Microfluidics Microbial Activity MicroAssay: An Automated In Situ Microbial Metabolic Detection System

With no direct extant-life detection instrumentation included in a space mission since the 1970s, the advancement of new technologies to be included in future space missions is imperative. We developed, optimized, and tested a semi-automated prototype, the microfluidics Microbial Activity MicroAssay (μMAMA). This system metabolically characterizes and detects extant microbial life by way of metabolism-indicator redox dyes. We first evaluated the robustness and sensitivity of six redox dye/buffer combinations, and we then tested their responses to metabolic activity in astrobiological analog high-Arctic samples. We determined that the Biolog Inoculating Fluid (IF)-C and AlamarBlue buffered in IF-0a (aB-IF0a) dye/buffer combinations were optimal, as they detected metabolic activity from the fewest microbial cells (10² cells/mL) while maintaining efficacy over a broad physiochemical range of pH (0-13), temperature (-10°C to 37°C), salinity and perchlorate (tested up to 30%), and in the presence of a Mars regolith simulant (MMS-2). The μMAMA, which incorporated these redox dyes, detected extant active cold-adapted microbial life from high Arctic analog sites, including samples amended with substrates targeting chemolithoautotrophic metabolisms. Given μMAMA's small size (we estimate a complete planetary instrument could occupy as little as 3 L) and potential for automation, it could easily be incorporated into almost any landed platform for life detection missions.