Collaborative Research: IDBR: Type A: The Nanosizer: A New Tool For the Photochemical Fabrication of Bioactive Nanoarrays

Overview This is a TYPE A proposal to develop a nanolithography instrument, whose purpose is to construct nanoarrays of peptides, oligonucleotides, or glycans with features below 100 nm and where the chemical composition and position at each feature across a 1 cm2 area can be controlled precisely. The proposed research will benefit the following biological research communities: proteomics, genomics, diagnostics, transcriptomics, embryology, and glycobiology. Intellectual merit of the proposed activity An instrument to create ultradense nanopatterns of biologically active molecules will be developed to model the chemical complexity of biological systems or create arrays for determining gene, protein, or glycan expression at the single-cell level. The goal of the proposed activity is to combine new surface chemistries with new instrument capabilities to make The Nanosizer the ideal tool for the in situ synthesis of combinatorial arrays of oligonucleotides, oligopeptides, and oligosaccharides with feature size and shape control over cm2 areas while maintaining nanometer scale resolution. The Nanosizer will provide orders of magnitude improvements over current instrumentation in terms of minimum feature size, feature production rate, and potentially, cost (Type A proposal). Fundamentally, the Nanosizer is enabled by two breakthrough advances to occur in the Mirkin and Braunschweig groups during a collaborative IDBR award, namely 1) the development of massively parallel scanning probe arrays that allow one to simultaneously direct tens of thousands of photochemical reactions in well-defined nano- to microscale regions, and 2) new surface immobilization chemistries for rapid surface-based synthesis. This proposal leverages these two advances with the aim of developing an automated platform that can rapidly construct spatially encoded combinatorial arrangements of biologically active molecules on surfaces. To do this, we propose to develop new instrumentation and photochemistries that enable massively parallel in situ synthesis. Importantly, the Nanosizer is poised to be the first instrument capable of synthesizing arbitrary patterns encompassing square centimeter areas where the chemical composition of each sub-micrometer feature in the pattern can be controlled precisely. This instrument represents an entirely new approach to forming combinatorial arrays of biologically active small molecules based on the new technologies described above, and its development is a highly interdisciplinary effort which combines chemistry, materials science, engineering, and nanotechnology. Additionally, fundamental biological experiments enabled by this new instrument, including measuring gene, protein, or glycan expression at the single cell level, or studying cell adhesion and differentiation, are described. Broader impacts resulting from the proposed activity The proposed research program, based on an extensive body of preliminary and collaborative results, will be carried out at Northwestern University (NU) and the University of Miami (UM). Central to the completion of the proposed project is the training of graduate students and postdoctoral associates (PAs). Two PAs will be trained each year in nanoscience, scanning probe lithography, surface chemistry, and instrument development. Consequently, they will acquire skills that will make them competitive in academia and industry. Over 50% of personnel for both of the project PIs in the last NSF IDBR grant were women and underrepresented minorities while one of the PIs (Braunschweig) is a member