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Updated: May 21, 2025

Analysis of Astrocyte Territory Volume and Tiling in Thick Free-Floating Tissue Sections
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Super-resolution Analysis of Astrocyte Morphology Using Expansion Microscopy.

Gerald Seifert1, Erik Sommer1, Stefan Passlick1

  • 1Institute of Cellular Neurosciences I, Medical Faculty, University of Bonn, Bonn, Germany.

Methods in Molecular Biology (Clifton, N.J.)
|March 20, 2025
PubMed
Summary

Expansion microscopy (ExM) offers a powerful method to visualize astrocyte structures and their fine processes. This technique enhances light microscopy resolution, aiding in understanding astrocyte function and cell interactions.

Keywords:
Astrocyte morphologyExpansion microscopyPerisynaptic astrocytic processesSuper-resolution microscopy

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Area of Science:

  • Neuroscience
  • Cell Biology
  • Microscopy

Background:

  • Astrocyte morphology is crucial for cellular functions, signal propagation, and intercellular communication within the brain.
  • Diffraction-limited microscopy struggles to resolve fine astrocytic processes, hindering detailed structural analysis.
  • Electron microscopy provides high resolution but is complex; expansion microscopy (ExM) offers an accessible alternative.

Purpose of the Study:

  • To present a comprehensive protocol for visualizing astrocytes and their associated structures using expansion microscopy.
  • To detail the localization of synaptic and astrocytic proteins within the fixed brain tissue.
  • To overcome the limitations of conventional microscopy in resolving fine astrocytic processes.

Main Methods:

  • Utilizing expansion microscopy (ExM) to physically expand fixed brain tissue.
  • Preserving the relative spatial positioning of fluorescent labels marking cellular structures and proteins.
  • Employing confocal fluorescence microscopy for imaging the expanded samples.

Main Results:

  • Successfully visualized fine astrocytic processes and their perisynaptic arrangements with enhanced resolution.
  • Enabled detailed localization of synaptic structures and key astrocytic and synaptic proteins.
  • Demonstrated the efficacy of ExM for detailed astrocyte structural analysis.

Conclusions:

  • Expansion microscopy provides a cost-effective and accessible method for high-resolution imaging of astrocytes.
  • This protocol facilitates a deeper understanding of astrocyte morphology and its role in brain function.
  • ExM is a valuable tool for studying astrocyte-synapse interactions and cellular architecture.