Cryo-em
Cryo-em - Related Videos
05:05
Preparation of High-Temperature Sample Grids for Cryo-EM
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Cited by 5 •
2021
This paper provides a detailed protocol for preparing sample grids at temperatures as high as 70 °C, prior to plunge freezing for cryo-EM experiments.
09:06
Cryo-EM and Single-Particle Analysis with Scipion
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Cited by 8 •
2021
Single-particle analysis in cryo-electron microscopy is one of the main techniques used to determine the structure of biological ensembles at high resolution. Scipion provides the tools to create the whole pipeline to process the information acquired by the microscope and achieve a 3D reconstruction of the biological specimen.
13:43
A Robust Single-Particle Cryo-Electron Microscopy (cryo-EM) Processing Workflow with cryoSPARC, RELION, and Scipion
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Cited by 17 •
2022
This article describes how to effectively utilize three cryo-EM processing platforms, i.e., cryoSPARC v3, RELION-3, and Scipion 3, to create a single and robust workflow applicable to a variety of single-particle data sets for high-resolution structure determination.
04:52
Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon
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Cited by 6 •
2022
The present protocol describes a generalized and easy-to-implement scheme for tilted single-particle data collection in cryo-EM experiments. Such a procedure is especially useful for obtaining a high-quality EM map for samples suffering from preferential orientation bias due to adherence to the air-water interface.
09:49
Routine Collection of High-Resolution cryo-EM Datasets Using 200 KV Transmission Electron Microscope
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Cited by 13 •
2022
High-resolution cryo-EM maps of macromolecules can be also achieved by using 200 kV TEM microscopes. This protocol shows the best practices for setting accurate optics alignments, data acquisition schemes, and selection of imaging areas that are all essential for the successful collection of high-resolution datasets using a 200 kV TEM.
FAQs
Vitreous ice provides a glass-like frozen environment that preserves molecular shape after rapid freezing. Because the sample does not undergo conventional chemical fixation or crystallization, researchers can examine biological molecules in conditions that remain close to their native state. This preservation is particularly valuable for dynamic or medically relevant systems.
Computational reconstruction combines many two-dimensional particle images into a three-dimensional representation. Each recorded view contributes information about the same type of molecule or complex, while the combined dataset supports interpretation of its overall structure. This approach turns image measurements into a structural model that can be analyzed in biological research.
Cryo-EM is useful when a biological complex contains heterogeneous or flexible forms. Rapid freezing preserves particles without requiring conventional crystallization, so the resulting images can reflect structural variation present in the sample. Computational treatment of many particle images then supports three-dimensional analysis of complexes that may be difficult to represent as one uniform structure.
Compared with approaches based on chemical fixation or crystallization, Cryo-EM preserves samples through rapid freezing into vitreous ice. This avoids the need to chemically fix molecules or arrange them into a crystal before imaging. The distinction matters for biological systems whose medically relevant structure or flexibility may be difficult to capture under those requirements.
A basic workflow starts with a biological sample, which is rapidly frozen to form vitreous ice. The frozen material is then exposed to an electron beam, producing many two-dimensional particle images. Computational methods combine those images into a three-dimensional reconstruction. Thus, the experiment connects physical sample preservation with image collection and structural interpretation.
Cryo-EM can examine proteins, nucleic acids, viruses, membranes, and molecular machines. Studying this range lets biologists investigate structures from individual biological molecules to larger molecular complexes. The method is therefore relevant across cell biology and molecular mechanism research, where structural information helps relate biological components to their organization and molecular interactions.
In biology, Cryo-EM provides structural evidence for studying dynamic systems and medically relevant molecules or complexes. Its applications include cell biology, molecular mechanism research, and structure-guided drug development. By revealing the architecture of proteins, nucleic acids, viruses, membranes, or molecular machines, it connects structural analysis with broader biological and biomedical questions.