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

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 convent...

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Bibliographic Details
Published in:Science (American Association for the Advancement of Science) 2003-01, Vol.299 (5607), p.682-686
Main Authors: Levene, M. J., Korlach, J., Turner, S. W., Foquet, M., Craighead, H. G., Webb, W. W.
Format: Article
Language:English
Subjects:
Biochemistry
Biochemistry - instrumentation
Biochemistry - methods
Chemistry
Computer Simulation
Coumarins
Deoxycytosine Nucleotides - metabolism
Deoxyribonucleic acid
Diffusion
DNA
DNA - biosynthesis
DNA polymerases
DNA-Directed DNA Polymerase - metabolism
Dyes
Enzymes
Enzymes, Immobilized - metabolism
Exact sciences and technology
Finite Element Analysis
Fluorescence
Fluorescent Dyes
General and physical chemistry
Kinetics
Ligands
Light
Metal films
Methods
Molecular biology
Molecular microbiology
Molecular spectroscopy
Molecules
Nanotechnology - instrumentation
Observation
Optics and Photonics
Spectrometry, Fluorescence - instrumentation
Spectrometry, Fluorescence - methods
Theory of reactions, general kinetics
Theory of reactions, general kinetics. Catalysis. Nomenclature, chemical documentation, computer chemistry
Waveguides
Wavelengths
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Summary: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.
ISSN:0036-8075
1095-9203
DOI:10.1126/science.1079700