Development of automated biosensor technologies for future biological research beyond low Earth orbit

1 Introduction

With the recent announcement of Moon Base, NASA and its partners are paving the path to establish a long-term human presence on the Moon and in deep space. Therefore, it is critical to understand the consequences of different spaceflight stressors on biological organisms. Beyond low Earth orbit (BLEO), outside the protection of Earth’s magnetic field, astronauts are exposed to several spaceflight stressors or hazards. The main spaceflight hazards that impact humans include space radiation, altered gravity, isolation, a hostile environment, and distance from Earth itself (Afshinnekoo et al., 2020). Even though these stressors can exacerbate each other’s effects during spaceflight, they can also generate confounding effects on complex biological organisms like humans. With astronauts heading back to the lunar surface and eventually to Mars, understanding these impacts is crucial to ensuring the development of necessary countermeasures that shield astronauts from the harsh deep space environment.

BLEO space radiation is composed primarily of galactic cosmic radiation (GCR) and solar particle events (SPEs). Both are modulated by the 11-year solar cycle, where SPEs are more frequent during solar maximum and GCR intensifies during solar minimum. SPEs are sporadic, originate from solar coronal mass ejections, and consist primarily of protons and helium particles. On the other hand, GCR originates from outside our solar system and is composed of a steady flux of high-energy protons and highly energetic heavy (HZE) particles. Importantly, the many forms of ionizing radiation can damage both electronic components and biological systems (Höeffgen et al., 2020; Al-Qabandi and Alshammary, 2022).

Even though it is possible to replicate some aspects of the space radiation environment using ground-based facilities (Simonsen et al., 2020), it is very difficult to simulate the protracted and omnidirectional nature of the GCR environment in deep space. On the lunar surface, the radiation environment is further complicated due to the production of albedo neutrons when GCR interacts with regolith (Adams et al., 2007). Similarly, microgravity simulators are valuable tools for developing spaceflight experiments (Herranz et al., 2013); however, they all have limitations and are rarely used in combination with ionizing radiation sources. Thus, further experimentation in situ can provide valuable information on the biological response to the space environment and how to protect future space explorers.

The development of automated and/or autonomous technologies has allowed scientists and engineers the opportunity to perform fundamental and applied research in deep space and on the lunar surface. While manned experiments provide immediate access to the samples and direct troubleshooting, astronaut interaction is expensive and requires significant training, in addition to limited crew time and no current infrastructure for such activities beyond the ISS. Small satellites, or CubeSats, are ever-evolving platforms that give academic institutions, commercial groups, and government agencies relatively affordable access to the space environment (Poghosyan and Golkar, 2017). The popularization of CubeSats can be attributed to their standardized small size, with one unit (1U) consisting of a 10-cm cube weighing approximately 1kg, allowing for a variety of payload configurations (Zhang et al., 2024). Most payloads onboard CubeSats are preprogrammed with commands and can operate automatically or autonomously, while still allowing for ground-crew intervention. Thus, in the last 20 years, CubeSats have gained popularity as invaluable platforms to investigate the effects of the space environment on different model organisms in both LEO and beyond (Santa Maria, 2023). In particular, fluidic-based instrumentation enables the testing of different phenomena in space, including the response of microbes to antimicrobials or how the space environment affects their metabolism (Harandi et al., 2022). As Artemis missions rapidly unfold and with the recent announcement of NASA’s planned Moon Base, it is necessary to send automated and/or autonomous missions to deep space to gain a greater understanding of the long-term impacts on biological systems. Given that most fully automated biological experiments to date have been performed onboard small satellites or CubeSats, this Perspective article focuses on those missions.

2 CubeSat-based biosensor technologies in low Earth orbit (LEO)

Given their accessibility and versatility, NASA pioneered the development of biosensor-based payloads in CubeSats over 20 years ago (Massaro Tieze et al., 2023; Santa Maria, 2023). Since 2006, NASA Ames Research Center has developed and launched a variety of biological CubeSats to investigate the impact of the harsh space environment on model biological organisms (Figure 1), including five missions to LEO and the BioSentinel deep space mission that launched in 2022.

