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Updated: Feb 7, 2026

A Protocol for Real-time 3D Single Particle Tracking
Published on: January 3, 2018
Electrically tunable lens speeds up 3D orbital tracking.
Paolo Annibale1, Alexander Dvornikov1, Enrico Gratton2
1Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California, Irvine USA ; Authors contributed equally to this work.
We improved 3D orbital particle tracking for live cells by using an Electrically Tunable Lens (ETL) instead of mechanical stages. This enhances speed, accuracy, and tracking range for cellular structures and genomic loci.
Area of Science:
- Biophysics
- Cell Biology
- Microscopy
Background:
- 3D orbital particle tracking is crucial for observing fast-moving objects in live cells.
- Current methods using piezoelectric stages have limitations in speed, range, and accuracy.
- Laser scanning microscopy is commonly used for 3D shape reconstruction.
Purpose of the Study:
- To enhance the performance of 3D orbital particle tracking.
- To introduce a cost-effective upgrade for scanning microscopes.
- To achieve high-speed, high-resolution 3D tracking of intracellular components.
Main Methods:
- Replaced mechanical piezoelectric stages with an Electrically Tunable Lens (ETL) in a laser scanning microscope.
- Developed an ETL-based 3D orbital particle tracking system.
- Tracked fluorescently labeled genomic loci in living cells.
Main Results:
- Achieved significant improvements in tracking range (500 microns axial), speed, and accuracy.
- Enabled tracking with a temporal resolution of 8ms using a high numerical aperture objective.
- Demonstrated the ability to reconstruct complex 3D shapes of cellular structures.
Conclusions:
- Electrically Tunable Lens technology offers a superior alternative to mechanical stages for 3D orbital particle tracking.
- The ETL-based method provides unprecedented temporal resolution for studying dynamic cellular processes.
- This cost-effective upgrade facilitates advanced live-cell imaging and analysis.
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