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A Protocol for Real-time 3D Single Particle Tracking

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Published on: January 3, 2018

Single-Particle Tracking with Scanning Non-Linear Microscopy.

Théo Travers¹, Vincent G Colin¹, Matthieu Loumaigne¹

¹MOLTECH-Anjou Laboratory, UMR CNRS 6200, University of Angers, 2 bd Lavoisier, 49045 Angers CEDEX, France.

Nanomaterials (Basel, Switzerland)

|August 7, 2020

Summary

This summary is machine-generated.

This study adapts non-linear microscopy for single-particle tracking (SPT), optimizing parameters for accurate long trajectory analysis. The covariance-based estimator (CVE) effectively determines diffusion coefficients, accounting for motion blur and localization errors.

Keywords:

free diffusion non-linear microscopy particle size characterization particle tracking algorithms scanning microscopy single particle tracking two-photon fluorescence imaging

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Area of Science:

Optics and Spectroscopy
Biophysics
Microscopy Techniques

Background:

Single-particle tracking (SPT) is crucial in biology, typically using single-photon fluorescence.
Non-linear microscopy presents challenges for imaging dynamic processes due to its scanning acquisition method.

Purpose of the Study:

To adapt non-linear microscopy for effective single-particle tracking (SPT).
To optimize experimental parameters (objective, resolution, frame rate) for accurate trajectory recording.
To evaluate and adapt estimation methods for analyzing SPT data from non-linear microscopy.

Main Methods:

Development and adaptation of non-linear microscopy for SPT.
Optimization of imaging parameters including objective lens, resolution, and frame rate.
Application and adaptation of estimation methods, including the covariance-based estimator (CVE).

Main Results:

Successful adaptation of non-linear microscopy for imaging moving particles.
Identification of optimal parameter balances for high-accuracy, high-frame-rate trajectory acquisition.
Demonstration that the covariance-based estimator (CVE) is suitable for evaluating diffusion coefficients from SPT data.

Conclusions:

Non-linear microscopy can be effectively utilized for single-particle tracking (SPT) with careful parameter optimization.
The covariance-based estimator (CVE) provides a robust method for analyzing diffusion dynamics in SPT experiments using this technique.
This work advances microscopy capabilities for studying molecular dynamics in biological systems.