Self-Contained Device Isolates Biological Samples

Gene expression analysis measures the activity of genes and reveals valuable information about the internal states of living cells. NASA Ames has developed a novel system for conducting quantitative, real-time gene expression analysis that not only significantly improves the sample preparation procedures and provides a quick, cost-effective solution needed in a laboratory, but also minimizes the risk of RNA contamination and degradation. The invention, a novel assay methodology and suite of devices have been developed to isolate nucleic acids from Prokaryotic and Eukaryotic cells and prepare samples for Reverse Transcriptase - quantitative Polymerase Chain Reaction (RT-qPCR) analysis. The invention consists of a method to extract and purify nucleic acids from biospecimen that does not require organics or toxic chemicals, and provides containment for the liquids. The assay employs an aqueous-based non-alcohol method that yields robust RNA quality; and PCR reaction tubes that are pre-loaded with stabilized lyophilized reagents designed to perform RT-qPCR analysis - all in all enabling rapid, cost effective, and portable manual operation in the laboratory or remote field environments. This innovative technology greatly reduces preparation time to less than 10% of the time it would take using standard techniques. This technology could be used with any commercial quantitative Polymerase Chain Reaction (qPCR) machine.

Bacterial contamination of water systems used in microgravity is a major issue for NASA because biofilms can clog or interfere with water system functions and bacterial ingestion can be harmful to astronaut health. To address this problem, NASA innovators developed a microwave based technology to purify contaminated water by eradicating and eliminating bacteria that grows in systems that generate potable water, in equipment utilizing cooling loops and heat exchangers, and removing bacterial contamination that is present on a variety of surfaces. This decontamination system is chemical free and requires minimal to no consumables. Initial testing identified a specific microwave frequency band and exposure times for killing bacteria (Burkholderia cepacia) and biofilm. Test results show that exposing static water to microwave energy for 90 seconds can effectively kill waterborne bacteria and biofilm within a water filtration system. Additional testing, using a circulating water test bed, demonstrated that microwave energy at the selected frequency can effectively eradicate waterborne bacteria within 30 seconds. This technology could be further developed into a portable, lightweight system for use in remote locations as well as commercial space applications. The microwave decontamination system could also be added to existing water systems to extend the life of the system.

The innovative technology from Ames acts as a microfluidics switch array, using combinations of pressure and flow conditions to achieve specific logic states that determine the sequences of sample movement between microfluidic wells. This advancement will enable automation of complex laboratory techniques not possible with earlier microfluidics technologies that are designed to follow predetermined flow paths and well targets. This novel method also enables autonomous operations including changing the flow paths and targeting well configurations in situ based on measured data decision parameters. This microfluidic system can be reconfigured for use in various experimental applications, requiring only an adjustment of the programmed pressure sequencing, reducing the need for custom design and development for each new application. For example, this technology could provide the ability to selectively constrain or move biological specimens in the experimental wells, allowing evolutionary studies of model organisms in response to various stressors, evaluation of different growth conditions on biological production of antibodies or other small molecule therapeutics, among other potential applications. Likewise, this platform can be used to foster time-dependent, step-wise, chemical reactions, which could be used for novel chemical processes or in situ resource utilization.

The technology provides miniaturized techniques for extracting trace amounts of organic molecules (lipids) from natural samples. It operates as an autonomous, miniaturized fluidic system, integrating lab techniques for lipid analysis while minimizing reagent volumes and concentrating organics for analysis, thereby increasing signal-to-noise ratios by orders of magnitude. The non-aqueous fluidic system described herein for astrobiological and life-detection missions (either in situ or returned sample) is configured to extract lipid organics from regolith using (1) a fluidic sample processor made of materials compatible with organic solvents and (2) a machine-learning system to select processing steps and parameters to maximize lipid yield. A critical gap is bridged by integrating technologies into a system that replicates analytical lab procedures autonomously on a spaceflight instrument scale with fidelity to original lab techniques. Automated fluidic devices combine controlled handling of liquids with sequential operations and parallelization of replicate processes. By designing such systems to closely interface with both sample-delivery and analytical measurement systems, laboratory analyses are automated. The technology adapts best practice laboratory methods for lipid analysis, overcoming analytical challenges like low organic abundance, interference of minerals/salts, and degradation of origin-diagnostic molecular structures. The extraction and concentration techniques from rock/soil samples can be applied to any biomarkers by changing the solvent, temperature, and agitation.

The miniature bioreactor system was developed to provide the capabilities for NASA to perform cell studies in space and then provide results back to investigators on Earth with minimal tools and cost. The miniature bioreactor system has the potential to also be used on Earth as a laboratory bench-top cell culturing system without the need for expensive equipment and reagents. The system can be operated under computer control to reduce the operator handling and to reduce result variations. The system includes a bioreactor, a fluid-handling subsystem, a chamber wherein the bioreactor is maintained in a controlled atmosphere and temperature, and control subsystems. The system can be used to culture both anchorage dependent and suspension cells (prokaryotic or eukaryotic cell types). Cells can be cultured for extended periods of time in this system, and samples of cells can be extracted and analyzed at specified intervals. The miniature bioreactor system for cell culturing has applications in pharmaceutical drug screening and cell culture studies.