Light engineering module for fast high-resolution whole-cell imaging

摘要 该小型企业创新研究(SBIR)第一阶段项目专注于设计、开发和 一种突破性的多维多功能定量光学显微镜组件的测试 活的全细胞研究。该模块将利用光工程来实现快速的高灵敏度、低噪声、高性能 分辨率测量低至单分子水平，同时提供宽广的体积视场。 尽管光学显微镜正在进行革命，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，容易获得和负担得起。 所提出的仪器是基于照明、3D光学响应、数据 用于荧光成像的采集、重建算法。具体地说，工程3D光激发 在减少光损伤和光毒性的同时，限制背景噪音。三维点的工程化 扩展功能支持多路复用功能，包括扩展景深、高灵敏度3D 单分子或细胞异质性的定位、多色和3D成像。集成系统 将使重建成为可能，具有卓越的切片能力。因此，目标绩效指标、 在学术实验室最近的研究演示的支持下，在以下方面超越了最先进的水平 空间/时间分辨率、信噪比、视野和易用性。 此SBIR项目旨在将可轻松集成的高性价比解决方案商业化 用现有的科学显微镜。商业模块将从第一阶段原型开发，将 包括小占地面积体系结构、一组用于点扩展函数工程的新型光学相位掩模 以及激励整形、稳健的光学机械设计和实时实验控制软件。测试的结果 合作伙伴实验室解决重大生物医学问题的仪器将验证最终用户的接受度并提供 对商业化有价值的反馈。 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。 新的成像能力可以进一步帮助干细胞和大脑研究。 Double Helix Optics是一家初创公司，拥有成熟的记录和对Light的独家许可权 科罗拉多大学的工程技术，以及新颖的四足和多色PSF 斯坦福大学的本地化发展。该公司总部设在美国生物前沿研究所 Boulder是成功将该产品推向市场的最佳选择。

Summary This Small Business Innovation Research (SBIR) Phase I project is focused on the design, development, and testing of a ground-breaking multidimensional multifunctional quantitative optical microscopy module suitable for live whole cell studies. The module will utilize Light Engineering to achieve fast high-sensitivity, low-noise, high- resolution measurements down to the single-molecule level while providing wide volumetric field of view. In spite of the ongoing revolution in optical microscopy, the availability of state-of-the-art commercial solutions has been slow to market, lacking in flexibility, ease of access and affordability. The proposed 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 photo-damage and photo-toxicity. The engineering of the 3D point spread function enables multiplex functionality including an extended depth of field, high-sensitivity 3D localization of single-molecules or cellular heterogeneities, multi-color, and 3D imaging. The integrated system will enable reconstruction with superb sectioning capability. As a result, the target performance metrics, supported by recent research demonstrations in academic labs, outperform the state of the art in terms of spatial/temporal resolution, signal-to-noise ratio, field of view, and ease of use. This SBIR project is targeted towards commercialization of a cost-effective solution that can be easily integrated with existing scientific microscopes. The commercial module, to be developed from the Phase I 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 further assist in stem cell and brain research. Double Helix Optics is a startup company with a proven record and 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.