A practical guide to time-resolved fluorescence microscopy and spectroscopy
Benjamin S. Clark,
Irene Silvernail,
Kenya Gordon
et al.
Abstract:Time-correlated single photon counting (TCSPC) coupled with confocal microscopy is a versatile biophysical tool that enables real-time monitoring of biomolecular dynamics across many timescales. With TCSPC, Fluorescence correlation spectroscopy (FCS) and pulsed interleaved excitation-Forster resonance energy transfer (PIE-FRET) is collected simultaneously on diffusing molecules to extract diffusion characteristics and proximity information. This article is a guide to calibrating FCS and PIE-FRET measurements w… Show more
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