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Development of Two Color Fluorescent Imager and Integrated Fluidic System for Nanosatellite Biology Applications

Nanosatellites offer frequent, low-cost space access as secondary payloads on launches of larger conventional satellites. We summarize the payload science and technology of the Microsatellite in-situ Space Technologies (MisST) nanosatellite for conducting automated biological experiments. The payload (two fused 10-cm cubes) includes 1) an integrated fluidics system that maintains organism viability and supports growth and 2) a fixed-focus imager with fluorescence and scattered-light imaging capabilities. The payload monitors temperature, pressure and relative humidity, and actively controls temperature. C. elegans (nematode, 50 m diameter x 1 mm long) was selected as a model organism due to previous space science experience, its completely sequenced genome, size, hardiness, and the variety of strains available. Three strains were chosen: two green GFP-tagged strains and one red tdTomato-tagged strain that label intestinal, nerve, and pharyngeal cells, respectively. The integrated fluidics system includes bioanalytical and reservoir modules. The former consists of four 150 L culture wells and a 4x5 mm imaging zone the latter includes two 8 mL fluid reservoirs for reagent and waste storage. The fluidic system is fabricated using multilayer polymer rapid prototyping: laser cutting, precision machining, die cutting, and pressure-sensitive adhesives it also includes eight solenoid-operated valves and one mini peristaltic pump. Young larval-state (L2) nematodes are loaded in C. elegans Maintenance Media (CeMM) in the bioanalytical module during pre-launch assembly. By the time orbit is established, the worms have grown to sufficient density to be imaged and are fed fresh CeMM. The strains are pumped sequentially into the imaging area, imaged, then pumped into waste. Reagent storage utilizes polymer bags under slight pressure to prevent bubble formation in wells or channels. The optical system images green and red fluorescence bands by excitation with blue (473 nm peak) and amber (587 nm peak) LEDs it achieves 8 m lateral resolution using a CMOS imaging chip (as configured for serial data speeds) or 4 m resolution using USB imaging chips. The imager consists of a modified commercial off-the-shelf CMOS chip camera, amber, blue and white LEDs, as well as a relay lens and dual-band filters to obviate moving parts while supporting both fluorescence wavelengths.

Wu, Diana Terri↗

Active Flow Separation Control on a NACA 0015 Wing Using Fluidic Actuators

Results are presented from a recent set of wind tunnel experiments using sweeping jet actuators to control ow separation on the 30% chord trailing edge ap of a 30 deg. swept wing model with an aspect ratio (AR) of 4.35. Two sweeping jet actuator locations were examined, one on the flap shoulder and one on the trailing edge flap. The parameters that were varied included actuator momentum, freestream velocity, and trailing edge flap deflection (Delta f ) angle. The primary focus of this set of experiments was to determine the mass flow and momentum requirements for controlling separation on the flap, especially at large flap deflection angles which would be characteristic of a high lift system. Surface pressure data, force and moment data, and stereoscopic particle image velocimetry (PIV) data were acquired to evaluate the performance benefits due to applying active flow control. Improvements in lift over the majority of the wing span were obtained using sweeping jet actuator control. High momentum coefficient, Cu, levels were needed when using the actuators on the ap because they were located downstream of separation. Actuators on the flap shoulder performed slightly better but actuator size, orientation, and spacing still need to be optimized.

Melton, Latunia P.↗

The Plant Water Management Experiments: Soil

A simple means of watering plants in the low-g environment aboard orbiting spacecraft is not obvious. Since the beginning of spaceflight, numerous approaches have been pursued to water plants that seek to maximize plant viability and system reliability, while minimizing crew time and system complexity. We are not there yet. The Plant Water Management (PWM) Soil experiments seek to apply recent advances in low-g capillary fluidics phenomena to the challenges faced by plant growth operations aboard spacecraft. The primary challenge is to establish earth-like flows minimizing low-g specific adaptations required of the plants. This is difficult due to the ever-present fluid physics challenges of poorly-wetting multiphase inertial-visco-capillary flows in geometrically complex conduits and containers. In this paper, we present recent flight results for the PWM Soil experiments where arcillite ‘soil reservoirs’ are arranged in a non-wetting host soil that serves as an O2-breathing wetting barrier. In this way, a largely terrestrial water-soil environment is mimicked where, as liquid is evapo-transpired through the growing plant foliage, the effective water table passively ‘falls’ reducing viscous lengths and increasing water uptake for the plant. We present data from 6 days of 24-7 experiments on the ISS testing 3 different plant root models. We also present and correlate a capillary flow model which captures the primary features of the flow. Our summary is valued for the assessment of current and future low-g plant watering systems employing soil media.

