Optical Lock-In Detection of FRET Using Synthetic and Genetically Encoded Optical Switches

Optical Lock-In Detection of FRET Using Synthetic and Genetically Encoded Optical Switches

The Förster resonance energy transfer (FRET) technique is widely used for studying protein interactions within live cells. The effectiveness and sensitivity of determining FRET, however, can be reduced by photobleaching, cross talk, autofluorescence, and unlabeled, endogenous proteins. We present a...

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Veröffentlicht in:Biophysical journal 2008-06, Vol.94 (11), p.4515-4524
Hauptverfasser: Mao, Shu, Benninger, Richard K.P., Yan, Yuling, Petchprayoon, Chutima, Jackson, David, Easley, Christopher J., Piston, David W., Marriott, Gerard
Format: Artikel
Sprache:eng
Schlagworte:
Biomedical materials
Biophysics
Cells
Equivalence
Fluorescence
Fluorescence Resonance Energy Transfer - methods
Fluorescent Dyes - metabolism
Fretting
Imaging
In vivo testing
In vivo tests
Molecular biology
Optical switching
Optics and Photonics
Protein Engineering - methods
Protein Interaction Mapping - methods
Proteins
Recombinant Proteins - metabolism
Spectroscopy, Imaging, Other Techniques
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Zusammenfassung:The Förster resonance energy transfer (FRET) technique is widely used for studying protein interactions within live cells. The effectiveness and sensitivity of determining FRET, however, can be reduced by photobleaching, cross talk, autofluorescence, and unlabeled, endogenous proteins. We present a FRET imaging method using an optical switch probe, Nitrobenzospiropyran (NitroBIPS), which substantially improves the sensitivity of detection to
ISSN:0006-3495
1542-0086
DOI:10.1529/biophysj.107.124859