NASA launched the first biological CubeSat, GeneSat-1, in 2006 (Ricco et al., 2007). The 3U satellite was launched into LEO, approx. 450 km above Earth, on a US Air Force Minotaur rocket from NASA Wallops Flight Facility. It was composed of a 1U bus and a 2U fully automated and self-contained payload. The science objective was to investigate the effect of the microgravity environment on gene expression in the bacterium E. coli that was genetically modified to express green fluorescent protein (GFP). The Escherichia coli cells were housed inside a 12-well microfluidic card; each well had a dedicated optical detection system consisting of a fluorescence excitation LED light, excitation and emission filters, and an intensity-to-frequency photodetector (Ricco et al., 2007). Once in LEO, the experiment was initiated via remote activation. After temperature increased to 34 °C, growth medium was delivered into the fluidic wells to start cell growth. Both fluorescence and cell growth were successfully observed in all the experimental wells. This mission was the first technology demonstration of a biological experiment in a CubeSat and served as the basis for future, more sophisticated payloads (Table 1).

TABLE 1. Biological CubeSat missions.

PharmaSat launched onboard a Minotaur rocket from NASA Wallops in 2009 to an orbit similar to GeneSat-1. The main objective of PharmaSat was to study the response of the budding yeast Saccharomyces cerevisiae to antifungal treatment in microgravity (Ricco et al., 2011). The 3U CubeSat was also composed of a 2U payload and a 1U bus. The microfluidic system consisted of an improved and larger 48-well fluidic card divided into four banks. This allowed the delivery of growth media containing different doses of the antifungal voriconazole. Cell growth and metabolic activity were monitored via absorbance readings and using the metabolic redox dye alamarBlue or resazurin. Each fluidic well had a dedicated optical system with 3 LED lights (470, 525, and 615 nm) and a photodetector (Figure 1). After reaching its operational orbit, the experiment was activated remotely by increasing the card temperature to 27 °C. Once the temperature stabilized, growth medium and dye were delivered into each well, followed by delivery of different antifungal doses 12 h later. Cell growth and metabolic activity were tracked every 15 min for 3 days (Ricco et al., 2011). Thus, making PharmaSat the first fully automated biological payload to monitor dose-response on a free-flyer in LEO.

Subsequently, NASA launched O/OREOS (Organism/Organic Exposure to Orbital Stresses), the first astrobiology mission in a LEO CubeSat (Nicholson et al., 2011). The 3U CubeSat launched in 2010 onboard a Minotaur rocket from Kodiak Launch Complex in Alaska. In comparison to GeneSat-1 and PharmaSat, it was deployed at a higher orbit, approx. 650 km above Earth. This allowed for a significantly higher radiation dose rate exposure, approx. 12–15 times higher. Moreover, O/OREOS carried two 1U experimental payloads, SEVO (Space Environment Viability of Organics) and SESLO (Space Environment Survivability on Living Organisms). The main objective of SESLO was to investigate the effects of LEO radiation and microgravity on spore germination, growth, and metabolism in the bacterium Bacillus subtilis for up to 6 months. SESLO utilized three 12-well fluidic cards (Figure 1) or bioblocks that allowed activation at three different timepoints (2 weeks, 3 months, and 6 months) (Nicholson et al., 2011). Therefore, once a stable temperature at 37 °C was reached, the growth medium was delivered to each card (Figure 1). Similar to the previous CubeSats, optical measurements were used to monitor biological activity using a 3-color LED illumination system (470, 525, and 615 nm). An onboard radiation detector was used to measure radiation exposure throughout the mission. The O/OREOS mission demonstrated that desiccated biology enables the development of automated instrumentation to study the effect of cumulative radiation doses over multiple months.

In 2014, SporeSat launched from Cape Canaveral, Florida, on a SpaceX Falcon nine rocket as a secondary payload on the ISS resupply mission CRS-3. The 3U CubeSat was deployed during transit to the ISS at approx. 325 km above Earth. The primary objective of SporeSat was to measure germination of fern spores in Ceratopteris richardii as a function of gravity level using calcium ion-sensitive electrodes and minicentrifuges to simulate artificial gravity (Park et al., 2017). In comparison to the previous missions, SporeSat used three lab-on-a-chip devices or bioCDs instead of active fluidic systems. Interestingly, two bioCDs were rotated to simulate artificial gravity using 50-mm minicentrifuges. The bioCDs utilized LED illumination to initiate spore germination while ion-sensitive electrodes were employed to measure concentration of calcium in the fern spores. Unfortunately, the light source necessary to initiate germination failed to turn on and thus the science objective was not achieved (Park et al., 2017). However, SporeSat demonstrated the use of novel technologies on a CubeSat platform, including successful generation of artificial gravity. An upcoming CubeSat mission, SAGE (Swiss Artificial Gravity Experiment), is currently under development to perform aging studies in human cells by generating an artificial gravity environment in LEO (Wicki, 2024). SAGE will be equipped with a microfluidic chip to house the human cells and a fluorescence microscope to observe their behavior.

