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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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Overview
The basic structure of RNA consists of a five-carbon sugar and one of four nitrogenous bases. Although most RNA is single-stranded, it can form complex secondary and tertiary structures. Such structures play essential roles in the regulation of transcription and translation.
Different Types of RNA Have the Same Basic Structure
There are three main types of ribonucleic acid (RNA): messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). All three RNA types consist of a...

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Related Experiment Video

Updated: Oct 2, 2025

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

Published on: January 9, 2015

Cryo-EM advances in RNA structure determination.

Haiyun Ma¹, Xinyu Jia¹, Kaiming Zhang²

¹The State Key Laboratory of Biotherapy, Department of Geriatrics and National Clinical Research Center for Geriatrics, West China Hospital, Sichuan University, Chengdu, Sichuan, 610044, China.

Signal Transduction and Targeted Therapy

|February 24, 2022

Summary

This summary is machine-generated.

Cryo-electron microscopy (cryo-EM) now reveals atomic-level protein structures. This review explores cryo-EM for protein-free RNA structures, overcoming previous limitations.

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Area of Science:

Structural Biology
Biochemistry
Molecular Biology

Background:

Cryo-electron microscopy (cryo-EM) offers atomic resolution for protein structures.
It excels with samples unsuitable for X-ray crystallography or NMR.
Cryo-EM handles heterogeneous and conformationally flexible complexes under near-native conditions.

Purpose of the Study:

To review challenges and progress in determining protein-free RNA structures using cryo-EM.
To highlight advancements enabling RNA structure determination via single-particle analysis.
To discuss the integration of cryo-EM with other methods for RNA structural insights.

Main Methods:

Cryo-electron microscopy single-particle analysis (cryo-EM SPA).
Advancements in sample preparation techniques for RNA.
Integration of complementary structural and biochemical methods.

Main Results:

Cryo-EM is increasingly capable of resolving protein-free RNA structures.
Improvements in sample handling and data processing have been crucial.
Combined approaches yield deeper insights into RNA structure and function.

Conclusions:

Cryo-EM is a powerful tool for exploring RNA structures.
Overcoming challenges in RNA sample preparation is key to its application.
Integrating cryo-EM with other techniques expands structural biology frontiers for RNA.