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

多参数流式细胞仪（MPFC）系统是一种专门类型的流式细胞仪，是培训高素质人员（HQP）的重要组成部分，他们参与细胞，微生物或藻类培养的实验模型。达尔豪西大学农业校区（Dal-AC）位于NS的特鲁罗，没有MPFC系统，HQP必须将活的人类细胞（生物安全风险组2）运送到哈利法克斯校区（100多公里），才能使用流式细胞仪设施。因此，在Dal-AC安装MPFC系统不仅可以加强许多HQP的实践培训，还可以满足加拿大公共卫生署的安全要求。** MPFC系统允许测量哺乳动物和微生物细胞照明诱导的多种荧光和光散射。MPFC的独特之处在于它可以同时测量培养细胞的多个结构和功能参数。该系统对于我们正在进行的癌症化学保护功能和膳食类黄酮及其代谢物机制的研究至关重要（NSERC Discovery Grant 2016-2021），通过测量人类正常细胞的DNA损伤信号传导和DNA修复机制。具体测量包括细胞活力和细胞凋亡导致的细胞死亡;细胞计数、细胞增殖、细胞周期和DNA含量;细胞膜完整性和过氧化;细胞溶质活性氧; DNA双链断裂和信号传导;基因表达和半胱天冬酶激活。该设备还将用于研究与急性咽炎有关的膳食类黄酮抗炎功能的分子基础，通过定量激活的扁桃体上皮细胞和巨噬细胞中的各种细胞因子。其他研究人员将使用MPFC系统来发现细菌中核黄素的合成（S。Yurgel）和为生物燃料开发设计的生物反应器中生长的微藻的脂质含量（S. He）。** 在加拿大，与饮食直接或间接相关的氧化应激相关慢性疾病流行病正面临重大挑战。科学证据表明，富含水果和蔬菜的饮食可以大大降低患多种癌症的风险。具有抗氧化和细胞信号调节特性的膳食类黄酮现在被认为是水果中的关键成分，它提供了超越基本营养功能的特定健康益处。这种拟议的设备将大大推进我们对膳食类黄酮及其代谢物如何有助于降低慢性疾病风险的分子基础的理解，例如保护健康细胞免受致癌因素造成的DNA损伤。因此，这些基本知识将有助于我们为加拿大人开发有效的癌症化学保护饮食策略和新的功能性食品和天然健康产品。**

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. **