Self-propelled motion is an agnostic biosignature that is observed widely, yet motility of microbes in their natural environments is sparsely studied. In this study we use a Digital Holographic Microscope (DHM) for in situ imaging of aquatic samples in extreme environments to investigate motility and morphology as biosignatures. Samples were collected from glaciovolcanic ice caves, glacial runoff, hot springs, and mixed glacial and hot spring samples.The transport and deposition of materials and heat from the volcanic subsurface in glaciovolcanic caves may be similar in some respects to the eruption processes of the plumes of Enceladus. Through different tracking methods, we identified concentrations of organisms, morphologies, swimming patterns, speeds, and turn angles. In every type of sample we looked we were able to identify motile organisms. Methods for distinguishing active swimming from Browian motion and drift are considered. Field work was done over two deployments in collaboration with the Thermal High-voltage Ocean-penetrating Research platform (THOR) science team and EELS robotics team. This work and these collaborations intend to inform future off-world extant life detection missions of the utility of DHM and motility as an investigation tool and biosignature, respectively.