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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.

Three-Dimensional Microscopy in Microbiology

Three-Dimensional Microscopy in Microbiology

Three-dimensional imaging techniques are essential in cell biology, allowing researchers to visualize intricate cellular structures with high resolution. Two prominent methods, Differential Interference Contrast Microscopy (DIC) and Confocal Scanning Laser Microscopy (CSLM), provide distinct advantages for imaging live and thick specimens, respectively.Differential Interference Contrast MicroscopyDIC microscopy enhances contrast in transparent, unstained samples by converting phase...

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Updated: Jun 11, 2026

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

Published on: February 24, 2026

Molecules and methods for super-resolution imaging.

Michael A Thompson¹, Julie S Biteen, Samuel J Lord

¹Department of Chemistry, Stanford University, Stanford, California, USA.

Methods in Enzymology

|July 15, 2010

Summary

This summary is machine-generated.

Researchers developed new molecules and methods for super-resolution fluorescence imaging. This technique allows visualization of nanostructures beyond the optical diffraction limit, enabling detailed study of biological systems.

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Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection

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Super-Resolution Live Cell Imaging of Subcellular Structures

Super-Resolution Live Cell Imaging of Subcellular Structures

Published on: January 13, 2021

Area of Science:

Biophysics
Optical Imaging
Molecular Biology

Background:

The optical diffraction limit restricts the resolution of conventional microscopes.
Super-resolution fluorescence imaging overcomes this limit by observing individual molecules.

Purpose of the Study:

To develop novel fluorophores and methodologies for advanced super-resolution fluorescence imaging.
To extend the capabilities of single-molecule imaging techniques.

Main Methods:

Design and characterization of switchable fluorophores, including azido-DCDHF molecules and Cy3-Cy5 dimers.
Development of the first photoswitchable fluorescent protein, enhanced yellow fluorescent protein (EYFP).
Application of single-molecule photoswitching for imaging protein superstructures in living bacteria.
Implementation of a double-helix point-spread function for 3D super-resolution imaging.

Main Results:

Successful development of new photoswitchable fluorophores and a fluorescent protein for super-resolution imaging.
Demonstration of super-resolution imaging in living Caulobacter crescentus bacteria, revealing protein superstructure details.
Establishment of a 3D super-resolution imaging method using a double-helix point-spread function.

Conclusions:

The developed molecules and methods significantly advance super-resolution fluorescence imaging capabilities.
Single-molecule photoswitching provides unprecedented insights into nanoscale biological structures.
New techniques enable 3D super-resolution imaging, expanding the potential for biological discovery.