Houston Jessica P, Yang Zhihua, Sambrano Jesse, Li Wenyan, Nichani Kapil, Vacca Giacomo
Department of Chemical & Materials Engineering, New Mexico State University, MSC, PO Box 30001, Las Cruces, NM, 88003, USA.
Kinetic River Corp., 897, Independence Avenue, Suite 4A, Mountain View, CA, 94043-2357, USA.
Methods Mol Biol. 2018;1678:421-446. doi: 10.1007/978-1-4939-7346-0_18.
The focus of this chapter is time-resolved flow cytometry, which is broadly defined as the ability to measure the timing of fluorescence decay from excited fluorophores that pass through cytometers or high-throughput cell counting and cell sorting instruments. We focus on this subject for two main reasons: first, to discuss the nuances of hardware and software modifications needed for these measurements because currently, there are no widespread time-resolved cytometers nor a one-size-fits-all approach; and second, to summarize the application space for fluorescence lifetime-based cell counting/sorting owing to the recent increase in the number of investigators interested in this approach. Overall, this chapter is structured into three sections: (1) theory of fluorescence decay kinetics, (2) modern time-resolved flow cytometry systems, and (3) cell counting and sorting applications. These commentaries are followed by conclusions and discussion about new directions and opportunities for fluorescence lifetime measurements in flow cytometry.
本章的重点是时间分辨流式细胞术，其广义定义为测量通过细胞仪或高通量细胞计数及细胞分选仪器的激发荧光团的荧光衰减时间的能力。我们关注这个主题主要有两个原因：第一，讨论这些测量所需的硬件和软件修改的细微差别，因为目前还没有广泛使用的时间分辨细胞仪，也没有一种适用于所有情况的方法；第二，总结基于荧光寿命的细胞计数/分选的应用领域，因为最近对这种方法感兴趣的研究人员数量有所增加。总体而言，本章分为三个部分：（1）荧光衰减动力学理论，（2）现代时间分辨流式细胞术系统，以及（3）细胞计数和分选应用。这些评论之后是关于流式细胞术中荧光寿命测量的新方向和机会的结论与讨论。