microgravity↗

The Plant Water Management Experiments: Hydroponics 3 & 4

As humans consider longer-duration missions in space, NASA has identified production of fresh vegetables aboard spacecraft as beneficial for crew nutrition, mental wellbeing, and enabling bioregenerative life support (i.e., air, water, and waste processing). Current low-g plant growth techniques have successfully grown a variety of leafy and flowering plants. However, unique microgravity fluidics challenges to maintain plant moisture levels persist which hamper overall system reliability. The Plant Water Management (PWM) experiments seek to demonstrate low-cost, low-mass, reusable plant growth systems that leverage recent advances in low-g capillary fluidics phenomena to provide routine, largely passive, water delivery to plants. This paper presents findings from a series of PWM-Hydroponics 3 & 4 experiments, which were collected during three ISS flight operations that occurred in March, May, and July of 2021. Open hydroponic capillary channel flows with synthetic evapo-transpiring plant models were used. Tests demonstrated flow stability for single and parallel channel flow configurations across a range of flow rates, plant types, and plant arrangements. Technology demonstrations of both passive aeration and bubble phase separation are reported. We provide details of the data reduction and archive. Insights from the successful flight demonstrations provide a foundation from which follow-on PWM-Hydroponics 5 & 6 experiments on ISS, potentially incorporate living plants, are being considered. Xx Summary of positives and negatives..

Marc B. Wasserman↗

Plant Water Management (PWM) Experiment

Plant Water Management is a technology demonstration of recent advances in micro-g capillary fluidics research applied to plant growth systems. It has applications in long-term food production systems for missions to the Moon and Mars, as well as the immediate need for ISS food supplements to the crew diet. PWM will demonstrate the low-gravity role of surface tension, wetting, and system geometry to effectively replace the role of gravity in certain terrestrial plant growth systems.

Hatch, Tyler↗

Fluidic Chevrons for Jet Noise Reduction

Chevron mixing devices are used to reduce noise from commercial separate-flow turbofan engines. Mechanical chevron serrations at the nozzle trailing edge generate axial vorticity that enhances jet plume mixing and consequently reduces far-field noise. Fluidic chevrons generated with air injected near the nozzle trailing edge create a vorticity field similar to that of the mechanical chevrons and allow more flexibility in controlling acoustic and thrust performance than a passive mechanical design. In addition, the design of such a system has the future potential for actively controlling jet noise by pulsing or otherwise optimally distributing the injected air. Scale model jet noise experiments have been performed in the NASA Langley Low Speed Aeroacoustic Wind Tunnel to investigate the fluidic chevron concept. Acoustic data from different fluidic chevron designs are shown. Varying degrees of noise reduction are achieved depending on the injection pattern and injection flow conditions. CFD results were used to select design concepts that displayed axial vorticity growth similar to that associated with mechanical chevrons and qualitatively describe the air injection flow and the impact on acoustic performance.

Kinzie, Kevin↗

A Computational Study of a New Dual Throat Fluidic Thrust Vectoring Nozzle Concept

