VitroJet: new features and case studies

Rene JM Henderikx, Daniel Mann, Ausra Domanska, Jing Dong, Saba Shahzad, Behnam Lak, Aikaterini Filopoulou, Damian Ludig, Martin Grininger, Jeffrey Momoh, Elina Laanto, Hanna M Oksanen, Kyrylo Bisikalo, Pamela A Williams, Sarah Butcher, Peter J Peters, Bart W.A.M.M. Beulen

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Single-particle cryo-electron microscopy has become a widely adopted method in structural biology due to many recent technological advances in microscopes, detectors and image processing. Before being able to inspect a biological sample in an electron microscope, it needs to be deposited in a thin layer on a grid and rapidly frozen. The VitroJet was designed with this aim, as well as avoiding the delicate manual handling and transfer steps that occur during the conventional grid-preparation process. Since its creation, numerous technical developments have resulted in a device that is now widely utilized in multiple laboratories worldwide. It features plasma treatment, low-volume sample deposition through pin printing, optical ice-thickness measurement and cryofixation of pre-clipped Autogrids through jet vitrification. This paper presents recent technical improvements to the VitroJet and the benefits that it brings to the cryo-EM workflow. A wide variety of applications are shown: membrane proteins, nucleosomes, fatty-acid synthase, Tobacco mosaic virus, lipid nanoparticles, tick-borne encephalitis viruses and bacteriophages. These case studies illustrate the advancement of the VitroJet into an instrument that enables accurate control and reproducibility, demonstrating its suitability for time-efficient cryo-EM structure determination.
Original languageEnglish
JournalActa crystallographica. Section D, Structural biology
Volume80
Issue number4
Pages (from-to)232-246
Number of pages15
ISSN2059-7983
DOIs
Publication statusPublished - 2024
MoE publication typeA1 Journal article-refereed

Fields of Science

  • 1182 Biochemistry, cell and molecular biology
  • 11832 Microbiology and virology

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