## 2 Materials and Methods
### Missions: Common Procedures  
Pre-flight sample preparation was done at Kennedy Space Center in the Space Station Processing Facility (SSPF). Freshly prepared yeast streak plates, and any specialized reagents or equipment, were shipped overnight to the SSPF laboratory. These reagents were shipped in replicate on consecutive days to minimize any complications of potential transport delays. The research team arrived approximately 10 days prior to launch, navigated the necessary badging and training requirements within the site-specific scheduling and availability, received the shipments of biologics, and began experimental preparations forthwith. Flight and terrestrial control samples, with a predetermined and condition-specific sample excess (spares), were prepared in bulk whenever possible. Following an incubation period, the bulk-prepared samples were inspection for contamination and other anomalies (bubbles, volume discrepancies, etc. ), and removed from the population as necessary. From the remaining samples, flight and terrestrial control specimens were chosen and extra samples were deconstructed, cleaned, and sterilized should they be needed for reloading in the event of an extended launch delay. It should be noted that selection of flight and terrestrial control samples was essentially random other than verifying that flight hardware was imperfection free (no glass chips/cracks, no angled septa, etc.). Following launch, the terrestrial control samples were shipped overnight to the Montana-based laboratory at the appropriate temperature using phase change packaging as needed. Flight and terrestrial control experiments were conducted near synchronously; the offset time being determined before flight and allowing for communication of each procedure from the flight crew, to the Implementation Partner (BioServe Space Technologies, Boulder, CO, USA), and then to the research team. Mission-specific timing is included in Table 1. In addition, Figure 1A,B synopsize the hardware utilized in each mission as well as the general contents and construction of the experimental samples. Specific experimental design details will be provided in the payload-specific sections below and are summarized in Figure 1C. To clarify the payload nomenclature, flight experiments are provided with an organizational name, in our case by NASA Space Biology as part of their funded microbial series of studies (Micro-\(x\)), and the payloads become broadly referred to by these names. The payloads (experiments) are manifested on a spaceflight mission once preflight requirements have been met and flight resources (space on a launch vehicle, crew time, International Space Station (ISS)- based resources) become available. Our first set of experiments is referred to as Micro-6, which was flown on Space X Cargo Resupply Service (CRS)-1, with a reflight referred to as Micro-8, flown on Space X CRS-4. The next set of experiments (Micro-14) was split between two missions so is further defined, for the benefit of clarity herein, by the hardware used, specifically the Fluid Processing Apparatus (FPA) and Fluorinated Ethylene Propylene (FEP) bags. Micro-14/FPA was flown on Space X CRS-16 and Micro-14/FEP was flown on Space X CRS-17. The details of each payload/experiment are described in detail below. In each case, the _C. albicans_ inoculum was prepared and stored in sterile water to maintain stasis. Experiments were initiated, or activated, by the addition of the inoculum to medium to stimulate growth. Following the defined growth period, experiments were terminated by the addition of fixative or by placing the samples at a restricted temperature (\(-95\)\({}^{\circ}\)C or \(4\)\({}^{\circ}\)C, depending on the experiment). Upon completion of the mission and once the samples were received in the home laboratory, yeast were isolated from the hardware, which in many cases required deconstruction of the hardware components (de-integration), and processed as indicated for each of the specific outcome measures.