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Proceedings Paper

Searching the fluorescent protein color palette for new FRET pairs
Author(s): Kristin L. Hazelwood; Ericka B. Ramko; Anna P. Ozarowska; Scott G. Olenych; Patrice N. Worthy; Amy Guan; Christopher S. Murphy; Michael W. Davidson
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Paper 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.

Paper Details

Date Published: 13 February 2008
PDF: 12 pages
Proc. SPIE 6868, Small Animal Whole-Body Optical Imaging Based on Genetically Engineered Probes, 68680C (13 February 2008); doi: 10.1117/12.764560
Show Author Affiliations
Kristin L. Hazelwood, The Florida State Univ. (United States)
Ericka B. Ramko, The Florida State Univ. (United States)
Anna P. Ozarowska, The Florida State Univ. (United States)
Scott G. Olenych, The Florida State Univ. (United States)
Patrice N. Worthy, The Florida State Univ. (United States)
Amy Guan, The Florida State Univ. (United States)
Christopher S. Murphy, The Florida State Univ. (United States)
Michael W. Davidson, The Florida State Univ. (United States)


Published in SPIE Proceedings Vol. 6868:
Small Animal Whole-Body Optical Imaging Based on Genetically Engineered Probes
Alexander P. Savitsky; Robert E. Campbell; Robert M. Hoffman, Editor(s)

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