TECH Core

这项技术是U54肿瘤免疫微环境多参数成像中心(C-MITIE)的核心技术 将开发高级成像和数据分析的集成工具包，以增强量化、机械化 预后不良实体瘤的免疫微环境动力学研究有很大的需求 改进的成像方法可以促进对物理和分子机制的理解 控制天然肿瘤微环境中的免疫渗透、分布和功能。我们提出了一个 寻求确定两个研究试验台的多参数成像和计算方法的数量 有效的抗肿瘤免疫和免疫治疗的物理和分子障碍。的一个主要主题 技术方法是使用无标记成像方法，可以表征和量化相互作用 免疫细胞和肿瘤微环境之间的关系。这些无标签方法在很大程度上构建在 多光子显微镜的平台方法，可用于完整的细胞和组织模型，只需最少的 微扰。T细胞的识别和激活将通过使用新的荧光寿命的代谢谱来跟踪 (FLIM)和超维成像(HDIM)方法。这些代谢敏感的方法将是 辅以全场光学相干断层扫描(FFOCT)，揭示对代谢的新见解 相关架构。基于Flim的FRET可以用来对信号分子相互作用产生新的见解 与免疫微环境动力学相关的富含胶原的细胞外基质(ECM)将受到质疑 用二次谐波(SHG)成像测量胶原纤维的拓扑结构和胶原交叉 将测量与增强型后向散射光谱(EBS)联系起来。多光子激发(MPE) 可以使用光化学制造来创建体外细胞就绪的胶原纤维组织模型，该模型 直接基于人类数据蓝图。高级计算分析方法，包括算法和 将使用机器学习方法来检查所有多参数信号，并在 免疫和微环境的相互作用。所有成像和计算方法将不仅共享 在密歇根大学和密歇根大学研究团队中广泛存在，但重要的是与普通癌症成像社区一起使用 制定了硬件和开源软件传播协议。

This TECH core of the U54 Center for Multiparametric Imaging of Tumor Immune Microenvironments (C-MITIE) will develop an integrated toolkit of advanced imaging and data analysis to power quantitative, mechanistic investigations of immune-microenvironment dynamics in poor prognosis solid tumors. There is great need for improved imaging methods that can advance understanding of the physical and molecular mechanisms governing immune infiltration, distribution, and function in native tumor microenvironments. We propose a number of multiparametric imaging and computational methods for the two research test beds that seek to define the physical and molecular barriers to effective anti-tumor immunity and immunotherapies. A major theme of the TECH approach is to use label-free imaging approaches that can characterize and quantitate the interactions between immune cells and the tumor microenvironment. These label free methods are largely built on the platform method of multiphoton microscopy and can be used on intact cell and tissue models with minimal perturbation. T-cell identity and activation will be tracked by metabolic profiling using new fluorescence lifetime (FLIM) and hyperdimensional imaging (HDIM) approaches. These metabolically sensitive methods will be complemented by Full-Field Optical Coherence Tomography (FFOCT) to reveal new insight into metabolically relevant architecture. FLIM based FRET can be used to yield new insights into signaling molecular interactions relevant to immune-microenvironment dynamics The collagen rich extracellular matrix (ECM) will be queried with Second Harmonic Generation (SHG) imaging for collagen fiber topology measurement and collagen cross- linking measurements with Enhanced Backscattering Spectroscopy (EBS). Multiphoton Excitation (MPE) photochemistry fabrication can be used to create in vitro cell ready models of collagen fiber organization that are directly based on human data blueprints. Advanced computational analysis methods including algorithmic and machine learning approaches will be used to examine all multiparametric signals and make correlation between immune and microenvironment interactions. All imaging and computational methods will be shared not only widely within the UW and UMN research teams but importantly with the general cancer imaging community using established hardware and open source software dissemination protocols.