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Sergio R Santa Maria

Publications and source records attributed to Sergio R Santa Maria.

Evolution of Biological Satellites: From Low Earth Orbit to NASA’s BioSentinel Deep Space Mission

NASA has set its sights on human exploration in deep space with the Artemis missions, with an ambitious plan to put astronauts back on the Moon and to eventually land human missions on Mars. Such missions will require significant countermeasures, likely both technological and biomedical, to protect biology from chronic radiation exposure. Small satellites like CubeSats can inform these countermeasures by querying relevant space environments with model organisms over relevant durations. NASA has launched five biological CubeSats into low Earth orbit (LEO) from GeneSat in 2006 to EcAMSat in 2017. Each one of these missions increased our understanding of the effects of spaceflight, while refining technologies and imparting valuable lessons to the next generation of CubeSats. The Artemis I rocket will carry ten CubeSats, each of them with its own objective. One in particular, BioSentinel, will conduct the first study of the biological response to interplanetary space radiation beyond LEO since Apollo 17. Once it reaches its heliocentric orbit – after a short lunar fly-by – BioSentinel will measure the DNA damage response to ambient radiation in a model organism, the budding yeast Saccharomyces cerevisiae, which will be compared to information provided by an onboard radiation sensor and to data obtained in LEO (on ISS) and on Earth. Once in interplanetary space, fluidic cards containing desiccated yeast cells will be activated by growth medium addition at different time points throughout the mission. Growth and metabolic activity will be tracked continuously via optical density. BioSentinel is a complete, autonomous spacecraft capable of conducting experiments in deep space. Its 4U BioSensor payload is a fully automated and adaptable platform that can perform biological measurements with a range of microorganisms in multiple space environments, including the ISS, free flyers, and other platforms like the Lunar Gateway and lander vehicles.

Sergio R Santa Maria

Space Biology Research and Biosensor Technologies: Past, Present, and Future

In light of future missions beyond low Earth orbit (LEO) and the potential establishment of bases on the Moon and Mars, the effects of the deep space environment on biology need to be examined in order to develop protective countermeasures. Although many biological experiments have been performed in space since the 1960s, most have occurred in LEO and for only short periods of time. These LEO missions have studied many biological phenomena in a variety of model organisms, and have utilized a broad range of technologies. However, given the constraints of the deep space environment, upcoming deep space biological missions will be largely limited to microbial organisms and plant seeds using miniaturized technologies. Small satellites such as CubeSats are capable of querying relevant space environments using novel, miniaturized instruments and biosensors. CubeSats also provide a low-cost alternative to larger, more complex missions, and require minimal crew support, if any. Several have been deployed in LEO, but the next iterations of biological CubeSats will travel beyond LEO. They will utilize biosensors that can better elucidate the effects of the space environment on biology, allowing humanity to return safely to deep space, venturing farther than ever before.

space biology

Developing Technologies for Biological Experiments in Deep Space

NASA Ames Research Center is the leader in developing autonomous nanosatellites or CubeSats to address strategic knowledge gaps about the effects of space travel on biological organisms, including GeneSat, PharmaSat, and EcAMSat. Now that NASA has set its sights on human exploration in deep space, such missions require significant technological and biomedical countermeasures to protect astronauts from chronic radiation exposure. CubeSats can inform these countermeasures by querying relevant space environments with model organisms and/or biosensors. BioSentinel will be the first interplanetary CubeSat to study the biological response to space radiation outside low Earth orbit in almost 50 years. BioSentinel is an autonomous platform able to support biology and to investigate the effects of space radiation on a model organism in interplanetary deep space. It will fly onboard Artemis-1, from which it will be deployed on a lunar fly-by trajectory and into a heliocentric orbit. BioSentinel, a 6U CubeSat (1U = 10-cm cube), will measure the DNA damage response to ambient space radiation in a model organism, which will be compared to information provided by an onboard radiation spectrometer and to data obtained on the ISS and on Earth. Even though the primary objective of the mission is to develop an autonomous spacecraft capable of conducting biological experiments in deep space, the 4U BioSensor science payload contained within the free-flyer is an adaptable instrument that can perform bio measurements with different microorganisms and in multiple space environments, including the ISS, lunar gateway, and on the surface of the Moon. Thus, nanosatellites like BioSentinel can be used to study the effects of both reduced gravity and space radiation and can house different organisms to answer specific science questions. In addition to their flexibility, nanosatellites also provide a low-cost alternative to more complex and larger missions, and require minimal crew support, if any

biosenses

2020 - 2030: New Technologies for Microbial Research in Deep Space

This presentation is part of a series of workshops in preparation for the Decadal Survey in Biological & Physical Sciences, and sponsored by the American Society for Gravitational and Space Research (ASGSR). The goal is to highlight novel technologies for microbial research that could be used in the upcoming 10 years and to promote forward-thinking in the space biology community.

