PROBES FOR FLOW CYTOMETRY

该项目的长期目标是开发新的探测器， 流动分析单个细胞的方法和仪器 图像细胞术，适用于肿瘤诊断和/或预后； 开发的方法将应用于细胞周期和 细胞对抗肿瘤药物的敏感性。具体的目标是集中在 细胞凋亡和细胞增殖标记物的发展。 DNA原位断链标记方法的研究进展 利用外源末端转移酶将继续；该技术 将与细胞增殖分析相结合。细胞增殖性 潜力将通过两者来估计，分析表达的 细胞周期特异性蛋白、细胞周期蛋白E和细胞周期蛋白B1与DNA检测 复制。后者将通过一种新的方法进行检测，该方法 将新复制的DNA的光解与DNA检测相结合 链子断了。这种方法提供了比 用5-溴脱氧尿苷抗体常规检测DNA复制， 还将就其测量DNA修复的能力进行测试。一个 将选择荧光染料的组合，也将允许 细胞DNA含量的测量。DNA的多变量分析 内容、细胞死亡(细胞凋亡)和增殖(细胞周期蛋白E和B1 表达或DNA复制)将应用于人类 化疗前、化疗过程中和乳房中的白血病 癌症；将评估所测量参数的预后价值。 预计这样的综合分析，特别是涉及 对G1和G2周期蛋白的估计，将具有很高的预后价值 将在治疗过程中使用，以监测肿瘤反应。

The long term objective of this project is the development of new probes, methods and instrumentation for analysis of individual cells by flow or image cytometry, applicable for tumor diagnosis and/or prognosis; the developed methodologies will be applied In studIes of the cell cycle and cell sensitivity to antitumor drugs. The specific aims are focused on development of markers of apoptosis, and of cell proliferation. Development of the method based on DNA in situ strand break labeling that employs exogenous terminal transferase will be continued; the technique will be combined with analysis of cell proliferation. Cell proliferative potential will be estimated by both, the analysis of expression of the cell cycle specific proteins, cyclin E and B1 and measurement of DNA replication. The latter will be detected by a novel approach, which combines photolysis of the newly replicated DNA with the detection of DNA strand breaks. This approach, which offers several advantages over the conventional detection of DNA replication by 5-bromodeoxyuridine antibody, will also be tested with respect to its ability to measure DNA repair. A combination of fluorochromes will be selected that will also allow measurement of cellular DNA content. The multivariate analysis of DNA content, cell death (apoptosis) and proliferation (cyclin E and B1 expression or DNA replication) will be applied in studies of human leukemias, before and during the course of chemotherapy, and in breast cancer; the prognostic value of the parameters measured will be evaluated. it is expected that such a combined analysis, especially involving estimates of the G1 and G2 cyclins, will be of high prognostic value, and will be used during treatment, to monitor tumor response.