Intellectual Merit: Understanding the abundance, diversity and activity of microbes over various time and space scales in terrestrial, freshwater and marine ecosystems is essential to understanding and addressing critical issues such as climate change, water quality, ecosystem conservation and human and environmental health. In the context of environmental microbiology research, flow cytometry offers distinct advantages over other techniques including (1) ability to count and characterize large numbers of cells in short times (~50,000 cells/sec), (2) ability to simultaneously measure multiple properties (size, color or specific genetic (DNA/RNA) or protein constituents using probes) of individual cells and therefore populations and (3) ability to sort and collect individual cells or populations of cells for downstream analyses. Combined, these capabilities make flow cytometry the method of choice to characterize microbial populations or mixtures of cells from higher organisms. A recently developed high-speed sorting flow cytometer for multi-user environmental microbial research will be acquired. Research to be enabled by this instrument includes enumeration and genetic and biochemical characterization of (1) marine microbes including both phytoplankton and bacteria (Archaea and Eubacteria) (2) marine and freshwater microbes in the context of environmental pollutants including nanoparticles, antibiotics, and bio-solid contaminants including triclosan (a bactericide found in personal care products), nonylphenol (a detergent/surfactant), naproxen (pharmaceutically active compound a.k.a. Aleve) and brominated flame retardants (BFRs) among others (3) cells harvested from higher organisms of interest to conservation (e.g. fishes, turtles, corals) and (4) algae for the renewable and carbon-neutral production of biofuels and animal protein. The instrument, which will be unique at Duke University and in rural eastern North Carolina, will be located at the Duke University Marine Laboratory, and will be available as part of a core-molecular biology recharge facility to researchers at both Duke University and other near-by institutions (e.g. NOAA, University of North Carolina Institute for Marine Sciences (UNC-IMS) and North Carolina State University Center for Marine Sciences and Technologies). Rapid advances now occurring in molecular- and micro-biology, marine and freshwater microbial ecology, and flow cytometric technology will allow this instrument to facilitate major advances in oceanography, environmental engineering and environmental microbial.
Broader Impacts: Acquisition of this instrument will enable a satellite campus of Duke University and other satellite campuses of eastern North Carolina universities to perform modern environmental microbiology research and to be more competitive for federal, state, local and privately funded research. Beyond developing research infrastructure, the instrument facility will be integral to teaching at Duke University and neighboring institutions including UNC-IMS and members of the NC Marine Sciences Education Consortium (MSEC), allowing incorporation of cutting edge techniques in undergraduate, professional masters and graduate student education. A preliminary list of undergraduate and graduate courses at Duke University that will directly benefit from this facility includes Biological Oceanography, Genetics and Molecular Biology, Environmental Molecular Tools, and Analysis of Ocean Ecosystems among others. The facility will also be made available for undergraduate independent study research as well as thesis research by Masters students.