Vectra Polaris microscope imaging system

项目总结 这项提案寻求为Vectra®北极星™自动定量成像系统(北极星™)提供资金，这是一种州- 最先进的多光谱成像显微镜为分枝杆菌内的多个NIH支持的项目服务 科罗拉多州立大学(CSU)研究实验室(MRL)。我们的集体使命是开发有效的 影响全球数百万人的分枝杆菌疾病的化疗和疫苗。生物活性和 只有通过描述宿主的特征，才能完全了解候选疫苗和治疗方法的安全性 免疫反应，主要是通过流式细胞仪和 免疫组织化学。MRL研究人员正在寻求通过以下方式可视化免疫生物标记物的空间表达 从临床前分枝杆菌疾病模型中获得的完整组织样本的多重染色。到目前为止， 通过共聚焦和传统的免疫荧光对生物标记物的显微分析受到以下因素的严重限制 组织中的自发荧光(特别是福尔马林固定和患病的组织)和低信噪比。这个 北极星™系统具有一组独特的微观功能，与INFORM®工作站相结合，使 组织切片中多个目标及其空间关系的同时识别和量化 和微阵列。PolarisTM多光谱技术的一个关键方面是能够容易地区分 从阳性生物标志物信号中自动荧光，大大提高了信噪比。这允许一个 更灵敏、更准确的样品分析，并能够揭示真正的细胞间相互作用。这个 系统界面友好，自动扫描系统可实现高速数字扫描和全幻灯片扫描 在10-40倍的亮度或荧光值之间。照明系统的性质显著降低 有害的影响，如光漂白和光损伤，因此样品可以更长时间地成像 例如，与使用共焦显微镜相比，时间间隔更短。此外，先进的基于机器学习的 所包含的软件的方法自动分割和量化组织结构。北极星™系统 将被纳入我们现有的显微镜核心设施，以扩大对受训者和其他 科罗拉多州州立大学和北科罗拉多州的研究小组，同时优先考虑主要用户的接入。合并到 基础核心将维持长期运作，并提供一致的监督和培训。总而言之， 这种最先进的成像技术对化疗和疫苗的发展至关重要。 并将为CSU的其他多个由NIH支持的项目增值。

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.