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

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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.