Jenkins Patrick, Naivar Mark A, Houston Jessica P
Department of Chemical & Materials Engineering, New Mexico State University, MSC 3805 P.O. Box 30001, Las Cruces, NM 88003-8001, USA.
DarklingX, 1199 45th Street, Los Alamos, NM 87544, USA.
J Biophotonics. 2015 Nov;8(11-12):908-17. doi: 10.1002/jbio.201400115. Epub 2015 Feb 26.
Flow cytometry is a powerful means for in vitro cellular analyses where multi-fluorescence and multi-angle light scattering can indicate unique biochemical or morphological features of single cells. Yet, to date, flow cytometry systems have lacked the ability to capture complex fluorescence dynamics due to the transient nature of flowing cells. In this contribution we introduce a simple approach for measuring multiple fluorescence lifetimes from a single cytometric event. We leverage square wave modulation, Fourier analysis, and high frequency digitization and show the ability to resolve more than one fluorescence lifetime from fluorescently-labelled cells and microspheres. Illustration of a flow cytometer capable of capturing multiple fluorescence lifetime measurements; creating potential for multi-parametric, time-resolved signals to be captured for every color channel.
流式细胞术是一种用于体外细胞分析的强大手段，其中多荧光和多角度光散射可以指示单个细胞独特的生化或形态特征。然而，迄今为止，由于流动细胞的瞬态性质，流式细胞术系统缺乏捕获复杂荧光动力学的能力。在本论文中，我们介绍了一种从单个细胞计数事件测量多个荧光寿命的简单方法。我们利用方波调制、傅里叶分析和高频数字化，并展示了从荧光标记的细胞和微球中分辨出不止一种荧光寿命的能力。展示了一种能够捕获多个荧光寿命测量值的流式细胞仪；为每个颜色通道捕获多参数、时间分辨信号创造了潜力。
