A flow cytometer for investigating functions and mechanisms of plant-food flavonoids in biological systems

The Multi-parameter Flow Cytometer (MPFC) system, is a specialized type of flow cytometer and is an essential component for the training of highly qualified personnel (HQP), who are involved in experimental models of the cell, microbial or algal cultures. Dalhousie University Agricultural Campus (Dal-AC), which is located in Truro, NS does not have an MPFC system, and HQPs have to transport live human cells (Biosafety Risk Group 2) to Halifax campus (over 100 km) to access a flow cytometry facility. Therefore, the installation of an MPFC system at Dal-AC, will not only enhance the hands-on training of many HQPs but also fulfill the safety requirement of the Public Health Agency of Canada. *The MPFC system allows the measurement of multiple fluorescences and light scatter induced by illumination of mammalian and microbial cells. MPFC is unique in that it provides the simultaneous measurement of multiple structural and functional parameters of cultured cells. The system is vital for our ongoing investigations of cancer chemoprotective functions and mechanisms of dietary flavonoids and their metabolites (NSERC Discovery Grant 2016-2021) by measuring DNA damage signaling and DNA repair mechanisms of human normal cells. The specific measurements include cell viability and cell death by apoptosis; cell enumeration, cell proliferation, cell cycle, and DNA content; cellular membrane integrity and peroxidation; cytosolic reactive oxygen species; DNA double-strand breaks and signaling; gene expression and caspase activation. The equipment will also be used to investigate the molecular basis of anti-inflammatory functions of dietary flavonoids in relation to acute pharyngitis by quantifying various cytokines in activated tonsil epithelial cells and macrophages. Other researchers will use the MPFC system for discovering riboflavin synthesis in bacteria (Dr. S. Yurgel) and lipid content of microalgae grown in bioreactors designed for biofuel development (Dr. S. He). *In Canada, significant challenges are being faced from oxidative stress-linked chronic disease epidemics, which are directly or indirectly linked to diet. Scientific evidence reveals that diets high in fruits and vegetables substantially reduce risks of several cancers. Dietary flavonoids with antioxidant and cell signaling modulating properties are now known as the crucial components in fruits that provide specific health benefits beyond the basic nutritional functions. This proposed equipment will significantly advance our understanding of molecular basis of how dietary flavonoids and their metabolites contribute to reducing the risk of chronic diseases such as the protection of healthy cells against DNA damage due to carcinogenic factors. Thus, this fundamental knowledge will contribute us to develop effective cancer chemoprotective dietary strategies and novel functional foods and natural health products for Canadians. *