Cryo-TEM infrastructure for the visualisation of nanoparticles, exosomes, and virus-like particles

点击翻译按钮获取中文摘要

Cryogenic transmission electron microscopy (cryo-TEM) instruments and methodologies have developed at an exponential rate in the last two decades, and several microscopy "nodes" equipped with high-resolution and state-of-the-art expensive equipment have been installed. In Canada, these cryo-TEM infrastructures are located in less than 10 universities and research centres, providing analytical services that are crucial for high-resolution proteomics, virology, and extracellular vesicle and nanoparticle microscopy studies. For researchers located in other, geographically remote institutions, access to these instruments is limited and thus problematic. Specifically, the preparation of cryogenic samples for TEM requires time, practice, and optimization, therefore adequate training of highly qualified personnel (HQP; graduate students, postdoctoral fellows, research associates, and researchers) is impossible in such conditions. The absence of a "satellite" cryo-TEM facility at Université Laval (and in all of Eastern Québec, including Québec City) thus has a dramatic impact on the number of cryo-TEM samples that can be effectively prepared by our research teams and subsequently observed in the high-end cryo-TEM nodes. In Canada, U.Laval is an academic hub in virology and infectiology, vision health, regenerative medicine, and nanotechnology applied to the biomedical sciences. Consequently, this absence of a local cryo-TEM infrastructure is extremely problematic. This project aims at purchasing a coherent group of instruments and services which, together, will complete a much-needed facility for cryo-TEM sample preparation and screening at the Centre de Recherche du Centre Hospitalier Universitaire de Québec (CR-CHUQ-UL). A group of six (6) researchers specialized in nanotechnology, virology, vision health, regenerative medicine, and biochemistry will establish a robust workflow focused on the high-resolution preparation and observation of both engineered and biological (extracellular vesicles, viruses, virus-like particles) nanoparticle samples in their biological context. An already installed TEM infrastructure that includes a cryo-holder and a vitrificator will be completed with an ultracryomicrotome capacity, cryogenic transport and storage equipment, and a glow discharge instrument for the preparation of TEM grids. This project will train at least 10 HQP per year on cryo-TEM sample preparation in Eastern Québec which, in turn, will allow the selection of optimal grids for further analysis in the high-resolution cryo-TEM nodes. The infrastructure will be accessible to the academic research community and more precisely through U.Laval's Research Centre for Advanced Materials (CERMA; >100 graduate students), Centre de Recherche en Infectiologie (CRI; >50 graduate students), and the Laboratoire d'Organogénèse Expérimentale (LOEX, >50 graduate students).