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

Science. 2003 Jan 31;299(5607):682-6. doi: 10.1126/science.1079700.


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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biochemistry / instrumentation
  • Biochemistry / methods*
  • Computer Simulation
  • Coumarins
  • DNA / biosynthesis
  • DNA-Directed DNA Polymerase / metabolism*
  • Deoxycytosine Nucleotides / metabolism*
  • Diffusion
  • Enzymes, Immobilized / metabolism
  • Finite Element Analysis
  • Fluorescence
  • Fluorescent Dyes
  • Ligands
  • Light
  • Nanotechnology* / instrumentation
  • Optics and Photonics
  • Spectrometry, Fluorescence / instrumentation
  • Spectrometry, Fluorescence / methods*


  • Coumarins
  • Deoxycytosine Nucleotides
  • Enzymes, Immobilized
  • Fluorescent Dyes
  • Ligands
  • 2'-deoxycytidine 5'-triphosphate
  • DNA
  • coumarin
  • bacteriophage T7 induced DNA polymerase
  • DNA-Directed DNA Polymerase