Light Engineering Module for Fast High-Resolution Whole-Cell Imaging

摘要 该SBIR第二阶段项目的重点是设计改进、开发和测试一种开创性的 适用于活全细胞的多维多功能定量光学显微镜模块系统 学习。拟议的光工程模块化系统满足了对灵活成像的迫切需求 对整个活细胞进行成像的技术具有低的光损伤和光毒性，同时提供高空间和 时间分辨率以及大的体积视场。 尽管光学显微镜取得了非凡的进步，但最先进的商业解决方案的可用性 推向市场的速度很慢，缺乏灵活性和准入的便利性。该模块化仪器基于 照明、3D光学响应、数据采集和重建算法的集成设计 荧光成像。具体地说，设计的3D光激励限制了背景噪音，同时降低了 光损伤和光毒性。经过设计的3D点扩展函数可实现多路复用功能 包括扩展景深成像、单分子或细胞的高灵敏度3D定位 异质、多色和3D成像。因此，目标性能超过了最先进的水平 在时空分辨率、信噪比、视场、单分子定位精度、 和易用性。 该项目旨在将经济高效的模块化解决方案商业化，该解决方案可以很容易 与现有的科学显微镜相结合。准备投入商业使用的原型将包括一个很小的占地面积 结构，一组用于点扩展函数工程和激励整形的新型光学相位掩模，一种 强大的光学机械设计和实时实验控制软件。该仪器的测试具有重要意义 合作伙伴实验室的生物医学问题将验证最终用户的接受度，并提供有价值的反馈 商业化。 这对生物医学成像的影响是深远的。例如，该仪器将有助于研究 肿瘤发生，由于其对检测受体和受体的空间定位的分子敏感性程度 肿瘤/细胞外基质内的其他信号分子。这也将使退行性疾病的研究成为可能。 该仪器可以帮助揭示疾病的分子来源并开发新的治疗策略。 新的成像能力还可以促进干细胞、癌症和大脑研究。 Double Helix Optics是一家创业公司，拥有来自 科罗拉多大学，以及新的四足动物和多色PSF的本地化发展 斯坦福大学。该公司总部设在博尔德的BioFrontiers Institute，处于最佳位置 成功地将这种产品推向市场。

Summary This SBIR Phase II project is focused on the design refinement, development, and testing of a ground-breaking multidimensional multifunctional quantitative optical microscopy modular system suitable for live whole cell studies. The proposed Light Engineering modular system addresses the critical need for flexible imaging techniques to image live whole cells with low photodamage and phototoxicity while providing high spatial and temporal resolution as well as a large volumetric field of view. Despite extraordinary advances in optical microscopy, the availability of state-of-the-art commercial solutions has been slow to market, lacking in flexibility and ease of access. The modular instrument is based on an integrated design of the illumination, 3D optical response, data collection, and reconstruction algorithms for fluorescence imaging. Specifically, engineered 3D light excitation limits the background noise while reducing photodamage and phototoxicity. The engineered 3D point spread functions enable multiplex functionality including an extended depth of field imaging, high-sensitivity 3D localization of single-molecules or cellular heterogeneities, multi-color, and 3D imaging. As a result, the target performance outperforms the state of the art in terms of spatial/temporal resolution, signal-to-noise ratio, field of view, single-molecule localization precision, and ease of use. This project is targeted towards commercialization of a cost-effective modular solution that can be easily integrated with existing scientific microscopes. The commercial-ready prototype, will include a small footprint architecture, a set of novel optical phase masks for point spread function engineering and excitation shaping, a robust optomechanical design, and real-time experiment control software. Tests of the instrument in significant biomedical problems at partners’ labs will validate end-user acceptance and provide valuable feedback towards commercialization. The implications in biomedical imaging are far-reaching. For instance, the instrument would benefit the study of oncogenesis, owing to its degree of molecular sensitivity for detecting the spatial localization of receptors and other signaling molecules within the tumor/extracellular matrix. It would also empower the study of degenerative diseases where the instrument can help reveal their molecular origin and develop novel therapeutic strategies. The new imaging capabilities could also advance stem cell, cancer and brain research. Double Helix Optics is a startup company with exclusive licensing rights to the Light Engineering technology from the University of Colorado, as well as the novel Tetrapod and Multicolor PSF localization developments from Stanford University. The company, headquartered in the BioFrontiers Institute in Boulder, is optimally positioned to successfully bring this product to market.