EcAMSat (E. coli Anti-Microbial Satellite) launched in 2017 onboard an Antares rocket from NASA Wallops bound to the ISS and deployed into an approx. 400-km orbit. EcAMSat was the first 6U biological satellite and the first 6U CubeSat to be deployed from the ISS. The mission objective was to measure cell growth and study antibiotic resistance in uropathogenic E. coli under microgravity conditions (Matin et al., 2017; Padgen et al., 2020). EcAMSat employed a microfluidic system similar to the one in PharmaSat. Once in LEO, temperature was increased to 37 °C and bacteria were grown to stationary phase followed by exposure to different doses of the antibiotic gentamicin. Metabolic activity was then measured using the redox dye alamarBlue (Padgen et al., 2020). Optical measurements were taken every 15 min throughout the experiment. EcAMSat demonstrated the value of the 6U CubeSat configuration for a biological payload in LEO.

Beyond NASA, there are a series on international CubeSat missions of interest. These include the SpacePharma DIDO-2 and DIDO-3 satellites that launched in 2017 and 2020, respectively. SpacePharma, a biotechnology company based in Israel, focuses on microgravity research in LEO to develop new drugs. These 3U CubeSat missions investigated enzymatic reactions and antibiotic resistance in bacteria under microgravity conditions (Krakos, 2024; Piscon et al., 2023). Optical density measurements at 600 nm demonstrated that successful conjugation and dissemination of antibiotic resistance (tetracycline and kanamycin) genes can occur in LEO (Piscon et al., 2023).

LabSat launched on 12 January 2022, onboard the SpaceX Falcon 9 Transporter 3 mission and deployed from a D-Orbit ION platform 2 weeks later. LabSat was developed by a partnership between Wroclaw University of Science and Technology and the SatRevolution company (Śniadek et al., 2025). LabSat was the first CubeSat that contained two different organisms, plants and fungi, within the same payload. Its main objective was to monitor cress (Lepidium sativum) seed germination and fungi (Fusarium culmorum) mycelium growth in glass and 3D-printed lab-chip devices (Śniadek et al., 2025). Unfortunately, due to issues with communications and power supply, the planned biological experiments could not be carried out.

On 1 April 2022, the 3U PlantSat satellite launched on the SpaceX Falcon 9 Transporter 4 mission and deployed from a D-Orbit ION platform into a 550 km orbit (Garrido et al., 2023). The main mission objective of PlantSat was to monitor the behavior of a plant in a microgravity environment. The plant genus Tillandsia, which can be found across the neotropics, the Andes, and the Atacama Desert, was selected for this mission based on its hardiness and versatility. An additional experimental payload carrying extremophile organisms was added late in its development (Garrido et al., 2023). Developed by the Space and Planetary Exploration Laboratory at the University of Chile, an important educational goal was to train students and engineers about satellite technologies.

Two 3U biological satellites, GreenCube and the AstroBio CubeSat, launched onboard the maiden flight of the Vega-C rocket on 13 July 2022, and deployed into a Medium Eart Orbit (MEO) at approximately 6,000 km of altitude. The primary goal of GreenCube was to demonstrate an autonomous biological laboratory for plant cultivation onboard a CubeSat platform (Marzioli et al., 2023). GreenCube carried microgreen seeds of garden cress (L. sativum) inside a pressurized vessel that autonomously monitored air composition and pressure, humidity, air recirculation, and lighting conditions. Plant germination and growth was monitored optically using VIS and IR cameras. AstroBio, on the other hand, carried a fully autonomous lab-on-a-chip platform for performing chemiluminescence-based biological assays (Nascetti et al., 2023). It employs an origami-like microfluidic paper-based analytical device that carries all the reagents in dried form. After substrate rehydration, detection is performed via a photodiode sensor array. Both satellites were managed by the Sapienza University of Rome and the Italian Space Agency.

