Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Super-resolution Fluorescence Microscopy

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.

Two-Dimensional Microscopy in Microbiology

Two-Dimensional Microscopy in Microbiology

Two-dimensional (2D) microscopy encompasses a range of optical techniques that capture images within a single focal plane, offering detailed representations of microscopic structures. These techniques are essential in biological and medical research, enabling the visualization of cellular and subcellular structures with different levels of contrast and specificity.There are several major types of 2D microscopy, each with strengths and applications.Bright-Field MicroscopyBright-field microscopy...

Confocal Fluorescence Microscopy

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,...

Protein Dynamics in Living Cells

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

Condensin I but not Condensin II is crucial for mitotic chromosome mechanics.

Nature communications·2026

Same author

Mechanistic basis for relaxation of DNA supercoils by human topoisomerase IIIα-RMI1-RMI2.

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

Same author

RIF1 orchestrates a multi-step restoration of post-replicative H3K9me3.

bioRxiv : the preprint server for biology·2025

Same author

Calyculin A Induces Premature Chromosome Condensation and Chromatin Compaction in G<sub>1</sub>-Phase HeLa Cells without Histone H1 Phosphorylation.

bioRxiv : the preprint server for biology·2025

Same author

Sensory stimuli and cilium trafficking defects trigger the release of ciliary extracellular vesicles from multiple ciliary locations.

bioRxiv : the preprint server for biology·2025

Same author

Delivery of intraflagellar transport proteins to the ciliary base and assembly into trains.

Science advances·2025

Search research articles

Related Experiment Video

Updated: May 29, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

A brief introduction to single-molecule fluorescence methods.

Siet M J L van den Wildenberg¹, Bram Prevo, Erwin J G Peterman

¹Department of Physics and Astronomy, VU University Amsterdam, Amsterdam, The Netherlands.

Methods in Molecular Biology (Clifton, N.J.)

|September 13, 2011

Summary

Single-molecule fluorescence microscopy offers high sensitivity for biological studies. This technique enables precise measurements of biomolecular processes, including protein counting, tracking, and distance determination.

More Related Videos

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

Published on: November 2, 2009

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Published on: July 5, 2021

Related Experiment Videos

Last Updated: May 29, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

Automated System for Single Molecule Fluorescence Measurements of Surface-immobilized Biomolecules

Published on: November 2, 2009

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

Published on: July 5, 2021

Area of Science:

Biological Sciences
Biophysics
Biochemistry

Background:

Single-molecule fluorescence microscopy is a powerful technique for observing biological processes at the molecular level.
Fluorescence methods offer the necessary sensitivity and access to measurable parameters relevant to the biomolecular world.

Purpose of the Study:

To provide a comprehensive overview of single-molecule fluorescence microscopy.
To discuss the fundamental principles, historical development, and applications of this technique.

Main Methods:

General principles of fluorescence.
History and evolution of single-molecule fluorescence microscopy.
Detailed review of fluorescent probes and essential equipment for single-molecule visualization.

Main Results:

Discussion of measurable parameters, including protein counting and tracking.
Explanation of distance measurements using Förster Resonance Energy Transfer (FRET).
Description of orientation measurements via fluorescence polarization.

Conclusions:

Single-molecule fluorescence microscopy is a versatile tool for quantitative biological research.
The technique allows for detailed insights into molecular dynamics and interactions.
Advancements in fluorescent probes and instrumentation continue to expand its capabilities.