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Related Experiment Video

Updated: May 26, 2025

Microcrystal Electron Diffraction of Small Molecules
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Fast event-based electron counting for small-molecule structure determination by MicroED.

Niko Vlahakis1, Songrong Qu1, Logan S Richards1

  • 1Department of Chemistry and Biochemistry, UCLA-DOE Institute for Genomics and Proteomics, STROBE, NSF Science and Technology Center, University of California, Los Angeles (UCLA), Los Angeles, CA 90095, USA.

Acta Crystallographica. Section C, Structural Chemistry
|February 21, 2025
PubMed
Summary
This summary is machine-generated.

Fast electron counting with the Apollo camera enables rapid MicroED structure determination, even at low doses. This technique minimizes errors and accelerates the analysis of small molecules like biotin and thiostrepton.

Keywords:
EBECMicroEDcrystal structureelectron countingelectron diffraction

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Area of Science:

  • Crystallography and Materials Science
  • Electron Microscopy and Imaging

Background:

  • Electron counting has revolutionized cryo-electron microscopy (cryoEM) and is now advancing Microcrystal Electron Diffraction (MicroED) structure determination.
  • New direct electron detectors with fast (kilohertz) event-based electron counting (EBEC) minimize coincidence loss (CL) for accurate and rapid structure analysis.

Purpose of the Study:

  • To evaluate the effectiveness of fast EBEC technology using the Direct Electron Apollo camera for MicroED data collection.
  • To assess the capability of EBEC for rapid, low-dose structure determination of small organic molecules.

Main Methods:

  • Utilized the Direct Electron Apollo camera for MicroED data collection, leveraging its single-electron counting capability.
  • Collected data from organic small-molecule crystals (salen ligand, biotin, thiostrepton) under various low incident electron beam flux densities.
  • Analyzed data using fast EBEC strategies, with and without coincidence loss adjustment, to determine crystal structures.

Main Results:

  • High-quality MicroED data were obtained from small organic crystals at electron beam flux densities as low as 0.01-0.045 e-/Ų/s.
  • Ab initio structures of a salen ligand and biotin were determined from single crystals in as little as 50 seconds with a fluence of ~0.5 e-/Ų.
  • A 1.5 Å structure of thiostrepton was achieved with a fluence of 2.25 or 3.33 e-/Ų; coincidence loss adjustment showed marginal impact on refinement, indicating high detector accuracy.

Conclusions:

  • Fast EBEC data collection strategies deliver low-dose, structure-worthy data, enabling rapid MicroED analysis.
  • This approach accelerates the determination of small molecule structures, opening avenues for high-throughput MicroED.
  • Modern kilohertz-rate detectors provide high counting accuracy, minimizing the need for extensive coincidence loss correction in MicroED.