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首页> 外文会议>Conference on Optical Diagnostics of Living Cells Ⅳ Jan 24-25, 2001, San Jose, USA >Flow cytometric, time-resolved measurements by frequency heterodyning of fluorescence emission signals
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Flow cytometric, time-resolved measurements by frequency heterodyning of fluorescence emission signals

机译：流式细胞术，时间分辨测量，通过荧光发射信号的频率异染色质

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摘要

A first generation phase-sensitive flow cytometer has been developed that combines flow cytometry (FCM) and fluorescence lifetime spectroscopy measurement principles to provide unique capabilities for making time-resolved fluorescence measurements in the frequency-domain on particles/cells labeled with fluorescent probes. Cells are analyzed as they flow through a chamber and intersect a high-frequency, intensity-modulated (sine-wave) laser excitation beam. Fluorescence emission signals are currently processed by analog homodyne methods to quantify lifetimes and resolve heterogeneous fluorescence based on differences in lifetimes expressed as phase shifts, while maintaining the capability to make conventional FCM measurements. In this study we report the current status of our phase flow cytometer using homodyne signal processing, including recent applications, along with the description of a flow cytometric method for quantifying fluorescence lifetimes by frequency heterodyning techniques. Lifetimes are determined from homodyned and heterodyned (1 MHz difference frequency) signals using analog signal processing electronics. These signals are then digitized and displayed as frequency distribution histograms in real time.
机译：已开发出第一代相敏流式细胞仪，该仪结合了流式细胞仪（FCM）和荧光寿命光谱测量原理，提供了独特的功能，可在频域上对用荧光探针标记的颗粒/细胞进行时间分辨的荧光测量。当细胞流过腔室并与高频强度调制（正弦波）激光激发光束相交时，对细胞进行分析。当前，通过模拟零差方法来处理荧光发射信号，以量化寿命并基于表示为相移的寿命差异来分辨异质荧光，同时保持进行常规FCM测量的能力。在这项研究中，我们报告了使用零差信号处理技术的流式细胞仪的当前状态，包括最近的应用，以及流式细胞术方法的描述，该方法通过频率异染色质技术定量了荧光寿命。使用寿命是使用模拟信号处理电子设备根据同质和异质（1 MHz差频）信号确定的。然后将这些信号数字化并实时显示为频率分布直方图。

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