Recent advances in digital technologies, such as high-speed computers and large-format digital imagers, have led to a burgeoning interest in the science and engineering of digital holographic microscopy (DHM). Here we report on a novel off-axis DHM, based on a twin-beam optical design, which avoids the limitations of prior systems, and provides many advantages, including compactness, intrinsic stability, robustness against misalignment, ease of use, and cost. These advantages are traded for a physically constrained sample volume, as well as a fixed fringe spacing. The first trade is not overly restrictive for most applications, and the latter provides for a pre-set assembly alignment that optimizes the spatial frequency sampling. Moreover, our new design supports use in both routine laboratory settings as well as extreme environments without any sacrifice in performance, enabling ready observation of microbial species in the field. The instrument design is presented in detail here, along with a demonstration of bacterial video imaging at sub-micrometer resolution at temperatures down to -15 °C. 1268-1270 (1997). 28. S. K. Jericho, P. Klages, J. Nadeau, E. M. Dumas, M. H. Jerico, and J. J. Kreuzer, "In-line digital holographic microscopy for terrestrial and exobiological research," Planet. Space Sci. 58(4), 701-705 (2010). 29.

Sea ice is an analog environment for several of astrobiology’s near-term targets: Mars, Europa, Enceladus, and perhaps other Jovian or Saturnian moons. Microorganisms, both eukaryotic and prokaryotic, remain active within brine channels inside the ice, making it unnecessary to penetrate through to liquid water below in order to detect life. We have developed a submersible digital holographic microscope (DHM) that is capable of resolving individual bacterial cells, and demonstrated its utility for immediately imaging samples taken directly from sea ice at several locations near Nuuk, Greenland. In all samples, the appearance and motility of eukaryotes were conclusive signs of life. The appearance of prokaryotic cells alone was not sufficient to confirm life, but when prokaryotic motility occurred, it was rapid and conclusive. Warming the samples to above-freezing temperatures or supplementing with serine increased the number of motile cells and the speed of motility; supplementing with serine also stimulated chemotaxis. These results show that DHM is a useful technique for detection of active organisms in extreme environments, and that motility may be used as a biosignature in the liquid brines that persist in ice. These findings have important implications for the design of missions to icy environments and suggest ways in which DHM imaging may be integrated with chemical life-detection suites in order to create more conclusive life detection packages.

SummaryA variety of ecologically important processes are driven by bacterial motility and taxis, yet these basic bacterial behaviours remain understudied in cold habitats. Here, we present a series of experiments designed to test the chemotactic ability of the model marine psychrophilic bacterium Colwellia psychrerythraea 34H, when grown at optimal temperature and salinity (88C, 35 ppt) or its original isolation conditions (-18C, 35 ppt), towards serine and mannose at temperatures from 288C to 278C (above its upper growth temperature of 188C), and at salinities of 15, 35 and 55 ppt (at 88C and 218C). Results indicate that C. psychrerythraea 34H is capable of chemotaxis at all temperatures tested, with strongest chemotaxis at the temperature at which it was first grown, whether 88C or 218C. This model marine psychrophile also showed significant halotaxis towards 15 and 55 ppt solutions, as well as strong substrate-specific chemohalotaxis. We suggest that such patterns of taxis may enable bacteria to colonize sea ice, position themselves optimally within its extremely cold, hypersaline and temporally fluctuating microenvironments, and respond to various chemical signals therein.

The Arctic Ocean is more susceptible to ocean acidification than other marine environments due to its weaker buffering capacity, while its cold surface water with relatively low salinity promotes atmospheric CO2 uptake. We studied how sea‐ice microbial communities in the central Arctic Ocean may be affected by changes in the carbonate system expected as a consequence of ocean acidification. In a series of four experiments during late summer 2018 aboard the icebreaker Oden, we addressed microbial growth, production of dissolved organic carbon (DOC) and extracellular polymeric substances (EPS), photosynthetic activity, and bacterial assemblage structure as sea‐ice microbial communities were exposed to elevated partial pressures of CO2 (pCO2). We incubated intact, bottom ice‐core sections and dislodged, under‐ice algal aggregates (dominated by Melosira arctica) in separate experiments under approximately 400, 650, 1000, and 2000 μatm pCO2 for 10 d under different nutrient regimes. The results indicate that the growth of sea‐ice algae and bacteria was unaffected by these higher pCO2 levels, and concentrations of DOC and EPS were unaffected by a shifted inorganic C/N balance, resulting from the CO2 enrichment. These central Arctic sea‐ice microbial communities thus appear to be largely insensitive to short‐term pCO2 perturbations. Given the natural, seasonally driven fluctuations in the carbonate system of sea ice, its resident microorganisms may be sufficiently tolerant of large variations in pCO2 and thus less vulnerable than pelagic communities to the impacts of ocean acidification, increasing the ecological importance of sea‐ice microorganisms even as the loss of Arctic sea ice continues.

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