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Related Concept Videos

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Related Experiment Video

Updated: Jul 6, 2026

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
10:20

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

Bayesian inference for improved single molecule fluorescence tracking.

Ji Won Yoon1, Andreas Bruckbauer, William J Fitzgerald

  • 1Department of Engineering and Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.

Biophysical Journal
|March 15, 2008
PubMed
Summary
This summary is machine-generated.

This study introduces a Bayesian inference method for single molecule tracking. The novel algorithm accurately tracks molecule trajectories in living cells, even with low signal and fluorophore issues.

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Last Updated: Jul 6, 2026

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Published on: September 5, 2019

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Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes
08:26

Automated Two-dimensional Spatiotemporal Analysis of Mobile Single-molecule FRET Probes

Published on: November 23, 2021

Area of Science:

  • Cellular and Molecular Biology
  • Biophysics
  • Imaging Science

Background:

  • Single molecule tracking is crucial for observing molecular dynamics in living cells.
  • Challenges include low signal-to-noise ratio, unknown molecule counts, and fluorophore artifacts like blinking and photobleaching.
  • These limitations hinder accurate, long-term trajectory analysis.

Purpose of the Study:

  • To develop an advanced algorithm for robust single molecule tracking.
  • To improve the extraction of molecular positional information from fluorescence image sequences.
  • To overcome limitations of current tracking methods in challenging experimental conditions.

Main Methods:

  • A Bayesian-based inference approach utilizing a trans-dimensional sequential Monte Carlo method.
  • Integration of spatial and temporal information from fluorescence image sequences.
  • Validation using simulated data with known trajectories and real experimental data.

Main Results:

  • The developed method enables accurate tracking of molecules over extended trajectories.
  • Effective performance demonstrated even with low signal-to-noise ratios.
  • Robustness shown against common issues like fluorophore blinking and photobleaching.
  • Successful application to real experimental single molecule tracking data.

Conclusions:

  • The Bayesian trans-dimensional sequential Monte Carlo method significantly enhances single molecule tracking accuracy.
  • This approach provides a powerful tool for analyzing molecular dynamics in living cells under difficult imaging conditions.
  • It offers a solution to extract maximum information from challenging fluorescence microscopy data.