Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A Unified Analytical Method Greenness Score (<i>uAMGS</i>) Quantifies How Microscopic Imaging Is Greener Than Conventional Liquid Chromatography.

ACS sustainable chemistry & engineering·2026
Same author

The non-linear effects of the number of stochastic single-molecule adsorption events on ensemble elution profiles.

The Analyst·2026
Same author

Polyethylene Glycol Induced Condensation Leads to an Anomalous FRET Response from a Flexible Linker-Fluorescent Protein Crowding Sensor.

ACS sensors·2026
Same author

Enhancing Interfacial Charge Transport in Gold Nanoparticle@Polyaniline Hybrids via N-Heterocyclic Carbene Linkers.

Angewandte Chemie (International ed. in English)·2026
Same author

A Trojan horse pathogen breaking through partner-choice barriers in the insect gut.

Proceedings of the National Academy of Sciences of the United States of America·2026
Same author

Solvated Electron Generation from Coupled Plasmon Modes of Gold Nanoparticles Using Visible Light.

Nano letters·2026

Related Experiment Video

Updated: May 16, 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

Improved analysis for determining diffusion coefficients from short, single-molecule trajectories with photoblinking.

Bo Shuang1, Chad P Byers, Lydia Kisley

  • 1Department of Chemistry and Rice Quantum Institute, Rice University, Houston, Texas 77251-1892, United States.

Langmuir : the ACS Journal of Surfaces and Colloids
|December 11, 2012
PubMed
Summary

We developed two maximum likelihood estimation (MLE) methods to improve single-molecule trajectory analysis, especially for short, noisy data with photobleaching. These optimized methods enhance the study of molecular machines like nanocars.

More Related Videos

A Simple, Robust, and High Throughput Single Molecule Flow Stretching Assay Implementation for Studying Transport of Molecules Along DNA
12:05

A Simple, Robust, and High Throughput Single Molecule Flow Stretching Assay Implementation for Studying Transport of Molecules Along DNA

Published on: October 1, 2017

Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy
12:15

Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy

Published on: April 9, 2019

Related Experiment Videos

Last Updated: May 16, 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

A Simple, Robust, and High Throughput Single Molecule Flow Stretching Assay Implementation for Studying Transport of Molecules Along DNA
12:05

A Simple, Robust, and High Throughput Single Molecule Flow Stretching Assay Implementation for Studying Transport of Molecules Along DNA

Published on: October 1, 2017

Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy
12:15

Image Processing Protocol for the Analysis of the Diffusion and Cluster Size of Membrane Receptors by Fluorescence Microscopy

Published on: April 9, 2019

Area of Science:

  • Biophysics
  • Physical Chemistry
  • Nanotechnology

Background:

  • Single-molecule spectroscopy generates complex trajectory data.
  • Experimental artifacts like noise and photobleaching complicate analysis.
  • Existing methods may not optimally handle short trajectories or specific artifacts.

Purpose of the Study:

  • To develop and optimize maximum likelihood estimation (MLE) methods for analyzing single-molecule trajectories.
  • To address challenges including experimental noise, photobleaching, and out-of-plane motion.
  • To compare the performance of new MLE methods against existing techniques.

Main Methods:

  • Development of two novel maximum likelihood estimation (MLE) algorithms.
  • Application to simulated single-molecule trajectory data, focusing on short trajectories with photoblinking.
  • Comparative analysis against established analytical techniques.
  • Validation using real experimental data from single-molecule diffusion studies.

Main Results:

  • Established the optimal MLE method for various experimental conditions.
  • Demonstrated superior performance of the optimized MLE methods for short, noisy trajectories.
  • Successfully applied the optimized MLE method to analyze the diffusion of nanocars.

Conclusions:

  • The developed MLE methods provide a robust framework for analyzing challenging single-molecule trajectory data.
  • Optimized MLE significantly improves the accuracy and reliability of single-molecule experiments.
  • This approach is valuable for studying the dynamics of molecular systems, including fluorescent molecular machines.