The planktonic communities within our oceans represent one of the most diverse and understudied ecosystems on the planet. A major hurdle in describing these systems is the sheer scale of the oceans along with logistical and economic constraints associated with their sampling. This is due to the limited amount of scientifically equipped fleets and affordable equipment. Here we demonstrate a modular approach for building a versatile, re-configurable imaging platform that can be adapted to a number of field applications, specifically focusing on oceanography. By using a modular hardware/software approach for building microscopes, we demonstrate high-throughput imaging of lab and field samples while enabling rapid device reconfiguration in order to match diverse applications and the evolving needs of the sampler. The presented versions of PlanktonScope are capable of autonomously imaging 1.7 ml per minute with a 1.5 µm resolution, and are built with under $400 in parts. This low cost enables new applications in laboratory settings such as the continuous imaging of suspension cultures, and in-field settings with the ability to scale up for long-term deployment on an international fleet of sailing boats enabling citizens based oceanographic research
The oceans represent 97% of all water on Earth and contain microscopic, drifting life, plankton, which drives global biogeochemical cycles. A major hurdle in assessing marine plankton is the planetary scale of the oceans and the logistical and economic constraints associated with their sampling. This difficulty is reflected in the limited amount of scientifically equipped fleets and affordable equipment. Here we present a modular hardware/software open-source strategy for building a versatile, re-configurable imaging platform - the PlanktoScope - that can be adapted to a number of applications in aquatic biology and ecology. We demonstrate high-throughput quantitative imaging of laboratory and field plankton samples while enabling rapid device reconfiguration to match the evolving needs of the sampler. The presented versions of PlanktoScope are capable of autonomously imaging 1.7 ml per minute with a 2.8 µm/px resolution and can be controlled from any WiFi-enabled device. The PlanktoScope’s small size, ease of use, and low cost - under $1000 in parts - enable its deployment for customizable monitoring of laboratory cultures or natural micro-plankton communities. This also paves the way toward consistent and long-term measurement of plankton diversity by an international fleet of citizen vessels at the planetary scale.
Living cells evolve under specific ecological conditions. The mass of our planet setting a constant gravitational pull and its daily rotation on its axis setting a circadian rhythm are two examples of universal ecological parameters that shape the physiology of every living creature on earth. In the ocean, gravity plays a critical role in shaping the life history of phytoplankton with vast diversity of size and cell geometries all depending on light to survive while gravity is pulling them down. In order to survive, these organisms must evolve means to navigate a vertically stratified ocean. Here we present how cellular topology can enable sinking, rising, and levitation dynamics of seemingly non-motile cells in the open ocean. By studying the bio-luminescent dinoflagellate Pyrocystis noctiluca, we discover how rapid inflation dynamics regulates cell size and buoyancy in coordination to its cell cycle. Live-cell light-sheet imaging reveals a (n-genus) toroidal cytoplasm that enables this rapid inflation of overall cell volume over 5 fold in 15 minutes without dilution of cytoplasmic contents. Here we present combined field and lab measurements with system modeling to build a general framework for how non-motile plankton can escape the gravitational sedimentation trap. Our work on cellular levitation emphasizes the critical role of studying cell and evolutionary biology in its planetary context.
Understanding key biophysical phenomena in the ocean often requires one to simultaneously focus on microscale entities, such as motile plankton and sedimenting particles, while maintaining the macroscale context of vertical transport in a highly stratified environment. This poses a conundrum: How to measure single organisms, at microscale resolution, in the lab, while allowing them to freely move hundreds of meters in the vertical direction? We present a solution in the form of a scale-free, vertical tracking microscope based on a circular "hydrodynamic-treadmill".Our technology allows us to transcend physiological and ecological scales, tracking organisms from marine zooplankton to single-cells over vertical scales of meters while resolving microflows and behavioral processes. We demonstrate measurements of sinking particles, including marine snow as they sediment tens of meters while capturing sub-particle-scale phenomena. We also demonstrate depth-patterned virtual-reality environments for novel behavioral analyses of microscale plankton. This technique offers a new experimental paradigm in microscale ocean biophysics by combining physiological-scale imaging with free movement in an ecological-scale patterned environment.One sentence summary: Scale-free vertical tracking microscopy captures, for the first time, untethered behavioral dynamics at cellular resolution for marine plankton.Our oceans represent the largest habitable ecosystem on the planet. With an average 1 depth of 4 kilometers, this unique ecosystem is highly vertically stratified with physical pa-2 rameters such as light, temperature, salinity and pressure varying dramatically as a function 3 of depth [1]. For example, only the first 200 meters of the ocean receives all the sunlight, 4 while the deeper parts of the ocean are effectively dark. For every 10 meters in depth, the 5 pressure increases by 1 atmosphere. Despite being only a few hundredths of the biomass 6 of terrestrial ecosystems [2], the oceans are responsible for half of the carbon fixed on our 7 planet [3]. Remarkably, this primary production in the ocean comes mostly from minuscule 8 plankton [4], the majority of whom are invisible to the naked eye. Although we have known 9 since the work of Haeckel [5] that the ocean abounds with microscopic plankton, only re-10 cently have we begun to realize their critical role in our planetary cycles [4, 6]. Despite their 11 importance, understanding the key biophysical mechanisms at the scale of planktonic single 12 cells and organisms, which help them navigate the ocean's complex vertical landscape, and in 13 turn influence planetary-scale processes, remains a major hurdle in biological oceanography. 14 A significant challenge in studying these biophysical processes is bridging the vast length 15 scales (from microns to kilometers) and time scales (from millisecond to days). Conven-16 tionally, vertical fluxes in the ocean are measured using sedimentation traps and sampling 17 at different depths [7]. Although crucial, the data is expensive t...
A multiplexed enzyme-linked immunosorbent assay (ELISA) that simultaneously measures antibody binding to multiple antigens can extend the impact of serosurveillance studies, particularly if the assay approaches the simplicity, robustness, and accuracy of a conventional single-antigen ELISA. Here, we report on the development of multiSero, an open-source multiplex ELISA platform for measuring antibody responses to viral infection. Our assay consists of three parts: (1) an ELISA against an array of proteins in a 96-well format; (2) automated imaging of each well of the ELISA array using an open-source plate reader; and (3) automated measurement of optical densities for each protein within the array using an open-source analysis pipeline. We validated the platform by comparing antibody binding to Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) antigens in 217 human sera samples, showing high sensitivity (0.978), specificity (0.977), positive predictive value (0.978), and negative predictive value (0.977) for classifying seropositivity, a high correlation of multiSero determined antibody titers with commercially available SARS-CoV-2 antibody tests, and antigen-specific changes in antibody titer dynamics upon vaccination. The open-source format and accessibility of our multiSero platform can contribute to the adoption of multiplexed ELISA arrays for serosurveillance studies, for SARS-CoV-2 and other pathogens of significance.
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