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A Protocol for Real-time 3D Single Particle Tracking
Published on: January 3, 2018
Lab-on-Chip for fast 3D particle tracking in living cells
Houssam Hajjoul1, Silvia Kocanova, Imen Lassadi
1CNRS, LAAS, 7 avenue du colonel Roche, F-31077 Toulouse, France.
Lab on a Chip
|October 14, 2009
Summary
We developed a new Lab-on-Chip technology using V-shaped micro-mirrors for precise 3D particle tracking in living cells. This innovation allows detailed study of chromatin dynamics with high temporal and spatial resolution.
Area of Science:
- Biophysics
- Cell Biology
- Microfluidics
Background:
- Accurate 3D tracking of intracellular components is crucial for understanding cellular processes.
- Existing methods often lack the necessary temporal or spatial resolution for dynamic studies.
Purpose of the Study:
- To introduce a novel Lab-on-Chip (LOC) technology for high-resolution 3D particle tracking in living cells.
- To demonstrate its application in studying chromatin dynamics.
Main Methods:
- Development of a LOC device featuring V-shaped micro-mirrors for multi-angle observation.
- Integration with standard fluorescence microscopy and an EMCCD camera.
- 3D reconstruction of fluorescently tagged gene loci (GFP) in yeast.
Main Results:
- Simultaneous stereo-images acquired from multiple vantage points.
- Successful 3D tracking of yeast gene loci with 27 nm error per axis.
- High temporal resolution achieved with inter-frame intervals up to 10 ms.
Conclusions:
- The V-shaped micro-mirror LOC technology enables precise 3D particle tracking in living cells.
- It provides quantitative insights into chromatin dynamics with unprecedented spatio-temporal resolution.
- The technology is compatible with standard fluorescence microscopes.
