Flow cytometry for high throughput microbial experimentation

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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. *****