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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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Related Experiment Video

Updated: Dec 23, 2025

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Single-Molecule Diffusion and Assembly on Polymer-Crowded Lipid Membranes

Published on: July 19, 2022

Wide-field subdiffraction imaging by accumulated binding of diffusing probes.

Alexey Sharonov¹, Robin M Hochstrasser

¹Department of Chemistry, University of Pennsylvania, Philadelphia, PA 19104-6323, USA.

Proceedings of the National Academy of Sciences of the United States of America

|December 5, 2006

Summary

This summary is machine-generated.

This study presents a new subdiffraction imaging technique using fluorescent probes to achieve nanoscale resolution. The method allows for rapid, high-precision imaging of biological structures without requiring fluorescent labeling.

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Super-resolution Imaging of the Bacterial Division Machinery

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Published on: January 21, 2013

Area of Science:

Biophysics
Nanotechnology
Microscopy

Background:

Conventional microscopy is limited by the Rayleigh criterion, hindering nanoscale imaging.
Existing super-resolution techniques often require specific fluorescent labeling, which can be complex and invasive.

Purpose of the Study:

To develop a novel subdiffraction imaging method for achieving high spatial resolution.
To demonstrate the capability of imaging nanoscale structures without fluorescent labeling.

Main Methods:

Utilizes collisional flux of diffusing fluorescent probes to target object surfaces.
Precisely locates immobilized probes by replacing their point-spread function with a centroid.
Accumulates probe locations over time to build high-resolution images.

Main Results:

Achieved a spatial resolution of approximately 25 nm.
Successfully imaged lipid bilayers, their contours, and large unilamellar vesicles.
Demonstrated rapid nanoscale imaging capabilities.

Conclusions:

The developed method enables sub-diffraction imaging with high precision.
The technique overcomes the limitations of the Rayleigh criterion for nanoscale imaging.
Proven effective for imaging without the necessity of fluorescent probes.