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Zero-Mode Waveguides for Single-Molecule Analysis at High Concentrations

Science  31 Jan 2003:
Vol. 299, Issue 5607, pp. 682-686
DOI: 10.1126/science.1079700

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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.

  • * Present address: Nanofluidics Incorporated, 17 Sheraton Drive, Ithaca, NY 14850, USA.

  • To whom correspondence should be addressed. E-mail: www2{at}cornell.edu

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