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Published March 2004 | public
Journal Article

Feedback controller design for tracking a single fluorescent molecule

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.

Additional Information

© 2004 Springer-Verlag. Received: 10 December 2003; Published online: 23 March 2004. We thank J. Stockton and R. van Handel for technical advice. This work was supported by the U.S. Army Research Office through the Institute for Collaborative Biotechnologies, grant DAAD19-03-D-0004, and by the National Science Foundation through grants 0323542 and 0113443. Any opinions, findings, and conclusions or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the National Science Foundation.

Additional details

Created:
August 19, 2023
Modified:
October 19, 2023