MRI: Track 1 Acquisition of a System for Integrated Confocal Microscopy and Mechanical Interrogation

This Major Research Instrumentation (MRI) award supports the acquisition of three commercially-available instruments that will be combined to create a new instrument. The three instruments to be acquired are a confocal microscope, an atomic force microscope (AFM), and a laser cutter. The confocal microscope will be able to do fast, high-resolution optical imaging of living cells and tissues as well as soft gels and other soft materials. The AFM will be able to apply well-controlled forces at well-defined locations on these samples, to measure their mechanical properties. Additionally, the AFM can also be used to do higher-resolution imaging of sample surfaces than the confocal light microscope. The laser cutter will be used to cut biological tissues or fiber networks while they are under tension, so that the amount of tension in the structure can be measured by measuring the recoil following the cut. Working together, the combination of these instruments will allow determination of the mechanical properties that develop in biological cells and tissues and how biological cells and tissues respond to a range of mechanical inputs from their environments. Although it is well-known that the interplay between biology and mechanics is important in a wide range of essential biological processes, specific tools for probing the different types of interplay have been limited. Therefore, the acquisition of this instrument will substantially advance basic scientific understanding of biological systems and of non-biological systems that resemble biological systems in composition, structure, and/or function. Acquisition of this instrument will also advance the engineering of materials that interface with biological systems to tune biological response in the desired way. This will benefit society through advances in multiple areas of biomedicine. The instrument will offer new training and research opportunities to scientists and engineers at all stages of their education and career. Specific efforts will be focused on benefiting early-career researchers and those who are members of under-represented groups. A high-resolution, high-speed confocal microscope, an AFM designed for work with “wet” biological and soft-matter samples, and a laser cutter will be combined to create one instrument, termed the “mechanoscope.” This instrument will be used to investigate mechanobiology, including the fiber networks that characterize the cellular cytoskeleton and the extracellular matrix, single-cell eukaryotic and prokaryotic mechanobiology (with a particular emphasis on signaling and differentiation), and tissue- and organism-level mechanobiology (with a particular emphasis on connective tissue and morphogenesis). This instrument will also be used to investigate interactions at the biotic-abiotic interface, focusing on photo-responsive dynamic hydrogels. Further, the instrument will be used in the development of new soft materials, including active soft gels, peptide fibrils, and stimuli-responsive soft materials that incorporate genetically engineered bacteria as a functional component for tuning gel properties. Confocal microscopy will be used for imaging and Forster/fluorescence resonance energy transfer (FRET) measurements. AFM will be used for controlled force application and for measurement of Young’s moduli and storage and loss moduli. The laser cutter will be used for ablation of fiber networks, cellular cytoskeletons, and tissues, and as a photo-stimulus for crosslinking dynamic hydrogels. This project is jointly funded by the Major Instrumentation Research Program (MRI) and the Biomechanics and Mechanobiology Program (BMMB) in the division of Civil, Mechanical and Manufacturing Innovation (CMMI). This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.