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Related Experiment Video

Updated: Nov 10, 2025

Super-resolution Imaging of Proteus mirabilis Biofilm by Expansion Microscopy

Super-resolution Imaging of Proteus mirabilis Biofilm by Expansion Microscopy

Published on: July 18, 2025

The Expandables: Cracking the Staphylococcal Cell Wall for Expansion Microscopy.

Tobias C Kunz¹, Marcel Rühling¹, Adriana Moldovan¹

¹Department of Microbiology, Julius-Maximilians-Universität Würzburg, Würzburg, Germany.

Frontiers in Cellular and Infection Microbiology

|April 2, 2021

Summary

Expansion Microscopy (ExM) now visualizes Gram-positive bacteria like Staphylococcus aureus. This new method uses enzymatic treatments for 4× expansion, enabling high-resolution imaging of bacteria.

Keywords:

Staphylococcus aureus autophagosomes endosomes expansion microscopy high-resolution imaging host-pathogen interaction

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Area of Science:

Microscopy
Bacteriology
Biotechnology

Background:

Expansion Microscopy (ExM) enhances fluorescence microscopy resolution by hydrogel embedding and physical expansion.
Previous ExM applications focused on intracellular Gram-negative pathogens.
Gram-positive bacteria present challenges for ExM due to their rigid cell walls.

Purpose of the Study:

To adapt and develop an ExM approach for imaging Gram-positive bacteria.
To achieve high-resolution visualization of Staphylococcus aureus using ExM.
To demonstrate the applicability of the developed ExM technique for both planktonic and intracellular bacteria.

Main Methods:

Developed a novel ExM protocol involving enzymatic treatments tailored for Gram-positive bacteria.
Applied the method to Staphylococcus aureus, achieving isotropic 4× physical expansion.
Utilized conventional confocal laser scanning microscopy for imaging.

Main Results:

Successfully achieved isotropic 4× expansion of Staphylococcus aureus using enzymatic treatments.
Enabled imaging of planktonic and intracellular Staphylococcus aureus.
Reached a spatial resolution of approximately 60 nm.

Conclusions:

The developed ExM approach effectively overcomes the challenges of imaging Gram-positive bacteria.
This technique significantly improves the spatial resolution for visualizing Staphylococcus aureus.
The method is suitable for studying bacteria in various states, including intracellularly.