Here, we present a method that can improve the z-tracking accuracy of the recently invented TSUNAMI (Tracking of Single particles Using Nonlinear And Multiplexed Illumination) microscope. This method utilizes a maximum likelihood estimator (MLE) to determine the particle's 3D position that maximizes the likelihood of the observed time-correlated photon count distribution. Our Monte Carlo simulations show that the MLE-based tracking scheme can improve the z-tracking accuracy of TSUNAMI microscope by 1.7 fold. In addition, MLE is also found to reduce the temporal correlation of the z-tracking error. Taking advantage of the smaller and less temporally correlated z-tracking error, we have precisely recovered the hybridization-melting kinetics of a DNA model system from thousands of short single-particle trajectories in silico. Our method can be generally applied to other 3D single-particle tracking techniques.
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12 October 2015
Research Article|
October 12 2015
Improving z-tracking accuracy in the two-photon single-particle tracking microscope
C. Liu;
C. Liu
a)
Department of Biomedical Engineering,
University of Texas at Austin
, Austin, Texas 78712, USA
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Y.-L. Liu;
Y.-L. Liu
a)
Department of Biomedical Engineering,
University of Texas at Austin
, Austin, Texas 78712, USA
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E. P. Perillo;
E. P. Perillo
Department of Biomedical Engineering,
University of Texas at Austin
, Austin, Texas 78712, USA
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N. Jiang;
N. Jiang
Department of Biomedical Engineering,
University of Texas at Austin
, Austin, Texas 78712, USA
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A. K. Dunn;
A. K. Dunn
b)
Department of Biomedical Engineering,
University of Texas at Austin
, Austin, Texas 78712, USA
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C. Liu
a)
Y.-L. Liu
a)
E. P. Perillo
N. Jiang
A. K. Dunn
b)
H.-C. Yeh
b)
Department of Biomedical Engineering,
University of Texas at Austin
, Austin, Texas 78712, USA
a)
C. Liu and Y.-L. Liu contributed equally to this work
b)
Authors to whom correspondence should be addressed. Electronic addresses: [email protected] and [email protected]
Appl. Phys. Lett. 107, 153701 (2015)
Article history
Received:
July 08 2015
Accepted:
September 22 2015
Citation
C. Liu, Y.-L. Liu, E. P. Perillo, N. Jiang, A. K. Dunn, H.-C. Yeh; Improving z-tracking accuracy in the two-photon single-particle tracking microscope. Appl. Phys. Lett. 12 October 2015; 107 (15): 153701. https://doi.org/10.1063/1.4932224
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