Quantification of physical and chemical characteristics of cells and bioparticles using flow cytometry

使用流式细胞术量化细胞和生物颗粒的物理和化学特性

• 批准号: RTI-2022-00315
• 负责人: Tabrizian, Maryam
• 金额: $ 4.36万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2021
• 资助国家: 加拿大
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=743053
• 关键词: Quantification; physical; chemical; characteristics; cells

The flow cytometer (FC) is a `convivial', yet a very powerful tool with unique capabilities that need to be accessible to laboratories interested in physical and chemical characterization of cells. It provides information and quantitative data on multiple physical characteristics, such as the size and granularity of a single cell/bioparticle as small as 100 nanometers to a few micrometers. Depending on the light scattering and fluorescence emission features, which are derived from dyes or monoclonal antibodies targeting either membrane-bound or intracellular molecules of the cells/bioparticles under investigation, a complex population can be sorted, counted, and analyzed in a short period of time. In addition to whole cell analysis, cellular components such as organelles, nuclei, DNA, RNA, chromosomes, cytokines, hormones, and protein content can also be independently investigated by flow cytometry. Analysis of cell proliferation, differentiation, and cell cycle dynamics, including measurements of calcium flux and membrane potentials, are the other commonly used examples of methods developed for flow cytometry. Interestingly, flow cytometry analyses can complement, and sometimes replace, techniques such as qPCR, fluorescent microscopy, ELISA, and Western blotting, quickly yielding statistically robust, multiparametric data on large collections of individual cells and bioparticles. These capabilities allow the FC to be used in various contexts of our research. Thus, in the framework of this Research Tools and Instruments Grant application, we request to acquire a FC to support and sustain our research in regenerative medicine, cell- and molecular-based therapies, and nanomedicine. Collectively, the PIs' research involves the development of biomaterials, biointerfaces, engineered nanoplexes for drug delivery, non-viral nanoparticle systems for genome editing, molecular profiling of extracellular vesicles merged as new therapeutics at the single vesicle-level, and lab-on-a-chip devices along with understanding the mechanical environment influencing skeletal maturation and aging. All projects involve extensive in-vitro studies, such as cell immunophenotyping and sorting, assessment of cellular activities (proliferation, differentiation, apoptosis), cytokine, and gene and protein expression. Due to limited access to a handful FC on campus and affiliated hospitals, at this stage, it is crucial that the applicants benefit from an accessible and user-friendly flow cytometer to maintain and enhance their innovative research capacity in regenerative medicine, nanomedicine, and diagnostics. In addition to enhance our research capacity, the availability of a flow cytometer in the PIs' laboratories will provide a transdisciplinary training to group members and students in which problem-based learning and teamwork are scaffolded to help them learn to cross inter- and multidisciplinary borders, thus better training them for diverse future careers.

流式细胞仪（FC）是一种“欢乐”，但一个非常强大的工具，具有独特的能力，需要有兴趣的细胞的物理和化学特性的实验室。它提供关于多种物理特性的信息和定量数据，例如小至100纳米至几微米的单个细胞/生物颗粒的大小和粒度。根据来自染料或单克隆抗体的光散射和荧光发射特征，可以在短时间内对复杂群体进行分选、计数和分析，所述染料或单克隆抗体靶向所研究的细胞/生物颗粒的膜结合或细胞内分子。除全细胞分析外，细胞组分如细胞器、细胞核、DNA、RNA、染色体、细胞因子、激素和蛋白质含量也可通过流式细胞术进行独立研究。细胞增殖、分化和细胞周期动力学的分析，包括钙通量和膜电位的测量，是为流式细胞术开发的方法的其他常用实例。有趣的是，流式细胞术分析可以补充，有时取代，技术，如qPCR，荧光显微镜，ELISA和蛋白质印迹，快速产生统计学上强大的，多参数数据的大量收集的单个细胞和生物颗粒。这些功能允许FC在我们研究的各种背景下使用。因此，在这个研究工具和仪器资助申请的框架内，我们要求获得一个FC，以支持和维持我们在再生医学，细胞和分子为基础的治疗和纳米医学的研究。总的来说，PI的研究涉及开发生物材料，生物界面，用于药物递送的工程纳米复合物，用于基因组编辑的非病毒纳米颗粒系统，在单个囊泡水平上作为新疗法合并的细胞外囊泡的分子分析，以及芯片实验室设备沿着理解影响骨骼成熟和衰老的机械环境。所有项目都涉及广泛的体外研究，如细胞免疫表型和分选，细胞活性（增殖，分化，凋亡），细胞因子，基因和蛋白质表达的评估。由于在校园和附属医院获得少数FC的机会有限，在这个阶段，申请人从一个可访问的和用户友好的流式细胞仪中受益，以保持和提高他们在再生医学，纳米医学和诊断方面的创新研究能力是至关重要的。除了提高我们的研究能力外，PI实验室的流式细胞仪的可用性将为小组成员和学生提供跨学科培训，其中以问题为基础的学习和团队合作是脚手架，以帮助他们学习跨越跨学科和多学科的边界，从而更好地培训他们未来的职业生涯。