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首页> 外文会议>Optical Investigations of Cells In Vitro and In Vivo >Progress toward combined phase and amplitude demodulation fluorescence lifetime measurements by flow cytometry
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Progress toward combined phase and amplitude demodulation fluorescence lifetime measurements by flow cytometry

机译：流式细胞术在相和幅相解调荧光寿命测量中的应用进展

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Abstract: A phase-sensitive flow cytometer has been developed to quantify fluorescence lifetimes directly as a parameter in real time on particles and cells labeled with fluorophores. This instrument combines flow cytometry and frequency-domain fluorescence lifetime spectroscopy measurement principles to provide unique features for making excited-state lifetime measurements. Cells are analyzed as they intersect a high- frequency, intensity-modulated (sine wave) laser excitation beam. Fluorescence signals are processed by analog phase comparator measurement electronics to provide output signals proportional to the sine and cosine of the phase difference between the modulated signal input and a steady state reference signal. The ratios, which are proportional to the lifetimes, are displayed as frequency distributions histograms. In this study we present the underlying theory to measure fluorescence lifetimes by both phase shift and amplitude demodulation. Prototype analog amplitude demodulation electronics have been constructed and are being tested using simulated signals, fluorescent microspheres, and cells labeled with fluorescent probes. !27
机译：摘要：已经开发出一种相敏流式细胞仪，可以直接实时定量荧光寿命，将其作为参数对荧光团标记的颗粒和细胞进行定量。该仪器结合了流式细胞仪和频域荧光寿命光谱测量原理，为进行激发态寿命测量提供了独特的功能。当细胞与高频，强度调制（正弦波）激光激发光束相交时，对细胞进行分析。荧光信号由模拟相位比较器测量电子设备处理，以提供与调制信号输入和稳态参考信号之间的相位差的正弦和余弦成比例的输出信号。与寿命成正比的比率显示为频率分布直方图。在这项研究中，我们提出了通过相移和幅度解调来测量荧光寿命的基础理论。原型模拟幅度解调电子设备已经构建，正在使用模拟信号，荧光微球和带有荧光探针标记的细胞进行测试。 ！27

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