Spectrally resolved fluorescence lifetime imaging: new developments and applications

A. Rueck; F. Dolp; B. v. Einem; C. A. F. v. Arnim; D. Strat

doi:10.1117/12.841782

26 February 2010 Spectrally resolved fluorescence lifetime imaging: new developments and applications

A. Rueck, F. Dolp, B. v. Einem, C. A. F. v. Arnim, D. Strat

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 75690Y (2010) https://doi.org/10.1117/12.841782
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

The fluorescence lifetime of different molecular species is calculated from the measured fluorescence intensity decrease following short pulsed laser excitation, by a multi-channel fitting procedure. In a FRET (Förster Resonant Energy Transfer) experiment the time dependent behaviour of the donor profile is assumed in a first view mono-exponential and the acceptor decay profile is solved analytically. A global minimization fitting algorithm has increased information content than a single channel fitting routine. In a normal FRET-FLIM experiment, the efficiency of FRET is calculated only by considering the kinetics of the donor. However, as will be shown, a considerable improvement could be achieved when time-resolved and spectral-resolved techniques are simultaneously incorporated.

Citation Download Citation

A. Rueck, F. Dolp, B. v. Einem, C. A. F. v. Arnim, and D. Strat "Spectrally resolved fluorescence lifetime imaging: new developments and applications", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 75690Y (26 February 2010); https://doi.org/10.1117/12.841782

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