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Science. 2003 Jan 31;299(5607):682-6.

Zero-mode waveguides for single-molecule analysis at high concentrations.

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  • 1Applied and Engineering Physics, Graduate Program in Biochemistry, Molecular, and Cell Biology, Cornell University, Clark Hall, Ithaca, NY 14853, USA.

Abstract

Optical approaches for observing the dynamics of single molecules have required pico- to nanomolar concentrations of fluorophore in order to isolate individual molecules. However, many biologically relevant processes occur at micromolar ligand concentrations, necessitating a reduction in the conventional observation volume by three orders of magnitude. We show that arrays of zero-mode waveguides consisting of subwavelength holes in a metal film provide a simple and highly parallel means for studying single-molecule dynamics at micromolar concentrations with microsecond temporal resolution. We present observations of DNA polymerase activity as an example of the effectiveness of zero-mode waveguides for performing single-molecule experiments at high concentrations.

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PMID:
12560545
[PubMed - indexed for MEDLINE]
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