Imaging of single molecule diffusion.
- 2 April 1996
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 93 (7), 2926-2929
- https://doi.org/10.1073/pnas.93.7.2926
Abstract
In recent years observations at the level of individual atoms and molecules became possible by microscopy and spectroscopy. Imaging of single fluorescence molecules has been achieved but has so far been restricted to molecules in the immobile state. Here we provide methodology for visualization of the motion of individual fluorescent molecules. It is applied to imaging of the diffusional path of single molecules in a phospholipid membrane by using phospholipids carrying one rhodamine dye molecule. For this methodology, fluorescence microscopy was carried to a sensitivity so that single fluorescent molecules illuminated for only 5 ms were resolvable at a signal/noise ratio of 28. Repeated illuminations permitted direct observation of the diffusional motion of individual molecules with a positional accuracy of 30 nm. Such capability has fascinating potentials in bioscience--for example, to correlate biological functions of cell membranes with movements, spatial organization, and stoichiometries of individual components.Keywords
This publication has 17 references indexed in Scilit:
- Automated detection and tracking of individual and clustered cell surface low density lipoprotein receptor moleculesBiophysical Journal, 1994
- Direct imaging of reptation for semiflexible actin filamentsNature, 1994
- Single molecule detection and photochemistry on a surface using near-field optical excitationPhysical Review Letters, 1994
- Single Molecules Observed by Near-Field Scanning Optical MicroscopyScience, 1993
- Confined lateral diffusion of membrane receptors as studied by single particle tracking (nanovid microscopy). Effects of calcium-induced differentiation in cultured epithelial cellsBiophysical Journal, 1993
- Microscopic versus macroscopic diffusion in one-component fluid phase lipid bilayer membranesBiophysical Journal, 1991
- Single pentacene molecules detected by fluorescence excitation in ap-terphenyl crystalPhysical Review Letters, 1990
- Single-molecule fluorescence detection: autocorrelation criterion and experimental realization with phycoerythrin.Proceedings of the National Academy of Sciences of the United States of America, 1989
- Tracking kinesin-driven movements with nanometre-scale precisionNature, 1988
- Atomic Force MicroscopePhysical Review Letters, 1986