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.

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,...

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...

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

Visualizing intraorganellar ultrastructures, dynamics, and interactions with open-access background-free Lock-in-SIM.

Nature communications·2025

Same author

Single-shot X-ray and near-infrared (NIR) dual-mode fusion imaging based on bifunctional NIR scintillators.

Light, science & applications·2025

Same author

Analytical model of a microlens array homogenizer based on an angular spectrum diffraction method.

Applied optics·2025

Same author

Cellular optical imaging techniques: a dynamic advancing frontier.

Science China. Life sciences·2025

Same author

Enhancing precision for simultaneous 3D localization and 3D orientation with structured illumination.

Optics letters·2025

Same author

In situ fully vectorial tomography and pupil function retrieval of tightly focused fields.

Nature communications·2025

Search research articles

Related Experiment Video

Updated: May 13, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Breaking the diffraction barrier using fluorescence emission difference microscopy.

Cuifang Kuang¹, Shuai Li, Wei Liu

¹State Key Laboratory of Modern Optical Instrumentation, Zhejiang University, Hangzhou 310027, China. cfkuang@zju.edu.cn

Scientific Reports

|March 15, 2013

Summary

We introduce fluorescence emission difference microscopy (FED), a new technique that overcomes the diffraction limit for clearer nanoscale imaging. This method achieves super-resolution below a quarter of the light wavelength, enabling detailed investigations of biological samples.

More Related Videos

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope

Published on: October 9, 2014

Visualizing Intracellular Sialylation with Click Chemistry and Expansion Microscopy

Visualizing Intracellular Sialylation with Click Chemistry and Expansion Microscopy

Published on: February 7, 2025

Related Experiment Videos

Last Updated: May 13, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope

From Fast Fluorescence Imaging to Molecular Diffusion Law on Live Cell Membranes in a Commercial Microscope

Published on: October 9, 2014

Visualizing Intracellular Sialylation with Click Chemistry and Expansion Microscopy

Visualizing Intracellular Sialylation with Click Chemistry and Expansion Microscopy

Published on: February 7, 2025

Area of Science:

Optics and Photonics
Microscopy
Nanotechnology

Background:

The diffraction limit restricts the resolution of conventional far-field microscopes.
Super-resolution microscopy techniques aim to overcome this fundamental limitation.
Existing methods often involve complex sample preparation or slow imaging speeds.

Purpose of the Study:

To propose and validate a novel physical mechanism for breaking the diffraction barrier in far-field microscopy.
To introduce fluorescence emission difference microscopy (FED) as a new super-resolution technique.
To demonstrate the practical applicability and high resolution achievable with FED.

Main Methods:

Development of a novel physical mechanism based on the intensity difference between two acquired images.
Application of fluorescence saturation to enhance resolving ability.
Theoretical analysis and simulation tests to validate the approach.
Experimental demonstration using fluorescent nanoparticles and biological cells.

Main Results:

Achieved a spatial resolution below λ/4, surpassing the diffraction limit.
Demonstrated the effectiveness of fluorescence saturation in further enhancing resolution.
Validated the FED technique through theoretical, simulation, and experimental studies.
Confirmed the potential for high imaging speed.

Conclusions:

Fluorescence emission difference microscopy (FED) offers a viable method for breaking the diffraction barrier.
FED provides super-resolution imaging with potential for high speed, suitable for nanoscale investigations.
The technique shows promise for wide application in biological and materials science.