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Cryo-electron Microscopy

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...

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Updated: Jul 16, 2026

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

Cryo-Structured Illumination Microscopic Data Collection from Cryogenically Preserved Cells

Published on: May 28, 2021

Transforming cryo-lift-out one section at a time.

Gaia Perone¹, Claire Overly Cottom¹, Philipp Sebastian Erdmann¹

¹Human Technopole, Milan, Italy.

Current Opinion in Structural Biology

|July 14, 2026

Summary

Cryo-electron tomography (cryo-ET) allows molecular-level cell imaging. Cryo-lift-out sample preparation is advancing in situ imaging of complex multicellular structures like organoids and tissues.

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Area of Science:

Cellular and Molecular Imaging
Biotechnology and Nanotechnology

Background:

Understanding cellular complexity requires in situ imaging techniques across scales.
Cryo-electron tomography (cryo-ET), enhanced by focused ion beam (FIB) milling, offers molecular resolution for cell visualization.

Purpose of the Study:

To review advances in cryo-lift-out for preparing multicellular specimens for cryo-ET.
To discuss challenges and future perspectives for in situ imaging of complex biological samples.

Main Methods:

Focused ion beam (FIB) milling for cryo-electron tomography (cryo-ET) sample preparation.
Cryo-lift-out technique for accessing deep cellular regions within multicellular specimens.

Main Results:

Cryo-lift-out enables cryo-ET analysis of complex multicellular samples like organoids and tissues.
Recent methodological developments are improving the accessibility and resolution of in situ cellular imaging.

Conclusions:

Cryo-lift-out is a critical technique for advancing in situ cryo-ET of complex biological systems.
Continued development is essential for achieving more holistic, molecular-resolution imaging of cellular processes within their native environments.