deep space

Lunar BioSensor: An Autonomous Instrument to Study the Effects of the Lunar Environment on Biological Organisms

One of the major challenges to long-duration space travel and habitation in deep space is an in-depth understanding of the biological effects of space radiation, often convoluted by the impact of reduced gravity. Nonetheless, due to the near impossibility of simulating prolonged exposure to these combined effects in terrestrial facilities, actual missions are needed to characterize the radiobiological hazards of this environment. NASA Ames has been the leader in developing autonomous bio nanosatellites to address strategic knowledge gaps about the effects of space travel on biological organisms, including GeneSat, PharmaSat, EcAMSat, and BioSentinel. BioSentinel will be the first interplanetary bio nanosatellite or CubeSat to study the biological response to space radiation outside Low Earth Orbit (LEO). BioSentinel is an autonomous platform able to support biology and to investigate the effects of space radiation on a model organism in interplanetary deep space. It will fly onboard NASA’s Artemis-1, from which it will be deployed on a lunar fly-by trajectory and into a heliocentric orbit. The BioSentinel nanosatellite, a 6U deep space CubeSat (1U = 10-cm cube), will measure the DNA damage and response to ambient space radiation in a model biological organism, the budding yeast S. cerevisiae, which will be compared to information provided by an onboard physical radiation sensor and to data obtained in LEO (on the ISS) and on Earth. Even though the primary objective of the mission is to develop an autonomous spacecraft capable of conducting biological experiments in deep space, the 4U BioSensor science payload contained within the 6U free-flyer is an adaptable instrument platform that can perform biological measurements with different microorganisms and in multiple space environments, including the ISS, lunar gateway, and on the surface of the Moon. The proposed 4U instrument will leverage the payload design of the 6U free-flyer, utilizing the lunar lander or vehicle for power and data relay. Thus, nanosatellites like BioSentinel (and Lunar BioSensor) can be used to study the effects of both reduced gravity and space radiation and can house different bio organisms to answer specific science questions. In addition to their flexibility, nanosatellites also provide a low-cost alternative to more complex and larger missions, and require minimal crew support, if any.

space biosensors

RadBREAD: Radiation Biology Research at an Elevated Altitude through Dosimetry – A student-designed payload

NASA uses extreme environment platforms (ground testing facilities, high-altitude balloons and aircraft, and CubeSats) to provide greater understanding of the conditions and limitations of extra-terrestrial environments. As part of a two-week flight planned for summer 2021, RadBREAD (Radiation Biology Research at an Elevated Altitude through Dosimetry) will fly as a secondary payload consisting of a M-42C (German Aerospace Center, DLR) ionizing radiation dosimeter, UV micro-logger, and multiple desiccated yeast samples. The platform is a novel high-altitude solar-powered aircraft: the Swift Engineering High-Altitude samples. The platform is a novel high-altitude solar-powered aircraft: the Swift Engineering High-Altitude Long-Endurance Unmanned Aircraft System (HALE UAS), which offers significantly longer flight durations than other high-altitude platforms. The yeast Saccharomyces cerevisiae will provide meaningful biological correlation for the sensor readings, due to its resistance to extremely low temperature and pressure when desiccated, ease of genetic manipulation, and homology to human genes. The RadBREAD team comprises the 2020 cohort of NASA’s Space Life Sciences Training Program (SLSTP) research associates as well as NASA scientists, engineers and radiation experts from NASA and the DLR. Yeast survival, metabolic, and transcriptomic changes will be correlated with environmental data collected during long-term exposure to the upper atmosphere. Additionally, the team will evaluate the upper atmospheric environment (radiation, pressure, and temperature) provided by the HALE UAS platform as a Mars surface analog for biological payloads. We hypothesize that exposure to upper atmospheric conditions during the HALE UAS flight will alter the survival, metabolism, and transcriptome of desiccated wild-type S. cerevisiae upon rehydration compared to sensitive and tolerant yeast strains exposed to the same conditions, and between the flight samples compared to asynchronous ground controls.

