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 Experiment Video

Updated: Dec 28, 2025

A Protocol for Real-time 3D Single Particle Tracking
10:16

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

15.2K

Multiplexed Nanometric 3D Tracking of Microbeads Using an FFT-Phasor Algorithm.

Thomas B Brouwer1, Nicolaas Hermans1, John van Noort1

  • 1Biological and Soft Matter Physics, Huygens-Kamerlingh Onnes Laboratory, Leiden University, Leiden, the Netherlands.

Biophysical Journal
|February 14, 2020
PubMed
Summary
This summary is machine-generated.

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

Topological Investigation of Protein Folding and Intrinsic Disorder.

The journal of physical chemistry. B·2026
Same author

Modulation of Archaeal Hypernucleosome Structure and Stability by Mg<sup>2</sup>.

Journal of molecular biology·2025
Same author

Long-read nucleosome mapping of single chromatin fibers using DNA methylation and nanopore sequencing.

Biophysical journal·2025
Same author

Phosphorylation-Regulated Conformational Diversity and Topological Dynamics of an Intrinsically Disordered Nuclear Receptor.

The journal of physical chemistry. B·2025
Same author

Single-molecule parallel analysis for rapid exploration of sequence space.

Nature protocols·2025
Same author

Establishment and maintenance of embryogenic cell fate during microspore embryogenesis.

The Plant journal : for cell and molecular biology·2025
Same journal

Tau protein differentially affects Piezo1 and Kir2.1 channels in brain capillary endothelial cells.

Biophysical journal·2026
Same journal

Emergent Intercellular Junction Stability during Cyclic Tissue Loading.

Biophysical journal·2026
Same journal

Enhanced-Sampling Simulations Reveal Distinct Intermediates in SARS-CoV-2 FSE Pseudoknot Interconversion.

Biophysical journal·2026
Same journal

Structure-based simulations of the full Flock House virus capsid reveal pathways and energetics of an infection-critical peptide externalization event.

Biophysical journal·2026
Same journal

Quantifying the Peripheral Surface Information Entropy from Conformational Ensembles of Globular Protein-Peptide Complexes.

Biophysical journal·2026
Same journal

Anisotropic unbinding and location-dependent hovering of a kinesin motor head over microtubule.

Biophysical journal·2026
See all related articles

A new 3D phasor algorithm offers efficient, nanometer-accurate tracking of microbeads for single-molecule biophysics. This method enhances high-throughput analysis of biomolecular mechanical properties, even with imaging challenges.

Area of Science:

  • Biophysics
  • Single-molecule techniques
  • Nanotechnology

Background:

  • Single-molecule biophysics often uses microbead tracking for biomolecular mechanical property analysis.
  • Holographic analysis of diffraction patterns provides 3D bead localization but is computationally intensive.
  • Efficient real-time tracking methods are crucial for multiplexed applications.

Purpose of the Study:

  • To introduce a computationally efficient 3D phasor algorithm for nanometric bead tracking.
  • To provide robust bead localization accuracy over a significant z-range under challenging imaging conditions.
  • To enable high-throughput analysis in biophysical experiments.

Main Methods:

  • Developed a 3D phasor algorithm utilizing 2D cross-correlation and fast Fourier transforms.

More Related Videos

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
11:28

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

Published on: October 1, 2014

10.6K
Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles
11:54

Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles

Published on: March 13, 2017

9.7K

Related Experiment Videos

Last Updated: Dec 28, 2025

A Protocol for Real-time 3D Single Particle Tracking
10:16

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

15.2K
3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles
11:28

3D Orbital Tracking in a Modified Two-photon Microscope: An Application to the Tracking of Intracellular Vesicles

Published on: October 1, 2014

10.6K
Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles
11:54

Microfluidic Platform with Multiplexed Electronic Detection for Spatial Tracking of Particles

Published on: March 13, 2017

9.7K
  • Employed computer-generated reference images for tracking.
  • Implemented the algorithm in magnetic tweezers experiments for real-time 3D tracking of multiple beads.
  • Validated accuracy against Lorenz-Mie scattering simulations and a look-up table approach.

    • Main Results:

    • The 3D phasor algorithm achieves nanometric localization accuracy over a 10 μm z-range.
    • Achieved processing rates of up to 10,000 regions of interest per second.
    • Successfully tracked over 100 beads in real time on a standard CPU.
    • Demonstrated robustness and accuracy comparable to computationally expensive methods, even with image artifacts.

    Conclusions:

    • The 3D phasor algorithm offers an efficient and robust alternative for nanometric bead tracking.
    • Its speed and accuracy improve multiplexed biophysical applications requiring high throughput.
    • The method is easily implemented and suitable for challenging imaging conditions.