ArticleBiophysical journal2020
Microscale Fluid Behavior during Cryo-EM Sample Blotting.
Article in Biophysical journal, 2020. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed, 69 citations in OpenAlex.
- Control of ice thickness in cryo-EM via confinement.Nature methods · 2026Article
- EasyGrid: a versatile platform for automated cryo-EM sample preparation and quality control.Nature methods · 2026Article
- PARP1-HPF1 structure and dynamics on nicked DNA suggest a mechanism for acute and localized ADP-ribosylation.Nature communications · 2026Article
- Passage of ribosomes through microsprayer increases functional activity - Implications for activity assays in time-resolved cryo-EM.Journal of structural biology · 2025Article
- The graphene-based affinity cryo-EM grid for the endogenous protein structure determination.bioRxiv : the preprint server for biology · 2025Article
- Unravelling complex mechanisms in materials processes with cryogenic electron microscopy.Chemical science · 2025Review
- In situ crystalline structure of the human eosinophil major basic protein-1.bioRxiv : the preprint server for biology · 2024Article
- Article
- Nanoscale architecture of synaptic vesicles and scaffolding complexes revealed by cryo-electron tomography.Proceedings of the National Academy of Sciences of the United States of America · 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
- Applications of microscopy and small angle scattering techniques for the characterisation of supramolecular gels.Beilstein journal of organic chemistry · 2024Review
- CryoEM Data Analysis of Membrane Proteins. Practical Considerations on Amphipathic Belts, Ligands, and Variability Analysis.Methods in molecular biology (Clifton, N.J.) · 2024Review
- Handling Difficult Cryo-ET Samples: A Study with Primary Neurons from Drosophila melanogaster.Microscopy and microanalysis : the official journal of Microscopy Society of America, Microbeam Analysis Society, Microscopical Society of Canada · 2023Article
- Copper Oxide Spike Grids for Enhanced Solution Transfer in Cryogenic Electron Microscopy.Molecules and cells · 2023Article
- Overview of Membrane Protein Sample Preparation for Single-Particle Cryo-Electron Microscopy Analysis.International journal of molecular sciences · 2023Review
- Role of surfactants in electron cryo-microscopy film preparation.Biophysical journal · 2023Article
- Cryo-Electron Tomography of Reconstituted Biomolecular Condensates.Methods in molecular biology (Clifton, N.J.) · 2023Article
- cryoWriter: a blotting free cryo-EM preparation system with a climate jet and cover-slip injector.Faraday discussions · 2022Article
- Polyelectrolyte coating of cryo-EM grids improves lateral distribution and prevents aggregation of macromolecules.Acta crystallographica. Section D, Structural biology · 2022Article
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
Abstract
Blotting has been the standard technique for preparing aqueous samples for single-particle electron cryo-microscopy for over three decades. This technique removes the excess solution from a transmission electron microscope grid by pressing absorbent filter paper against the specimen before vitrification. However, this standard technique produces vitreous ice with inconsistent thickness from specimen to specimen and from region to region within the same specimen, the reasons for which are not understood. Here, high-speed interference contrast microscopy is used to demonstrate that the irregular pattern of fibers in the filter paper imposes tortuous, highly variable boundaries during the removal of excess liquid from a flat, hydrophilic surface. As a result, aqueous films of nonuniform thickness are formed while the filter paper is pressed against the substrate. This pattern of nonuniform liquid thickness changes again after the filter paper is pulled away, but the thickness still does not become completely uniform. We suggest that similar topographical features of the liquid film are produced during the standard technique used to blot EM grids and that these manifest in nonuniform ice after vitrification. These observations suggest that alternative thinning techniques, which do not rely on direct contact between the filter paper and the grid, may result in more repeatable and uniform sample thicknesses.
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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.