SBIR Phase II: Widefield Three-Dimensional Superresolution Microscopy Module

这个小型企业创新研究(SBIR)第二阶段项目专注于设计、开发和测试一种突破性的三维(3D)多功能定量光学显微镜模块，该模块具有单分子灵敏度，适用于活细胞研究和纳米级传感。尽管目前光学显微镜发生了革命，但实现超过衍射极限的分辨率的商业解决方案上市缓慢，缺乏灵活性，而且成本高昂。此外，大多数商业产品太适合正在研究的系统，很难推广到广泛使用。将要开发的仪器是基于荧光成像的照明、光学、数据采集和重建算法的集成设计。这些技术能够以极高的精度确定发射器的3D位置，从而使3D成像具有比竞争方法更好的分辨率和景深。实验结果一直显示出分辨率低于20 nm的3D能力，比大多数研究机构目前使用的光学显微镜提高了一个数量级。该项目的更广泛的影响/商业潜力解决了不断扩大的显微镜市场的重大机遇。该模块的独特特性和可获得的价位将为美国和世界各地的数千家实验室提供多功能3D超分辨率成像能力。为了加快商业化进程，该项目寻求实现一个具有成本效益的、灵活的模块化子系统，该子系统可以与现有的商业科学显微镜相结合。凭借其广泛的可及性，该仪器将影响包括生物物理、细胞生物学和生物医学科学与工程在内的多个科学和工程领域。从更广泛的角度来看，该项目将通过为日常生物研究提供纳米级光学成像来提高科学和技术能力，从而加强美国在全球光学显微镜行业的存在。

This Small Business Innovation Research (SBIR) Phase II project is focused on the design, development, and testing of a ground-breaking three-dimensional (3D) multifunctional quantitative optical microscopy module with single-molecule sensitivity suitable for live cell studies and nanometer scale sensing. In spite of the current revolution in optical microscopy, commercial solutions to achieve resolution beyond the diffraction limit have been slow to market, lacking in flexibility, and costly. Furthermore, most commercial products are too tailored to the systems being investigated and are difficult to generalize for widespread use. The instrument to be developed is based on an integrated design of the illumination, optics, data collection, and reconstruction algorithms for fluorescence imaging. These techniques enable the determination of the 3D position of emitters with great precision leading to 3D imaging with better resolution and depth of field than competing methods. Experimental results have consistently shown 3D capability with resolutions below 20 nm, representing one order of magnitude improvement over optical microscopes currently in use at most research institutions.The broader impact/commercial potential of this project addresses a major opportunity in the expanding microscopy market. The module's unique characteristics and accessible price point will offer multifunctional 3D superresolution imaging capability to thousands of laboratories in the US and worldwide. In an effort to accelerate commercialization, the project seeks to implement a cost-effective, flexible modular subsystem that can be integrated with existing commercial scientific microscopes. With its widespread accessibility, the instrument will impact multiple fields of science and engineering including biophysics, cellular biology, and biomedical science and engineering. From a broader perspective, this project will increase scientific and technical capabilities by providing nanoscale optical imaging for everyday biological research, thereby strengthening the US presence in the worldwide optical microscopy industry.