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Proceedings Paper

Single-molecule microscopy using tunable nanoscale confinement
Author(s): Christopher M. J. McFaul; Jason Leith; Bojing Jia; François Michaud; Adriel Arsenault; Andrew Martin; Daniel Berard; Sabrina Leslie
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Paper Abstract

We present the design, construction and implementation of a modular microscopy device that transforms a basic inverted fluorescence microscope into a versatile single-molecule imaging system. The device uses Convex Lens- Induced Confinement (CLIC) to improve background rejection and extend diffusion-limited observation time. To facilitate its integration into a wide range of laboratories, this implementation of the CLIC device can use a standard flow-cell, into which the sample is loaded. By mechanically deforming the flow-cell, the device creates a tunable, wedge-shaped imaging chamber which we have modeled using finite element analysis simulations and characterized experimentally using interferometry. A powerful feature of CLIC imaging technology is the ability to examine single molecules under a continuum of applied confinement, from the nanometer to the micrometer scale. We demonstrate, using freely diffusing λ-phage DNA, that when the imposed confinement is on the scale of individual molecules their molecular conformations and diffusivity are altered significantly. To improve the flow-cell stiffness, seal, and re-usability, we have innovated the fabrication of thin PDMS-bonded flow-cells. The presented flow-cell CLIC technology can be combined with surface-lithography to provide an accessible and powerful approach to tune, trap, and image individual molecules under an extended range of imaging conditions. It is well-suited to tackling open problems in biophysics, biotechnology, nanotechnology, materials science, and chemistry.

Paper Details

Date Published: 11 September 2013
PDF: 12 pages
Proc. SPIE 8811, Physical Chemistry of Interfaces and Nanomaterials XII, 881102 (11 September 2013); doi: 10.1117/12.2021768
Show Author Affiliations
Christopher M. J. McFaul, McGill Univ. (Canada)
Jason Leith, McGill Univ. (Canada)
Bojing Jia, McGill Univ. (Canada)
François Michaud, McGill Univ. (Canada)
Adriel Arsenault, McGill Univ. (Canada)
Andrew Martin, McGill Univ. (Canada)
Daniel Berard, McGill Univ. (Canada)
Sabrina Leslie, McGill Univ. (Canada)

Published in SPIE Proceedings Vol. 8811:
Physical Chemistry of Interfaces and Nanomaterials XII
Natalie Banerji; Carlos Silva, Editor(s)

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