GFP-based FRET analysis in live cells Export

@article{citeulike:2760620, 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.}, author = {Takanishi, Christina L. and Bykova, Ekaterina A. and Cheng, Wei and Zheng, Jie}, citeulike-article-id = {2760620}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/j.brainres.2006.01.119}, citeulike-linkout-1 = {http://www.sciencedirect.com/science/article/B6SYR-4JFGFHG-4/1/9dd8e450c9ebc41e786b75ecf9c02006}, day = {26}, doi = {10.1016/j.brainres.2006.01.119}, journal = {Brain Research}, keywords = {fret, gfp}, month = {May}, number = {1}, pages = {132--139}, posted-at = {2008-05-06 12:01:19}, priority = {2}, title = {GFP-based FRET analysis in live cells}, url = {http://dx.doi.org/10.1016/j.brainres.2006.01.119}, volume = {1091}, year = {2006} }

TY - JOUR ID - citeulike:2760620 L3 - citeulike-article-id:2760620 N2 - 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. IS - 1 JF - Brain Research EP - 139 TI - GFP-based FRET analysis in live cells VL - 1091 SP - 132 KW - fret KW - gfp AU - Takanishi, Christina AU - Bykova, Ekaterina AU - Cheng, Wei AU - Zheng, Jie PY - 2006/05/26/ UR - http://dx.doi.org/10.1016/j.brainres.2006.01.119 ER -