CORE--FLUORESCENCE ACTIVATED CELL SORTING (FACS)

FACS核心设施为 该计划项目的调查人员。实验室已经提供了 并开发了单细胞分析的尖端技术。 由于这笔赠款的重点是细胞凋亡，TdT-b-dUTP检测 建立并修改了细胞凋亡率(TUNEL) DNA和BUDR相结合的多参数分析 测量(细胞周期、倍体)、免疫表型分析 祖细胞和血统受限的细胞群体， 与细胞凋亡相关的细胞内蛋白(bcl2、p53、Rb)，以及 细胞表面抗原包括Fas和mdr1。最近，绑定了 膜联蛋白V到磷脂酰丝氨酸(PS)被认为是一种检测 在改变之前的膜脂结构的改变 TUNEL法检测细胞凋亡情况。量化： 细胞抗原现在是可用的，使我们能够确定 抗体结合能力(ABC)。细胞动力学变化可以是 宫内节育器和BUDR输注后患者的研究 流式细胞仪分析。荧光原位杂交 (FISH)也与细胞凋亡检测相结合，允许 美国用来区分正常细胞和白血病细胞的凋亡。这个 残留白血病细胞的数量、表型和增殖 (MRD)可进行FISH分析的异常情况 仅在30,000名正常人群中检测出1例白血病 细胞。这项检测可以预测缓解期，并将 适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.