Live microscopy and cytometry in vascular biology

描述(由申请者提供)：这是一个多学科的协作研究项目，汇集了来自多个机构的研究人员，跨越技术和学科边界开展工作。该小组在血管生物学方面有着共同的兴趣，特别是在应用现代光学技术回答与血管生物学相关的关键问题方面。该技术平台将基于谢彭斯眼科研究所和威尔曼光医学实验室之前开发的扫描激光检眼镜和实时在体共聚焦显微镜。现有的技术将随着新的发展而得到加强，以提高活体动物成像的图像分辨率、对比度、灵敏度、量化方法和灵活性。需要解决的具体问题包括： 1.控制正常血管发育和稳定的细胞过程是什么？ 2.调控血管生成、淋巴管生成和免疫细胞迁移的因素是什么？3.镰状细胞和糖尿病视网膜病变的细胞机制是什么？4.我们能否在活体内非侵入性地观察视网膜色素上皮的早期变化？ 5.我们可以在不抽血的情况下检测体内的循环细胞吗？循环中肿瘤细胞的数量能很好地预测肿瘤负担和治疗反应吗？ 细胞水平的成像将使生物学家能够随着时间的推移研究活动物中的问题，获得经典静态测量(组织学、免疫细胞化学等)所无法获得的生理学见解，从而大大减少回答这些关键问题所需的动物数量。

DESCRIPTION (provided by applicant): This is a multidisciplinary, collaborative research program bringing together investigators from several institutions to work across technology and disciplinary boundaries. The group shares a common interest in vascular biology, particularly in the eye, and specifically in the application of modern optical technology to answer critical questions related to vascular biology. The technology platform will be based on the scanning laser ophthalmoscope and the real-time in vivo confocal microscope previously developed at Schepens Eye Research Institute and at the Wellman Laboratories of Photomedicine. The existing technology will be enhanced with new development to improve image resolution, contrast, sensitivity, methods for quantification, and flexibility of imaging in living animals. Specific questions to be addressed include: 1. What are the cellular processes governing normal vascular development and stabilization? 2. What are the factors governing angiogenesis, Iymphangiogenesis, and immune cell trafficking? 3. What are the cellular mechanisms for the development of sickle cell and diabetic retinopathy? 4. Can we visualize early changes in the retinal pigment epithelium noninvasively in vivo? 5. Can we detect circulating cells in vivo without drawing blood? Is the number of circulating tumor cells a good predictor for tumor burden and response to therapy? Imaging at the cellular level will enable biologists to study problems in living animals over time, gaining physiological insights beyond what can be obtained by classic static measurement (histology, immunocytochemistry, etc.), substantially reducing the number of animals required to answer these critical questions.