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Electron-counting MicroED data with the K2 and K3 direct electron detectors.

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Direct electron detectors, commonly used in cryo-electron microscopy (cryo-EM), can now be effectively used for Microcrystal Electron Diffraction (MicroED) data collection. This advancement allows for macromolecular structure determination using widely available equipment.

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Area of Science:

  • Structural Biology
  • Biophysics
  • Materials Science

Background:

  • Microcrystal Electron Diffraction (MicroED) is a powerful technique for determining the 3D structure of small crystals.
  • Advances in hardware and methods have significantly improved MicroED data quality, enabling *ab initio* structure determination of macromolecules.
  • Direct electron detectors offer high sensitivity and fast readout, ideal for electron diffraction, but concerns about dynamic range and coincidence loss have limited their use.

Purpose of the Study:

  • To evaluate the suitability of K2 and K3 direct electron detectors for MicroED data collection in electron-counting mode.
  • To demonstrate the application of these detectors for solving macromolecular structures using MicroED.
  • To highlight the potential for utilizing existing cryo-EM facilities for MicroED studies.

Main Methods:

  • Instrument setup for low-exposure MicroED data collection using K2 and K3 direct electron detectors.
  • Data collection in electron-counting mode with continuous sample rotation.
  • Structure determination of two macromolecules using the collected diffraction data.

Main Results:

  • Integrated intensities from MicroED data collected on K2 and K3 detectors were successfully used to solve macromolecular structures.
  • Resolutions between 1.2 Å and 2.8 Å were achieved for the determined structures.
  • No significant camera damage was observed, even without a beam stop during K3 detector studies.

Conclusions:

  • K2 and K3 direct electron detectors are effective for MicroED data collection and macromolecular structure determination.
  • This approach democratizes MicroED by enabling its use in facilities already equipped with common cryo-EM detectors.
  • The findings expand access to MicroED for researchers without dedicated MicroED instrumentation.