PROBES FOR FLOW CYTOMETRY

该计划的主要长期目标是继续发展 新的细胞化学，生物物理和分子探针和技术， 测量个体的各种成分的含量和/或构象 细胞 这些探针（技术）将被设计用于细胞分析， 流式细胞术和细胞分选，并预计具有临床意义。 在癌症筛查、诊断、分类和预后中的应用， 在肿瘤药物敏感性评价和治疗监测中的应用 方面的影响. 本研究目前的具体目标（项目）是：1） 评价染色质结构差异可能对 结肠和乳腺肿瘤中的DNA含量估计; 2）RNA含量估计： a）开发适用于以下的RNA测量的灵敏方法： 从实体瘤分离的细胞核; B）负责 核糖体中rRNA与多聚核糖体原位差异染色， 派洛宁Y及其在新淋巴细胞中的应用 激发试验：（3）共振能量的研究与应用 派洛宁Y与罗丹明123之间的转移作为一种新的线粒体探针; (4)DNA原位变性检测方法的扩展 分离自结肠、乳腺、肺和膀胱肿瘤;（5）分析 具有亲和性的抗肿瘤药物选择性释放的核蛋白 核酸，并测试针对这些蛋白质开发的抗体， 药物对靶细胞作用的可能探针。 第二个目标（具体目标或项目6）是应用已开发的 研究细胞生长，有丝分裂周期进展， 细胞分化和对不同抗肿瘤药物的反应。 的 数据将与这些过程的动力学相关，并将提供一个 代谢变化和细胞异质性的更完整描述 在细胞周期中。 第三个目标（具体目标或项目7）是，利用生物物理学， 和生物化学方法，以研究探针之间的分子相互作用 以及各种细胞成分，主要是核酸。 因为很多 探针是抗肿瘤药物本身或药物类似物，这些研究 除了有助于开发新的诊断技术外， 预计将揭示药物作用于细胞的机制的信息， 分子水平。 将这些数据与全细胞药物研究相关联 将提供细胞内药物靶点，机制， 参与药物结合和与细胞动力学相关的细胞毒性，或 代谢状态 这反过来将有助于评估细胞特征 预测对特定药物的敏感性并帮助新药设计。

The primary long term objective of this program is to continue development of new cytochemical, biophysical and molecular probes and techniques to measure content and/or conformation of various constituents of individual cells. These probes (techniques) will be designed for cell analysis by flow cytometry and cell sorting and are expected to have clinical applications in cancer screening, diagnosis, classification and prognosis, in evaluation of drug sensitivity of tumors and in monitoring treatment effects. Specific current aims (projects) of this study are: 1) evaluation of the possible effect of differences in chromatin structure on DNA content estimates in colon and breast tumors; 2) RNA content estimates: a) development of sensitive method(s) of RNA measurement applicable to nuclei isolated from solid tumors; b) studies of mechanisms responsible for differential staining of rRNA in ribosomes vs polyribosomes in situ with pyronin Y, and application of this phenomenon to a new lymphocyte stimulation assay; (3) studies and application of the resonance energy transfer between pyronin Y and rhodamine 123 as a new mitochondrial probe; (4) extension of the method for assay of DNA denaturation in situ to nuclei isolated from colon, breast, lung and bladder tumors; (5) analysis of nuclear proteins selectively released by antitumor drugs having affinity to nucleic acids, and testing antibodies developed against these proteins as a possible probe of drug effects on target cells. The second objective (specific aim or project 6) is to apply the developed probes to studies of cell growth, progression through the mitotic cycle, differentiation and response of cells to different antitumor drugs. The data will be correlated with kinetics of those processes and will provide a more complete description of metabolic changes and cell heterogeneity during the cell cycle. The third objective (specific aim or project 7) is, by using biophysical and biochemical methods, to study molecular interactions between the probes and various cell constituents, predominantly nucleic acids. Because many probes are either antitumor drugs themselves or drug analogs, these studies in addition to being helpful in developing new diagnostic techniques, are expected to reveal information on mechanisms of drug action on cells at the molecular level. Correlating these data with drug studies on whole cells will provide a comprehensive view on intracellular drug targets, mechanisms involved in drug binding, and cytotoxicity as related to cell kinetics or metabolic state. This in turn will help assess the cellular features predicting susceptibility to particular drugs and aid in new drug design.