A Passaged 3-species Synthetic Microbial Community

We constructed de novo a community of three model bacterial species, Vibrio cholerae, Staphylococcus aureus, and Escherichia coli, which all constitutively produce a distinct fluorescent protein. This fluorescence permitted hourly fluorescence tracking over three daily passages.

Cells were quantified using a 10μm hemocytometer, imaged in a Nikon ELCIPSE Ti microscope, using a combination of brightfield and fluorescence microscopy. The filters used were GFP for green, YFP for yellow, and dsRed for red fluorescent proteins. Counts from four hemocytometer squares were averaged to quantify each replicate.