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

6.8K
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...
6.8K
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

13.0K
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,...
13.0K
Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

2.1K
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...
2.1K

You might also read

Related Articles

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

Sort by
Same author

Enhancing biomedical optical volumetric imaging via self-supervised orthogonal learning.

Science advances·2026
Same author

Orthogonally Dispersed Spectroscopic Single-Molecule Localization Microscopy.

Nanophotonics (Berlin, Germany)·2026
Same author

A bright and regenerative array tag with fluorogenic ligands for long-term live-cell nanoscopy and single-molecule tracking.

Nature communications·2025
Same author

Real-time self-supervised denoising for high-speed fluorescence neural imaging.

Nature communications·2025
Same author

Parallel Three-Dimensional Tracking of Quantum Rods Using Polarization-Sensitive Spectroscopic Photon Localization Microscopy.

ACS photonics·2025
Same author

Super resolution reconstruction of fluorescence microscopy images by a convolutional network with physical priors.

Biomedical optics express·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: May 27, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
00:10

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

8.1K

Enabling real-time reconstruction for large field-of-view single-molecule localization microscopy using discrete

Jun Lu1, Lei Xu1, Shuyao Liao1

  • 1Academy for Engineering and Technology, Yiwu Research Institute, Fudan University, Shanghai 200433, China.

Biomedical Optics Express
|February 17, 2025
PubMed
Summary
This summary is machine-generated.

Optical aberrations limit the field-of-view in 3D single-molecule localization microscopy (SMLM). This study introduces discrete field-dependent point-spread functions (PSFs) to enable precise localization across larger areas, improving SMLM applications.

More Related Videos

A Protocol for Real-time 3D Single Particle Tracking
10:16

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

14.8K
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

8.4K

Related Experiment Videos

Last Updated: May 27, 2025

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules
00:10

Single-Molecule Tracking Microscopy - A Tool for Determining the Diffusive States of Cytosolic Molecules

Published on: September 5, 2019

8.1K
A Protocol for Real-time 3D Single Particle Tracking
10:16

A Protocol for Real-time 3D Single Particle Tracking

Published on: January 3, 2018

14.8K
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

8.4K

Area of Science:

  • Biophysics
  • Optical Microscopy
  • Super-resolution Imaging

Background:

  • Single-molecule localization microscopy (SMLM) achieves super-resolution by localizing individual fluorescent molecules.
  • High numerical aperture objectives essential for SMLM exhibit distorted point-spread functions (PSFs) off-axis, limiting the field-of-view (FOV).
  • These aberrations introduce significant 3D localization errors, restricting SMLM to small FOVs (e.g., 50 µm × 50 µm).

Purpose of the Study:

  • To systematically investigate the impact of optical aberrations on large FOV 3D SMLM.
  • To propose and validate a method for expanding the effective FOV in 3D SMLM.
  • To enable real-time, high-precision molecular localization in large imaging areas.

Main Methods:

  • Evaluated localization accuracy using unmodified, astigmatic, and double-helix PSFs under varying optical aberrations.
  • Developed and tested discrete field-dependent PSFs for aberration correction.
  • Employed GPU acceleration for high-speed image reconstruction of simulated and biological samples.

Main Results:

  • Optical aberrations significantly degrade localization precision and accuracy at the edges of the FOV.
  • Discrete field-dependent PSFs effectively correct for off-axis aberrations, enabling precise 3D localization across larger areas.
  • GPU-accelerated discrete PSF models allow for real-time SMLM image reconstruction.

Conclusions:

  • Optical aberrations are a critical limitation for large FOV 3D SMLM.
  • Discrete field-dependent PSFs offer a viable strategy to overcome these limitations, enhancing FOV.
  • Real-time reconstruction using GPU acceleration makes large FOV SMLM more practical for biological studies.