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Super-resolution Fluorescence Microscopy

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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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Updated: Mar 30, 2026

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

High-Throughput Total Internal Reflection Fluorescence and Direct Stochastic Optical Reconstruction Microscopy Using a Photonic Chip

Published on: November 16, 2019

Superresolution imaging with optical fluctuation using speckle patterns illumination.

MinKwan Kim¹, ChungHyun Park^2,3, Christophe Rodriguez^2,3

¹Graduate School of Nanoscience and Technology, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Republic of Korea.

Scientific Reports

|November 18, 2015

Summary

This summary is machine-generated.

This study introduces a new superresolution fluorescence microscopy technique that works with any fluorophore. It achieves subdiffraction resolution by controlling temporal emission fluctuations using speckle pattern illumination.

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Last Updated: Mar 30, 2026

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Published on: October 28, 2018

Area of Science:

Biophysics
Optical Microscopy
Cell Biology

Background:

Superresolution fluorescence microscopy enables subdiffraction resolution studies of cellular processes.
Current methods often rely on specific fluorophore properties like stochastic blinking, limiting their application.
A need exists for superresolution techniques applicable to a broader range of fluorophores.

Purpose of the Study:

To develop a superresolution fluorescence microscopy method applicable to general fluorophores.
To overcome the limitations of methods requiring specific intrinsic fluorophore emission properties.
To achieve subdiffraction resolution imaging using controlled temporal emission fluctuations.

Main Methods:

Utilized speckle pattern illumination to induce and control temporal emission fluctuations of fluorophores.
Employed statistical properties of these fluctuations to reconstruct superresolution images.
Validated the method theoretically and experimentally across various samples and objective lenses.

Main Results:

Demonstrated the capability to produce subdiffraction resolution images using general fluorophores.
Achieved spatial resolutions of 500 nm, 300 nm, and 140 nm with different numerical aperture (NA) objective lenses.
Observed an enhancement factor of 1.6 compared to conventional fluorescence microscopy.

Conclusions:

The developed speckle-based superresolution method broadens the applicability of superresolution microscopy.
It allows imaging with common fluorophores, overcoming previous limitations.
This technique offers a versatile approach for high-resolution biological imaging.