Real-Time Measurement of the Liquid Amount in Cryo-Electron Microscopy Grids Using Laser Diffraction of Regular 2-D Holes of the Grids

Jinsook Ahn, Dukwon Lee, Inseong Jo, Hyeongseop Jeong, Jae Kyung Hyun, Jae Sung Woo, Sang Ho Choi, Nam Chul Ha

Research output: Contribution to journalArticlepeer-review

Abstract

Cryo-electron microscopy (cryo-EM) is now the first choice to determine the high-resolution structures of huge protein complexes. Grids with two-dimensional arrays of holes covered with a carbon film are typically used in cryo-EM. Although semi-automatic plungers are available, notable trial-and-error is still required to obtain a suitable grid specimen. Herein, we introduce a new method to obtain thin ice specimens using real-time measurement of the liquid amounts in cryo-EM grids. The grids for cryo-EM strongly diffracted laser light, and the diffraction intensity of each spot was measurable in real-time. The measured diffraction patterns represented the states of the liquid in the holes due to the curvature of the liquid around them. Using the diffraction patterns, the optimal time point for freezing the grids for cryo-EM was obtained in real-time. This development will help researchers rapidly determine highresolution protein structures using the limited resource of cryo-EM instrument access.

Original languageEnglish
Pages (from-to)298-303
Number of pages6
JournalMolecules and cells
Volume43
Issue number3
DOIs
Publication statusPublished - 2020 Mar 31

Keywords

  • cryo electron microscopy
  • grid preparation
  • laser diffraction
  • real-time measurement
  • vitreous ice

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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