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Intracellular Dynamics of Extracellular Vesicles by Segmented Trajectory Analysis
Kaisa Rautaniemi1, Thomas John2, Maximilian Richter3,4
1Chemistry and Advanced Materials, Faculty of Engineering and Natural Sciences, Tampere University, Korkeakoulunkatu 8, 33720Tampere, Finland.
Analytical Chemistry
|December 13, 2022
Summary
This study compares nanoparticle (NP) movement in cells using video tracking. Extracellular vesicle (EV) transport was faster than commercial NPs, suggesting motor protein involvement.
Area of Science:
- Cell Biology
- Nanotechnology
- Biophysics
Background:
- Video tracking analysis of nanoparticle (NP) dynamics in live cells offers detailed insights into cellular interactions and trafficking.
- While not yet routine, the necessary equipment for such experiments is widely available in research laboratories.
Purpose of the Study:
- To compare trajectory patterns, diffusion coefficients, and velocities of different nanoparticles (NPs) within A549 cells.
- To investigate the impact of recording frame rate and analysis parameters on trajectory data using commercial NPs.
- To analyze the intracellular transport of both commercial NPs and bio-derived extracellular vesicles (EVs).
Main Methods:
- Utilized a fluorescence microscope and open-source trajectory analysis software for live-cell imaging.
- Studied commercial fluorescent polymeric particles and two subpopulations of PC-3 cell-derived EVs.
- Examined the effect of varying recording frame rates and analysis parameters on trajectory results.
Main Results:
- Trajectory classification and apparent particle velocities were influenced by the recording frame rate.
- Diffusion coefficients remained comparable across different frame rates.
- All NP types exhibited similar trajectory patterns, resembling intracellular vesicular transport.
- Extracellular vesicles (EVs) demonstrated faster movement than commercial NPs, despite size differences.
Conclusions:
- Recording frame rate significantly affects nanoparticle trajectory classification and velocity measurements, but not diffusion constants.
- Intracellular transport patterns of commercial NPs and EVs are similar to vesicular transport.
- Faster EV movement suggests a potentially greater reliance on motor proteins for their intracellular trafficking compared to synthetic NPs.
