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）是一个“欢乐”，但具有非常强大的工具，具有独特的功能，需要对细胞的物理和化学表征感兴趣的实验室可以使用。它提供了有关多种物理特征的信息和定量数据，例如单个细胞/生物颗粒的大小和粒度，至少100粒微米。取决于光散射和荧光发射特征，这些特征源自所研究的细胞/生物粒子的染料或单克隆抗体，靶向膜结合的膜或细胞内分子，可以在短时间内对，计算和分析复杂的种群。除全细胞分析外，细胞组成部分，例如细胞器，核，DNA，RNA，染色体，细胞因子，激素和蛋白质含量，也可以通过流式细胞仪独立研究。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，是用于流式细胞仪开发的其他常用方法的示例。有趣的是，流式细胞术分析可以补充，有时会取代QPCR，荧光显微镜，ELISA和Western印迹等技术，从而迅速产生统计上强大的，有关大量单个细胞和生物颗粒的多参数数据。这些功能允许FC在我们的研究的各种情况下使用。因此，在此研究工具和工具授予应用程序的框架内，我们要求获得FC，以支持和维持我们在再生医学，基于细胞和分子的疗法以及纳米医学方面的研究。总体而言，PIS的研究涉及生物材料的发展，生物界面，用于药物输送的工程纳米纳经，用于基因组编辑的非病毒纳米颗粒系统，在单个囊泡级别的环境中，在单个囊泡级别和实验室环境中合并的细胞外囊泡的分子谱图，并与该机制相动。所有项目都涉及广泛的体外研究，例如细胞免疫表型和分选，评估细胞活性（增殖，分化，凋亡），细胞因子以及基因和蛋白质表达。由于在校园和附属医院获得少数足球俱乐部的机会有限，因此在此阶段，至关重要的是，申请人从可访问且用户友好的流式细胞仪中受益，以维持和增强其在再生医学，纳米医学和诊断方面的创新研究能力。除了提高我们的研究能力外，PIS实验室中的流式细胞仪的可用性还将为小组成员和学生提供跨学科培训，其中基于问题的学习和团队合作被脚手架，以帮助他们学会跨越跨越的多学科边界，从而更好地培训他们为多元化的未来职业培训。