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Cryo-Electron Microscopic Grid Preparation for Time-Resolved Studies using a Novel Robotic System, Spotiton
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Tidy up cryo-EM sample grids with 3D printed tools.

Tasuku Hamaguchi1, Koji Yonekura2

  • 1Biostructural Mechanism Laboratory, RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo, Hyogo 679-5148, Japan.

Journal of Structural Biology
|November 8, 2019
PubMed
Summary

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Cryo-electron microscopy (Cryo-EM) sample preparation is streamlined with new 3D-printed tools. These CryoGridTools simplify grid handling and organization for high-throughput structure determination.

Area of Science:

  • Structural Biology
  • Biophysics
  • Biochemistry

Background:

  • High-throughput cryo-electron microscopy (Cryo-EM) enables rapid structure determination of biological macromolecules.
  • Screening numerous cryo-EM sample grids under diverse conditions is often necessary for challenging targets.
  • Efficient sample handling is crucial for maintaining grid integrity and data quality.

Purpose of the Study:

  • To design and fabricate novel tools for manipulating and organizing cryo-EM sample grids.
  • To improve the efficiency and accessibility of sample handling during cryo-EM workflows.
  • To provide open-access blueprints for custom-made laboratory tools.

Main Methods:

  • 3D printing (fused deposition modeling) of plastic fiber tools.
Keywords:
3D printerCRYO ARMCryo-EMGrid caseSingle-particle analysisSurface polishing

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  • Acetone vapor polishing for tool surface finishing.
  • Development of a grid case stacker for organizing multiple cryo-EM grids.
  • Design of tools for handling grid cases under liquid nitrogen and within electron microscopes.

    • Main Results:

    • Successfully designed and fabricated functional tools, named CryoGridTools.
    • Developed a grid case stacker that fits within standard laboratory consumables (50 mL centrifuge tubes).
    • Created specialized tools for safe and efficient manipulation of cryo-EM grids in cryogenic environments.
    • Made blueprints for CryoGridTools publicly available on GitHub.

    Conclusions:

    • The developed CryoGridTools significantly enhance the manipulation and organization of cryo-EM sample grids.
    • These tools facilitate high-throughput screening and structure determination by improving workflow efficiency.
    • The open-access nature of the tool designs promotes wider adoption and customization in cryo-EM laboratories.