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...

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,...

You might also read

Related Articles

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

Sort by

Same author

Galvanometer-scanning transient phase microscopy with balanced detection and arbitrary pump polarization.

Journal of the Optical Society of America. B, Optical physics·2026

Same author

Microtubules guide Aurora B substrate geometries for accurate chromosome segregation.

Science advances·2026

Same author

Tissue type and tissue depth dependency of pulse broadening in three-photon microscopy: impact on third harmonic generation.

Biomedical optics express·2025

Same author

Confinement in fibrous environments positions and orients mitotic spindles.

PNAS nexus·2025

Same author

Introducing the Special Issue on Label-Free Quantitative Phase Imaging Honoring Prof. Gabriel Popescu, a Pioneer in Biomedical Optics.

Journal of biomedical optics·2024

Same author

Inhibition of p38-MK2 pathway enhances the efficacy of microtubule inhibitors in breast cancer cells.

bioRxiv : the preprint server for biology·2024

Same journal

Tau protein as a regulator of mitochondrial function and dynamics.

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

Same journal

A scalable, dividing cell model for the robust propagation and quantification of human sporadic Creutzfeldt-Jakob disease prions.

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

Same journal

Epigenetic regulation of mesenchymal BMP signaling directs postnatal organ innervation.

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

Same journal

Single-shot wide-field biochemical imaging at 1 kHz frame rate.

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

Same journal

Morphogenesis and topological evolution of a frustrated nematic liquid crystal under confinement.

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

Same journal

B cell-intrinsic CXCR3 drives efficient generation of ectopic pulmonary germinal center responses to influenza A virus infection.

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

See all related articles

Search research articles

Related Experiment Video

Updated: Mar 20, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Superresolved multiphoton microscopy with spatial frequency-modulated imaging.

Jeffrey J Field¹, Keith A Wernsing², Scott R Domingue²

¹Department of Electrical and Computer Engineering, Colorado State University, Fort Collins, CO 80523; Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523; Microscope Imaging Network Foundational Core Facility, Colorado State University, Fort Collins, CO 80523; Institute for Genome Architecture and Function, Colorado State University, Fort Collins, CO 80523; jjfield@rams.colostate.edu.

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

|May 28, 2016

Summary

This summary is machine-generated.

A new superresolution technique enables imaging below the diffraction limit using any contrast mechanism, including virtual energy states. This multiphoton technique offers enhanced resolution for biological and inorganic samples.

Keywords:

harmonic generation multiphoton microscopy superresolution

More Related Videos

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Published on: February 24, 2026

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Published on: April 9, 2014

Related Experiment Videos

Last Updated: Mar 20, 2026

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

Published on: December 9, 2013

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Published on: February 24, 2026

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Highly Resolved Intravital Striped-illumination Microscopy of Germinal Centers

Published on: April 9, 2014

Area of Science:

Optics and Photonics
Microscopy
Biophysics

Background:

Superresolution microscopy achieves resolutions beyond the diffraction limit of light.
Existing techniques primarily use real energy states of fluorescent molecules.
Contrast mechanisms via virtual energy states, like harmonic generation, are typically excluded from superresolution imaging.

Purpose of the Study:

To develop a superresolution technique applicable to any contrast mechanism, including those involving virtual energy states.
To enable superresolved nonlinear microscopy with single-pixel detection.
To demonstrate multimodal superresolved imaging using different contrast mechanisms.

Main Methods:

Spatial Frequency-Modulated Imaging (SPIFI) was adapted for nonlinear microscopy.
The technique, termed Multiphoton SPIFI (MP-SPIFI), was applied to two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG).
Single-pixel detection was employed for image acquisition.

Main Results:

MP-SPIFI achieved superresolved imaging with both TPEF and SHG contrast mechanisms.
High-resolution images were obtained from biological and inorganic samples.
Spatial resolution up to 2η below the diffraction limit was demonstrated, where η is the nonlinear response power.

Conclusions:

MP-SPIFI overcomes the limitation of real energy states in superresolution microscopy.
The technique allows superresolved imaging with any contrast mechanism, including nonlinear optical processes.
MP-SPIFI offers potential for enhanced resolution in optically thin and scattering media.