ArticleJournal of microscopy2019
Current outcomes when optimizing 'standard' sample preparation for single-particle cryo-EM.
Article in Journal of microscopy, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed.
- Two datasets are better than one: method of double moments for 3D reconstruction in cryo-EM.Inverse problems · 2026Article
- Diversifying particle-orientation distributions in cryo-EM with LEA protein additives.bioRxiv : the preprint server for biology · 2026Article
- Systematic scale-up and enhanced purification of marine cyanophage P-SSP7.Frontiers in microbiology · 2026Article
- NMR in the Age of Modern Biomedical Research and Drug Discovery.Journal of molecular biology · 2025Review
- Structural Biology for Target Identification and Validation.Methods in molecular biology (Clifton, N.J.) · 2025Article
- High-resolution single-particle imaging at 100-200 keV with the Gatan Alpine direct electron detector.Journal of structural biology · 2024Article
- Self-assembled superstructure alleviates air-water interface effect in cryo-EM.Nature communications · 2024Article
- Cryo-EM phase-plate images reveal unexpected levels of apparent specimen damage.bioRxiv : the preprint server for biology · 2024Article
- An immobilized antibody-based affinity grid strategy for on-grid purification of target proteins enables high-resolution cryo-EM.Communications biology · 2024Article
- Electrospray-assisted cryo-EM sample preparation to mitigate interfacial effects.Nature methods · 2024Article
- VitroJet: new features and case studies.Acta crystallographica. Section D, Structural biology · 2024Article
- Mapping Protein-Protein Interactions at Birth: Single-Particle Cryo-EM Analysis of a Ribosome-Nascent Globin Complex.ACS central science · 2024Article
- High-resolution single-particle imaging at 100-200 keV with the Gatan Alpine direct electron detector.bioRxiv : the preprint server for biology · 2024Article
- Overcoming resolution attenuation during tilted cryo-EM data collection.Nature communications · 2024Article
- Structural Analysis of Protein Complexes by Cryo-Electron Microscopy.Methods in molecular biology (Clifton, N.J.) · 2024Review
- Streptavidin-Affinity Grid Fabrication for Cryo-Electron Microscopy Sample Preparation.Journal of visualized experiments : JoVE · 2023Article
- Cryo-Electron Microscopy Screening Automation across Multiple Grids using Smart Leginon.Journal of visualized experiments : JoVE · 2023Article
- A large expert-curated cryo-EM image dataset for machine learning protein particle picking.Scientific data · 2023Article
- Role of surfactants in electron cryo-microscopy film preparation.Biophysical journal · 2023Article
- CryoPPP: A Large Expert-Labelled Cryo-EM Image Dataset for Machine Learning Protein Particle Picking.bioRxiv : the preprint server for biology · 2023Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
Although high-resolution single-particle cryo-electron microscopy (cryo-EM) is now producing a rapid stream of breakthroughs in structural biology, it nevertheless remains the case that the preparation of suitable frozen-hydrated samples on electron microscopy grids is often quite challenging. Purified samples that are intact and structurally homogeneous - while still in the test tube - may not necessarily survive the standard methods of making extremely thin, aqueous films on grids. As a result, it is often necessary to try a variety of experimental conditions before finally finding an approach that is optimal for the specimen at hand. Here, we summarize some of our collective experiences to date in optimizing sample preparation, in the hope that doing so will be useful to others, especially those new to the field. We also hope that an open discussion of these common challenges will encourage the development of more generally applicable methodology. Our collective experiences span a diverse range of biochemical samples and most of the commonly used variations in how grids are currently prepared. Unfortunately, none of the currently used optimization methods can be said, in advance, to be the one that ultimately will work when a project first begins. Nevertheless, there are some preferred first steps to explore when facing specific problems that can be more generally recommended, based on our experience and that of many others in the cryo-EM field.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.