Cryogenic Electron Microscopy
Cryogenic Electron Microscopy - Related Videos
06:32
Optimizing Sample Preparation for Cryogenic Electron Microscopy
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2025
This protocol presents a practical method using Methanocaldococcus jannaschii (MjsHSP16.5) as an example to address uneven particle distribution through sample preparation optimization, providing a reference for researchers to efficiently elucidate macromolecular structures using cryogenic electron microscopy (cryo-EM).
02:03
Preparation of Legionella pneumophila for Cryogenic Electron Tomography
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2025
Source: Chetrit, D., et al. Applying Live Cell Imaging and Cryo-Electron Tomography to Resolve Spatiotemporal Features of the Legionella pneumophila Dot/Icm Secretion System. J. Vis. Exp. (2020)The video describes how to prepare Legionella pneumophila samples for cryo-electron tomography by combining bacterial culture, gold nanoparticle marking, and vitrification techniques. The process preserves native cellular structures and enables high-resolution imaging of membrane-associated secretion...
08:29
Workflow Using a Cryogenic Coincident Fluorescence, Electron, and Ion Beam Microscope for Targeted Milling of Cells
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Cited by 2 •
2025
This workflow enables lamella production targeting fluorescently labeled biological structures that are small (<1 μm in axial extent) and rare (1 copy per cell) using a cryogenic tri-coincident imaging platform. This platform integrates fluorescence microscopy, focused ion beam milling, and scanning electron microscopy at a single focal position and enables simultaneous fluorescence microscopy while milling.
08:16
Strategies for Optimization of Cryogenic Electron Tomography Data Acquisition
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Cited by 8 •
2021
The increasing demand for large-scale data collection in cryogenic electron tomography requires high-throughput image acquisition routines. Described here is a protocol that implements the recent developments of advanced acquisition strategies aimed at maximizing the time-efficiency and throughput of tomographic data collection.
11:16
Correlative Light- and Electron Microscopy Using Quantum Dot Nanoparticles
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Cited by 12 •
2016
A method is described whereby quantum dot (QD) nanoparticles can be used for correlative immunocytochemical studies of epoxy embedded human pathology tissue. We employ commercial antibody fragment conjugated QDs that are visualized by widefield fluorescence light microscopy and transmission electron microscopy.
FAQs
Rapid cooling converts the sample water into vitreous ice rather than allowing crystals to form. This preserves the organization of biological molecules and materials in a near-native state, so observed structures are less dependent on crystallization. The resulting preservation is especially relevant when bioengineers need to examine proteins, nucleic acids, membranes, nanoparticles, or engineered biomaterials as organized specimens.
An electron beam passes through the vitrified specimen, and detectors record images of the transmitted electrons. Computational methods then combine the captured image information to reconstruct a three-dimensional structure. This workflow links direct imaging with analysis, enabling researchers to move from observations of a specimen to structural information that can support studies of molecular function and engineered materials.
Maintaining a near-native state helps retain the organization of molecules and materials without relying on crystallization. That distinction matters because crystallization is avoided during preservation, allowing cryo-EM to examine structures such as membranes, nucleic acids, and engineered biomaterials in forms that reflect their organized sample state. The structural information can clarify relationships between molecular arrangement and function.
A basic workflow begins by rapidly cooling the specimen to form vitreous ice. The preserved sample is then exposed to an electron beam, while detectors capture the resulting images. Finally, computational reconstruction combines image data to generate three-dimensional structural information. Each stage contributes a different function: preservation, signal capture, and structural interpretation.
Bioengineers may choose cryo-EM when they need structural information about proteins, nucleic acids, membranes, nanoparticles, or engineered biomaterials in near-native states. The method is useful for connecting molecular or material organization with function. In applied research, those insights can guide therapeutic design and support development of diagnostic and regenerative technologies.
Three-dimensional structural information can reveal how biological molecules or engineered materials are organized, helping researchers understand molecular function. In bioengineering, that understanding provides a basis for guiding therapeutic design and improving diagnostic or regenerative technologies. The same imaging approach also supports structural biology by connecting preserved specimen organization with broader questions about biological and material performance.