Flow Cytometry Core

说明（申请人提供） 这一至关重要的核心为脾细胞、颈淋巴结细胞、PBL和CNS来源的炎性细胞提供了常规和一致的分析。正如在个别项目中所指出的，对感染期间浸润CNS的细胞以及驻留CNS细胞（少突胶质细胞、星形胶质细胞和小胶质细胞）的批判性分析是整个项目的重要组成部分。 通过活化状态和CNS驻留细胞的身份，用于精确确定浸润CNS的细胞类型的表面标志物的差异表达对于所提出的项目都是至关重要的。 流式细胞术上级于免疫组织化学，因为可以通过荧光强度以及多种表面或细胞内标记来区分细胞类型。最好的例子是通过CD45染色的不同强度将小胶质细胞与浸润的血管周围巨噬细胞分离。这种区别不能通过组织学检查进行。不能通过组织学检查分析驻留CNS细胞类型。因此，分析驻留的CNS细胞类型的MHC和共刺激分子的表达有望是高度敏感的。此外，通过分选获得的纯化群体需要对其精确身份进行严格评估。 所有这些功能都由这个核心来执行。 核心技术员和核心总监均接受了BD关于配备2个激光器进行四色分析的FACS Calibur流式细胞仪操作的培训。

Description (provided by applicant) This critically important core provides routine and consistent analysis of spleen cells, cervical lymph node cells, PBL and inflammatory cells derived from the CNS. As noted in the individual projects, critical analysis of the cells infiltrating the CNS during infection, as well as the resident CNS cells (oligodendroglia, astrocytes and microglia) are important components of the overall program. Differential expression of surface markers for precise determination of the cell types infiltrating the CNS, thru activation state and the identity of CNS resident cells are all critical to the proposed projects. Flow cytometry is superior to immunohistochemistry in that cell types can be distinguished by intensity of fluorescence as well as by multiple surface or intracellular markers. The best example is the separation of microglia from infiltrating perivascular macrophages via their differential intensity of CD45 staining. This distinction cannot be made by histological examination. Analysis of resident CNS cell types cannot be made by histological examination. Analysis of resident CNS cell types for expression of MHC and co-stimulatory molecules thus promises to be highly sensitive. Furthermore, purified populations to be obtained by sorting, require a critical evaluation of their precise identity. All of these functions are carried out by this core. Both the core technician and the core director have been trained by BD in the operation of the FACS Calibur flow cytometer equipped with 2 lasers for 4-color analysis.