A computational investigation of a two-dimensional nozzle was completed to assess the use of fluidic injection to manipulate flow separation and cause thrust vectoring of the primary jet thrust. The nozzle was designed with a recessed cavity to enhance the throat shifting method of fluidic thrust vectoring. Several design cycles with the structured-grid, computational fluid dynamics code PAB3D and with experiments in the NASA Langley Research Center Jet Exit Test Facility have been completed to guide the nozzle design and analyze performance. This paper presents computational results on potential design improvements for best experimental configuration tested to date. Nozzle design variables included cavity divergence angle, cavity convergence angle and upstream throat height. Pulsed fluidic injection was also investigated for its ability to decrease mass flow requirements. Internal nozzle performance (wind-off conditions) and thrust vector angles were computed for several configurations over a range of nozzle pressure ratios from 2 to 7, with the fluidic injection flow rate equal to 3 percent of the primary flow rate. Computational results indicate that increasing cavity divergence angle beyond 10 is detrimental to thrust vectoring efficiency, while increasing cavity convergence angle from 20 to 30 improves thrust vectoring efficiency at nozzle pressure ratios greater than 2, albeit at the expense of discharge coefficient. Pulsed injection was no more efficient than steady injection for the Dual Throat Nozzle concept.

Deere, Karen A.↗

Active Flow Control on Vertical Tail Models

Active flow control (AFC) subscale experiments were conducted at the Lucas Wind Tunnel of the California Institute of Technology. Tests were performed on a generic vertical tail model at low speeds. Fluidic oscillators were used at the trailing edge of the main element (vertical stabilizer) to redirect the flow over the rudder and delay or prevent flow separation. Side force increases in excess of 50% were achieved with a 2% momentum coefficient (C(sub μ)) input. The results indicated that a collective C(sub μ) of about 1% could increase the side force by 30–50%. This result is achieved by reducing the spanwise flow on the swept back wings that contributes to early flow separation near their tips. These experiments provided the technical backdrop to test the full-scale Boeing 757 vertical tail model equipped with a fluidic oscillator system at the National Full-scale Aerodynamics Complex 40-by 80-foot Wind Tunnel, NASA Ames Research Center. The C(sub μ) is shown to be an important parameter for scaling a fluidic oscillator AFC system from subscale to full-scale wind tunnel tests. The results of these tests provided the required rationale to use a fluidic oscillator AFC configuration for a follow-on flight test on the Boeing 757 ecoDemonstrator.

Andino, Marlyn Y.↗

A Heritage BioSensor for Lunar Biology Experiments

Introduction: Automated biological experiments on small spacecraft missions have gained prominence over the past decade due to their simplicity, accessibility, and small mass, volume, and power needs. Most recently, the BioSensor microfluidic CubeSat payload aboard BioSentinel used an automated microfluidic cell culture system to study the effects of environmental stressors like deep space radiation and microgravity on yeast growth and metabolism. BioSentinel’s successor, the Lunar Explorer Instrument for space biology Applications (LEIA), will study the effects of lunar gravity and radiation using an improved version of the BioSensor microfluidic platform. The BioSensor payload has great adaptability to host a diverse range of biological experiments with single- and multi-celled organisms in both crewed and uncrewed missions, making it a compelling candidate for future space biology studies in a lunar surface environment. BioSensor Instrumentation on BioSentinel: The first spaceflight mission with the BioSensor, BioSentinel’s biology experiments occurred at three locations -- deep space, ISS and ground. The payload contained 18 microfluidic cards, each featuring 16 growth wells (a total of 288 growth wells). Each well was loaded before launch with desiccated yeast. In space, liquid culture medium (nutrients) was automatically introduced to batches of wells at a time to initiate a series of biology experiments. Temperature was maintained by thin film heaters on both sides of each card. Each well was equipped with three LEDs emitting at 570 nm, 630 nm, and 850 nm, paired with photodetectors to measure cell concentration and the alamarBlue (metabolic indicator dye) color transition from blue to pink. Phenotypic parameters like cell viability, metabolic rate, and generation time can be derived from these measurements. The sequence and timing of fluid fills, optical measurements, and thermal control were stored onboard, but could be updated asynchronously via ground communication. LEIA: LEIA is slated for launch no earlier than 2026 on a CLPS lander. BioSentinel’s BioSensor has been modified for use in LEIA. These improvements include: (a) storage for multiple culture medium types, (b) additional LED color (465 nm) for a new biological assay for antioxidant (carotenoid) production, (c) housing modifications for later biology load before launch, (d) improved isolation between electronic and fluidic components, and (e) improved humidity control for prolonged organism viability in case of post-load launch delay. Future Prospects: The consistent and successful demonstration of complex fluidics platforms alongside reliable instrument operations in a space environment is poised to create strong momentum for BioSensor-based biological experiment payloads. Planned future developments with the BioSensor include extending compatibility to a broader range of organisms and assays. Preliminary work has already demonstrated successful growth of Arabidopsis seedlings in fluidic cards. With a few modifications to the optical assembly, the setup could easily measure photosynthetic traits in plants and cyanobacteria. The addition of fluorescence measurements and generation of novel luminescent assays will elevate BioSensor’s functionality further. Beyond the BioSensor’s potential uses on free-flyer missions, ISS and Gateway, and CLPS landers, deploying the BioSensor to the lunar surface or in an artificial habitat on crewed missions could enable pioneering research on both how life responds to lunar conditions and future bioproduction capabilities making the BioSensor an indispensable tool for future space biology research.

