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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 developed.

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Related Experiment Video

Updated: May 11, 2026

High-Accuracy Correction of 3D Chromatic Shifts in the Age of Super-Resolution Biological Imaging Using Chromagnon

High-Accuracy Correction of 3D Chromatic Shifts in the Age of Super-Resolution Biological Imaging Using Chromagnon

Published on: June 16, 2020

Correcting chromatic offset in multicolor super-resolution localization microscopy.

Miklos Erdelyi¹, Eric Rees, Daniel Metcalf

¹Department of Chemical Engineering and Biotechnology, University of Cambridge, Pembroke Street, Cambridge CB2 3RA, UK. meerdelyi@gmail.com

|May 15, 2013

Summary

This summary is machine-generated.

Precise chromatic aberration correction is essential for multi-color super-resolution microscopy. This study quantifies chromatic offset effects and presents a calibration protocol for accurate colocalization analysis.

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Last Updated: May 11, 2026

High-Accuracy Correction of 3D Chromatic Shifts in the Age of Super-Resolution Biological Imaging Using Chromagnon

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Published on: June 16, 2020

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Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy

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Published on: December 9, 2013

Area of Science:

Microscopy
Optical Physics
Biophysics

Background:

Super-resolution microscopy demands precise drift correction due to near-limit performance.
Multi-color imaging faces challenges from static, spatially-dependent chromatic offsets.
These chromatic offsets, negligible in conventional microscopy, impede super-resolution colocalization.

Purpose of the Study:

To quantify the impact of chromatic offset on multi-color super-resolution imaging.
To demonstrate the necessity of chromatic correction for accurate object colocalization.
To present a practical calibration protocol for correcting chromatic optical offset.

Main Methods:

Computer simulations to model and quantify chromatic offset effects.
Experimental validation using fluorescent beads at multiple wavelengths.
Application of a novel calibration protocol to biological samples.

Main Results:

Simulated chromatic offset effects accurately predicted experimental observations.
Multi-color super-resolution colocalization is impossible without chromatic correction.
The developed protocol effectively corrects chromatic offset in biological imaging.

Conclusions:

Chromatic optical offset is a critical factor limiting multi-color super-resolution microscopy.
Accurate colocalization requires robust correction of chromatic aberrations.
The presented calibration method enables reliable multi-color super-resolution analysis.