Research Techniques Made Simple: Methodology and Applications of Forster Resonance Energy Transfer (FRET) Microscopy

[摘要] Classical biochemical techniques have contributed a great deal to our understanding of the mechanisms regulating fundamental biological processes. However, these approaches are typically end-point, population-based assays and are often insufficient in examining transient molecular events. Forster resonance energy transfer (FRET) microscopy is a powerful technique capable of investigating dynamic interactions between proteins and a plethora of biochemical signaling events based on the development of specific biosensors. This technique exploits the principle that when FRET occurs, energy from a donor fluorophore is transferred to an acceptor fluorophore only when certain conditions are met. These include dependence on both distance and fluorophore orientation. In this article, applications of FRET microscopy to protein interactions and modifications are discussed, and examples are given of the types of biosensors that can be developed. There are a number of methods to measure FRET. The most common modalities and specific advantages and shortcomings for each are reviewed. Finally, general considerations and guidelines for choosing a method are discussed.

[发布日期] 2017-11-01