Moreover, there are several other biological CubeSats under development that aim to study the survival of microbes, plants, and other model organisms in extreme conditions. Importantly, many of these missions are being designed and developed at academic institutions throughout the world, which again highlights the versatility of small satellites to perform automated and/or autonomous biological experiments. In addition, they provide incredible educational and training value for upcoming generations of scientists and engineers.

3 Automated biosensor technologies beyond low Earth orbit (BLEO)

Each CubeSat mission in LEO refined spaceflight-proven technologies and imparted valuable lessons to the following generation of CubeSats (Table 1). This can be seen clearly in the development of the BioSentinel deep space CubeSat and the upcoming LEIA lunar surface mission.

BioSentinel is the first CubeSat developed to support biological experiments in deep space and NASA’s first biological mission outside of Earth’s magnetic field since Apollo 17 in 1972. It launched as a secondary payload onboard the Artemis I rocket in 2022. After deployment and a ∼400-km lunar fly-by, BioSentinel reached a stable heliocentric orbit (Lingam et al., 2025). After more than 3 years, it is currently located at 95+ million kms from the Earth. It had two primary objectives: to investigate the response of a model biological organism to the deep space environment, and to characterize the radiation environment using a charged particle detector (Massaro Tieze et al., 2023; Rahmanian et al., 2025).

BioSentinel had many key innovations and technological advancements compared to previous biological CubeSats (Lingam et al., 2025). The 6U CubeSat is composed of a 4U instrument payload and a 2U spacecraft bus (Ricco et al., 2020; Padgen et al., 2023). At its core is the 4U BioSensor, an automated microfluidic-spectroscopy instrument originally validated on the ISS in 2021–2022. This versatile system can be accommodated into different platforms, including the ISS, lunar landers, and free-flying satellites or CubeSats.

The 4U BioSensor instrument payload increases the number of samples and potential experiments compared to previous missions. This is a result of the eighteen microfluidic cards it contains, where each is composed of sixteen wells (Figure 1) (Padgen et al., 2023). The fluidic cards are divided into two independent fluidic manifolds with embedded desiccant chambers to maintain low humidity inside the cards. The model organism selected for BioSentinel is the budding yeast S. cerevisiae, which can be desiccated for long durations (Santa Maria et al., 2023). The delivery of the growth medium and the alamarBlue redox dye is controlled by a series of individual valves and bubble traps integrated into the manifolds. The growth medium and dye are mixed by a system of pumps and valves, which are integrated into two smaller manifolds that are contained in a separate enclosure together with the reagent bags (Padgen et al., 2023). Once the CubeSat reached a stable heliocentric orbit, the temperature in the fluidic cards was raised to 23 °C before delivery of the mixed growth medium. Cell growth and metabolic activity was monitored using a 3-color LED optical detection system (570, 630, and 850 nm) with measurements every 2 minutes. Two of the LEDs aid in the detection of the oxidized (630 nm) and reduced (570 nm) forms of the alamarBlue dye, while a third one (850 nm) measures light scattering produced by cell growth. Moreover, each fluidic card has a dedicated thermal control system that allows for active cards to be kept at the growth temperature (23 °C), while keeping the remaining cards and fluidic components at ∼5 °C. This allowed for the activation of two cards at a time at increasing distances from Earth, up to nine times throughout the entire mission duration. The BioSentinel nominal mission was supposed to last 6 months. However, the BioSensor instrument had to be turned off after a few weeks due to issues with electronics inside the 4U enclosure, likely due to the high relative humidity inside (Lingam et al., 2025). Importantly, all the subsystems inside the BioSensor (fluidics, optics, mixing, thermal control, data telemetry) were validated both on ISS and in deep space.

BioSentinel also carries a second scientific instrument, a Timepix-based particle detector or linear energy transfer (LET) spectrometer, which was mounted outside the BioSensor enclosure. This radiation detector reports cumulative radiation dose measurements and LET particle spectra (Pinsky and Pospisil, 2020). To date, the BioSentinel LET spectrometer continues to function without major anomalies and has reported dozens of solar particle events or SPEs through solar maximum, deepening our understanding of the radiation environment outside the Earth-Moon system and its potential effects on future crewed missions to the Moon and beyond (Rahmanian et al., 2025).

