Introduction

Fluorescence resonance energy transfer (FRET) is a distance-dependent physical process by which energy is transferred nonradiative from an excited molecular fluorophore (the donor) to another fluorophore (the acceptor) by means of intermolecular long-range dipole-dipole coupling. The theory supporting energy transfer is based on the concept of treating an excited fluorophore as an oscillating dipole that can undergo an energy exchange with a second dipole having a similar resonance frequency. Unlike radiative mechanisms, resonance energy transfer can yield a significant amount of structural information concerning the donor-acceptor pair.

FRET can be an accurate measurement of molecular proximity at angstrom distances and highly efficient if the donor and acceptor are positioned within the Förster radius. FRET now is widely used in cell biology, among all the applications, there are five general FRET approaches have proven particularly useful:

5. Fluorescence Polarization Imaging - Measure polarization parallel and perpendicular to excitation with high signal-to-noise

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