University of Regina Cellular Impacts Facility Microscopes and Cell Culture

里贾纳大学细胞影响设施显微镜和细胞培养

• 批准号: RTI-2023-00561
• 负责人: Dahms, Tanya
• 金额: $ 10.93万
• 依托单位: University of Regina
• 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=748446
• 关键词: University; Regina; Cellular; Impacts; Facility

Cell sorting and microscopy are well established methods for understanding the physiology of cellular systems, their response to xenobiotics and host-pathogen interactions, shared challenges of the applicants. The UR Cell Impacts Facility (CIF) in the Institute for Environmental Change and Society houses a platform for these studies. For example, the study of host-pathogen interactions first requires BL2 pathogens and host cells to be cultured, pathogen or host cells to be assessed/separated based on biomarkers using a high end flow cytometer/cell sorter, and finally host-pathogen interactions imaged (atomic force, confocal, fluorescence microscopies) of as a function of phenotypic and physiological changes. The proposed components and equipment would restore to full function this shared platform, essential to our NSERC-funded research programs, comprehensive HQP training, in-house and external collaborations and for retaining our national and international reputations. As the CIF equipment ages, a number of the instruments associated with this platform are in need of upgrading or replacement. The BL2 labs used to prepare samples for cell sorting/flow cytometry, mass spectral analysis and microscopy, are currently inoperable, necessitating an incubator-shaker for fungal cell culture and a microscope to assess human cell viability, confluence and quality. Validation of host cell status (cytometry) and enrichment of infected host cells (sorting) based on fluorescent biomarkers is not currently possible, requiring a new 488 nm main laser line to operate, and a 405 nm laser that excites dyes in the violet-blue. Host-pathogen interactions are then screened by fluorescence microscopy followed by confocal or correlative imaging to produce high-resolution 3D publication quality images of their interaction including the subcellular co-localization of labeled protein targets. The Zeiss 780 laser scanning confocal microscope currently has five continuous wave laser lines (458, 488, 514, 543, 594 nm) and a 63 x objective, precluding ultra-high resolution (100 x) imaging of dyes that excite in the violet-blue (405 nm). The research projects of our HQP are currently limited without a fully functioning platform, with certain experiments at a standstill and others having to be modified accordingly, ultimately slowing progress. The ability to use this platform to its full capacity is thus critically required to enable the co-applicants and their HQP to address ambitious research questions. Our applicant team and HQP emerge from a range of backgrounds that is reflective of current Canadian diversity, fueling our creativity, discovery and innovation. The equipment upgrades and replacements are indispensable tools that will allow this platform to operate at maximum capacity for many years, supporting research programs that impact both environmental and health sciences.

细胞分选和显微镜检查是用于理解细胞系统的生理学、它们对外源性物质的响应和宿主-病原体相互作用的成熟方法，这是申请人的共同挑战。环境变化和社会研究所的UR细胞影响设施（CIF）为这些研究提供了一个平台。例如，宿主-病原体相互作用的研究首先需要培养BL 2病原体和宿主细胞，使用高端流式细胞仪/细胞分选仪基于生物标志物评估/分离病原体或宿主细胞，最后根据表型和生理变化成像（原子力、共聚焦、荧光显微镜）宿主-病原体相互作用。拟议的组件和设备将恢复这个共享平台的全部功能，这对我们的NSERC资助的研究计划，全面的HQP培训，内部和外部合作以及保持我们的国家和国际声誉至关重要。 随着CIF设备的老化，与该平台相关的许多仪器需要升级或更换。用于制备细胞分选/流式细胞术、质谱分析和显微镜检查样品的BL 2实验室目前无法使用，需要一个用于真菌细胞培养的培养箱-摇床和一个用于评估人类细胞活力、汇合度和质量的显微镜。基于荧光生物标志物的宿主细胞状态验证（细胞计数）和感染宿主细胞富集（分选）目前尚不可能，需要新的488 nm主激光线运行，以及激发紫蓝色染料的405 nm激光。然后通过荧光显微镜筛选宿主-病原体相互作用，然后进行共聚焦或相关成像，以产生其相互作用的高分辨率3D出版质量图像，包括标记的蛋白质靶标的亚细胞共定位。蔡司780激光扫描共聚焦显微镜目前有五个连续波激光线（458，488，514，543，594 nm）和一个63倍物镜，排除了超高分辨率（100倍）成像的染料激发的紫蓝色（405 nm）。 我们的HQP的研究项目目前受到限制，没有一个功能齐全的平台，某些实验处于停顿状态，其他实验必须进行相应的修改，最终减缓了进展。因此，充分利用这个平台的能力是至关重要的，以使共同申请人及其HQP能够解决雄心勃勃的研究问题。 我们的申请人团队和HQP来自一系列背景，反映了当前加拿大的多样性，激发了我们的创造力，发现和创新。设备升级和更换是必不可少的工具，使该平台能够以最大容量运行多年，支持影响环境和健康科学的研究项目。