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Related Experiment Video

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A Protocol for Real-time 3D Single Particle Tracking

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

Real-time multi-parameter spectroscopy and localization in three-dimensional single-particle tracking.

Christian Hellriegel¹, Enrico Gratton

¹Laboratory for Fluorescence Dynamics and Department of Biomedical Engineering, University of California, Irvine, 3120 Natural Sciences II, Irvine, CA 92697, USA.

Journal of the Royal Society, Interface

|August 30, 2008

Summary

This summary is machine-generated.

This study introduces an advanced "on-the-fly" single-particle tracking method in optical microscopy. This technique offers superior resolution and data acquisition for dynamic processes in materials and biological systems.

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Area of Science:

Optical Microscopy
Biophysics
Material Sciences

Background:

Single-particle tracking circumvents ensemble averaging, revealing mechanistic details of dynamic processes.
Conventional tracking analyzes images post-acquisition, limiting temporal resolution and data richness.

Purpose of the Study:

To describe the principle and advantages of the 'on-the-fly' tracking technique.
To demonstrate the capabilities of this method using experimental data.

Main Methods:

Implementation of 'on-the-fly' tracking on a two-photon laser scanning microscope.
Utilizing a feedback algorithm to maintain the observation volume centered on moving particles.
Acquiring data on particle dynamics, including translational and rotational motion, spectral dynamics, and fluorescence properties.

Main Results:

Achieved superior spatio-temporal resolution (2-50nm, 1-32ms).
Enabled inference of additional data (fluorescence lifetime, spectrum, polarization, intensity dynamics).
Successfully tracked dye-labeled particles in liquid and protein assemblies in living cells.

Conclusions:

'On-the-fly' tracking significantly enhances the study of dynamic processes at the single-particle level.
The technique provides a powerful tool for high-resolution, multi-modal characterization of moving objects in various scientific fields.