Center for Fluorescence Spectroscopy, Department of Biochemistry, University of Maryland at Baltimore School of Medicine, 108 North Greene Street, 21201, Baltimore, Maryland.
J Fluoresc. 1994 Mar;4(1):117-36. doi: 10.1007/BF01876666.
Time-resolved fluorescence spectroscopy is presently regarded as a research tool in biochemistry, biophysics, and chemical physics. Advances in laser technology, the development of long-wavelength probes, and the use of lifetime-based methods, are resulting in the rapid migration of timeresolved fluorescence to the clinical chemistry lab, the patient's bedside, and even to the doctor's office and home health care. Additionally, time-resolved imaging is now a reality in fluorescence microscopy and will provide chemical imaging of a variety of intracellular analytes and/or cellular phenomena. Future horizons of state-of-the-art spectroscopy are also described. Two photon-induced fluorescence provides an increased information content to time-resolved data. Two photoninduced fluorescence, combined with fluorescence microscopy and time-resolved imaging, promises to provide detailed three-dimensional chemical imaging of cells. Additionally, it has recently been demonstrated that the pulses from modern picosecond lasers can be used to quench and/or modify the excited-state population by stimulated emission since the stimulated photons are directed along the quenching beam and are not observed. The phenomenon of light quenching should allow a new class of multipulse time-resolved fluorescence experiments, in which the excited-state population is modified by additional pulses to provide highly oriented systems.
时间分辨荧光光谱学目前被认为是生物化学、生物物理学和化学物理学的研究工具。激光技术的进步、长波长探针的开发以及基于寿命的方法的应用，使得时间分辨荧光迅速迁移到临床化学实验室、患者床边，甚至医生办公室和家庭医疗保健领域。此外，时间分辨成像现在已经成为荧光显微镜的现实，将提供各种细胞内分析物和/或细胞现象的化学成像。还描述了最先进光谱学的未来发展前景。双光子激发荧光为时间分辨数据提供了更高的信息量。双光子激发荧光与荧光显微镜和时间分辨成像相结合，有望提供细胞的详细三维化学成像。此外，最近已经证明，现代皮秒激光器的脉冲可以通过受激辐射来猝灭和/或修饰激发态种群，因为受激光子沿着猝灭光束传播，并且不会被观察到。光猝灭现象应该允许一类新的多脉冲时间分辨荧光实验，其中通过附加脉冲来修饰激发态种群，以提供高度定向的系统。