Chinmayee Govinda Raj↗

Impact of Azimuthally Controlled Fluidic Chevrons on Jet Noise

The impact of azimuthally controlled air injection on broadband shock noise and mixing noise for single and dual stream jets was investigated. The single stream experiments focused on noise reduction for low supersonic jet exhausts. Dual stream experiments included high subsonic core and fan conditions and supersonic fan conditions with transonic core conditions. For the dual stream experiments, air was injected into the core stream. Significant reductions in broadband shock noise were achieved in a single jet with an injection mass flow equal to 1.2% of the core mass flow. Injection near the pylon produced greater broadband shock noise reductions than injection at other locations around the nozzle periphery. Air injection into the core stream did not result in broadband shock noise reduction in dual stream jets. Fluidic injection resulted in some mixing noise reductions for both the single and dual stream jets. For subsonic fan and core conditions, the lowest noise levels were obtained when injecting on the side of the nozzle closest to the microphone axis.

Henderson, Brenda S.↗

Improving Satellite Compatible Microdevices to Study Biology in Space

The technology for biology in space lags far behind the gold standard for biological experiments on Earth. To remedy this disparity, the Rothschild lab works on proof of concept, prototyping, and developing of new sensors and devices to further the capabilities of biology research on satellites. One such device is the PowerCell Payload System. One goal for synthetic biology in aiding space travel and colonization is to genetically engineer living cells to produce biochemicals in space. However, such farming in space presupposes bacteria retain their functionality post-launch, bombarded by radiation, and without the 1G of Earth. Our questions is, does a co-culture of cyanobacteria and protein-synthesizing bacteria produce Earth-like yields of target proteins? Is the yield sensitive to variable gravitational forces? To answer these questions, a PowerCell Payload System will spend 1 year aboard the German Aerospace Center's Euglena and Combined Regenerative Organic-food Production In Space (Eu:CROPIS) mission satellite. The PowerCell system is a pair of two 48-well microfluidic cards, each well seeded with bacteria. The system integrates fluidic, thermal, optical, electronic, and control systems to germinate bacteria spores, then measure the protein synthesized for comparison to parallel experiments conducted on the Earth. In developing the PowerCell Payload, we gained insight into the shortcomings of biology experiments on satellites. To address these issues, we have started three new prototyping projects: 1) The development of an extremely stable and radiation resistant cell-free system, allowing for the construction of proteins utilizing only cell components instead of living cells. This can be lyophilized on a substrate, like paper. (2) Using paper as a microfluidic platform that is flexible, stable, cheap, and wicking. The capillary action eliminates the need for pumps, reducing volume, mass, and potential failing points. Electrodes can be printed on the paper to sense for biochemicals. (3) Developing a modular, semi-autonomous microfluidic device that can be easily adapted for a variety of common biological experiments. This versatility will allow for quicker and cheaper experimentation. These improvements to satellite experiment platforms have the potential to radically increase the return from NASA's biological and field studies with reduced development time, mass, and cost with increased robustness data and interpretation.

biology in spac↗

Computational Characterization and Boundary Condition Models of a Jet Interaction Fluidic Oscillator

