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Automated vitrification of cryo-EM samples with controllable sample thickness using suction and real-time optical inspection

Author

Listed:
  • Roman I. Koning

    (Leiden University Medical Center)

  • Hildo Vader

    (Linkam Scientific Instruments Ltd)

  • Martijn Nugteren

    (Linkam Scientific Instruments Ltd)

  • Peter A. Grocutt

    (Linkam Scientific Instruments Ltd)

  • Wen Yang

    (Leiden University)

  • Ludovic L. R. Renault

    (Leiden University)

  • Abraham J. Koster

    (Leiden University Medical Center)

  • Arnold C. F. Kamp

    (Linkam Scientific Instruments Ltd)

  • Michael Schwertner

    (Linkam Scientific Instruments Ltd)

Abstract

The speed and efficiency of data collection and image processing in cryo-electron microscopy have increased over the last decade. However, cryo specimen preparation techniques have lagged and faster, more reproducible specimen preparation devices are needed. Here, we present a vitrification device with highly automated sample handling, requiring only limited user interaction. Moreover, the device allows inspection of thin films using light microscopy, since the excess liquid is removed through suction by tubes, not blotting paper. In combination with dew-point control, this enables thin film preparation in a controlled and reproducible manner. The advantage is that the quality of the prepared cryo specimen is characterized before electron microscopy data acquisition. The practicality and performance of the device are illustrated with experimental results obtained by vitrification of protein suspensions, lipid vesicles, bacterial and human cells, followed by imaged using single particle analysis, cryo-electron tomography, and cryo correlated light and electron microscopy.

Suggested Citation

  • Roman I. Koning & Hildo Vader & Martijn Nugteren & Peter A. Grocutt & Wen Yang & Ludovic L. R. Renault & Abraham J. Koster & Arnold C. F. Kamp & Michael Schwertner, 2022. "Automated vitrification of cryo-EM samples with controllable sample thickness using suction and real-time optical inspection," Nature Communications, Nature, vol. 13(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:13:y:2022:i:1:d:10.1038_s41467-022-30562-7
    DOI: 10.1038/s41467-022-30562-7
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    References listed on IDEAS

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    1. Ka Man Yip & Niels Fischer & Elham Paknia & Ashwin Chari & Holger Stark, 2020. "Atomic-resolution protein structure determination by cryo-EM," Nature, Nature, vol. 587(7832), pages 157-161, November.
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