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Super-resolution Fluorescence Microscopy

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Related Experiment Video

Updated: Nov 16, 2025

Lensfree On-chip Tomographic Microscopy Employing Multi-angle Illumination and Pixel Super-resolution

Lensfree On-chip Tomographic Microscopy Employing Multi-angle Illumination and Pixel Super-resolution

Published on: August 16, 2012

Direct supercritical angle localization microscopy for nanometer 3D superresolution.

Anindita Dasgupta^1,2,3, Joran Deschamps¹, Ulf Matti¹

¹Cell Biology and Biophysics, European Molecular Biology Laboratory, Heidelberg, Germany.

Nature Communications

|February 20, 2021

Summary

This summary is machine-generated.

Supercritical Angle Localization Microscopy (SALM) achieves over four-fold improved z-resolution for 3D super-resolution microscopy. This advancement enables nanometer precision imaging of cellular structures, crucial for structural cell biology.

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Related Experiment Videos

Last Updated: Nov 16, 2025

Lensfree On-chip Tomographic Microscopy Employing Multi-angle Illumination and Pixel Super-resolution

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Published on: August 16, 2012

Three-dimensional Super Resolution Microscopy of F-actin Filaments by Interferometric PhotoActivated Localization Microscopy iPALM

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Published on: December 1, 2016

Three-dimensional Imaging of Bacterial Cells for Accurate Cellular Representations and Precise Protein Localization

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Published on: October 29, 2019

Area of Science:

Biophysics
Cell Biology
Microscopy

Background:

3D single molecule localization microscopy (SMLM) offers high-resolution imaging of cellular structures.
Supercritical Angle Localization Microscopy (SALM) determines z-positions based on fluorescence intensity, but z-resolution is limited.

Purpose of the Study:

To significantly enhance the z-resolution of SALM for improved 3D super-resolution microscopy.
To achieve nanometer localization precision for cellular structures using SALM.

Main Methods:

Directly splitting supercritical and undercritical emission using an ultra-high numerical aperture (NA) objective.
Applying advanced fitting routines to precisely extract single emitter intensities.
Utilizing supercritical angle fluorescence for z-position determination.

Main Results:

Achieved over a four-fold improvement in z-resolution compared to existing SALM methods.
Demonstrated nanometer isotropic localization precision on DNA origami structures.
Successfully imaged clathrin-coated vesicles and microtubules in cells with high precision.

Conclusions:

The enhanced SALM technique significantly advances 3D super-resolution microscopy capabilities.
This method provides unprecedented precision for visualizing nanoscale structures in cell biology.
SALM shows great potential for detailed structural analysis in cellular environments.