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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: Oct 8, 2025

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope

Published on: March 16, 2022

A modular platform for automated cryo-FIB workflows.

Sven Klumpe¹, Herman Kh Fung², Sara K Goetz^2,3

¹Department Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany.

|December 24, 2021

Summary

This summary is machine-generated.

Automating lamellar micromachining using focused ion beam milling (cryo-FIB) speeds up sample preparation for cellular cryo-electron tomography. The open-source SerialFIB software streamlines this process, enabling advanced imaging techniques.

Keywords:

S. cerevisiae automation chlamydomonas reinhardtii correlative light and electron microscopy cryo-FIB-SEM cryo-electron tomography cryo-lift-out drosophila melanogaster electron microscopy emiliania huxleyi human in situ structural biology molecular biophysics structural biology

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Last Updated: Oct 8, 2025

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Expression and Purification of the Human Lipid-sensitive Cation Channel TRPC3 for Structural Determination by Single-particle Cryo-electron Microscopy

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Published on: December 1, 2023

Area of Science:

Cryo-electron microscopy and tomography
Biophysics and structural biology
Materials science and nanotechnology

Background:

Focused ion beam (FIB) milling at cryogenic temperatures (cryo-FIB) is crucial for preparing biological samples for cryo-electron tomography.
Current cryo-FIB protocols are often slow and labor-intensive due to low ion beam currents needed to preserve sample integrity.
Advancements in cryo-FIB instrumentation now allow for automated operations.

Purpose of the Study:

To introduce SerialFIB, an open-source software tool designed to automate and customize cryo-FIB lamellar preparation protocols.
To provide a user-friendly interface and scripting capabilities for advanced sample preparation workflows.
To facilitate integration with correlative light and electron microscopy (CLEM) techniques.

Main Methods:

Development of an open-source software, SerialFIB, featuring a graphical user interface and Python scripting module.
Integration of SerialFIB with existing three-dimensional correlative light and electron microscopy (CLEM) tools.
Implementation of advanced cryo-FIB protocols including multi-modal imaging and in situ lamella lift-out.

Main Results:

SerialFIB enables automated and customizable lamellar micromachining for cryo-FIB applications.
The software streamlines complex preparation workflows, including CLEM-guided procedures.
It supports advanced techniques like multi-modal imaging and in situ lamella lift-out.

Conclusions:

SerialFIB significantly enhances the efficiency and flexibility of cryo-FIB sample preparation.
The software provides a robust foundation for developing novel cryogenic imaging and sample preparation strategies.
It empowers researchers to perform more sophisticated analyses in cellular cryo-electron tomography.