In addition to the BioSensor instrument and the LET spectrometer, the BioSentinel CubeSat is equipped with subsystems developed for the deep space environment, including solar panels, rechargeable batteries, star tracker navigation, micropropulsion, transponder, antennas, and command and data handling systems (Ricco et al., 2020). All the data are telemetered back to Earth via the Deep Space Network (DSN). Since the Artemis I launch in November 2022, the BioSentinel Mission Operations team at NASA Ames Research Center continues to communicate, upload commands, and download data once a week via the DSN.

Another example of an automated BLEO payload is the Deep Space Radiation Genomics (DSRG) experiment (Zea et al., 2022). Similar to BioSentinel, the DSRG payload launched on Artemis I in November 2022, but it was located inside the Orion spacecraft and thus returned to the ground 25 days later. The main science objective of the DSRG experiment was to identify the metabolic and genomic pathways in yeast affected by microgravity, space radiation, and their combination in the BLEO environment. Barcoded genome-wide knockout and overexpression collections of S. cerevisiae were used in this experiment (Zea et al., 2022). The desiccated yeast collections were rehydrated once outside the Earth’s magnetic field using an automated system. Once on the ground, the samples were processed and analyzed. More recently, in April 2026, AVATAR performed a unique biological experiment inside Orion as part of the Artemis II lunar fly-by mission. AVATAR, short for A Virtual Astronaut Tissue Analog Response, used organ chips grown from the Artemis II astronauts’ own bone marrow stem and progenitor cells. Since the organ chips were exposed to the same trajectory as the astronauts onboard Orion, they were also exposed to the same deep space radiation and microgravity environment. The goal is to understand the effects of this environment on bone marrow. This is the first time organ chips have been utilized beyond LEO, and additional AVATAR chips are expected to be used in future crewed missions.

A variety of science concepts have been proposed for BLEO biological CubeSat missions in recent years. One interesting example is the MINERVA satellite mission, which is being proposed by Mahidol University, Thailand, as a 6U CubeSat on a cislunar orbit for >4 months (Klomchitcharoen et al., 2022). Its primary objective is to investigate the radioresistance capability of genetically engineered Caenorhabditis elegans. By expressing Dsup proteins from tardigrades, it is expected that transgenic C. elegans can tolerate the cislunar radiation environment better than non-modified organisms. DNA damage monitoring will be performed via expression of fluorescent proteins and an optical detection system. MINERVA will house C. elegans inside fluidic cards with dedicated thermal control and an automated monitoring system. A Timepix-based radiation detector, similar to the one on BioSentinel, will be used to characterize the radiation environment in cislunar orbit.

4 Lunar lander-based biosensor technologies

In 2022, NASA selected LEIA through the Payload and Research Investigations on the Surface of the Moon (PRISM) solicitation. LEIA, short for Lunar Explorer Instrument for space biology Applications, is NASA’s first biological payload to be delivered to the Moon via the Commercial Lunar Payload Applications (CLPS) Program and NASA’s first biological mission on the Moon since 1972. The LEIA science suite is composed of three instruments: a 4U BioSensor, a LET spectrometer, and a fast neutron detector (Lambacher et al., 2025). Since most of the bus components from BioSentinel are not needed in LEIA, power and communications will be managed through the CLPS lander. This mission is projected to launch no earlier than October 2027. LEIA’s primary objectives are: to study the biological response to the reduced gravity and radiation environment on the lunar surface, to test the feasibility of synthetic biology countermeasures for human exploration, and to characterize the radiation environment using the two onboard detectors.

LEIA includes design improvements that address the challenges encountered by the BioSentinel free-flyer mission (Lambacher et al., 2025; Lingam et al., 2025). LEIA’s BioSensor, for example, will have sixteen fluidic cards (instead of 18) mounted on two manifolds, thus freeing space for two large desiccant chambers to improve humidity control. The reagent manifolds containing the pumps and valves for reagent mixing are separated from the reagent bags, which are contained in a separate enclosure. Additionally, the LEIA BioSensor enclosure has been redesigned and now includes an open top to allow accessibility for late load or swapping of components prior to launch without compromising the fluidic and electronics systems. LEIA’s cards will carry desiccated yeast cells and will have dedicated thermal control and a 3-LED optical system per fluidic well.

