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

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.

You might also read

Related Articles

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

Sort by
Same author

Modulation of Oncogenic KRAS Signaling by Branched Actin-driven Cell Membrane Protrusions.

Research square·2026
Same author

Axially Swept Light-Sheet Microscopy using scattering and fluorescence contrast mechanisms.

Proceedings of SPIE--the International Society for Optical Engineering·2026
Same author

Machine learning assisted wavefront sensor.

Proceedings of SPIE--the International Society for Optical Engineering·2026
Same author

Altair-dvOPM: an open-access platform for large-field three-dimensional tissue imaging.

bioRxiv : the preprint server for biology·2026
Same author

Modulation of Oncogenic KRAS Signaling by Branched Actin-driven Cell Membrane Protrusions.

bioRxiv : the preprint server for biology·2026
Same author

Unraveling Subcellular Ultrastructure with Cyclically Multiplexed Expansion Microscopy.

bioRxiv : the preprint server for biology·2026
Same journal

Dynamic changes in OTULIN and progranulin levels in experimental myocardial infarction and cardiac remodeling.

Histochemistry and cell biology·2026
Same journal

Eosinophil-associated matrix remodeling in a sterile granulomatous inflammation model: a temporal histopathological analysis.

Histochemistry and cell biology·2026
Same journal

Cellular accumulation of lipofuscin in the heart: implications in health and disease.

Histochemistry and cell biology·2026
Same journal

From lipofuscin accumulation to cellular dysfunction: a focus on liver pathophysiology.

Histochemistry and cell biology·2026
Same journal

Immunohistochemical study of α-keratin, loricrin, filaggrin-like protein, and transglutaminase-1 expression in orthokeratinized and parakeratinized epithelium of the tongue of domestic goose (Anser anser f. domestica) during embryonic development.

Histochemistry and cell biology·2026
Same journal

Replication stress induced exposure to methotrexate in root meristem cells of Vicia faba.

Histochemistry and cell biology·2026
See all related articles

Related Experiment Video

Updated: Jun 8, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Widefield fluorescence microscopy with extended resolution.

Andreas Stemmer1, Markus Beck, Reto Fiolka

  • 1Nanotechnology Group, Department of Mechanical and Process Engineering, ETH Zurich, Tannenstrasse 3, 8092, Zurich, Switzerland. astemmer@ethz.ch

Histochemistry and Cell Biology
|September 24, 2008
PubMed
Summary
This summary is machine-generated.

Widefield fluorescence microscopy achieves 100 nm resolution using non-uniform light patterns and advanced reconstruction. Further improvements down to 10 nm are possible by utilizing non-linear fluorophore responses.

More Related Videos

Expansion Microscopy: High-Resolution Fluorescent Imaging with a Conventional Microscope
08:53

Expansion Microscopy: High-Resolution Fluorescent Imaging with a Conventional Microscope

Published on: December 19, 2025

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection
07:42

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

Published on: February 24, 2026

Related Experiment Videos

Last Updated: Jun 8, 2026

Conducting Multiple Imaging Modes with One Fluorescence Microscope
08:32

Conducting Multiple Imaging Modes with One Fluorescence Microscope

Published on: October 28, 2018

Expansion Microscopy: High-Resolution Fluorescent Imaging with a Conventional Microscope
08:53

Expansion Microscopy: High-Resolution Fluorescent Imaging with a Conventional Microscope

Published on: December 19, 2025

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection
07:42

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

Published on: February 24, 2026

Area of Science:

  • Optics and Photonics
  • Microscopy Techniques
  • Biophysical Imaging

Background:

  • Widefield fluorescence microscopy resolution is limited by diffraction.
  • Traditional Köhler illumination is being replaced by advanced illumination strategies.
  • Non-linear responses of fluorophores offer potential for super-resolution.

Purpose of the Study:

  • To review image formation and reconstruction in widefield microscopy using patterned illumination.
  • To discuss methods for achieving super-resolution beyond 100 nm.
  • To explore extensions of these techniques to other microscopy modalities.

Main Methods:

  • Application of non-uniform excitation light patterns (sinusoidal intensity variations).
  • Utilizing computational image reconstruction techniques.
  • Generating harmonic light patterns via interfering laser beams to form standing waves.

Main Results:

  • Achieving 100 nm resolution in all three dimensions.
  • Further resolution enhancement down to several 10 nm by exploiting non-linear fluorophore behavior.
  • Demonstrating image formation and reconstruction principles for patterned illumination.

Conclusions:

  • Patterned illumination combined with reconstruction significantly enhances widefield microscopy resolution.
  • Non-linear fluorophore responses are key to achieving super-resolution.
  • The described principles are extendable to total internal reflection, non-linear, and 3D microscopy.