cross-sections of the autofluorescent proteins
- Author
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Gregory S. Harms, Gerhard A. Blab, Laurent Cognet, Piet H.M. Lommerse, and Thomas Schmidt
- Subjects
Autofluorescence ,Two-photon excitation microscopy ,Absorption spectroscopy ,Chemistry ,Cyan ,Biophysics ,Analytical chemistry ,General Physics and Astronomy ,Flavin group ,Physical and Theoretical Chemistry ,Absorption (electromagnetic radiation) ,Fluorescence ,Blueshift - Abstract
We report on the values of the two-photon excitation action cross-sections of commercially available enhanced cyan, green, yellow, and red fluorescent proteins. The two-photon absorption spectra are very similar in shape to those measured for one-photon absorption. However, they exhibit a significant blueshift, which is attributed to the participation of a vibrational mode in the two-photon absorption process. The two-photon spectra are compared to that of flavine mononucleotide, which constitutes the main source of autofluorescence in mammalian cells. The definition of a relative detection yield between the autofluorescent proteins and flavine allows us to quantify the applicability of autofluorescent proteins in two-photon single-molecule studies in living cells.
- Published
- 2001
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