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

用于纳米颗粒、外泌体和病毒样颗粒可视化的冷冻 TEM 基础设施

• 批准号: RTI-2023-00202
• 负责人: Fortin, MarcAndré
• 金额: $ 10.93万
• 依托单位: Université Laval
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=746653
• 关键词: Cryo; TEM; infrastructure; visualisation; nanoparticles

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).

在过去的二十年里，低温透射电子显微镜（cryo-TEM）仪器和方法以指数级速度发展，并且已经安装了几个配备高分辨率和最先进的昂贵设备的显微镜“节点”。在加拿大，这些cryo-TEM基础设施位于不到10所大学和研究中心，为高分辨率蛋白质组学、病毒学、细胞外囊泡和纳米颗粒显微镜研究提供至关重要的分析服务。对于位于其他地理位置偏远的机构的研究人员来说，使用这些仪器是有限的，因此是有问题的。具体而言，TEM低温样品的制备需要时间，实践和优化，因此在这种条件下，高素质人员（HQP;研究生，博士后研究员，研究员和研究人员）的充分培训是不可能的。因此，拉瓦尔大学（以及魁北克省东部的所有地区，包括魁北克市）缺乏“卫星”低温TEM设施，这对我们的研究团队可以有效制备并随后在高端低温TEM节点中观察到的低温TEM样品的数量产生了巨大影响。在加拿大，U.拉瓦尔是病毒学和感染学，视力健康，再生医学和应用于生物医学科学的纳米技术的学术中心。因此，缺乏局部低温TEM基础设施是非常成问题的。该项目旨在购买一组连贯的仪器和服务，这些仪器和服务将共同完成魁北克大学中心医院研究中心（CR-CHIQ-UL）急需的低温TEM样品制备和筛选设施。由六（6）名专门从事纳米技术、病毒学、视觉健康、再生医学和生物化学的研究人员组成的小组将建立一个强大的工作流程，重点关注工程和生物（细胞外囊泡、病毒、病毒样颗粒）的高分辨率制备和观察。）纳米颗粒样品在其生物背景下。一个已经安装的TEM基础设施，包括一个低温保持器和一个玻璃化器将完成一个ultracyomicrotome能力，低温运输和存储设备，和辉光放电仪器的TEM网格的准备。该项目每年将在魁北克东部培训至少10名HQP进行低温TEM样品制备，这反过来将允许选择最佳网格，以便在高分辨率低温TEM节点中进行进一步分析。该基础设施将提供给学术研究界，更准确地说，是通过U.拉瓦尔的先进材料研究中心（CERMA; >100名研究生）、生物学研究中心（CRI; >50名研究生）和实验有机材料实验室（LOEX，>50名研究生）。