Abstract
The technique of Fluorescence Resonance Energy Transfer (FRET) has been extensively used in optical microscopy for the study of bio-molecular interactions. As most transient and weak intermolecular interactions only occur in intact cells under physiological conditions, it is essential to be able to record FRET from live cells. This chapter introduces a new spectra FRET approach that has recently been developed and tested. It provides the spatial resolution for separation of membrane and intracellular signals. Its spectrum-based procedure is easy to carry out and is powerful in separating donor and acceptor emissions as well as detecting the presence of other background light. Moreover, spectra FRET allows for a fine correction for common contaminations in FRET experiments, such as cross-talk and bleed-through, therefore contributing to the reduction of errors in FRET analysis.
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