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

流式细胞仪（FC）是一种“快乐的”，但它是一种非常强大的工具，具有独特的功能，需要对细胞的物理和化学特性感兴趣的实验室使用。它提供了多种物理特性的信息和定量数据，例如单个细胞/生物颗粒的大小和粒度，小到100纳米到几微米。根据来自染料或单克隆抗体的光散射和荧光发射特征，可以在短时间内对细胞/生物颗粒的膜结合或细胞内分子进行分类、计数和分析。除了全细胞分析，细胞器、细胞核、DNA、RNA、染色体、细胞因子、激素和蛋白质含量等细胞成分也可以通过流式细胞术独立研究。细胞增殖、分化和细胞周期动力学分析，包括钙通量和膜电位的测量，是其他常用的流式细胞术方法的例子。有趣的是，流式细胞术分析可以补充，有时甚至取代qPCR、荧光显微镜、ELISA和Western blotting等技术，快速产生大量单个细胞和生物颗粒的统计可靠的多参数数据。这些功能允许FC在我们研究的各种环境中使用。因此，在这项研究工具和仪器拨款申请的框架下，我们要求获得一个FC来支持和维持我们在再生医学、细胞和分子治疗以及纳米医学方面的研究。总体而言，pi的研究涉及生物材料、生物界面、用于药物输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、作为单囊泡水平新疗法合并的细胞外囊泡分子谱、芯片实验室设备的开发，以及影响骨骼成熟和衰老的机械环境的理解。所有项目都涉及广泛的体外研究，如细胞免疫分型和分选、细胞活动（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，申请人必须受益于易于使用和用户友好的流式细胞仪，以保持和增强他们在再生医学，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，在pi的实验室中使用流式细胞仪将为小组成员和学生提供跨学科的培训，在这种培训中，基于问题的学习和团队合作是基础，帮助他们学习跨越跨学科和多学科的边界，从而更好地培养他们未来多样化的职业。

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.