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

Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

7.6K
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...
7.6K
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

14.3K
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,...
14.3K

You might also read

Related Articles

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

Sort by
Same author

Separation estimation of two freely rotating dipole emitters near the quantum limit.

Physical review. A·2026
Same author

Tetraspanin CD82 shapes EGFR signaling outcomes through nanoscale receptor organization.

The Journal of cell biology·2026
Same author

A MATLAB-based Instrument Control (MIC) package for fluorescence imaging.

Journal of open source software·2025
Same author

Calibration-free estimation of field dependent aberrations for single molecule localization microscopy across large fields of view.

bioRxiv : the preprint server for biology·2024
Same author

SMITE: Single Molecule Imaging Toolbox Extraordinaire (MATLAB).

Journal of open source software·2024
Same author

Axial de-scanning using remote focusing in the detection arm of light-sheet microscopy.

Nature communications·2024
Same journal

Generalizable framework for multi-site bone density prediction using non-dominant wrist optical biomarkers.

Biomedical optics express·2026
Same journal

Erratum: Review of dynamic optical coherence tomography for intracellular motility [Invited]: errata.

Biomedical optics express·2026
Same journal

Digital-micromirror-device-based illumination strategies for background suppression in single-molecule localization microscopy.

Biomedical optics express·2026
Same journal

Synergistic combination of convective self-assembly and hollow core fiber for sensitive SERS detection of glucose molecules.

Biomedical optics express·2026
Same journal

Multimodal diagnostic network integrating infrared and mass spectra for lung cancer.

Biomedical optics express·2026
Same journal

Multimodal Optical Biosensing for Precision Medicine and Healthcare: Introduction to the feature issue.

Biomedical optics express·2026
See all related articles

Related Experiment Video

Updated: Sep 11, 2025

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle
15:06

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle

Published on: January 3, 2016

12.9K

Enhanced supercritical angle localization microscopy through point spread function modeling.

Sajjad A Khan1,2, Keith A Lidke1, Sheng Liu1

  • 1Department of Physics and Astronomy, University of New Mexico, Albuquerque, New Mexico, USA.

Biomedical Optics Express
|August 14, 2025
PubMed
Summary
This summary is machine-generated.

This study introduces a simplified single-channel supercritical angle fluorescence (SAF) method for single-molecule localization microscopy (SMLM). The new approach enhances axial localization precision and accuracy for imaging cellular structures.

More Related Videos

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

8.6K
Super-resolution Imaging of the Bacterial Division Machinery
08:47

Super-resolution Imaging of the Bacterial Division Machinery

Published on: January 21, 2013

11.9K

Related Experiment Videos

Last Updated: Sep 11, 2025

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle
15:06

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle

Published on: January 3, 2016

12.9K
Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
11:06

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells

Published on: June 30, 2018

8.6K
Super-resolution Imaging of the Bacterial Division Machinery
08:47

Super-resolution Imaging of the Bacterial Division Machinery

Published on: January 21, 2013

11.9K

Area of Science:

  • Biophysics
  • Microscopy
  • Cell Biology

Background:

  • Single-molecule localization microscopy (SMLM) precisely localizes molecules in cells.
  • Supercritical angle fluorescence (SAF) in SMLM estimates axial positions using fluorophore-coverslip distance.
  • Conventional SAF-SMLM uses complex two-channel systems, reducing light efficiency.

Purpose of the Study:

  • To simplify SAF-SMLM by using a single-channel detection system.
  • To improve axial localization precision and accuracy in SMLM.
  • To develop a stage-tilt correction algorithm for enhanced field-of-view imaging.

Main Methods:

  • Developed a single-channel SAF-SMLM approach by directly detecting all fluorescence.
  • Utilized accurate point spread function (PSF) modeling incorporating SAF.
  • Implemented a stage-tilt correction algorithm within the PSF model.

Main Results:

  • Simulations showed single-channel SAF-SMLM offers better localization precision than two-channel systems.
  • The stage-tilt correction algorithm improved axial precision across the field of view.
  • Experimental imaging of F-actin filaments in HeLa cells demonstrated enhanced axial localization.

Conclusions:

  • The simplified single-channel SAF-SMLM method efficiently utilizes SAF information.
  • This approach achieves superior axial localization precision and accuracy compared to traditional methods for single-channel systems.
  • The developed technique offers a more efficient and precise tool for nanoscale imaging in cell biology.