High-resolution, single-molecule measurements of biomolecular motion.
Many biologically important macromolecules undergo motions that are essential to their function. Biophysical techniques can now resolve the motions of single molecules down to the nanometer scale or even below, providing new insights into the mechanisms that drive molecular movements. This review outlines the principal approaches that have been used for high-resolution measurements of single-molecule motion, including centroid tracking, fluorescence resonance energy transfer, magnetic tweezers, atomic force microscopy, and optical traps. For each technique, the principles of operation are outlined, the capabilities and typical applications are examined, and various practical issues for implementation are considered. Extensions to these methods are also discussed, with an eye toward future application to outstanding biological problems.
Greenleaf
William J
WJ
Department of Applied Physics, Stanford University, Stanford, California 94305-5030, USA.
Woodside
Michael T
MT
Block
Steven M
SM
D001703:Biophysics / Q000379:methods*; D002627:Chemistry, Physical / Q000379:methods*; D004867:Equipment Design; D031541:Fluorescence Resonance Energy Transfer / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes / Q000494:pharmacology; D046911:Macromolecular Substances; D008280:Magnetics; D018625:Microscopy, Atomic Force / Q000295:instrumentation / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation; D008968:Molecular Conformation; D009038:Motion; D052898:Optical Tweezers; D012107:Research Design
D005456:Fluorescent Dyes; D046911:Macromolecular Substances