Microscopy & Microtechniques

DEVELOPMENT OF A TWO PHOTON BURST INTEGRATED FLUORESCENCE LIFETIME SYSTEM FOR SINGLE-PAIR FRET TO ANALYSE INTERACTING PROTEIN POPULATIONS

Nov 05 2008

Author: Senthila Pathmananthan James Hunt Daniel R. Matthews Andrew J. Beavil Tony Ng Simon M. Ameer-Beg

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Fluorescence detection of single molecules represents the ultimate in sensitivity for fluorescence-based assays in biology and medicine by eliminating ensemble-averaging. We describe the development of a time-resolved burst integrated fluorescence lifetime (BiFL) methodology to observe single-molecule dynamics and measure fluorescence lifetime of these molecules as they diffuse through a femtoliter volume probed by two photon excitation.

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