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

Crowdsourcing enables robust cell annotation for breast cancer pathology.

Scientific reports·2026

Same author

Tumour-reactive heterotypic CD8 T cell clusters from clinical samples.

Nature·2025

Same author

Final clinical analysis of pre-operative ipilimumab and nivolumab in locally advanced urothelial cancer and exploration of tumor-draining lymph node composition: The NABUCCO trial.

European journal of cancer (Oxford, England : 1990)·2025

Same author

Cytidine diphosphate diacylglycerol synthase 2 is a synthetic lethal target in mesenchymal-like cancers.

Nature genetics·2025

Same author

SRBD1, a highly conserved gene required for chromosome individualization.

Cell reports·2025

Same author

A novel lineage-tracing tool reveals that hypoxic tumor cells drive tumor relapse after radiotherapy.

Radiotherapy and oncology : journal of the European Society for Therapeutic Radiology and Oncology·2024

Search research articles

Related Experiment Video

Updated: Oct 26, 2025

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

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

Published on: June 30, 2018

Multicolor Localization-Based Super Resolution Microscopy.

Leila Nahidiazar¹, Rolf Harkes²

¹Netherlands Cancer Institute, Amsterdam, Netherlands. l.nahidi@nki.nl.

Methods in Molecular Biology (Clifton, N.J.)

|July 31, 2021

Summary

This summary is machine-generated.

This study presents a step-by-step method to optimize super-resolution microscopy, specifically single-molecule localization microscopy (SMLM), for improved multicolor imaging of cellular structures.

Keywords:

Fluorescence Imaging buffer Microscopy Multicolor OxEA SMLM Super resolution

More Related Videos

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 of the Bacterial Division Machinery

Super-resolution Imaging of the Bacterial Division Machinery

Published on: January 21, 2013

Related Experiment Videos

Last Updated: Oct 26, 2025

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

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

Published on: June 30, 2018

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 of the Bacterial Division Machinery

Super-resolution Imaging of the Bacterial Division Machinery

Published on: January 21, 2013

Area of Science:

Cell Biology
Microscopy Techniques
Biophysics

Background:

Super Resolution (SR) microscopy offers nanometer-scale insights into cellular architecture.
Single Molecule Localization Microscopy (SMLM) relies on fluorophore blinking for precise localization.
Image quality in SMLM is contingent on label density and localization accuracy.

Purpose of the Study:

To present a detailed, step-by-step method for optimizing SMLM.
To enhance the facilitation of multicolor imaging in SMLM.
To address key factors influencing SMLM image quality.

Main Methods:

Development of a comprehensive protocol for SMLM optimization.
Focus on factors influencing fluorophore blinking and localization accuracy.
Implementation of strategies for multicolor imaging acquisition.

Main Results:

A refined methodology for SMLM data acquisition.
Improved control over fluorophore behavior for enhanced localization.
Successful application of the method for multicolor SMLM imaging.

Conclusions:

The presented method provides a robust framework for optimizing SMLM.
This facilitates higher quality multicolor imaging of complex biological samples.
Optimized SMLM protocols are crucial for advancing nanoscale cell biology research.