Nanaerobic growth enables direct visualization of dynamic cellular processes in human gut symbionts
Leonor García-Bayona, Michael J. Coyne, Noam Hantman, Paula Montero Llopis, Salena S Von, Takeshi Ito, Michael H. Malamy, Marek Basler, Blanca Barquera, Laurie E. Comstock
Abstract
Significance Despite recent technological advances to study the human gut microbiota, we still lack a facile system to image dynamic cellular processes in most abundant gut species due to the requirement of oxygen for chromophore maturation of commonly used fluorescent proteins. Here, we took advantage of the ability of anaerobes of the gut microbiota to respire aerobically and grow robustly at 0.10 to 0.14% oxygen. This physiologic concentration of oxygen is sufficient for fluorescent proteins to mature, allowing for visualization of biological processes never before imaged in these bacteria. This advance will allow for numerous types of analyses in actively growing “nanaerobic” gut bacteria including subcellular protein localizations, single-cell analyses, biofilm imaging, and protein interactions with other microbes and the host.