Robust real-time 3D single-particle tracking using a dynamically moving laser spot
- 14 June 2017
- journal article
- research article
- Published by Optica Publishing Group in Optics Letters
- Vol. 42 (12), 2390-2393
- https://doi.org/10.1364/ol.42.002390
Abstract
Real-time three-dimensional (3D) single-particle tracking uses optical feedback to lock on to freely diffusing nanoscale fluorescent particles, permitting precise 3D localization and continuous spectroscopic interrogation. Here we describe a new method of real-time 3D single-particle tracking wherein a diffraction-limited laser spot is dynamically swept through the detection volume in three dimensions using a two-dimensional (2D) electro-optic deflector and a tunable acoustic gradient lens. This optimized method, called 3D dynamic photon localization tracking (3D-DyPLoT), enables high-speed real-time tracking of single silica-coated non-blinking quantum dots ( diameter) with diffusive speeds exceeding at count rates as low as 10 kHz, as well as YFP-labeled virus-like particles. The large effective detection area () allows the system to easily pick up fast-moving particles, while still demonstrating high localization precision (, , and ). Overall, 3D-DyPLoT provides a fast and robust method for real-time 3D tracking of fast and lowly emitting particles, based on a single excitation and detection pathway, paving the way to more widespread application to relevant biological problems.
Keywords
Funding Information
- Duke University
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