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

2.1K
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...
2.1K

You might also read

Related Articles

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

Sort by
Same author

Effects of Remote Web-Based Interventions on the Physiological and Psychological States of Patients With Cancer: Systematic Review With Meta-Analysis.

JMIR mHealth and uHealth·2025
Same author

The Prognostic Value and Immunomodulatory Role of Spsb2, a Novel Immune Checkpoint Molecule, in Hepatocellular Carcinoma.

Genes·2025
Same author

Unraveling the Structural Evolution of Cobalt Sulfides in Electrocatalytic NO<sub>3</sub>RR: the Inescapable Influence of Cl<sup></sup>.

Inorganic chemistry·2025
Same author

Synergistic engineering of heteroatom doping and heterointerface construction in V-doped Ni(OH)<sub>2</sub>/FeOOH to boost both oxygen evolution and urea oxidation reactions.

Journal of colloid and interface science·2023
Same author

Unveiling the Structural Self-Reconstruction and Identifying the Reactive Center of a V, Fe Co-Doped Cobalt Precatalyst toward Enhanced Overall Water Splitting by Operando Raman Spectroscopy.

Inorganic chemistry·2023
Same author

Characterizing the metabolites and the microbial communities of the soy sauce mash affected by temperature and hydrostatic pressure.

Food research international (Ottawa, Ont.)·2019

Related Experiment Video

Updated: May 23, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
00:10

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

Published on: September 5, 2019

8.1K

Single-molecule tracking in living microbial cells.

Xiaomin Chen1, Qianhong Guo1,2, Jiexin Guan1,3

  • 1Institute of Fundamental and Frontier Sciences, University of Electronic Science and Technology of China, Chengdu 611731, China.

Biophysics Reports
|March 12, 2025
PubMed
Summary
This summary is machine-generated.

Researchers can now study molecular mechanisms in living cells using single-molecule imaging. This protocol details single particle tracking photoactivated localization microscopy (sptPALM) for high-resolution intracellular analysis.

Keywords:
Diffusion coefficientsLiving microbial cellsPALMSingle-molecule trackingTrajectory

More Related Videos

Visualizing Protein-DNA Interactions in Live Bacterial Cells Using Photoactivated Single-molecule Tracking
16:21

Visualizing Protein-DNA Interactions in Live Bacterial Cells Using Photoactivated Single-molecule Tracking

Published on: March 10, 2014

17.7K
Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy
11:26

Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy

Published on: September 8, 2009

9.3K

Related Experiment Videos

Last Updated: May 23, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
00:10

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

Published on: September 5, 2019

8.1K
Visualizing Protein-DNA Interactions in Live Bacterial Cells Using Photoactivated Single-molecule Tracking
16:21

Visualizing Protein-DNA Interactions in Live Bacterial Cells Using Photoactivated Single-molecule Tracking

Published on: March 10, 2014

17.7K
Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy
11:26

Visualizing Single Molecular Complexes In Vivo Using Advanced Fluorescence Microscopy

Published on: September 8, 2009

9.3K

Area of Science:

  • Microbiology
  • Cell Biology
  • Biophysics

Background:

  • Model organisms are crucial for studying fundamental biological processes due to their stability.
  • Understanding intracellular environments requires high spatiotemporal resolution techniques.
  • Traditional ensemble methods lack the detail needed for single-molecule analysis.

Purpose of the Study:

  • To present a detailed protocol for single-molecule tracking in living cells.
  • To enable researchers to study molecular and cellular mechanisms with high precision.
  • To facilitate the application of sptPALM in biological research.

Main Methods:

  • Utilizing single particle tracking photoactivated localization microscopy (sptPALM).
  • Employing stochastic photoactivation of fluorophores for imaging.
  • Recording image series to identify and track individual molecule positions.
  • Linking positions to generate molecular trajectories.

Main Results:

  • Achieving tens of nanometers spatial and millisecond temporal resolution in vivo.
  • Generating quantitative kinetic and spatial data (reaction rates, diffusion coefficients).
  • Providing insights into complex intracellular dynamics unattainable by ensemble methods.

Conclusions:

  • The presented sptPALM protocol offers a powerful tool for biological research.
  • Directly visualizing molecular mechanisms in living cells is now more accessible.
  • This method enhances understanding of essential biological processes at the molecular level.