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

这个i-Corps项目的更广泛的影响/商业潜力是开发一种超分辨率显微镜方法，旨在对整个细胞和组织样本进行详细的3D成像。大多数生物实验室使用荧光显微镜，大多数是共焦的，分辨率极限在300 nm左右，这对许多亚细胞结构来说是不够的。虽然扫描电子显微镜(SEM)和低温电子显微镜等其他技术提供了更高的分辨率，但它们也有自己的局限性，例如无法区分蛋白质种类。该系统建立在超分辨率显微镜的基础工作基础上，与活细胞兼容，并使用自适应光学、同时双色成像和实时、无伪影的超分辨率图像重建。这项拟议的技术可能具有广泛的影响，促进细胞生物学、生物医学工程和治疗学的进步，并提供对生物过程的细粒度见解，并潜在地改进诊断和治疗。这项I-Corps项目基于为3D组织成像定制的集成单分子定位显微镜(SMLM)的开发。虽然传统的荧光显微镜，如共焦，由于光衍射达到了分辨率在~300 nm以上的极限，但SMLM提供了一种独特的方法。这项拟议的技术利用了单个荧光团的“闪烁”现象，并在多帧图像中精确定位这些事件，从而产生超清晰的超分辨率图像。此外，这项技术将人工智能驱动的自适应光学技术集成到该过程中，便于对样本诱导的像差进行瞬时调整，从而释放了深层次组织超分辨率成像的能力，最高可切割250微米的脑切片。这项拟议的技术可能服务于不同的研究领域，特别是研究细胞生物学、神经科学和发育生物学的实验室环境。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.