VitroJet: new features and case studies

Acta Crystallographica Section D Structural Biology, vol. 80, pp. 232–246

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.

Authors 17

1. René Henderikx

   Maastricht University

   Affiliation as printed

   Maastricht Multimodal Molecular Imaging Institute (M4i), Division of Nanoscopy, Maastricht University, Maastricht, The Netherlands

   CryoSol-World, Weert, The Netherlands

2. Daniel Mann

   Forschungszentrum Jülich · Ernst Ruska Centre

   Affiliation as printed

   Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C-3): Structural Biology, Forschungszentrum Jülich, Jülich, Germany

   Institute of Biological Information Processing (IBI-6): Structural Cell Biology, Forschungszentrum Jülich, Jülich, Germany

3. Aušra Domanska

   University of Helsinki

   Affiliation as printed

   Molecular and Integrative Bioscience Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland

   Helsinki Life Science Institute–Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland

4. Jing Dong
   Affiliation as printed

   Astex Pharmaceuticals, 436 Cambridge Science Park, Milton Road, Cambridge CB4 0QA, United Kingdom

5. Saba Shahzad

   Forschungszentrum Jülich · Ernst Ruska Centre

   Affiliation as printed

   Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C-3): Structural Biology, Forschungszentrum Jülich, Jülich, Germany

   Institute of Biological Information Processing (IBI-6): Structural Cell Biology, Forschungszentrum Jülich, Jülich, Germany

6. Behnam Lak

   University of Helsinki

   Affiliation as printed

   Molecular and Integrative Bioscience Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland

   Helsinki Life Science Institute–Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland

7. Aikaterini Filopoulou

   Forschungszentrum Jülich · Ernst Ruska Centre

   Affiliation as printed

   Institute of Biological Information Processing (IBI-6): Structural Cell Biology, Forschungszentrum Jülich, Jülich, Germany

   Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons (ER-C-3): Structural Biology, Forschungszentrum Jülich, Jülich, Germany

8. Damian L. Ludig

   Goethe University Frankfurt

   Affiliation as printed

   Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt am Main, Germany

9. Martin Grininger

   Goethe University Frankfurt

   Affiliation as printed

   Institute of Organic Chemistry and Chemical Biology, Goethe University Frankfurt, Frankfurt am Main, Germany

10. Jeffrey Momoh Aachen Department of Nanomedicine and Theranostics Institute for Experimental Molecular Imaging

    RWTH Aachen University

    Affiliation as printed

    Department of Nanomedicine and Theranostics, Institute for Experimental Molecular Imaging, RWTH Aachen University, Aachen, Germany

11. Elina Laanto

    University of Jyväskylä

    Affiliation as printed

    Department of Biological and Environmental Science, Nanoscience Center, University of Jyväskylä, 40014 Jyväskylä, Finland

12. Hanna M. Oksanen

    University of Helsinki

    Affiliation as printed

    Molecular and Integrative Bioscience Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland

13. Kyrylo Bisikalo

    University of Helsinki

    Affiliation as printed

    Molecular and Integrative Bioscience Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland

    Helsinki Life Science Institute–Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland

14. Pamela A. Williams
    Affiliation as printed

    Astex Pharmaceuticals, 436 Cambridge Science Park, Milton Road, Cambridge CB4 0QA, United Kingdom

15. Sarah J. Butcher

    University of Helsinki

    Affiliation as printed

    Molecular and Integrative Bioscience Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland

    Helsinki Life Science Institute–Institute of Biotechnology, University of Helsinki, 00014 Helsinki, Finland

16. Peter J. Peters

    Maastricht University

    Affiliation as printed

    Maastricht Multimodal Molecular Imaging Institute (M4i), Division of Nanoscopy, Maastricht University, Maastricht, The Netherlands

17. Bart W. A. M. M. Beulen
    Affiliation as printed

    CryoSol-World, Weert, The Netherlands