BROWN UNIVERSITY CONSORTIUM - PHENOTYPING CORE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 这一核心专注于鉴定骨髓细胞移植后皮肤和骨骼肌中细胞的性质。总体计划是通过共聚焦和去卷积荧光显微镜来确定供体细胞及其特定谱系并进行共定位。在这个计划中，通过绿色荧光蛋白、β-半乳糖苷酶的表达或Make DNA的存在，可以在组织切片或细胞悬液中追踪供体细胞。供体细胞的谱系可以通过标准的形态、地理位置和免疫组织化学来确定。使用共聚焦或去卷积荧光显微镜可以确认供体和谱系标记的共定位。核心可以为这些方法提供高水平的专业知识。核心包括两个蔡司荧光电机驱动的显微镜，一个直立的Axioplan 2和一个倒置的Axiovert 200M，以及一个能够进行四色成像的蔡司510共焦激光扫描显微镜。该核心能够进行标准的去卷积荧光显微镜、3D重建和共聚焦显微镜。后者加上光映射可用于同时显示供体标记和细胞类型特定标记。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This core focuses on identifying the nature of cells in skin and skeletal muscle after marrow cells transplantation. The overall plan has been for donor cells and their specific lineages to be ascertained and colocalized by confocal and deconvolution fluorescent microscopy. In this plan, donor cells can be tracked in tissue sections or cell suspensions by expression of GFP, beta-galactosidase or the presence of make DNA. The lineage of identified donor cells can be ascertained by standard morphology, geographical location and immunohistochemistry. Colocalization of donor and lineage markers canbe confirmed using confocal or deconvolution fluorescent microscopy. The core canprovide high-level expertise for these approaches. The core contains two Zeiss fluorescent motor-driven microscopes, an upright Axioplan 2 and an inverted Axiovert 200M, and a Zeiss 510 confocal laser-scanning microscope capable of four-color imaging. The core is capable of standard deconvolution fluorescent microscopy, 3D reconstructions and confocal microscopy. These latter, plus photomapping can be used to simulateously show a donor marker and a cell type specific marker.