Abstract：
:Fluorescence resonance energy transfer (FRET) is a widely utilized optical technique for measuring small distances of 1-10 nm in live cells. In recent years, its application has been greatly popularized by the discovery of green fluorescent protein (GFP) and many improved variants which make good donor-acceptor fluorophore pairs. GFP-based proteins are structurally stable, relatively inert, and can be reliably attached to points of interest. The combination of easy access to the GFP-based FRET technique and its obvious usefulness in many applications can lead to complacency. Potential problems such as light contaminants, e.g., bleed-through and cross-talk, and inconsistent donor and acceptor concentrations are easily overlooked and can lead to errors in FRET calculation and data interpretation. In this article, we outline possible pitfalls of GFP-based FRET and approaches that address these issues, including a "Spectra FRET" technique that can be easily applied to live cell studies.
journal_name
Brain Resjournal_title
Brain researchauthors
Takanishi CL,Bykova EA,Cheng W,Zheng Jdoi
10.1016/j.brainres.2006.01.119subject
Has Abstractpub_date
2006-05-26 00:00:00pages
132-9issue
1eissn
0006-8993issn
1872-6240pii
S0006-8993(06)00323-4journal_volume
1091pub_type
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