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Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 17, Iss. 26 — Dec. 21, 2009
  • pp: 23975–23985

Novel perspectives for the application of total internal reflection microscopy

Giovanni Volpe, Thomas Brettschneider, Laurent Helden, and Clemens Bechinger  »View Author Affiliations

Optics Express, Vol. 17, Issue 26, pp. 23975-23985 (2009)

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Total Internal Reflection Microscopy (TIRM) is a sensitive non-invasive technique to measure the interaction potentials between a colloidal particle and a wall with femtonewton resolution. The equilibrium distribution of the particle-wall separation distance z is sampled monitoring the intensity I scattered by the Brownian particle under evanescent illumination. Central to the data analysis is the knowledge of the relation between I and the corresponding z, which typically must be known a priori. This poses considerable constraints to the experimental conditions where TIRM can be applied (short penetration depth of the evanescent wave, transparent surfaces). Here, we introduce a method to experimentally determine I(z) by relying only on the distance-dependent particle-wall hydrodynamic interactions. We demonstrate that this method largely extends the range of conditions accessible with TIRM, and even allows measurements on highly reflecting gold surfaces where multiple reflections lead to a complex I(z).

© 2009 Optical Society of America

OCIS Codes
(120.0120) Instrumentation, measurement, and metrology : Instrumentation, measurement, and metrology
(120.5820) Instrumentation, measurement, and metrology : Scattering measurements
(180.0180) Microscopy : Microscopy
(240.0240) Optics at surfaces : Optics at surfaces
(240.6690) Optics at surfaces : Surface waves
(260.6970) Physical optics : Total internal reflection

ToC Category:
Instrumentation, Measurement, and Metrology

Original Manuscript: September 29, 2009
Revised Manuscript: November 19, 2009
Manuscript Accepted: November 20, 2009
Published: December 16, 2009

Virtual Issues
Vol. 5, Iss. 1 Virtual Journal for Biomedical Optics

Giovanni Volpe, Thomas Brettschneider, Laurent Helden, and Clemens Bechinger, "Novel perspectives for the application of total internal reflection microscopy," Opt. Express 17, 23975-23985 (2009)

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