LEIA will carry up to sixteen engineered S. cerevisiae strains. Half of the strains will be used to study the DNA damage and stress response to the lunar surface environment. The fluidic wells carrying these strains will have the same LED lights used on BioSentinel (570, 630, 850 nm) to monitor cell growth and metabolic activity using a redox dye. The remaining strains will be bioengineered to produce dietary antioxidants in the form of carotenoids as potential countermeasures for maintaining astronaut health in future deep space missions (Ball et al., 2021). The production of carotenoids can be monitored using light absorbance at lower wavelengths. Thus, carotenoid-producing yeast cells will be housed in fluidic wells that use a different 3-LED light combination where the 570-nm LED light is replaced with a 465-nm LED (Lambacher et al., 2025). Understanding the response of these strains on the Moon will allow scientists to evaluate and improve upon the capability for biomanufacturing in BLEO space.

Another lunar lander-based biological mission is LEAF. LEAF, short for Lunar Effects on Agricultural Flora, was selected by NASA as part of the Artemis III Deployed Instrument Program in 2024. Even though LEAF was selected as a payload on Artemis III, it is highly likely that it will now fly on Artemis IV after NASA’s recent announcement that Artemis III will not land on the Moon in preparation for Moon Base. LEAF will investigate how the lunar environment affects the physiological responses of model plant crops, including the effects on germination, morphology, growth, carbon dioxide consumption, and oxygen production (Jeseritz et al., 2025). Once on the lunar surface, water will be delivered to the plants using capillary channels for gravity independent hydroponic hydration. The payload will autonomously regulate its internal environment to maintain optimal conditions for plant biomass growth, including temperature, humidity, carbon dioxide levels, and lighting. Cameras inside the chamber will regularly capture images, enabling analysis of plant morphology and development over time. After 3–5 days on the lunar surface, astronauts will collect a subset of seedling samples for return to Earth for subsequent advanced systems biology studies. The surface-stay plants will continue to grow for several more days, providing valuable data on early plant growth and photosynthetic gas exchange. The surface experiment will conclude after approximately 14 Earth days, when solar power is lost due to the lunar night. LEAF is being developed by Space Lab Technologies in Boulder, Colorado.

It is important to mention that the first plant growth experiment on the lunar surface was performed as part of China’s Chang’e−4 mission in 2019 (Li et al., 2021). Chang’e−4 was the first lander/rover mission on the far side of the Moon and carried multiple science instruments. One instrument contained a biosphere experiment led by Chongqing University (Castelvecchi and Tatalovic, 2019). The sealed habitat contained a variety of organisms, including cotton, potato, and Arabidopsis seeds, fruit fly eggs, and yeast, with the objective of generating a mini ecosystem. Even though the experiment only lasted for a short period of time, images from the lander showed cotton seed germination. The biosphere experiment was terminated before the lander entered its first lunar night.

5 What are the main constraints and challenges when planning biological BLEO missions?

Undoubtedly, one of the major constraints of biological missions to deep space is the long pre-launch duration. Currently, BLEO flight opportunities require payloads to be delivered for integration months prior to launch, and the lead-up time could be up to 1 year between the time that the biological samples are prepared and the time mission operations start. For NASA’s BioSentinel mission, for example, the CubeSat was integrated into the Orion Stage Adapter 14 months prior to launch, while LEIA will be delivered to the CLPS lander provider 8 months prior to launch, at the earliest. Because of this, biological organisms must be able to remain in stasis for long periods of time while maintaining viability, thus limiting the type of candidate organisms or even the type of assay that can be performed. Researchers must plan for long-term experiments to test the condition of the biological samples over time. Since most automated biological experiments rely on microfluidic systems, the reagents must also undergo long-term stability testing to account for degradation or loss of activity of certain chemicals.

To make these long-term tests even more complicated, the research teams must estimate the environmental conditions the payload (and biological samples) will experience from the time the samples are prepared until the experiment is initiated in space. For example, payload integration facilities are not always controlled to the required specifications of a biological mission. Temperature and humidity, and fluctuations of both, can significantly affect the viability of the samples. Moreover, after integration into the vehicle/lander, it is usually not possible to turn on the payload to monitor the conditions inside the payload (e.g., temperature, relative humidity, pressure). It is thus very important to maintain good communication between the research team, the payload and vendor integration teams, and the mission operations team. A good strategy involves communicating the requirement to have environmental monitoring of the facility where the payload will be kept during integration and before launch. A simple data logger that provides temperature and relative humidity data over time can provide valuable information to assess mission success even before launch, and to be able to perform a proper ground control experiment. If permitted, a battery-operated data logger inside the payload might be preferred.

