MRI: Acquisition of an Imaging Flow Cytometer for Multidisciplinary Organic and Inorganic Particle Research and Education

该项目的主要目的是促进国家的最先进的流式细胞仪与成像和荧光触发能力，这将是位于密西西比大学（UM），生物系的收购和使用。流式细胞仪和显微镜，或FlowCAM，利用高分辨率CCD相机，多个物镜和图像分析软件，允许计数，测量和图像捕获的单个悬浮的有机和无机颗粒的大小范围从0.002到2毫米。该仪器是为浮游生物评估而设计的，但它可用于这一尺寸范围内的任何流体悬浮颗粒。因此，除了浮游生物生态学研究外，该仪器的计划用途还包括共生、真菌学、生物地球化学、水文学和野生生物学研究。FlowCAM的主要特点是前所未有的样品处理速度（每分钟高达10，000个颗粒）和为每个颗粒生成数字参考图像。处理速度提高了群落数据的统计鲁棒性，自动化导致浮游生物识别和计数中的人类偏见显着减少。由于基于显微镜的群落分析往往是浮游生物研究的限制因素，这一工具将使调查人员能够在前所未有的空间和时间尺度上解决与生态有关的问题，包括实时监测浮游生物动态，如水生污染事件和有害藻类大量繁殖。基于Web的数据访问门户将使远程用户能够快速接收发送到UM的样品的数据。如果科学家能够轻松识别、计数和记录微观物体，如单细胞藻类、真菌孢子和原生动物寄生虫，那么生物学中的许多科学问题都可以得到更快速、更有效的回答; FlowCAM将做到这一点。 该工具的多学科性质将鼓励不同的科学家群体使用，大大扩展UM现有的研究和培训能力，并鼓励整个地区的研究人员之间的合作。该工具支持的项目将包括评估藻类在生态系统服务中的作用，例如受农业影响的溪流中的营养保留;测量入侵的、以浮游生物为食的亚洲鲤鱼对密西西比河流域本地鱼类食物资源的影响;将寄生虫负载与野生火鸡的交配选择联系起来;了解菌根和珊瑚-藻类共生中的共生关系和共同进化;并了解本地和入侵贻贝幼虫的生存和分布模式。图像数据库将通过Morphbank（一个开放的生物图像网络数据库）供公众使用，详细的社区数据库通过生物复杂性知识网络共享。PI将为高中到大学的夏桥项目提供教育示范和研究机会，该项目旨在吸引少数民族学生进入数学、科学和工程领域。在大学一级，该仪器将被纳入几个实验室课程（水生生物学、微生物学原理、湖沼学方法），并将用于教师监督的独立研究项目（例如，在生物科学指导研究，夏季REU计划）。研究数据将在主要科学文献中传播。

The main purpose of this project is to facilitate acquisition and use of a state-of-the-art flow cytometer with imaging and fluorescence triggering capabilities, which will be located at the University of Mississippi (UM), Department of Biology. The Flow Cytometer and Microscope, or FlowCAM, utilizes a high resolution CCD camera, multiple objectives, and image analysis software to allow enumeration, measurement, and image capture of individual suspended organic and inorganic particles ranging from 0.002 to 2 millimeters in size. This instrument was designed for plankton assessment, but it can be used for any fluid-suspended particle in this size range. Therefore, in addition to studies in plankton ecology, planned uses of the instrument include research in symbiosis, mycology, biogeochemistry, hydrology, and wildlife biology. The key features of the FlowCAM are an unprecedented speed of sample processing (up to 10,000 particles per minute) and production of a digital reference image for each particle. The processing speed improves statistical robustness of community data, and the automation leads to a significant reduction in human bias in plankton identification and enumeration. As microscope-based community analysis is often the limiting factor in plankton research, this instrument will allow the investigators to address ecologically relevant questions at a spatial and temporal scale not before possible, including real-time monitoring of plankton dynamics such as aquatic pollution events and harmful algal blooms. A web-based data access portal will enable remote users to quickly receive data from samples sent to UM. Many scientific questions in biology could be more rapidly and efficiently answered if scientists could easily identify, count, and document microscopic objects such as unicellular algae, fungal spores, and protozoan parasites; the FlowCAM will do just that. The multi-disciplinary nature of this instrument will encourage use by a diverse group of scientists, greatly expand UM existing research and training capabilities, and encourage collaborations among researchers across the region. Projects supported by the instrument will include assessing algal roles in ecosystem services such as nutrient retention in agriculturally influenced streams; measuring the impact of the invasive, planktivorous Asian carp on the food resources of native fish species in the Mississippi River Basin; relating parasite load to mate choice in wild turkeys; understanding symbiotic relationships and co-evolution in mycorrhizal and coral-algal symbioses; and understanding survival and distribution patterns of native and invasive mussel larvae. Image databases will be made accessible for public use through Morphbank, an open web database of biological images, and detailed community databases shared through the Knowledge Network for Biocomplexity. The PIs will provide educational demonstrations and research opportunities for a high school-to-college summer bridge program designed to attract minority students to the fields of math, science, and engineering. At the university level, the instrument will be integrated into several laboratory-based courses (Aquatic Biology, Principles of Microbiology, Limnological Methods) and will be used in faculty-supervised independent research projects (e.g., Directed Study in Biological Sciences, summer REU programs). Research data will be disseminated in the primary scientific literature.