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Determination and Correction of Position Detection Nonlinearity in Single Particle Tracking and Three-Dimensional Scanning Probe Microscopy

Published online by Cambridge University Press:  01 August 2004

Christian Tischer ,
Arnd Pralle  and
Ernst-Ludwig Florin
Christian Tischer
Affiliation:
Cell Biology and Biophysics Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany
Arnd Pralle
Affiliation:
Department of Molecular Cell Biology, University of California–Berkeley, Berkeley, CA 94720, USA
Ernst-Ludwig Florin
Affiliation:
Cell Biology and Biophysics Programme, European Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg, Germany
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Abstract

A general method is presented for determining and correcting nonlinear position detector responses in single particle tracking as used in three-dimensional scanning probe microscopy based on optical tweezers. The method uses locally calculated mean square displacements of a Brownian particle to detect spatial changes in the sensitivity of the detector. The method is applied to an optical tweezers setup, where the position fluctuations of a microsphere within the optical trap are measured by an interferometric detection scheme with nanometer precision and microsecond temporal resolution. Detector sensitivity profiles were measured at arbitrary positions in solution with a resolution of approximately 6 nm and 20 nm in the lateral and axial directions, respectively. Local detector sensitivities are used to reconstruct the real positions of the particle from the measured position signals.

Keywords

single particle tracking (SPT)three-dimensional position detectiondetection nonlinearitiesoptical tweezersoptical trapthermal noise imagingthree-dimensional scanning probe microscopyBrownian motionlocal mean square displacement (MSD)
Type
Instrumentation and Techniques
Information
Microscopy and Microanalysis , Volume 10 , Issue 4 , August 2004 , pp. 425 - 434
Copyright
© 2004 Microscopy Society of America

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