Microscopy-Image Analysis Core

显微镜和图像核心一直是PPG历史上的关键核心资源。 由于我们对细胞命运的建立和基因的相互作用越来越感兴趣， 在发育中的肺中表达，我们预期继续大量使用这种有价值的资源。此外，本发明还提供了一种方法， 为了提供组织学技术和分析的帮助，我们增加了流式细胞术和细胞计数。 将组件排序到此核心。这利用了标记的单克隆抗体的广泛可用性， 抗体，承认这种技术的力量，现在是可能的，因为一个机构的高度 快速流式细胞仪设备。此外，我们还发展了必要的专门知识，以方便我们使用 一种新的激光捕获显微切割仪器 该核心旨在专门提供：1）在各种性能方面的专业知识和咨询 基于组织学和流式细胞术的技术2）PPG的正式和实际操作培训 免疫染色，原位杂交， 图像分析和流式细胞仪数据挖掘3）关键设备的管理及其预算 4）这些程序所需的一般用品（石蜡、载玻片等）5)一个 经验丰富的技术人员管理核心设备，教授组织学程序，帮助标准化新的 方法，并在我们的部门计算机服务器上维护实验室协议，以及6） 个体研究者使用机构仪器（高速细胞分选仪和共聚焦显微镜）。 该PPG具有成功采用免疫组织化学和原位杂交的悠久传统， 以及发展新的组织学技术来研究肺的发育。这一成功归功于 方案、试剂、设备的有效管理，以及核心人员的日常监督 其主任。在核心中，我们现在有3个显微镜，基于计算机软件成像， 明场、相位、暗场和荧光显微镜的分析能力。此外，还有2 我们将使用的机构共聚焦仪器，我们已经准备好使用高速细胞分选仪， 分析细胞仪装置。总体而言，我们预计这一核心将继续成为确保 将实现各个项目的科学目标。

A Microscopy and Image Core has been a key, central resource throughout the history of this PPG. Since we have become more interested in the interplay between establishment of cell fate and gene expression in the developing lung, we anticipate continued heavy use of this valuable resource. In addition, to providing assistance for histological techniques and analysis, we have added a flow cytometry and cell sorting component to this Core. This takes advantage of the wide availability of tagged monoclonal antibodies, acknowledges the power of this technique, and is now possible because of an institutional high speed flow cytometry facility. Further, we have also developed the necessary expertise to facilitate our use of a newly available laser capture micro-dissection intsrument. This Core is designed to specifically provide: 1) expertise and consultation in the performance of various histological and flow-cytomtery based techniques 2) formal and practical hands-on training for PPG personnel and post-doctoral trainees in relevant techniques such as immuno-staining, in situ hybridization, image analysis, and flow cytometry data mining 3) management of key equipment and a budget for their maintenance 4) common supplies required for these procedures (paraffin, glass slides etc.) 5) an experienced technician to manage core equipment, teach histological procedures, help standardize new methods, and maintain lab protocols on our department computer server and 6) budgetary support for individual investigator use of institutional instruments (high speed cell sorter and confocal microscope). This PPG has a long tradition of successfully employing immunohistochemistry and in situ hybiridization, and in developing new histological techniques for studies of lung development. This success is due to the efficient management of protocols, reagents, equipment, and the fact that there is daily oversight by the Core by its Director. Within the Core, we now have 3 microscopes with computer software based imaging and analytical capabilities for brighfield, phase, darkfield, and fluorescence microscopy. Further, there are 2 institutional confocal instruments that we will use, and we have ready access to a high-speed cell sorter and analytical cytometry device. Overall, we expect this Core to continue to be a key factor ensuring that the scientific goals of the individual projects will be achieved.