With the recent announcement of Moon Base by NASA and the proposed increase in lunar surface activities using commercial landers and vehicles, researchers should remain hopeful that these long lead-up times will get reduced over time. In such case, researchers should not need to perform long-term tests and would allow the use of additional types of organisms, assays, and technologies. We can also envision the option to carry more biological experiments inside pressurized vehicles like Orion (e.g., AVATAR and DSRG), which do not require long waiting times and allow sample return for more sophisticated studies back on Earth.

Once in space, payloads will be exposed to extreme conditions like significant shifts in external temperature and constant omnidirectional ionizing radiation exposure. Thus, the payload instrumentation should have the capacity to mitigate such conditions. This, of course, depends heavily on the type of platform, the specific space environment, and the duration of the mission. BLEO free-flyers, for example, should be able to maintain a proper thermal environment throughout the mission. The most used strategy is self-powering via solar panels and rechargeable batteries. Such strategy allows for long duration experiments in deep space. Payloads mounted on lunar landers and/or vehicles, on the other hand, get power and data handled by the lander, and experiments are currently limited to a single lunar day (∼14 Earth days). Even though not available yet, upcoming CLPS landers should be able to provide survive-the-night capability, thus potentially increasing the duration for biological missions and allowing longer exposures to both reduced lunar gravity and the lunar surface radiation.

One question that always comes up during mission planning and requirement development is whether radiation tolerant components are necessary. Even though the logical answer might be yes, radiation hardened electronics are expensive and sometimes might not even be available. In BioSentinel, for example, a radiation hardened LEON-3 microprocessor was selected due to the complex radiation environment and the original nominal mission duration (i.e., 6–18 months). LEIA, on the other hand, will carry a commercial-of-the-shelf (COTS) computer that has been flight validated and used in the BioSentinel-ISS mission. An on-board COTS computer was selected due to its reliability and the short duration of the LEIA mission. All the components inside the BioSensor enclosure for both BioSentinel and LEIA are COTS, including the pumps, valves, electronic boards, heaters, sensors, etc.

Once a microfluidic-based payload is hermetically sealed prior to launch, an increase in humidity can become problematic for both electronics and the biological samples. Strategies to prevent this include physical separation of reagents/fluids, presence of desiccant inside the payload, use of impermeable materials, potting of electronic connectors, desiccation and/or lyophilization of the biological samples, etc. As mentioned before, many of these strategies were implemented for the design of the LEIA lunar surface mission based on lessons learned from the BioSentinel free-flyer, which experienced electronic malfunctions a few weeks after deployment (Lingam et al., 2025). In addition to these strategies, the BioSensor fluidic manifolds in both missions contain small desiccant chambers that are in vapor-gas contact with the fluidic cards, which maintain a low humidity inside the wells where the biological samples are loaded.

Other common fluidic issues are material biocompatibility, sterilization method, and bubble formation. During material selection for the different fluidic components (cards, manifolds), it is important to select materials that are both biocompatible and that can be sterilized via a benign method. Some sterilization methods like ethylene oxide gas exposure and electron-beam irradiation, while highly effective, can be readily absorbed into plastic polymers (ethylene oxide) or can modify the polymer surface structure (e-beam), potentially generating compatibility issues with both biology and reagents over long durations. Polycarbonate, for example, is a commonly used polymer for manifold manufacturing because it is both biocompatible and autoclavable. Unfortunately, most electronic components cannot be autoclaved, and we must rely on methods like ethylene oxide or e-beam sterilization followed by a bakeout process to remove remaining toxic volatiles. To prevent bubble formation during the delivery of reagents to the biological samples, a common strategy is the use of bubble traps. In both BioSensor-based missions (BioSentinel and LEIA), for example, the fluidic manifolds carry two types of bubble traps, a larger trap at the entry point to remove bubbles formed during fluid movement, and smaller bubble traps connected to each fluidic card to prevent bubbles from entering the cards. Importantly, reagent mixing is improved by the presence of bubble traps in the manifolds.

