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Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 9, Iss. 5 — Apr. 29, 2014

Three-dimensional tracking of a single fluorescent nanoparticle using four-focus excitation in a confocal microscope

James A. Germann and Lloyd M. Davis  »View Author Affiliations

Optics Express, Vol. 22, Issue 5, pp. 5641-5650 (2014)

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We report high sensitivity detection and tracking of a single fluorescent nanoparticle in solution by use of four alternately pulsed laser diodes for fluorescence excitation in a confocal microscope. Slight offsets between the centers of the overlapping laser foci together with time-resolved photon counting enable sub-micron precision position measurements. Real-time correction for diffusional motion with a xyz-piezo stage then enables tracking of a nanoparticle with diffusivity up to ~12 μm2 s−1. Fluorescence correlation spectroscopy and calibration measurements indicate a net fluorescence photon detection efficiency of ~6–9%, comparable to that of an optimized single-molecule microscope.

© 2014 Optical Society of America

OCIS Codes
(180.1790) Microscopy : Confocal microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:

Original Manuscript: January 30, 2014
Revised Manuscript: February 24, 2014
Manuscript Accepted: February 24, 2014
Published: March 4, 2014

Virtual Issues
Vol. 9, Iss. 5 Virtual Journal for Biomedical Optics

James A. Germann and Lloyd M. Davis, "Three-dimensional tracking of a single fluorescent nanoparticle using four-focus excitation in a confocal microscope," Opt. Express 22, 5641-5650 (2014)

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