ArticleProceedings of the National Academy of Sciences of the United States of America2021
Benchmarking the ideal sample thickness in cryo-EM.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed.
- Advances of 3D microcrystals electron diffraction for transmembrane protein structure determination.Biophysical journal · 2026Review
- Electrons and X-rays for diffraction and imaging.Acta crystallographica. Section D, Structural biology · 2026Review
- A cost-effective 200 kV cryo-EM core facility for high resolution single particle analysis.Methods in microscopy · 2026Article
- Single-cell structural biology with intracellular electron crystallography.Nature communications · 2026Article
- Direct from the seed: an atomic resolution protein structure by ab initio MicroED.Nature communications · 2026Article
- PhaseT3M: 3D imaging at 1.6 Å resolution via electron cryo-tomography with nonlinear phase retrieval.Nature communications · 2025Article
- Standards for MicroED.Acta crystallographica. Section C, Structural chemistry · 2025Article
- Recovering high-resolution information using energy filtering in MicroED.Structural dynamics (Melville, N.Y.) · 2025Article
- Comprehensive microcrystal electron diffraction sample preparation for cryo-EM.Nature protocols · 2025Review
- Review
- Energy filtering enables macromolecular MicroED data at sub-atomic resolution.Nature communications · 2025Article
- Unraveling atomic complexity from frozen samples.Structural dynamics (Melville, N.Y.) · 2025Article
- A strategic approach for efficient cryo-EM grid optimization using design of experiments.Journal of structural biology · 2025Article
- Cryo-electron tomography pipeline for plasma membranes.Nature communications · 2025Article
- From Beam Damage to Massive Reaction Amplification under the Electron Microscope: An Ionization-Induced Chain Reaction in Crystals of a Dewar Benzene.ACS central science · 2024Article
- VMXm - A sub-micron focus macromolecular crystallography beamline at Diamond Light Source.Journal of synchrotron radiation · 2024Article
- An Updated Structure of Oxybutynin Hydrochloride.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2024Article
- Energy filtering enables macromolecular MicroED data at sub-atomic resolution.bioRxiv : the preprint server for biology · 2024Article
- Doses for X-ray and electron diffraction: New features in RADDOSE-3D including intensity decay models.Protein science : a publication of the Protein Society · 2024Article
- Cryo-electron tomography pipeline for plasma membranes.bioRxiv : the preprint server for biology · 2024Article
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5 authors.
Funding
Abstract
The relationship between sample thickness and quality of data obtained is investigated by microcrystal electron diffraction (MicroED). Several electron microscopy (EM) grids containing proteinase K microcrystals of similar sizes from the same crystallization batch were prepared. Each grid was transferred into a focused ion beam and a scanning electron microscope in which the crystals were then systematically thinned into lamellae between 95- and 1,650-nm thick. MicroED data were collected at either 120-, 200-, or 300-kV accelerating voltages. Lamellae thicknesses were expressed in multiples of the corresponding inelastic mean free path to allow the results from different acceleration voltages to be compared. The quality of the data and subsequently determined structures were assessed using standard crystallographic measures. Structures were reliably determined with similar quality from crystalline lamellae up to twice the inelastic mean free path. Lower resolution diffraction was observed at three times the mean free path for all three accelerating voltages, but the data quality was insufficient to yield structures. Finally, no coherent diffraction was observed from lamellae thicker than four times the calculated inelastic mean free path. This study benchmarks the ideal specimen thickness with implications for all cryo-EM methods.
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