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

You might also read

Related Articles

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

Sort by
Same author

Substrate recognition, not sequestration, drives the engagement of an H3K9 methyltransferase in living cells.

bioRxiv : the preprint server for biology·2026
Same author

Single-molecule tracking of RNA-DNA hybrid removal enzymes important for lagging-strand replication.

Biophysical journal·2026
Same author

Polyphosphate modulates the stress-responsive formation of functional RNA-protein condensates in bacteria and mammalian cells.

PLoS biology·2026
Same author

Polyphosphate acts as an architectural regulator of carbon fixation and nucleoid structure in cyanobacteria.

bioRxiv : the preprint server for biology·2026
Same author

The biophysical properties of the bacterial nucleoid are dynamic, heterogeneous, and responsive to perturbations of cellular processes.

bioRxiv : the preprint server for biology·2026
Same author

Dps binds and protects DNA in starved Escherichia coli with minimal effect on chromosome accessibility, dynamics, and organization.

Nucleic acids research·2026

Related Experiment Video

Updated: May 5, 2026

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

2.2K

Top-hat and asymmetric Gaussian-based fitting functions for quantifying directional single-molecule motion.

David J Rowland1, Julie S Biteen

  • 1Department of Chemistry, University of Michigan, Ann Arbor, MI 48109 (USA).

Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry
|December 7, 2013
PubMed
Summary

New fitting functions enable super-resolution microscopy to track fast-moving molecules by accounting for motion. This improves localization precision and allows for measurement of instantaneous velocity and direction in live bacteria cells.

Keywords:
directed motionfluorescenceinstantaneous velocitysingle-molecule studiessuper-resolution

More Related Videos

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
09:52

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

Published on: September 15, 2020

2.4K
Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends
12:20

Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends

Published on: March 15, 2014

13.9K

Related Experiment Videos

Last Updated: May 5, 2026

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

2.2K
An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics
09:52

An In Vitro Single-Molecule Imaging Assay for the Analysis of Cap-Dependent Translation Kinetics

Published on: September 15, 2020

2.4K
Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends
12:20

Preparation of Segmented Microtubules to Study Motions Driven by the Disassembling Microtubule Ends

Published on: March 15, 2014

13.9K

Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Super-resolution microscopy relies on localizing single fluorescent molecules.
  • Traditional methods assume molecules are stationary, limiting tracking of dynamic biological processes.
  • Accurate localization is crucial for nanometer-scale precision in imaging.

Purpose of the Study:

  • To develop novel fitting functions for single-molecule localization that account for directed motion.
  • To achieve nanometer-scale localization precision comparable to traditional methods for moving emitters.
  • To enable the measurement of instantaneous velocity and direction of fast-moving biological targets.

Main Methods:

  • Developed two new fitting functions for single-molecule fluorescence data.
  • Utilized Fisher information analysis to assess localization precision.
  • Validated methods through simulations and experimental imaging of live bacteria cells.
  • Applied new functions to measure instantaneous velocity and direction of bacterial motion.

Main Results:

  • The new fitting functions provide nanometer-scale localization precision for moving emitters.
  • Methods successfully measure instantaneous velocity and direction of live bacteria.
  • Achieved similar precision to traditional methods while explicitly modeling motion.
  • Increased information content from single-molecule images of fast-moving targets.

Conclusions:

  • Novel fitting functions enhance single-molecule imaging by accommodating molecular motion.
  • These methods allow for precise tracking and velocity measurement of dynamic biological systems.
  • The approach enables slower imaging speeds without loss of localization accuracy.
  • Improved tracking algorithms can be developed by calculating motion parameters during image acquisition.