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Science 11 November 1994:
Vol. 266. no. 5187, pp. 1018 - 1021
DOI: 10.1126/science.7973650
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Articles

Science, Vol 266, Issue 5187, 1018-1021
Copyright © 1994 by American Association for the Advancement of Science

articles

Probing individual molecules with confocal fluorescence microscopy

S Nie, DT Chiu, and RN Zare

Department of Chemistry, Stanford University, CA 94305.

Confocal fluorescence microscopy coupled with a diffraction-limited laser beam and a high-efficiency detection system has been used to study the diffusive movement and emission process of individual fluorescent molecules in the liquid phase at room temperature. The high detection sensitivity achieved at fast data acquisition speeds (greater than 1 kilohertz) allows real-time observation of single-molecule fluorescence without statistical analysis. The results show fluorescence-cycle saturation at the single-molecule level and multiple recrossings of a single molecule into and out of the probe volume as well as the triplet state.


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