Cell vitrification to enable the study of structure and dynamics of macromolecular complexes within their native environment

The project aimed to develop enabling technology for optimal sample preparation of entire cells by jet vitrification in a consistent and automated manner. This technology enables the study of macromolecules within their native cellular environment, which provides fundamental knowledge to develop novel medicines. The VitroJet for Cells technology builds upon extensive experience within Maastricht University on sample preparation for cryo-electron microscopy (cryo-EM) for single particle analysis. Within this project, a next generation device was developed. The expected societal contribution is the enhanced and accelerated development of targeted medicines, rational drugs and structural vaccinology (e.g. to combat antibiotic resistance). The VitroJet for Cells technology enables the fundamental precondition to analyse pathologic vs healthy cells and gain 3D insight in the targets these drugs will act upon. During the project, we performed fundamental research to study smartly chosen parameters that influence cell vitrification and generated optimal preparation protocols. These protocols were validated by executing the entire workflow of cell analysis both by state-of-the-art cryo-electron microscopy (cryo-EM) as well as advanced synchrotron X-ray diffraction techniques. Complementary to this, we performed industrial research and developed a TRL 6 functional model starting from the current basic mathematical model of jet vitrification and we improved this complex system based on the results of the fundamental part. The resulting TRL 6 functional model serves to engage in business development activities such as organising demonstrations, marketing the product and initiating close customer contact. After this project, we want to set up beta-testing collaborations, in which a minimal viable product (TRL 7-8), will be offered to launching customers by CryoSol World BV. The cryo-EM community is very well organized and can easily be reached by our lead partner.