Fluidically oscillating jet actuators that generate sweeping jets when supplied with a pressurized fluid have been used to mitigate separation and reduce drag in a range of flow control applications. Their implementation in future flight vehicles would require fundamental understanding and accurate predictive techniques of the physics of their internal flow and jet formation. The present investigations focus on high-fidelity, time-accurate simulations to characterize the flow physics of the actuator in quiescent conditions. An important element of the present simulations is to demonstrate the ability of the computational fluid dynamics (CFD) solver to predict the jet characteristics and provide a basis for the development of improved boundary conditions (BC) without entirely resolving the geometrical features of the fluidic device. The CFD-predicted oscillation frequencies of the engendered jets were found to be in excellent agreement with experiments, even on two-dimensional meshes. The study revealed that three-dimensional simulations are required to capture some of the flow features of the sweeping jet such as the double peak in time-averaged velocity distributions downstream of the actuator’s orifice that were measured in experiments. Furthermore, several approaches for modeling the actuator were implemented and assessed in quiescent conditions. The evaluation of a boundary condition at the device throat, based on the phase-averaged flow variables, provides the basis for devising surface-based boundary conditions. The influence and necessity of including turbulent characteristics as part of the boundary conditions have also been identified.

42 ENGINEERING↗

Surface Tension Containment Experiment (STCE) - Increasing Science Throughput on ISS

The Microgravity Science Glovebox (MSG) on the International Space Station (ISS) is used for fluid transfer in many types of experiments. Reagents are handled in the MSG to prevent their accidental release into the cabin. However, the MSG is currently over-subscribed, creating a backlog of users in flight. As a recourse, current experiments are underway to assess the possibility of moving certain science operations from the MSG into the open cabin of the ISS. The experiments are designed to assess the efficacy of exploiting surface tension as a control to prevent the unwanted release of liquids. Dyed water currently serves as an ersatz for potentially more hazardous liquids. Common wet-lab operations such as de/mating wetted Luer-Lok fittings, liquid-bearing container lid removal, and pipetting between well plates are performed illustrating the facility and challenges imposed by the microgravity environment. Concerning the latter, various pipette cannula sizes are deployed at various injection, withdrawal, and translations rates to map the existence, size, velocity, and trajectory of satellite droplets expected to form when breaking contact between the water surface and the pipette tip. Though such drops frequently form in terrestrial operations, they are nearly imperceptible and inconsequential—due in part to their speed and because gravity quickly returns them to the well plate from which they came. The use of airflow to capture and collect such satellite droplets is demonstrated. The dynamic stability of the liquid-filled well plates is quantified in response to a variety of crew-imparted disturbances. From a safety perspective, the results from the STCE are of immediate practical value. If such routine low-gravity capillary fluidic operations can be established as mundane, their performance may be moved out of the MSG and into the cabin, significantly increasing the efficiency of experiments performed on ISS.

Ungar, Eugene K.↗

EcAMSat: Small Satellite to Examine E. coli's Response in Microgravity to the Antibiotic Gentamicin

We have successfully flown the EcAMSat (Escherichia coli Antimicrobial Satellite) free-flyer mission. This was a 6U small satellite that autonomously conducted an experiment in low Earth orbit to explore the impact of the space environment on antibiotic resistance in uropathogenic E. coli (UPEC) and the role a particular sigma factor plays in the response. After being held in stasis during transport to orbit, two strains†a wildtype UPEC and an isogenic mutant with a deleted gene that encodes a sigma factor†were grown to stationary phase in a fluidic card inside EcAMSat's payload, then incubated with three concentrations of the antibiotic gentamicin. The payload then administered alamarBlue®, a redox indicator, into all wells of the fluidic card. The cells were then incubated for 144 hours and metabolic activity was measured optically using the payloads' LED and detector system. Data were then telemetered to the ground and compared to a control experiment conducted in an identical satellite in a lab. The results of this experiment will help us better understand important therapeutic targets for treating bacterial infections on Earth and in space. Such targets are particularly relevant to deep-space and long-duration missions where crew may be more susceptible to infection and treatments for them may work differently.

