Description: In our previous NASA SBIR Phase II (80NSSC18C0171) we built a single-molecule detection instrument for search of life in outer space. The instrument reveals the molecule size and concentration from unique electrical fingerprints. Our cm-scale instrument relies on fast electronics and thin (20-nm-thick) solid-state nanopores in low-noise glass chips. Our work identified electrical signatures of pure samples (microRNA, DNA, bovine serum albumin (BSA) and polylysine) and environmental samples (synthetic sea water and Mars analogue soils), and compiled a database of results. DNA at concentrations as low as 1.5 ng/g from Mars analogue soil were detected using our instrument. To improve this instrument, we propose two modifications. First, to fabricate advanced, thinner (< 5-nm-thick) nanopore membranes that increase the overall spatial resolution and sensitivity of the nanopore sensors for improved precision to extraterrestrial life search missions. These chips allow detection of molecules as small as ~ 1 nm in size. And second, to integrate nanopores into optimized solid-state nanochannel-nanopore devices that will enable the DNA linearization for improved DNA sequencing, and can be used for linearization and detection of other informational polymers molecules. Nanochannel-linearized DNA can help accuracy of solid-state nanopores for nucleotide resolution by geometrically constraining and guiding long strands of DNA to nanopores for improved threading and reading. Compared to the biological nanopores, solid-state pores offer the promise of robustness and stability, which can survive long flight missions. Furthermore, they own the advantage of being an enzyme-less system, offering a faster, highly controllable, more flexible and robust sequencing platform than the biological pores. We further propose to optimize the nanochannel-nanopore device developed for reading of nucleotides using high-bandwidth electronics.

Benefits: This project directly aligns with the SeqLOW COLDTech goals for the Development of Nanopore Sequencing for Automated Ocean World Life Detection led by Program Officer Christopher McKay at NASA Ames Research. The technology also has potential insertions as an agnostic life detection instrument and small molecule sensor within the scientific payload for both Europa Lander and Enceladus Orbiter mission concepts and for possible follow-on submersible missions. The technology would also be well suited for searching for extant life on Mars.

The proposed nanopore sensor architecture, with its miniaturized and robust design, has potential in a wide variety of terrestrial applications ranging from DNA sequencing, point-of-care diagnostics, human pathogen surveillance to agricultural water monitoring. Additionally, the small molecule analysis capability can be applied to EPA and USDA needs for measuring water quality.