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Super-resolution Fluorescence Microscopy01:37

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 21, 2026

Super-Resolution Imaging and Shared Management: A Protocol for Confocal Microscopy with Multiplex Detection
07:42

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Published on: February 24, 2026

Three-dimensional, tomographic super-resolution fluorescence imaging of serially sectioned thick samples.

Siddharth Nanguneri1, Benjamin Flottmann, Heinz Horstmann

  • 1Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany.

Plos One
|June 5, 2012
PubMed
Summary

We developed tomoSTORM, a novel super-resolution microscopy technique, to achieve near-molecular resolution 3D imaging in thick brain tissue. This advance enables detailed visualization of cellular structures like synapses and mitochondria.

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Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
08:04

Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography

Published on: March 12, 2017

Area of Science:

  • Neuroscience
  • Biophysics
  • Microscopy

Background:

  • Achieving near-molecular resolution 3D imaging in thick biological tissues is a significant challenge.
  • Existing methods often struggle with resolution, penetration depth, or structural integrity.

Purpose of the Study:

  • To develop a robust method for high-resolution 3D imaging of intact brain tissue.
  • To enable detailed visualization of synaptic structures and organelles within large tissue volumes.

Main Methods:

  • Developed tomoSTORM, integrating single-molecule localization microscopy with array tomography.
  • Serially imaged consecutive tissue sections with high lateral (28 nm) and axial (40 nm) resolution.
  • Applied the technique to structurally intact brain tissue, including the calyx of Held synapse.

Main Results:

  • Achieved near-molecular resolution 3D imaging in tissue volumes up to 50 µm×50 µm×2.5 µm.
  • Successfully delineated the membrane and fine structure of mitochondria at the calyx of Held.
  • Demonstrated multiplexed super-resolution imaging with resolution three orders of magnitude better than confocal microscopy.

Conclusions:

  • tomoSTORM overcomes limitations in 3D super-resolution microscopy for thick tissues.
  • This technique provides unprecedented detail for studying neural circuits and synaptic organization.
  • Enables advanced investigations into the molecular architecture of the brain.