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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 20, 2025

Preparation of High-Temperature Sample Grids for Cryo-EM

Preparation of High-Temperature Sample Grids for Cryo-EM

Published on: July 26, 2021

Copper Oxide Spike Grids for Enhanced Solution Transfer in Cryogenic Electron Microscopy.

Dukwon Lee^1,2, Hansol Lee^3,4,2, Jinwook Lee¹

¹Research Institute of Agriculture and Life Sciences, Center for Food and Bioconvergence, Department of Agricultural Biotechnology, Interdisciplinary Programs in Agricultural Genomics, College of Agriculture and Life Sciences (CALS), Seoul National Un.

Molecules and Cells

|August 2, 2023

Summary

Controlling vitreous ice thickness for cryogenic electron microscopy (cryo-EM) is challenging. This study introduces a novel copper oxide spike grid method to improve ice homogeneity and reproducibility for better cryo-EM data.

Keywords:

copper oxide spike grids cryo-electron microscopy vitrification transmission electron microscopy grids

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Published on: July 26, 2021

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Published on: February 25, 2021

Area of Science:

Cryo-electron microscopy sample preparation
Materials science for microscopy

Background:

Uniform vitreous ice formation is critical for high-quality cryo-EM data.
Classical blotting methods struggle with excess water, leading to heterogeneous ice thickness.
Reproducible control of ice thickness remains a significant challenge in cryo-EM.

Purpose of the Study:

To develop a novel method for controlling vitreous ice thickness and homogeneity in cryo-EM sample preparation.
To address the limitations of classical blotting techniques in water removal.
To enhance the reproducibility of vitrified ice formation for improved cryo-EM imaging.

Main Methods:

A novel approach combining nanowire self-wicking with classical blotting.
Generation of copper oxide spike (COS) grids by inducing COSs on commercial copper grids.
Utilizing COS grids to effectively remove excess water during blotting without damaging the carbon membrane.

Main Results:

Achieved effective control over vitreous ice thickness and homogeneity.
Demonstrated good reproducibility in ice thickness control compared to non-oxidized grids.
COS grids efficiently removed excess water during the blotting procedure.

Conclusions:

The developed COS grid modification method offers an effective solution for reproducible vitreous ice formation.
This technique enhances data quality in cryo-EM imaging.
The method is adaptable for various electron microscopy techniques requiring vitrified samples.