Vectra Polaris microscope imaging system

项目摘要 该提案旨在为Vectra® Polaris™自动定量成像系统（Polaris™）提供资金，该系统是一个国家级项目， 最先进的多光谱成像显微镜服务于分枝杆菌内的多个NIH支持的项目 研究实验室（MRL）在科罗拉多州立大学（CSU）。我们的集体使命是制定有效的 化学疗法和疫苗的分枝杆菌疾病影响数百万世界各地。生物活性及 候选疫苗和治疗的安全性特征只能通过表征宿主来充分了解 免疫应答，主要通过流式细胞术评估大型免疫生物标志物网络， 免疫组化MRL研究人员正在寻求通过以下方式可视化免疫生物标志物的空间表达： 从分枝杆菌疾病的临床前模型获得的完整组织样品的多重染色。到现在为止， 通过共聚焦和传统的免疫荧光进行的生物标志物的显微镜分析受到以下因素的极大限制： 组织中的自发荧光（特别是福尔马林固定的和患病的组织）和低信噪比。的 Polaris™系统具有一套独特的显微镜功能，与inForm®工作站相结合， 组织切片中多个靶标及其空间关系的同时鉴定和定量 和微阵列。PolarisTM多光谱技术的一个关键方面是能够轻松区分 通过检测来自阳性生物标志物信号的自体荧光，显著提高信噪比。这允许 更灵敏和准确的样品分析，并能够揭示真正的细胞间相互作用。的 系统是用户友好的，其自动扫描系统允许高速数字和全载玻片扫描 10- 40倍之间的明场或荧光。照明系统的性质显著降低 光漂白和光损伤等有害影响，因此样本可以成像更长时间 时间周期比，例如，共焦显微镜。此外，基于先进机器学习的 所包括的软件的方法自动分割和定量组织结构。Polaris™系统 将被纳入我们现有的显微镜核心设施，以扩大获得学员和其他 研究小组在科罗拉多州立大学和北方科罗拉多，同时优先访问的主要用户。纳入 基础核心将维持长期运作，并提供一致的监督和培训。总的来说， 这种最先进的成像技术对化学疗法和疫苗的发展至关重要 并将增加价值，以多个其他NIH支持的项目在CSU。

PROJECT SUMMARY This proposal seeks funding for a Vectra® Polaris™ Automated Quantitative Imaging System (Polaris™), a state- of-the-art multispectral imaging microscope to serve multiple NIH-supported projects within the Mycobacteria Research Laboratories (MRL) at Colorado State University (CSU). Our collective mission is to develop effective chemotherapies and vaccines for mycobacterial diseases affecting millions worldwide. The biological activity and safety profile of candidate vaccines and treatments can only be fully understood through characterizing the host immune response, primarily by evaluating large networks of immune biomarkers through flow cytometry and immunohistochemistry. MRL researchers are seeking to visualize the spatial expression of immune biomarkers by multiplex staining of intact tissue samples obtained from preclinical models of mycobacterial diseases. Until now, microscopic analysis of biomarkers through confocal and traditional immunofluorescence was drastically limited by autofluorescence in tissue (especially in formalin fixed and diseased tissue) and low signal-to-noise ratios. The Polaris™ system has a unique set of microscopic capabilities that combined with the inForm® workstation allows simultaneous identification and quantification of multiple targets and their spatial relationships in tissue sections and microarrays. A critical aspect of the PolarisTM multispectral technology is the ability to readily distinguish autofluorescence from positive biomarker signal, improving the signal-to-noise ratio substantially. This allows for a more sensitive and accurate analysis of samples and enables true cell-to-cell interactions to be revealed. The system is user-friendly, and its automated scanning system allows for high speed digital and whole-slide scanning between 10-40x in brightfield or fluorescence. The nature of the illumination system significantly decreases deleterious effects such as photobleaching and photodamage, and thus specimens can be imaged for longer periods of time than with, for example, a confocal microscope. Furthermore, the advanced machine-learning based approaches of the included software automatically segment and quantitate tissue structures. The Polaris™ system will be incorporated into our existing microscopy core facility in order to expand access to trainees and other research groups at CSU and in Northern Colorado, while prioritizing access for the major users. Incorporation into the foundational core will sustain long term operation and provide consistent oversight and training. In summary, this state-of-the-art imaging technology is of critical importance to development of chemotherapies and vaccines by the MRL and will add value to multiple other NIH-supported projects at CSU.