FLOW CYTOMETER FOR QUANTIFYING FLUORESCENCE LIFETIME

该项目的长期目标是开发先进的 用于测量相位分辨荧光的流式细胞术方法 与细胞结合的荧光染料的发射和激发态寿命 和染色体。 该项目的具体目标是：1）应用 将这项技术应用于广泛的生物系统， 这些独特的测量能力的优势; 2）确定 探测和测量低水平放射性物质技术的局限性 背景中荧光探针的发射信号， 细胞自发荧光，通过光谱发射重叠， 荧光检测通道，通过未结合/非特异性荧光团 标记，并通过拉曼/瑞利散射;和3）改善和提高 相位分辨荧光的制作技术， 使用单调制或双调制激光的寿命测量 激发 相敏流式细胞仪项目相对 新的，使其许多潜在的应用程序还没有得到充分 探索和发展。 然而，由于它可以分离荧光， 电子和光学发射，量化寿命 直接作为一个参数，也使传统的流式细胞仪 测量，它具有广泛的技术可能性 应用. 这项新技术将增加 可用于多重标记应用的荧光标记物; 通过提高测量精度获得更准确的结果， 灵敏度并减少背景干扰;并通过 生物医学研究，该技术将显着扩大 研究人员对细胞生物过程的理解， 亚细胞和分子水平。

The long-term goal of this project is the development of advanced flow cytometric methods for measuring phase-resolved fluorescence emissions and excited-stated lifetimes on fluorochromes bound to cells and chromosomes. The specific aims of this project are: 1) to apply the technology to a wide range of biological systems that take advantage of these unique measurement capabilities; 2) determine the limits of the technology for detecting and measuring low-level emission signals from fluorescent probes in backgrounds caused by cellular autofluorescence, by spectral emission overlap among fluorescence detection channels, by unbound/nonspecific fluorophore labeling, and by Raman/Rayleigh scatter; and 3) to improve and advance the technology for making phase-resolved multicolor fluorescence and lifetime measurements using single- or dual-modulated laser excitation. The phase-sensitive flow cytometer project is relatively new so that many of its potential applications have not been fully explored and developed. However, because it can separate fluorescence emissions both electronically and optically, quantify lifetime(s) directly as a parameter, and also make conventional flow cytometric measurements, it has a wide range of technically possible applications. This new technology will increase the range of fluorescent markers that can be used in multi-labeling applications; yield more accurate results by enhancing measurement precision and sensitivity and reducing background interferences; and through biomedical research, the technology will significantly expand the researchers' understanding of biological processes at the cellular, subcellular, and molecular level.