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Super-resolution Fluorescence Microscopy

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Updated: Jul 12, 2025

Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

Published on: October 31, 2020

Enhanced synaptic protein visualization by multicolor super-resolution expansion microscopy.

Janna Eilts¹, Sebastian Reinhard¹, Nikolas Michetschläger¹

¹University of Würzburg, Department of Biotechnology and Biophysics, Biocenter, Würzburg, Germany.

|October 27, 2023

Summary

This summary is machine-generated.

Researchers developed a novel 4-color super-resolution microscopy method for synaptic protein imaging. This technique achieves 20-30 nm resolution, offering new insights into neuronal network organization.

Keywords:

10-fold robust expansion microscopy expansion microscopy multicolor imaging structured illumination microscopy super-resolution microscopy synaptic proteins

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Last Updated: Jul 12, 2025

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Area of Science:

Neuroscience
Cell Biology
Microscopy

Background:

Understanding biomolecular complex organization and dynamics is vital for cellular functions, especially in neuronal synapses.
Super-resolution (SR) microscopy has advanced synapse imaging, but multicolor imaging at 20-30 nm resolution remains a challenge.

Purpose of the Study:

To develop a method enabling multicolor fluorescence imaging of synaptic proteins with 20-30 nm spatial resolution.
To overcome limitations in current SR microscopy for detailed synaptic molecular organization studies.

Main Methods:

Utilized post-expansion immunolabeling of eightfold expanded hippocampal neurons.
Combined expansion microscopy with Airyscan and structured illumination microscopy (SIM).

Main Results:

Achieved efficient labeling of synaptic proteins in crowded cellular compartments with minimal linkage error.
Demonstrated 4-color 3D fluorescence imaging with 20-30 nm lateral resolution.
Identified colocalization of synaptic vesicles (Synaptobrevin 2) with pre-synaptic fusion sites (RIM1/2).

Conclusions:

The optimized expansion microscopy approach enhances visualization and localization of synaptic proteins.
Provides invaluable insights into the spatial organization of proteins at neuronal synapses.
Facilitates a deeper understanding of synaptic function and dysfunction.