Even though not the main objective of this discussion, it is important to mention additional differences between LEO and BLEO CubeSats. Communications and telemetry are of particular importance given the nature of these missions (i.e., no Earth return, long distance, automated/autonomous operations). Once in deep space, satellites (and missions on different planetary bodies) rely primarily on the DSN, short for Deep Space Network. The DSN relies on large dish antennas located around the globe, and data can take several minutes to reach Earth depending on the distance and whether relay satellites are involved. LEO communications (including the ISS), on other hand, rely on direct-to-Earth communications or on relay satellites that then beam the data down to ground stations on Earth. BLEO satellites thus require more specialized radio transponders to send and receive data. BioSentinel uses the same Iris deep space transponder used on the MarCO (Mars Cube One) satellites. While expensive and highly sophisticated, the Iris radio has proven to be very reliable to this date. Nowadays, satellite communications are shifting from radio to optical communications or lasers, which can transmit data 10–100 times faster than traditional radio signals. BLEO satellites also rely heavily on more sophisticated navigation and propulsion technologies in order to self-power and to communicate efficiently; in many cases, BLEO satellites carry pre-programmed instructions in order to perform autonomous operations.

It is expected that upcoming and/or proposed biological missions to deep space will rely on observation methods that require and generate large amounts of data, including microscopy, high-resolution imaging, and genome sequencing. Such constraint can be alleviated with current AI/ML tools, which should allow for more efficient onboard data processing and identification of relevant data. In comparison, missions like BioSentinel and LEIA rely on simple optical density measurements and most (if not all) of the raw data can be send back to Earth without major restrictions.

Finally, with the increasing number of planned lunar and deep space missions, researchers, space agencies, commercial entities, and governments must account for planetary protection. Planetary protection is a set of guidelines and policies designed to prevent forward contamination of celestial bodies during exploration missions and to protect Earth from hazards associated with the return of extraterrestrial samples (Olsson-Francis et al., 2026). The Moon, for example, is classified as a category II body, where there are minimal contamination risks, but documentation and organic inventory tracking are required. By ensuring that we minimize the risk of contaminating other celestial bodies, scientific data become more reliable and thus reduce the potential of false-positive results. Importantly, international collaboration is essential to ensure the implementation of planetary protection controls and the sharing of best practices.

6 Future directions and conclusions

Thus far, most biological experiments using automated technologies have been designed to answer fundamental questions that allow scientists to investigate how biological organisms respond to different spaceflight stressors. With the introduction of missions like BioSentinel and LEIA, we can gain a better understanding of how cells can respond to the damage and stress caused by the space radiation environment both in deep interplanetary space and on the lunar surface, respectively. LEIA, in particular, will perform the first applied science experiment on the lunar surface via biomanufacturing of dietary antioxidants on the Moon. Importantly, these instruments have been designed to perform automated operations without crew interaction and are the result of a continuous evolution of previous technologies and procedures. Future long-term missions to deep space and the lunar surface will require such technologies to support the human explorers during scientific missions and to produce biomedical countermeasures, like the use of microbial organisms for in situ production of food and/or medicines.

The development of modular systems with multiple fluidic cards and timepoints in automated platforms, like CubeSats or lander-based payloads, allows long-term experimentation and the addition of different assays or biological organisms in the same instrument. Two limitations of the BioSensor instrument used in both BioSentinel and LEIA is the use of the same biological organism (budding yeast) and a single monitoring approach (optical absorbance). With this in mind, the BioSensor instrument and its different components are currently being tested to perform experiments with other organisms like cyanobacteria, algae, and microgreen crops for future missions to deep space. New assays include fluorescence and luminescence detection without major redesign of the other components inside the BioSensor. The development of miniaturized instruments also allows the introduction of more efficient and sensitive detection systems. We can now envision the use of automated platforms to carry more complex technologies like miniaturized DNA/RNA sequencers and microscopes or even human-based organ chips that can provide a more accurate analog system to understand human responses. Ultimately, the goal is to develop versatile and robust instruments that can be easily adapted to perform different types of experiments and to be compatible with different platforms and/or vehicles.

With the rapid advancements of the commercial space sector and the upcoming Moon Base, consistent payload delivery to the lunar surface has moved from a developmental goal to a more active, high-cadence reality. As long-term crewed missions BLEO advance, automated instruments will be invaluable tools for both fundamental and applied biological research to support sustained human operations on the Moon and beyond.

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Footnotes

Edited by: Marta Cortesão, Instituto de Astrofísica e Ciências do Espaço (IA), Portugal

Reviewed by: Stella Marie Koch, German Aerospace Center (DLR), Germany

Patrycja Śniadek, Wrocław University of Science and Technology, Poland