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

Total Internal Reflection Fluorescence Microscopy01:05

Total Internal Reflection Fluorescence Microscopy

Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been developed.
Fluorescence and Phosphorescence: Instrumentation01:25

Fluorescence and Phosphorescence: Instrumentation

Fluorometers and spectrofluorometers are two types of instruments used for measuring molecular fluorescence. These instruments differ in how they select excitation and emission wavelengths and the type of light sources they utilize. Fluorometers use absorption interference filters to choose excitation and emission wavelengths. The excitation source in a fluorometer is typically a low-pressure mercury vapor lamp that emits intense lines distributed throughout the ultraviolet and visible regions.
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

Geometry and dynamics of annealed optimization in the coherent Ising machine with hidden and planted solutions.

Physical review. E·2026
Same author

Ultra-broadband mid-infrared generation in dispersion-engineered thin-film lithium niobate.

Optics express·2022
Same author

Engineering a Kerr-Based Deterministic Cubic Phase Gate via Gaussian Operations.

Physical review letters·2020
Same author

Experimental investigation of performance differences between coherent Ising machines and a quantum annealer.

Science advances·2019
Same author

Measurement of Mesoscale Conformational Dynamics of Freely Diffusing Molecules with Tracking FCS.

Biophysical journal·2018
Same author

Single-Molecule Fluorescence Reveals Commonalities and Distinctions among Natural and in Vitro-Selected RNA Tertiary Motifs in a Multistep Folding Pathway.

Journal of the American Chemical Society·2017
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
Same journal

Kinesin-5/Cut7 C-terminal tail phosphorylation influence on motor regulation through multi-scale molecular modeling.

Biophysical journal·2026
Same journal

Dynamic conformations of fluorophores on self-labeling protein tags.

Biophysical journal·2026
Same journal

Different actions of RyR2 open and closed channel block explained by a multiscale Ca<sup>2+</sup> release model.

Biophysical journal·2026
Same journal

Membrane Environment Sets the Functional pK<sub>a</sub> of Ionizable Lipids.

Biophysical journal·2026
See all related articles

Related Experiment Video

Updated: Jun 10, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

Intramolecular fluorescence correlation spectroscopy in a feedback tracking microscope.

Kevin McHale1, Hideo Mabuchi

  • 1Edward L. Ginzton Laboratory, Stanford University, Stanford, California, USA. mchalek@gmail.com

Biophysical Journal
|July 27, 2010
PubMed
Summary
This summary is machine-generated.

Feedback tracking microscopy reveals molecular dynamics. This technique offers improved sensitivity for studying molecular size, diffusion, and heterogeneity compared to traditional methods.

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

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
11:24

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

Related Experiment Videos

Last Updated: Jun 10, 2026

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells
14:12

Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

Published on: December 11, 2021

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

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
11:24

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

Area of Science:

  • Biophysics
  • Physical Chemistry
  • Molecular Biology

Background:

  • Understanding molecular dynamics is crucial for various biological processes.
  • Traditional methods like fluorescence correlation spectroscopy have limitations in sensitivity and scope.
  • Shape fluctuations and intramolecular dynamics significantly influence molecular behavior.

Purpose of the Study:

  • To derive the statistical properties of signals from large molecules using feedback tracking microscopy.
  • To investigate the impact of intramolecular dynamics on tracking system response.
  • To develop a general expression for the fluorescence autocorrelation function applicable to linear intramolecular dynamics.

Main Methods:

  • Development of a theoretical framework for feedback tracking microscopy signal analysis.
  • Incorporation of intramolecular dynamics into the tracking system response model.
  • Derivation of a generalized fluorescence autocorrelation function.
  • Experimental validation using a 3D tracking microscope and labeled lambda-phage DNA molecules.

Main Results:

  • The study provides a method to analyze signals from shape fluctuations in large molecules.
  • A general expression for the fluorescence autocorrelation function was derived, accounting for linear intramolecular dynamics.
  • Feedback tracking microscopy demonstrated enhanced sensitivity to translational diffusion, molecular size, and heterogeneity.
  • The technique proved effective for studying long-timescale decays in molecular behavior.

Conclusions:

  • Feedback tracking microscopy offers superior sensitivity for characterizing molecular properties compared to traditional fluorescence correlation spectroscopy.
  • The derived theoretical framework enables detailed analysis of molecular dynamics and heterogeneity.
  • This approach advances the study of large biomolecules like DNA, providing insights into their behavior and structure.