ArticleJournal of structural biology2012
Spotiton: a prototype for an integrated inkjet dispense and vitrification system for cryo-TEM.
Article in Journal of structural biology, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 93 papers.
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Who cites it
93 citing papers in PubMed.
- EasyGrid: a versatile platform for automated cryo-EM sample preparation and quality control.Nature methods · 2026Article
- Overcoming the challenge of preferred particle orientation in cryo-EM.Nature reviews. Molecular cell biology · 2026Article
- A Surfactant Cocktail Overcomes Air-Water Interface Artifacts in Single-Particle CryoEM.bioRxiv : the preprint server for biology · 2026Article
- Advances in Crystallography and Cryo-Electron Microscopy for the Study of Membrane Proteins.Advances in experimental medicine and biology · 2026Review
- MISO: microfluidic protein isolation enables single-particle cryo-EM structure determination from a single cell colony.Nature methods · 2025Article
- Coming Clean and Avoiding Bubble Trouble-Using Detergents Wisely in the Purification of Membrane Proteins for Cryo-EM Studies.Biomolecules · 2025Review
- Passage of ribosomes through microsprayer increases functional activity - Implications for activity assays in time-resolved cryo-EM.Journal of structural biology · 2025Article
- Multiplexed stamp-transfer AFM deposition improves resolution of protein-DNA conformational states.Biophysical journal · 2025Article
- Foam film vitrification for cryo-EM.Nature communications · 2025Article
- Advancing Cryo-EM and Cryo-ET through Innovation in Sample Carriers: A Perspective.Analytical chemistry · 2025Review
- A novel "bio-tag" for cryo-EM studies based on the small, electron-dense protein Csp1.Biophysical journal · 2025Article
- Preparation of oxygen-sensitive proteins for high-resolution cryoEM structure determination using blot-free vitrification.Nature communications · 2025Article
- A strategic approach for efficient cryo-EM grid optimization using design of experiments.Journal of structural biology · 2025Article
- Review
- Small LEA proteins mitigate air-water interface damage to fragile cryo-EM samples during plunge freezing.Nature communications · 2024Article
- Factors affecting macromolecule orientations in thin films formed in cryo-EM.Acta crystallographica. Section D, Structural biology · 2024Article
- Tuning ice thickness using the chameleon for high-quality cryoEM data collection.bioRxiv : the preprint server for biology · 2024Article
- VitroJet: new features and case studies.Acta crystallographica. Section D, Structural biology · 2024Article
- Metallo-supramolecular branched polymer protects particles from air-water interface in single-particle cryo-electron microscopy.Communications biology · 2024Article
- B56δ long-disordered arms form a dynamic PP2A regulation interface coupled with global allostery and Jordan's syndrome mutations.Proceedings of the National Academy of Sciences of the United States of America · 2024Article
33 more citing papers are in PubMed but not listed here.
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Authors and funding
5 authors.
Funding
Abstract
Over the last three decades, Cryo-TEM has developed into a powerful technique for high-resolution imaging of biological macromolecules in their native vitrified state. However, the method for vitrifying specimens onto EM grids is essentially unchanged - application of ∼3 μL sample to a grid, followed by blotting and rapid plunge freezing into liquid ethane. Several trials are often required to obtain suitable thin (few hundred nanometers or less) vitrified layers amenable for cryo-TEM imaging, which results in waste of precious sample and resources. While commercially available instruments provide some level of automation to control the vitrification process in an effort to increase quality and reproducibility, obtaining satisfactory vitrified specimens remains a bottleneck in the Cryo-TEM pipeline. We describe here a completely novel method for EM specimen preparation based on small volume (picoliter to nanoliter) dispensing using inkjet technology. A first prototype system (Spotiton v0.5) demonstrates feasibility of this new approach for specimen vitrification. A piezo-electric inkjet dispenser is integrated with optical real-time cameras (100 Hz frame rate) to analyze picoliter to nanoliter droplet profiles in-flight and spreading dynamics on the grid, and thus provides a method to optimize timing of the process. Using TEM imaging and biochemical assays we demonstrate that the piezo-electric inkjet mechanism does not disrupt the structural or functional integrity of macromolecules. These preliminary studies provide insight into the factors and components that will need further development to enable a robust and repeatable technique for specimen vitrification using this novel approach.
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