antibiotic↗

The Promise of Macromolecular Crystallization in Micro-fluidic Chips

Micro-fluidics, or lab on a chip technology, is proving to be a powerful, rapid, and efficient approach to a wide variety of bio-analytical and microscale bio-preparative needs. The low materials consumption, combined with the potential for packing a large number of experiments in a few cubic centimeters, makes it an attractive technique for both initial screening and subsequent optimization of macromolecular crystallization conditions. Screening operations, which require equilibrating macromolecule solution with a standard set of premixed solutions, are relatively straightforward and have been successfully demonstrated in a micro-fluidics platform. More complex optimization methods, where crystallization solutions are independently formulated from a range of stock solutions, are considerably more complex and have yet to be demonstrated. To be competitive with either approach, a micro-fluidics system must offer ease of operation, be able to maintain a sealed environment over several weeks to months, and give ready access for the observation of crystals as they are grown.

vanderWoerd, Mark↗

BioSensor Users' Guide

The BioSensor is a fully autonomous 3-color LED-based spectrophotometer paired with a fluidics system that supports microbes in liquid culture. Originally developed for the BioSentinel CubeSat mission to study the response of a wild type and mutant strain of yeast to the deep space environment, the BioSensor consists of a series of 16-well, independently plumbed fluidics cards. The cards utilize filters to allow the fluidic system to administer reagents while constraining the microbes in their wells. Heaters on each card incubate to the appropriate growth temperature once the experiment begins. During the active experiment, the LED/detector system measures the transmission of light through each well at three specific wavelengths, similar to a standard benchtop spectrophotometer. The transmission/absorbance kinetics curves for each well are telemetered back to Earth, along with temperature data, for analysis on the ground. The BioSensor is being upgraded from its original CubeSat free-flyer interface to be a secondary payload on lunar landers, Gateway, and other BLEO applications, while maintaining the same functionality and science utility for future experiments.

Matthew Lera↗

BioSensor Users' Guide

The BioSensor is a fully autonomous 3-color LED-based spectrophotometer paired with a fluidics system that supports microbes in liquid culture. Originally developed for the BioSentinel CubeSat mission to study the response of a wild type and mutant strain of yeast to the deep space environment, the BioSensor consists of a series of 16-well, independently plumbed fluidics cards. The cards utilize filters to allow the fluidic system to administer reagents while constraining the microbes in their wells. Heaters on each card incubate to the appropriate growth temperature once the experiment begins. During the active experiment, the LED/detector system measures the transmission of light through each well at three specific wavelengths, similar to a standard benchtop spectrophotometer. The transmission/ absorbance kinetics curves for each well are telemetered back to Earth, along with temperature data, for analysis on the ground. The BioSensor is being upgraded from its original CubeSat free-flyer interface to be a secondary payload on lunar landers, Gateway, and other BLEO applications, while maintaining the same functionality and science utility for future experiments.

Matthew Lera↗

EcAMSat: A Small Satellite Flown to Explore the Role a Sigma Factor Plays in E. coli's Response to the Antibiotic Gentamicin

We have successfully flown the EcAMSat (Escherichia coli Antimicrobial Satellite) free-flyer mission. This was a 6U (six unit - CubeSat) small satellite that autonomously conducted an experiment in low Earth orbit to explore the impact of the space environment on antibiotic resistance in uropathogenic E. coli (UPEC) and the role a particular sigma factor plays in the response. After being held in stasis during transport to orbit, two strains - a wildtype UPEC and an isogenic mutant with a deleted gene that encodes a sigma factor - were grown to stationary phase in a fluidic card inside EcAMSat's payload, then incubated with three concentrations of the antibiotic gentamicin. The payload then administered alamarBlue (registered trademark), a redox indicator, into all wells of the fluidic card. The cells were then incubated for 144 hours and metabolic activity was measured optically using the payloads' LED (Light-Emitting Diode) and detector system. Data were then telemetered to the ground and compared to a control experiment conducted in an identical satellite in a lab. The results of this experiment will help us better understand important therapeutic targets for treating bacterial infections on Earth and in space. Such targets are particularly relevant to deep-space and long-duration missions where crew may be more susceptible to infection and treatments for them may work differently.

small satellite↗