Direct Measurement of Single-Molecule Diffusion and Photodecomposition in Free Solution

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Science  21 Feb 1997:
Vol. 275, Issue 5303, pp. 1106-1109
DOI: 10.1126/science.275.5303.1106

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Continuous monitoring of submillisecond free-solution dynamics of individual rhodamine-6G molecules and 30-base single-stranded DNA tagged with rhodamine was achieved. Fluorescence images were recorded from the same set of isolated molecules excited either through the evanescent field at the quartz-liquid interface or as a thin layer of solution defined by micron-sized wires, giving diffraction-limited resolution of interconnected attoliter volume elements. The single-molecule diffusion coefficients were smaller and the unimolecular photodecomposition lifetimes were longer for the dye-DNA covalent complex as compared with those of the dye molecule itself. Unlike bulk studies, stochastic behavior was found for individual molecules of each type, and smaller diffusion coefficients were observed.

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