Intravital Imaging, Invasive Cell Collection and Expression Profiling

核心A将提供所需的专业知识、设备和解释技能，以定义 在体内分辨率，肿瘤细胞的侵袭表型，其相应的表达谱，相关 FISH，以及肿瘤细胞和包括巨噬细胞在内的间质细胞之间相互作用的特征， 均位于原发肿瘤和肺转移部位。这个核心是唯一可行的方法来使 多光子成像，以及来自活体肿瘤的侵袭细胞的相关收集，可供所有 项目，因为专业知识和设备独一无二，价格昂贵，不能重复用于个人用途。 多光子成像和侵入性细胞采集所需的技能要求这些方法必须 由相同的人对所有项目进行实践，以确保结果的一致性，以便可以比较结果 横跨动物模型。核心A还将作为表情分析和生物信息学的联络人 AECOM和AECC设施使这些技术在项目中的应用统一。最后， 核心A将支持使用高分辨率FISH进行原位表达谱分析。 将专门用于核心A的设施和设备包括： 1.体内肿瘤细胞和间质细胞的多光子显微镜 2.活体侵袭性肿瘤细胞和间质细胞的采集及体外侵袭实验研究 侵袭的自分泌和旁分泌刺激 3.利用抗体和聚合酶链式反应进行侵袭性细胞分型 4.FACS和微型FACS 5.RNA扩增、表达谱和生物信息学 6.具有相关免疫荧光的FISH显微镜

Core A will provide the expertise, equipment and interpretive skills needed to define, at cellular resolution in vivo, the invasive phenotype of tumor cells, their corresponding expression profiles, correlated FISH, and the characterization of interactions between tumor cells and stromal cells including macrophages, all within the primary tumor and at sites of lung metastasis. This Core is the only practical way to make multiphoton imaging, and the correlated collection of invasive cells from live tumors, available to all of the projects since the expertise and equipment is unique, expensive, and cannot be duplicated for individual use. The skill required for multiphoton imaging and invasive cell collection necessitates that these methods be practiced by the same people for all projects to ensure uniformity of results so that results can be compared across animal models. Core A will also act as the liaison for expression profiling and bioinformatics with the AECOM and AECC facilities making the application of these technologies across projects uniform. Finally, Core A will support the use of high resolution FISH for in situ expression profiling. The Facilities and equipment to be dedicated to Core A are: 1. Multiphoton microscopy of tumor cells and stromal cells in vivo 2. In vivo Invasion Assay for the collection of live invasive tumor cells and stromal cells and investigation of autocrine and paracrine stimulation of invasion 3. Invasive cell typing using antibodies and PCR 4. FACs and micro-FACs 5. RNA amplification, expression profiling and Bioinformatics 6. FISH microscopy with correlated immunofluorescence