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Updated: Dec 15, 2025

Applying Live Cell Imaging and Cryo-Electron Tomography to Resolve Spatiotemporal Features of the Legionella pneumophila Dot/Icm Secretion System

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Imaging Bacterial Colonies and Phage-Bacterium Interaction at Sub-Nanometer Resolution Using Helium-Ion Microscopy.

Miika Leppänen^1,2, Lotta-Riina Sundberg², Elina Laanto²

¹Nanoscience Center, Department of Physics, University of Jyvaskyla, P. O. Box 35, FI-40014, Jyväskylä, Finland.

Advanced Biosystems

|July 11, 2020

Summary

Helium ion microscopy (HIM) offers a novel method for imaging bacterial colonies and virus-bacteria interactions directly on agar plates. This advanced technique provides high-resolution, subsurface views, enhancing microbial interaction studies.

Keywords:

bioimaging focused ion beam milling helium ion microscopy phage-bacteria interactions

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Area of Science:

Microbiology
Microbial Interactions
Microscopy

Background:

Traditional electron microscopy methods limit in situ imaging of microbial interactions.
Studying interactions between bacteria and bacteriophages (phages) requires advanced imaging techniques.

Purpose of the Study:

To introduce and demonstrate Helium Ion Microscopy (HIM) for in situ imaging of bacterial colonies and phage-bacterium interactions on agar plates.
To explore the capabilities of HIM, including subsurface imaging and in situ sectioning, for microbial studies.

Main Methods:

Utilized Helium Ion Microscopy (HIM) for imaging bacterial colonies and phage-bacterium interactions on agar cultures.
Employed helium and neon ion milling for subsurface imaging and in situ cross-sectioning of bacterial colonies and individual bacteria.

Main Results:

Presented the first HIM images of bacterial colonies and phage-bacterium interactions at various infection stages.
Demonstrated the feasibility of ion milling for revealing subsurface structures of bacterial colonies and intracellular structures of bacteria.

Conclusions:

HIM provides significant advantages over traditional methods, including sub-nanometer resolution and enhanced surface sensitivity.
HIM is a powerful tool for advancing microbial imaging, particularly for studying virus-cell interactions in their native environment.