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Updated: Jun 8, 2026
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3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
Published on: October 1, 2014
Three-dimensional tracking of single fluorescent particles with submillisecond temporal resolution.
Manuel F Juette1, Joerg Bewersdorf
1Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06520 United States.
Nano Letters
|October 14, 2010
Summary
This study introduces a new instrument for ultrafast 3D tracking of nanoscale dynamics. It achieves submillisecond time resolution and nanometer precision, ideal for observing biological processes like virus movement.
Area of Science:
- Biophysics
- Optical Microscopy
- Nanotechnology
Background:
- Observing nanoscale dynamics in biological systems requires high temporal resolution.
- Acquiring three-dimensional (3D) trajectories of fluorescently labeled objects like viruses or vesicles necessitates advanced imaging techniques.
Purpose of the Study:
- To present a novel instrument for ultrafast 3D localization and tracking.
- To achieve submillisecond time resolution and nanometer precision for nanoscale dynamics.
Main Methods:
- Utilizing scanning-free multiplane detection at 3.2 kHz frame rate with single photon sensitivity.
- Implementing optimized beam-steering for focused excitation and avoiding confocal pinholes.
- Demonstrating 3D tracking of single fluorescent particles over large volumes and extended durations.
Main Results:
- Achieved ultrafast 3D localization with submillisecond time resolution and nanometer precision.
- Demonstrated 3D tracking of single fluorescent particles at speeds up to 150 nm/ms.
- Maximized detection efficiency with minimal laser irradiation through focused excitation.
Conclusions:
- The developed instrument enables high-speed, high-precision 3D tracking of nanoscale dynamics.
- The system's design offers high live-sample compatibility for future biomedical applications.
- This technology advances the study of dynamic biological processes at the nanoscale.
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