I-Corps: A super-resolution optical imaging system for whole cells and tissues

The broader impact/commercial potential of this I-Corps project is the development of a super-resolution microscopy approach designed for detailed 3D imaging in whole cell and tissue samples. Most biological laboratories utilize fluorescence microscopy, with the majority being confocal and having a resolution limit of around 300 nm, which is insufficient for many subcellular structures. While other techniques such as scanning electron microscopy (SEM) and cryogenic electron microscopy offer greater resolution, they come with their own limitations, such as the inability to differentiate protein species. The proposed system, built on the foundational work on super-resolution microscopy, is compatible with live cells, and uses adaptive optics, simultaneous dual-color imaging, and real-time, artifact-free super-resolution image reconstruction. The proposed technology may have broad implications, facilitating advancements in cell biology, biomedical engineering, and therapeutics, and offering granular insights into biological processes and potentially improving diagnostics and treatments. This I-Corps project is based on the development of an integrated single-molecule localization microscopy (SMLM) tailored for 3D tissue imaging. While conventional fluorescence microscopes like confocal reach their limit with a resolution above ~300 nm due to light diffraction, the SMLM provides a distinct methodology. The proposed technology capitalizes on the "blinking" phenomena of individual fluorophores, and pinpoints these events over multiple frames, resulting in an ultra-defined super-resolution image. In addition, this technology integrates artificial intelligence driven adaptive optics into the process facilitating instantaneous adjustments for sample-induced aberrations, which unlocks the capability for deep tissue super-resolution imaging up to 250 µm-cut brain sections. The proposed technology may serve varied research landscapes, specifically lab environments that work on cell biology, neuroscience, and developmental biology. 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.