VitroJet: new features and case studies

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

doi:10.1107/s2059798324001852

Title: VitroJet: new features and case studies

Journal Article · 14 March 2024 · Acta Crystallographica. Section D. Structural Biology

DOI: https://doi.org/10.1107/s2059798324001852 · OSTI ID:2471651

Henderikx, Rene M. ^[1];

^[2]; Domanska, Aušra ^[3]; Dong, Jing ^[4]; Shahzad, Saba ^[2]; Lak, Behnam ^[3]; Filopoulou, Aikaterini ^[2];

^[5]; Grininger, Martin ^[5]; Momoh, Jeffrey ^[6]; Laanto, Elina ^[7];

^[3];

^[4];

^[3];

^[8]; Beulen, Bart W. A. M. M. ^[9]

CryoSol-World, Weert (Netherlands); Maastricht University (Netherlands)
Forschungszentrum Juelich (Germany)
Univ. of Helsinki (Finland)
Astex Pharmaceuticals (United Kingdom)
Goethe Univ., Frankfurt (Germany)
RWTH Aachen Univ. (Germany)
Univ. of Jyvaskyla (Finland)
Maastricht University (Netherlands)
CryoSol-World, Weert (Netherlands)

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.

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Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States). Environmental Molecular Sciences Laboratory (EMSL)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC05-76RL01830

OSTI ID:: 2471651

Journal Information:: Acta Crystallographica. Section D. Structural Biology, Journal Name: Acta Crystallographica. Section D. Structural Biology Journal Issue: 4 Vol. 80; ISSN 2059-7983

Publisher:: International Union of CrystallographyCopyright Statement

Country of Publication:: United States

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
VitroJet
cryo-EM
ice thickness
jet vitrification
pin printing

Title: VitroJet: new features and case studies

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