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，是培养参与细胞、微生物或藻类培养实验模型的高素质人员（HQP）的重要组成部分。位于特鲁罗的达尔豪斯大学农业校区（Dal-AC）没有MPFC系统，HQPs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（超过100公里）以使用流式细胞仪设备。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系统允许测量由哺乳动物和微生物细胞的照明引起的多重荧光和光散射。MPFC的独特之处在于它可以同时测量培养细胞的多种结构和功能参数。该系统对于我们正在进行的通过测量人类正常细胞的DNA损伤信号和DNA修复机制来研究膳食类黄酮及其代谢物的癌症化学保护功能和机制（NSERC Discovery Grant 2016-2021）至关重要。具体指标包括细胞活力和细胞凋亡死亡；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；胞质活性氧；DNA双链断裂和信号传导；基因表达和半胱天冬酶活化。该设备还将用于通过定量激活扁桃体上皮细胞和巨噬细胞中的各种细胞因子来研究膳食黄酮类化合物抗炎作用与急性咽炎的分子基础。其他研究人员将使用MPFC系统来发现细菌中的核黄素合成（S. Yurgel博士）和为生物燃料开发而设计的生物反应器中生长的微藻的脂质含量（S. He博士）。******在加拿大，与氧化应激相关的慢性疾病流行病正面临着重大挑战，这些疾病与饮食直接或间接相关。科学证据表明，多吃水果和蔬菜的饮食可以大大降低患几种癌症的风险。膳食类黄酮具有抗氧化和细胞信号调节特性，现在被认为是水果中除了基本营养功能外，还提供特定健康益处的关键成分。这一提议的设备将大大提高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，如保护健康细胞免受致癌因素引起的DNA损伤。因此，这些基础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型功能食品和天然保健品。**

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. **