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Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 22, Iss. 15 — Jul. 28, 2014
  • pp: 18662–18667

Four-directional stereo-microscopy for 3D particle tracking with real-time error evaluation

R. F. Hay, G. M. Gibson, M. P. Lee, M. J. Padgett, and D. B. Phillips  »View Author Affiliations

Optics Express, Vol. 22, Issue 15, pp. 18662-18667 (2014)

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High-speed video stereo-microscopy relies on illumination from two distinct angles to create two views of a sample from different directions. The 3D trajectory of a microscopic object can then be reconstructed using parallax to combine 2D measurements of its position in each image. In this work, we evaluate the accuracy of 3D particle tracking using this technique, by extending the number of views from two to four directions. This allows us to record two independent sets of measurements of the 3D coordinates of tracked objects, and comparison of these enables measurement and minimisation of the tracking error in all dimensions. We demonstrate the method by tracking the motion of an optically trapped microsphere of 5 μm in diameter, and find an accuracy of 2–5 nm laterally, and 5–10 nm axially, representing a relative error of less than 2.5% of its range of motion in each dimension.

© 2014 Optical Society of America

OCIS Codes
(140.7010) Lasers and laser optics : Laser trapping
(170.4520) Medical optics and biotechnology : Optical confinement and manipulation
(350.4855) Other areas of optics : Optical tweezers or optical manipulation

ToC Category:

Original Manuscript: June 13, 2014
Manuscript Accepted: July 11, 2014
Published: July 24, 2014

Virtual Issues
Vol. 9, Iss. 9 Virtual Journal for Biomedical Optics

R. F. Hay, G. M. Gibson, M. P. Lee, M. J. Padgett, and D. B. Phillips, "Four-directional stereo-microscopy for 3D particle tracking with real-time error evaluation," Opt. Express 22, 18662-18667 (2014)

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