Searching the fluorescent protein color palette for new FRET pairs

Kristin L. Hazelwood; Ericka B. Ramko; Anna P. Ozarowska; Scott G. Olenych; Patrice N. Worthy; Amy Guan; Christopher S. Murphy; Michael W. Davidson

doi:10.1117/12.764560

13 February 2008 Searching the fluorescent protein color palette for new FRET pairs

Kristin L. Hazelwood, Ericka B. Ramko, Anna P. Ozarowska, Scott G. Olenych, Patrice N. Worthy, Amy Guan, Christopher S. Murphy, Michael W. Davidson

Author Affiliations +

Proceedings Volume 6868, Small Animal Whole-Body Optical Imaging Based on Genetically Engineered Probes; 68680C (2008) https://doi.org/10.1117/12.764560
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

One of the most promising imaging techniques for monitoring dynamic protein interactions in living cells with optical microscopy, universally referred to as FRET, employs the non-radiative transfer of energy between two closely adjacent spectrally active molecules, often fluorescent proteins. The use of FRET in cell biology has expanded to such a degree that hundreds of papers are now published each year using biosensors to monitor a wide spectrum of intracellular processes. Most of these sensors sandwich an environmentally active peptide between cyan and yellow fluorescent protein (CFP and YFP) derivatives to assay variables such as pH, calcium ion concentration, enzyme activity, or membrane potential. The availability of these sensitive indicators is growing rapidly, but many are hampered by a low dynamic range that often is only marginally detectable over the system noise. Furthermore, extended periods of excitation at wavelengths below 500 nm have the potential to induce phototoxic effects that can mask or alter the biological events under observation. Recent success in expanding the fluorescent protein color palette offers the opportunity to explore new FRET partners that may be suitable for use in advanced biosensors.

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Kristin L. Hazelwood, Ericka B. Ramko, Anna P. Ozarowska, Scott G. Olenych, Patrice N. Worthy, Amy Guan, Christopher S. Murphy, and Michael W. Davidson "Searching the fluorescent protein color palette for new FRET pairs", Proc. SPIE 6868, Small Animal Whole-Body Optical Imaging Based on Genetically Engineered Probes, 68680C (13 February 2008); https://doi.org/10.1117/12.764560

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