Novel Flow Cytometric Analysis of the Blood Brain Barrier

1. To develop a novel, flow cytometry based assay to study the effects of various treatments on the specialized brain microvascular endothelial cells of the blood brain barrier. 2. To examine the leukocyte-endothelial cell interactions at the blood brain barrier that mediate adhesion to the vasculature by a flow cytometric assay. Conclusion: This work describes the novel use of flow cytometry to assess the effects of various experimental agents on the specialized endothelial cells comprising the blood brain barrier, with an emphasis on leukocyte-endothelial cell interactions. Each experiment was performed in at least three replicate conditions, three independent times (n=3). Isotype matched negative controls were used for every detection antibody in the study. FMO controls were used in multicolor staining parameters.1. 开发一种基于流式细胞术（flow cytometry）的新型检测方法，用以研究不同处理因素对血脑屏障（blood brain barrier）特化脑微血管内皮细胞的影响。2. 采用流式细胞术检测血脑屏障部位的白细胞-内皮细胞相互作用，该相互作用可介导白细胞对血管的黏附过程。 结论： 本研究阐述了流式细胞术的创新性应用场景，用以评估各类实验干预因素对构成血脑屏障的特化内皮细胞的作用效果，研究重点聚焦于白细胞-内皮细胞相互作用。每项实验均设置至少3组重复条件，并独立重复3次（n=3）。本研究中所有检测抗体均采用同型匹配阴性对照；多色染色实验参数则使用荧光减一（FMO, fluorescence minus one）对照。