Preserving Anerobic Cells During Sorting

Preserving Anerobic Cells During Sorting

How Regulatory Genetic Diversity Drives Cellular Physiology

Anaerobic microorganisms are major components in numerous environments ranging from naturally-occurring subsurface ground waters, anaerobic hydrocarbon deposits, and anoxic waters and sediments to man-made environments such as wastewater treatment plants and anaerobic digesters. Defining the community structure and the functional ecology of these microbial communities is a key component of research campaigns within the ENIGMA project. Flow cytometry is a powerful tool for high-throughput single cell analysis and sorting of targeted individual cells or populations, however preserving the viability of anaerobic cells during cell sorting has remained a challenge. We are developing a technology, called Live Anaerobic Cell Sorting (LAnCS), to address this challenge. Modifications made to the BD Influx high-speed cell sorter create an enclosed path cell sorter that is capable of maintaining an anoxic environment from sample to sort collection tube. We tested LAnCS on a synthetic anaerobic community composed of two organisms and demonstrate that LAnCS was successful in preserving the viability of both members of this anaerobic community, Desulfovibrio vulgaris Hildenborough (DvH) and Methanococcus maripaludis (Mmp), by growing colonies from single-cell sorts of both organisms on agar plates.