Department of Chemical Engineering and Biotechnology, University of Applied Sciences Darmstadt, Haardtring 100, D-64295 Darmstadt, Germany.
Methods Appl Fluoresc. 2017 Oct 24;5(4):042002. doi: 10.1088/2050-6120/aa7c7a.
Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.
荧光寿命(FLT)是荧光物质的一种稳健的固有特性和材料常数。测量这一重要的物理指标已经从实验室的好奇心发展成为一种强大而成熟的技术,广泛应用于药物发现、医学诊断和基础生物学研究等领域。这种明显的趋势主要是由改进的、同时又负担得起的激光和检测仪器推动的,另一方面是合适的 FLT 探针和生物测定方法的发展。在这个过程中,出现了两种基本的工作方法。第一种方法主要集中在高通量应用上,采用生化体外测定,不需要高空间分辨率。第二种甚至更具活力的趋势是通过荧光寿命成像将高时间和空间分辨率的荧光数据结合起来的测定方法的显著扩展。后一种方法目前正在被用于不仅研究永生肿瘤细胞系,还研究活体动物的特定组织甚至器官。本文试图对基于 FLT 的生物测定的现状以及在生物医学和生命科学领域的广泛应用机会进行全面概述。此外,还将讨论 FLT 技术的未来趋势。