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Protein Dynamics in Living Cells

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Updated: Jul 3, 2026

Tracking Single Proteins in Lipid Bilayers Using Fluorescence Microscopy

Tracking Single Proteins in Lipid Bilayers Using Fluorescence Microscopy

Published on: December 12, 2025

Robust single-particle tracking in live-cell time-lapse sequences.

Khuloud Jaqaman¹, Dinah Loerke, Marcel Mettlen

¹Department of Cell Biology, The Scripps Research Institute, 10550 N. Torrey Pines Rd, La Jolla, California 92037, USA. kjaqaman@scripps.edu

|July 22, 2008

Summary

This summary is machine-generated.

A new algorithm improves single-particle tracking (SPT) in live-cell imaging by solving complex challenges like high particle density and disappearance. This enables better understanding of receptor organization and dynamics within the plasma membrane.

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Area of Science:

Cell Biology
Biophysics
Computational Biology

Background:

Single-particle tracking (SPT) is crucial for studying subcellular dynamics but is often limited by technical challenges.
Existing SPT methods struggle with high particle density, motion heterogeneity, and temporary particle loss, hindering accurate trajectory reconstruction.

Purpose of the Study:

To develop and validate a novel tracking algorithm that overcomes principal challenges in single-particle tracking.
To apply the algorithm to investigate the role of dynamin in endocytic kinetics and CD36 receptor aggregation.

Main Methods:

A new algorithm formulated as global combinatorial optimization problems for linking particles and track segments.
The method addresses high particle density, motion heterogeneity, temporary disappearance, and particle merging/splitting.
Application to analyze dynamin's effect on endocytic structures and CD36 receptor motion and aggregation.

Main Results:

The algorithm successfully reconstructs complete particle trajectories even in dense and dynamic cellular environments.
Demonstrated differential effects of GTPase dynamin on the kinetics of short- and long-lived endocytic structures.
Showed that CD36 receptor motion along linear tracks enhances their aggregation probability.

Conclusions:

The developed tracking algorithm provides robust and complete tracking of dense particle fields in live-cell imaging.
This advancement is essential for dissecting mechanisms of receptor organization at the plasma membrane.
Highlights the importance of accurate tracking for understanding complex cellular dynamics.