IDBR TYPE B: Development of a $100 high-throughput whole slide imaging kit

授予康涅狄格大学开发具有成本效益的全载玻片成像（WSI）附加套件。高通量全载玻片成像系统是生物学研究的重要工具。通过使用WSI系统，可以在几秒或几分钟内以高分辨率对大规模生物样品进行数字化处理，显著提高实验室效率，促进科学进步。该项目旨在开发一个100美元的WSI附加套件，用于高通量多模态显微镜成像。通过将此附加套件适配到现有的常规显微镜平台，可以将其转换为高通量，多模式WSI平台。WSI附加套件的结果是整个样品的全面数字渲染，尺寸为厘米级，在衍射极限分辨率下可见，二维或三维，并具有多种成像模式。通常在单细胞水平手动进行并一次解决有限视野的实验现在可以以自动化和智能的方式对整个样品进行。由于生物学和生物医学研究的许多领域都需要高内容的图像，因此在大约100美元的预算中传播WSI附加套件可能会导致小型研究实验室中的新型实验设计，并使各种与健康相关的应用受益。 该项目旨在通过集成三个不同的模块来开发WSI附加套件：1）自动对焦模块，可立即识别样品的最佳焦点位置，2）扫描模块，可高速驱动常规显微镜的载物台，以及3）照明模块，可将多种成像模式集成到平台中。对于自动对焦模块，两个针孔调制相机将连接到目镜端口，用于即时焦平面检测。通过分析从这两个目镜端口捕获的图像，可以推断出2D薄切片的最佳焦点位置和3D样品的断层扫描结构，而无需z扫描。不同的z采样策略可以用于更好的图像采集。对于扫描模块，步进电机和相关齿轮将用于控制显微镜载物台的运动。将测试不同的机械耦合方案，并根据扫描速度和位置重复性选择最佳方案。对于照明模块，将在聚光透镜的后焦平面处插入液晶显示器。通过在显示器上设置不同的二进制模式，该照明模块能够将多种成像模式集成到平台中，包括明场、暗场、相位对比、定量相位、偏振、3D断层扫描和超分辨率傅立叶重叠关联成像。该项目的教育/推广计划将包括康州大学研究生和本科生的课程开发，高中生和高中/中学教师的暑期实习。通过该项目产生的知识将通过会议介绍和讲习班、在科学期刊上发表文章以及提高公众科学素养的新闻编辑室文章传播。建立WSI套件的协议将在一个专门的网站上发布。将确定工业合作伙伴，以促进WSI附加套件的商业化。

An award is made to University of Connecticut to develop a cost-effective whole slide imaging (WSI) add-on kit. High-throughput whole slide imaging (WSI) system is an important tool for various biological research applications. By using WSI systems, large-scale biological samples can be digitalized with high resolution in seconds or minutes, significantly increasing laboratory efficiency and promoting the progress of science. This project aims to develop a $100 WSI add-on kit for high-throughput multimodal microscopy imaging. By adapting this add-on kit to an existing regular microscope platform, one can convert it into a high-throughput, multimodal WSI platform. The result from the WSI add-on kit is a comprehensive digital rending of the entire sample, on the order of centimeter in size, visible at diffraction-limited resolution, in two or three dimensions, and with multiple imaging modalities. Experiments that were typically carried out manually in single cell level and addressed a limited field of view at a time can now be done for the entire sample in an automated and intelligent manner. As high-content images are desired in many fields of biological and biomedical research, the dissemination of the WSI add-on kit in ~$100 budget could lead to new types of experimental designs in small research labs and benefit various health-related applications. This project aims to develop the WSI add-on kit by integrating three different modules: 1) an autofocusing module that instantly identifies the optimal focal position of the sample, 2) a scanning module that drives the stages of a regular microscope in high speed, and 3) an illumination module that integrates multiple imaging modalities into the platform. For the autofocusing module, two pinhole-modulated cameras will be attached to the eyepiece ports for instant focal plane detection. By analyzing the captured images from these two eyepiece ports, one can infer the optimal focal position for a 2D thin section and the tomography structure of a 3D sample without a z-scan. Different z-sampling strategies can then be used for better image acquisition. For the scanning module, stepper motors and the associated gears will be used to control the motion of the microscope stages. Different mechanical coupling schemes will be tested and the best scheme will be chosen in terms of the scanning speed and positional repeatability. For the illumination module, a liquid crystal display will be inserted at the back-focal plane of the condenser lens. By setting different binary patterns on the display, this illumination module is able to integrate multiple imaging modalities into the platform, including brightfield, darkfield, phase contrast, quantitative phase, polarization, 3D tomography, and super-resolution Fourier ptychographic imaging. The education/outreach program of this project will include course development for graduate and undergraduate students at UConn, summer internships to high-school students and high-school/middle-school teachers. The knowledge generated through this project will be disseminated through conference presentations and workshops, publications in scientific journals, and news-room articles that raise the scientific literacy of the public. The protocol of building the WSI kit will be posted on a dedicated website. Industrial partners will be identified to promote the commercialization of the WSI add-on kit.