Journal
JOURNAL OF FLUORESCENCE
Volume 17, Issue 6, Pages 775-783Publisher
SPRINGER/PLENUM PUBLISHERS
DOI: 10.1007/s10895-007-0214-0
Keywords
fluorescence spectroscopy; biological systems; fluorescence resonance energy transfer; single-molecule biophysics
Funding
- NIGMS NIH HHS [R01 GM 073104, R01 GM 066833] Funding Source: Medline
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This review provides an account of single-molecule fluorescence methodologies for freely diffusing molecules applied to a diverse array of biological problems pertaining to biomolecular folding and assembly. We describe the principles of confocal fluorescence microscopy to detect and analyze fluorescence bursts from diffusing single molecules. These methods including single-molecule fluorescence resonance energy transfer, coincidence, correlations and polarization offer a powerful means to uncover hidden information about conformational sub-populations and interconversion dynamics of biomolecular systems in a wide range of timescales. We offer several key examples to illustrate how these methodologies have been extremely useful in teasing out structural and dynamical aspects of many important biomolecular systems.
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