Flow cytometry for high throughput microbial experimentation

Rob Ness教授和Steven Short教授正在请求支持一种用于微生物细胞及其相关病毒的精确表型的流式细胞仪。内斯教授的研究项目结合了实地采样、实验进化和基因组学，以了解在基因水平上推动进化的力量。肖特教授研究了自然水生群落中宿主与其专门化病毒之间的复杂相互作用。这两个项目的共同点是它们都把藻类作为模式生物。藻类对生态系统服务至关重要，产生的氧气占地球总量的50%以上。此外，藻类作为实验室生物具有优势，因为它们体积小，便于进行基因操作，生长迅速，并可进行高通量实验。我们正在寻找微生物研究高通量诊断的关键基础设施元素--流式细胞仪。流式细胞仪将细胞流通过激光光束，光束与细胞的相互作用被用来在几分钟内描述数以万计的单个细胞的详细特征。利用流式细胞仪，奈斯教授将把自然发生的突变的适合度效应与这些突变改变的基因之间的相互联系联系起来。只有使用高通量的流式细胞仪才能精确测量数千个突变系的生长率。内斯教授还将使用流式细胞仪测量藻类产生可转化为生物燃料的脂肪的能力。利用流式细胞仪，奈斯教授将研究天然线条在脂肪产生方面如何以及为什么会发生变化，以及是什么基因导致了这种变化。流式细胞术一直是Short教授研究的关键工具，他在病毒裂解实验中通过精确的藻细胞计数和表型鉴定从淡水中分离出病毒。未来的实验计划对Short教授和他的HQP最近发现的新型藻类病毒宿主联合体进行表征，并需要一台现代化的高通量流式细胞仪。该仪器将用于对组合实验中产生的细胞和病毒进行快速表型和计数，这些组合实验旨在梳理环境条件与竞争同一宿主藻类的多个病毒和卫星病毒的相互作用。这项工作的结果可能会改变目前对藻类生态的理解，从而能够建立更准确的水生初级生产力预测模型。如果得到资助，这种流式细胞仪将对Ness和Short教授资助的研究项目的及时执行至关重要，有助于HQP的研究和培训。流式细胞仪将使他们能够利用现有的资金来促进我们对基因组进化和病毒生态学的知识。*****

Professors Rob Ness and Steven Short are requesting support for a flow cytometer for precision phenotyping of microbial cells and their associated viruses. Prof. Ness' research program combines field sampling, experimental evolution and genomics to understand the forces that drive evolution at the genetic level. Prof. Short investigates the complex interplay between hosts and their specialized viruses in natural aquatic communities. The commonality between these two programs is their focus on algae as model organisms. Algae are crucial to ecosystem services and produce over 50% of the earth's oxygen. In addition, algae have advantages as laboratory organisms because their small size facilities genetic manipulation, fast growth, and high-throughput experimentation. We are seeking a key infrastructural element of high-throughput diagnostics for microbial research, a flow cytometer. Flow cytometers pass a stream of cells through a laser beam and the interaction of the beam with the cells is used to describe detailed characteristic of tens of thousands of individual cells in minutes. Using the flow cytometer, Prof. Ness will link how the fitness effects of naturally occurring mutations relate to the interconnection of the genes those mutations alter. Precision measurement of the growth rate of thousands of mutated lines is only possible using a high-throughput flow cytometer. Prof. Ness will also use the flow cytometer to measure algae's ability to produce lipids that can converted into biofuel. Using the flow cytometer, Prof. Ness will investigate how and why natural lines vary in their lipid production and what genes underlie this variation. Flow cytometry has been a key tool for Prof. Short's research has isolated virus from freshwater by precision algal cell enumeration and phenotyping during viral lysis experiments. Future experiments are planned to characterize the novel algal virus-host consortium recently discovered by Prof. Short and his HQP, and require a modern, high-throughput flow cytometer. The instrument will be used to rapidly phenotype and enumerate cells and viruses generated in combinatorial experiments designed to tease apart the interactions of environmental conditions and multiple viruses and satellite viruses competing for the same host alga. The results of this work could transform current understanding of algal ecology enabling more accurate predictive models of aquatic primary production. If funded, this flow cytometer will be critical to the timely execution of Prof. Ness and Short's NSERC funded research programs, contributing to research and training of HQP. The flow cytometer will allow them to capitalize on existing funding to advance our knowledge of genome evolution and viral ecology. *****