radiation exposure

BioSentinel: NASA’s First Deep Space Biological Mission

Since Apollo 17 in 1972, NASA has sent no humans or other biological organisms outside of Earth’s protective magnetosphere. NASA’s current Artemis program plans to put astronauts back on the Moon and eventually land human missions on Mars. One of the major challenges to long-duration crewed travel and habitation in deep space is an in-depth understanding of the biological effects of space radiation, often convoluted by the impact of reduced gravity. Such missions will require significant countermeasures, likely both technological and biomedical, to protect organisms from chronic radiation exposure. Small satellite missions like CubeSats can inform these countermeasures by investigating model organisms in relevant space environments. The BioSentinel mission is comprised of four segments developed at NASA Ames Research Center: a 6U CubeSat (1U = 10-cm cube), an ISS payload launched in December 2021 and two ground units, one for the mission’s CubeSat and one for the ISS payload. The last three segments have been operational since January 2022 and serve as experimental controls. BioSentinel’s 6U CubeSat is planned to launch as a secondary payload on the Artemis-1 rocket. It will be deployed on a lunar fly-by trajectory and into a heliocentric orbit. BioSentinel will be the first interplanetary satellite to study the biological response to space radiation outside Low Earth Orbit (LEO) in almost 50 years. BioSentinel is a complete, autonomous spacecraft capable of conducting experiments in deep space. Its 4U BioSensor payload is a fully automated and adaptable platform that can perform biological measurements with a range of microorganisms in multiple space environments, including the ISS, free flyers, and other platforms like the Lunar Gateway and lander vehicles. Once it reaches its orbit, BioSentinel’s CubeSat will measure the DNA damage response to ambient radiation in a model organism, the budding yeast Saccharomyces cerevisiae, which will be compared to information provided by an onboard radiation sensor and to data obtained in LEO (on ISS) and on Earth. Once in interplanetary space, fluidic cards containing desiccated yeast will be activated by growth medium addition at different time points throughout the mission. Growth and metabolic activity will be tracked continuously via optical measurements. This paper describes BioSentinel’s objectives, science, data management, and preliminary results from the ISS segment.

BioSentinel

NASA's Biosentinel Mission: Lessons Learned and What's Next

In the last two years, two BioSentinel payloads were launched to space. The ISS mission launched in December 2021, and returned to the ground in August 2022 after successfully completing eight biological experiments while validating the different instruments. On the other hand, the deep space mission launched onboard Artemis I in November 2022, and is currently in a heliocentric orbit over 20 million kilometers away from the Earth. Even though all hardware subsystems were validated in deep space, the microfluidic subsystems experienced anomalies throughout the initial 6-month mission. The main goals of this presentation are (1) to present flight data from the deep space payload, including biology, fluidics, electronics, data processing, and mission operations, and (2) to discuss the lessons learned – what worked and what did not – from this unique complex mission, and how these lessons are aiding in the development of the Lunar Exploration Instrument for space biology Applications (LEIA) mission, launching to the lunar surface on a commercial lander in 2026. As of the writing of this abstract, the satellite continues to work nominally, communicating to Earth via the Deep Space Network (DSN) twice per week. Importantly, the mission received and extension to continue recording data on the deep space radiation environment on its way to solar maximum (i.e., higher probability of solar particle events). BioSentinel is supported by NASA Exploration Systems Development Mission Directorate (ESDMD).

BioSentinel

Single-Cell Analysis of Yeast (Saccharomyces cerevisiae) Using Hydrogel Encapsulation

Space radiation poses a major health risk to astronauts. To fulfill NASA’s mission of exploration beyond Earth, the biological effects of Galactic Cosmic Radiation and gamma radiation must be investigated to elucidate cellular damage mechanisms and inform countermeasure protocols to safely bring humans beyond Earth’s magnetosphere. Budding yeast (Saccharomyces cerevisiae) are commonly used in experiments as a model organism for studying the effects of radiation on eukaryotes. Radiobiology of yeast at the single cell level is poorly understood, yet crucial for informing models to aid in the design and interpretation of experiments. We are using a novel method of microencapsulation in hydrogel particles (PicoShells) to enable analysis of the distribution of radiation-induced damage among yeast cells at the single-cell level, in high throughput. Here we describe the development of methods for culturing, visualization, and quantification of encapsulated yeast. The encapsulated yeast are cultured in Yeast extract-Peptone-Dextrose (YPD) medium, fixed in formaldehyde or ethanol, and stained with DAPI or propidium iodide, then visualized using microscopy or enumerated using flow cytometry, with the aim of developing a protocol to enumerate the distribution of viable cells in each PicoShell. This will allow us to quantify how different forms of radiation can generate different distributions of damage across a population of cells, ultimately providing insight into the biological effects of space-relevant ionizing radiation.

