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A Protocol for Real-time 3D Single Particle Tracking
Published on: January 3, 2018
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Deep and high-resolution three-dimensional tracking of single particles using nonlinear and multiplexed illumination
Evan P Perillo1, Yen-Liang Liu1, Khang Huynh1
1Department of Biomedical Engineering, The University of Texas at Austin, 107 W Dean Keeton Street, C0800, Austin, Texas 78712, USA.
Nature Communications
|July 30, 2015
Summary
Scientists developed a new 3D tracking method to observe molecular transport in cells up to 200 μm deep. This technique offers unprecedented speed and precision for studying disease mechanisms.
Area of Science:
- Biophysics
- Cell Biology
- Optical Imaging
Background:
- Molecular trafficking is crucial for understanding diseases like cancer.
- Current 3D tracking methods have limited depth (∼15 μm) and speed.
- Need for advanced techniques to study molecular dynamics in complex biological systems.
Purpose of the Study:
- To develop a novel 3D molecular tracking method.
- To achieve deep tissue imaging (up to 200 μm) with high spatiotemporal resolution.
- To enable real-time quantification of molecular transport dynamics.
Main Methods:
- Spatiotemporally multiplexed two-photon excitation.
- Single-detector system for 3D particle tracking.
- High signal-to-noise ratio enabling fast temporal resolution (50 μs) and precise localization (35 nm).
Main Results:
- Demonstrated 3D tracking in highly scattering environments up to 200 μm depth.
- Achieved a system response time of 1 ms.
- Successfully tracked epidermal growth factor receptor complexes at ∼100 μm depth in tumor spheroids.
Conclusions:
- The new method overcomes limitations of existing 2D and shallow-depth 3D tracking techniques.
- Enables detailed study of molecular transport in complex biological models.
- Potential applications in cancer research and drug development.
