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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...

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The early pioneers of microscopy opened a window into the invisible world of microorganisms. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes that leveraged nonvisible light, such as fluorescence microscopy that uses an ultraviolet light source and electron microscopy that uses short-wavelength electron beams. These advances significantly improved magnification, image resolution, and contrast. By comparison, the...

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Updated: Jun 12, 2025

Conducting Multiple Imaging Modes with One Fluorescence Microscope

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Published on: October 28, 2018

Scanning Switch-off Microscopy for Super-Resolution Fluorescence Imaging.

Zhaoshuai Gao^1,2, Shangguo Hou^3,4, Suhui Deng^3,5

¹School of Chemistry and Chemical Engineering, Frontiers Science Center for Transformative Molecules and National Center for Translational Medicine, Shanghai Jiao Tong University, 200240 Shanghai, China.

|September 19, 2024

Summary

This summary is machine-generated.

Scanning switch-off microscopy (SSM) enables super-resolution imaging using standard confocal microscopes by utilizing fluorophore blinking. This technique achieves ~100 nm resolution for fixed and live samples, enhancing accessibility to super-resolution microscopy.

Keywords:

Confocal microscope Exposure dose Photon switch-off effects Super-resolution microscope

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Published on: August 22, 2019

Area of Science:

Optical Microscopy
Biophysics
Nanotechnology

Background:

Super-resolution (SR) microscopy breaks the diffraction limit, offering higher resolution than conventional light microscopy.
The complexity and cost of specialized equipment limit the widespread adoption of SR microscopy.
Fluorophore blinking, or switch-off response, is an inherent property of many fluorescent molecules.

Purpose of the Study:

To introduce a novel super-resolution imaging concept called scanning switch-off microscopy (SSM).
To demonstrate that SSM can achieve super-resolution using a commercial confocal microscope without specialized optics.
To validate the SSM method through theoretical calculations and experimental results.

Main Methods:

Developed the scanning switch-off microscopy (SSM) concept.
Utilized the inherent switch-off response of fluorophores for image reconstruction.
Performed imaging experiments on DNA origami nanostructures and cellular cytoskeletons.
Achieved super-resolution imaging of live cells using a dronpa fluorescent protein.

Main Results:

Obtained an imaging resolution of approximately 100 nm for various biological and nanoscale samples.
Successfully imaged fixed samples including DNA origami and cellular cytoskeletons.
Demonstrated the capability of SSM for super-resolution imaging of live cells.
Validated the theoretical model of SSM with experimental data.

Conclusions:

SSM provides a practical approach to super-resolution imaging using readily available confocal microscopes.
The method leverages the natural blinking behavior of fluorophores, simplifying instrumentation requirements.
SSM is adaptable to various laser scanning-based microscopes, broadening its potential applications.