CORE--FLUORESCENCE ACTIVATED CELL SORTING (FACS)

流式细胞仪核心设施提供细胞分析， 这个项目的研究人员。实验室提供了 并开发了单细胞分析的尖端技术。 随着这项资助的重点放在细胞凋亡上，TdT-b-dUTP检测 凋亡（TUNEL）的建立和修改， DNA和BUdR联合多参数分析 测量（细胞周期，倍性），免疫表型分析 祖细胞和谱系限制的细胞群， 细胞凋亡相关蛋白（bcl-2、p53、Rb）， 细胞表面抗原包括Fas和MDR 1。最近，绑定 膜联蛋白V与磷脂酰丝氨酸（PS）的结合被认为是检测 在变化之前的膜脂质结构的变化 通过TUNEL在经历凋亡的细胞中检测。定量 细胞抗原现在可以让我们确定 抗体结合能力（ABC）。细胞动力学变化可以是 在输注IUdR和BUdR的患者中研究， 通过流式细胞术分析。荧光原位杂交 （FISH）也已与细胞分裂试验相结合， 我们可以区分正常和白血病细胞的凋亡。的 残留白血病细胞的数量、表型和增殖 (MRD)可以用FISH分析的异常， 在30，000名正常人中， 细胞该测定可预测缓解持续时间， 用于诱导治疗后的患者和骨髓 移植

The FACS Core facility provides cellular analysis to the investigators of this program project. The laboratory has provided and developed cutting edge techniques in single cell analysis. With the focus of this grant on apoptosis, the TdT-b-dUTP assay for apoptosis (TUNEL) was established and modified for multiparameter analysis in combination with DNA and BUdR measurements (cell cycle, ploidy), immunophenotype to analyze progenitor cells and lineage restricted cell populations, intracellular proteins related to apoptosis (bcl-2, p53, Rb), and cell surface antigens including fas and MDR1. Recently, binding of Annexin V to phosphatidyl serine (PS) was recognized as a test for changes in the membrane lipid structure that precedes changes detected by TUNEL in cells undergoing apoptosis. Quantitation of cellular antigens is now available allowing us to determine the Antibody Binding Capacity (ABC). Cell kinetic changes can be studied in patients with infusion of IUdR and BUdR and subsequent analysis by flow cytometry. Fluorescence in situ hybridization (FISH) has also been combined with the apoptosis-assays, allowing us to discriminate apoptosis in normal and leukemic cells. The number, phenotype and proliferation of residual leukemic cells (MRD) with abnormalities amenable to FISH analysis can be determined at levels of as few as 1 leukemic in 30,000 normal cells. This assay is predictive of remission duration and will be applied to patients after induction therapy and bone marrow transplantation.