Novel perspectives for the application of total internal reflection microscopy

Date
2009-12
Authors
Volpe, Giovanni
Brettschneider, T.
Helden, L.
Bechinger, C.
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Source Title
Optics Express
Print ISSN
Electronic ISSN
1094-4087
Publisher
Optical Society of America
Volume
17
Issue
26
Pages
23975 - 23985
Language
English
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Abstract

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).

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