VitroJet : new features and case studies
Henderikx, R. J. M., Mann, D., Domanska, A., Dong, J., Shahzad, S., Lak, B., Filopoulou, A., Ludig, D., Grininger, M., Momoh, J., Laanto, E., Oksanen, H. M., Bisikalo, K., Williams, P. A., Butcher, S. J., Peters, P. J., & Beulen, B. W. A. M.M. (2024). VitroJet : new features and case studies. Acta Crystallographica Section D : Structural Biology, 80(4). https://doi.org/10.1107/s2059798324001852
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2024Copyright
© 2024 the Authors
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, tickborne 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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International Union of Crystallography (IUCr)ISSN Search the Publication Forum
2059-7983Keywords
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https://converis.jyu.fi/converis/portal/detail/Publication/207698934
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This research was funded by the Swedish Research Council, reference No. 2018-05851 (to SJB), and by the European Research Council (ERC) under the European Union’s Horizon Europe research and innovation programme, grant agreement No. 10117204 (to EL). A portion of this research was supported by NIH grant U24GM129547, performed at the PNCC at OHSU and accessed through EMSL (grid.436923.9), a DOE Office of Science User Facility sponsored by the Office of Biological and Environmental Research. ...License
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