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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics

| EXPLORING THE INTERFACE OF LIGHT AND BIOMEDICINE

  • Editor: Gregory W. Faris
  • Vol. 2, Iss. 1 — Jan. 19, 2007

Dual-color fluorescence cross-correlation spectroscopy in a single nanoaperture: towards rapid multicomponent screening at high concentrations

Jérôme Wenger, Davy Gérard, Pierre-François Lenne, Hervé Rigneault, José Dintinger, Thomas W. Ebbesen, Annie Boned, Fabien Conchonaud, and Didier Marguet  »View Author Affiliations


Optics Express, Vol. 14, Issue 25, pp. 12206-12216 (2006)
http://dx.doi.org/10.1364/OE.14.012206


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Abstract

Single nanometric apertures in a metallic film are used to develop a simple and robust setup for dual-color fluorescence crosscorrelation spectroscopy (FCCS) at high concentrations. If the nanoaperture concept has already proven to be useful for single-species analysis, its extension to the dual-color case brings new interesting specificities. The alignment and overlap of the two excitation beams are greatly simplified. No confocal pinhole is used, relaxing the requirement for accurate correction of chromatic aberrations. Compared to two-photon excitation, nanoapertures have the advantage to work with standard fluorophore constructions having high absorption cross-section and well-known absorption/emission spectra. Thanks to the ultra-low volume analysed within one single aperture, fluorescence correlation analysis can be performed with single molecule resolution at micromolar concentrations, resulting in 3 orders of magnitude gain compared to conventional setups. As applications of this technique, we follow the kinetics of an enzymatic cleavage reaction at 2 µM DNA oligonucleotide concentration.We also demonstrate that FCCS in nanoapertures can be applied to the fast screening of a sample for dual-labeled species within 1 s acquisition time. This offers new possibilities for rapid screening applications in biotechnology at high concentrations.

© 2006 Optical Society of America

OCIS Codes
(050.1220) Diffraction and gratings : Apertures
(170.2520) Medical optics and biotechnology : Fluorescence microscopy
(170.6280) Medical optics and biotechnology : Spectroscopy, fluorescence and luminescence

ToC Category:
Medical Optics and Biotechnology

History
Original Manuscript: October 6, 2006
Revised Manuscript: December 1, 2006
Manuscript Accepted: December 2, 2006
Published: December 11, 2006

Virtual Issues
Vol. 2, Iss. 1 Virtual Journal for Biomedical Optics

Citation
Jérôme Wenger, Davy Gérard, Pierre-François Lenne, Hervé Rigneault, José Dintinger, Thomas W. Ebbesen, Annie Boned, Fabien Conchonaud, and Didier Marguet, "Dual-color fluorescence cross-correlation spectroscopy in a single nanoaperture : towards rapid multicomponent screening at high concentrations," Opt. Express 14, 12206-12216 (2006)
http://www.opticsinfobase.org/vjbo/abstract.cfm?URI=oe-14-25-12206


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