yeast

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

Developing Autonomous Technologies for Biological Missions to Deep Space

In upcoming biological missions beyond low Earth orbit (LEO), the use of autonomous instrumentation will allow scientists to perform a variety of experiments, including the characterization of the response to different space environments (Moon, Mars, interplanetary space) using biological models like microbes, plants, organoids, and tissue chips. BioSentinel is an ongoing deep space mission, currently at over 50 million kilometers from Earth and the first instrument developed to perform biological experiments beyond LEO. Even though the primary objective of this CubeSat mission was to investigate the effects of the deep space radiation environment on budding yeast, the spacecraft bus (i.e., all the subsystems that support the biological payload like power, thermal, data telemetry, navigation, etc.) can accommodate a variety of biological (and physical) experiments and model organisms. LEIA, an upcoming CLPS mission to the lunar surface, uses a microfluidic and optical instrument based on BioSentinel to study the effects of the lunar environment on different cellular processes and on bioproduction of antioxidants. A new series of science mission concepts are being proposed to be accommodated into platforms like BioSentinel. These missions will investigate the response of a variety of organisms to the deep space environment, including but not limited to single-cell eukaryotes, cyanobacteria, plants (including crops), organoids, and tissue chips. In addition to optical absorbance measurements like the ones performed in BioSentinel (and LEIA), we are investigating the use of fluorescence detection, microscopy, sequencing devices, etc. Thus, instruments like the ones proposed here can be adapted to a variety of platforms like free-flyers, deployable payloads, landers, rovers, and the lunar Gateway. These technologies can be used as steppingstones for establishing a sustained human presence on the Moon and in deep space while providing knowledge for the development of potential countermeasures.

Sergio R Santa Maria

Development of Genetic Countermeasures for Enhancing Cellular Stress Tolerance on a Lunar Surface Mission

The Lunar Explorer Instrument for space biology Applications (LEIA) LEIA investigates the response to partial gravity and ionizing radiation of: Different DNA damage and stress response pathways and Bioproduction of antioxidants LEIA utilizes: Various strains of the yeast Saccharomyces cerevisiae, which will be desiccated in fluidic cards and rehydrated on the lunar surface LEIA develops: Genetic countermeasures to improve tolerance to the desiccation process and the constraints associated with long duration missions beyond low Earth orbit (LEO)

Neha Lingam

The Lunar Explorer Instrument for Space Biology Applications (LEIA) Pre-Flight Tests: Mechanical Considerations​

The BioSensor payload on the upcoming LEIA platform aboard the CLPS (CP-22) lander will carry yeast to the Moon's south pole to study the effects of lunar radiation and gravity. From Earth, BioSensor will transport desiccated yeast within 16 fluidic cards, each containing 16 fluidic wells of 100 microliters volume. At the start of the experiment, these microbes will be autonomously rehydrated on the Moon, and optical absorbance measurements at three wavelengths will be performed across all 256 wells. Absorbance measurements in fluidic wells are susceptible to noise and artifacts due to bubble formation during well rehydration and microbial growth. BioSensor, as a biofluidics system, faces significant data anomalies caused by payload vibrations, particularly from the operation of the co-located PROSPECT’s ProSEED drill, to assess lunar resources by penetrating the subsurface. To evaluate the impact of drill vibrations, fully assembled and filled fluidic cards were subjected to sine sweep signals in the x, y, z planes, ranging from 5 - 2560 Hz at g-loads of 0.25 and 0.5, with on-board accelerometer for localized vibration measurement. Wells with various fill conditions, including partial fills with different bubble sizes and completely filled wells, were tested. Results indicated that small bubbles adhered to well walls at high frequencies but were dislodged at low frequencies, causing data anomalies. Medium and large bubbles exhibited higher interference with optical data due to lower surface tension and variable bubble surface flexion. Overall, it was found that irrespective of the fill type, low-frequency vibrations were most detrimental to optical readings. Understanding the drill schedule and correlating it with BioSensor timestamps will help identify periods when BioSensor data is unreliable, a crucial step for accurately interpreting the microbial response data and ensuring the validity of the BioSensor's scientific measurements on the lunar surface.

Chinmayee Govinda Raj