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Feedback controller design for tracking a single fluorescent molecule

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Abstract

In this paper, we revisit the problem of tracking a single fluorescent molecule in a laser-scanning confocal microscope. We utilize optimal control theory to design a feedback controller and use numerical simulation to analyze its ability to track a molecule in two dimensions. A major theme in this paper is the inclusion of all relevant experimental limitations including moderate signal-to-noise fluorescence detection and the finite-bandwidth response of an electromechanical translation stage. The results presented here demonstrate the experimental feasibility of tracking single fluorescent molecules with diffusion coefficients as large as 0.1 μm2/ms. We show that for molecules that are even moderately confined along a third dimension (as, for example, by microscope cover slides), the two-dimensional tracking algorithm appears to be robust and effective. We expect this technology to enable single-molecule experiments with long observation times and high time-resolution.

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Authors and Affiliations

  1. Departments of Physics and Control and Dynamical Systems, MC 12-33, California Institute of Technology, 91125, Pasadena, CA, USA

    A.J. Berglund & H. Mabuchi

Authors
  1. A.J. Berglund
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  2. H. Mabuchi
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Correspondence to A.J. Berglund.

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PACS

87.64.Tt; 87.64.Ni; 87.15.Vv

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Berglund, A., Mabuchi , H. Feedback controller design for tracking a single fluorescent molecule. Appl Phys B 78, 653–659 (2004). https://doi.org/10.1007/s00340-004-1426-5

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  • DOI: https://doi.org/10.1007/s00340-004-1426-5

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