HIGH-RESOLUTION DUAL LASER FLOW CYTOMETRY
HIGH-RESOLUTION DUAL LASER FLOW CYTOMETRY
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DOI：
10.1177/26.8.357646
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发表时间：
1978-01-01
影响因子：
3.2
通讯作者：
PINKEL, D
中科院分区：
文献类型：
--
作者：
DEAN, PN;PINKEL, D
基于光学传感的流式细胞仪利用外部光源和荧光染料来测量液体悬浮液中单个细胞或亚细胞颗粒的一种或多种特定成分或特性。为了实现双染实验中两种染料的独立激发，构建了一种高分辨率流式细胞仪，该流式细胞仪使用2束激光提供2种波长的激发。这些光束在空间上分开，以便细胞按顺序流过它们，时间间隔约为20 μ s。由于染料是顺序激发的，因此它们的发射发生在不同的时间，它们的发射光谱可以重叠而不会造成分析困难。新的光收集光学系统被开发出来，允许在每个电池上进行多达4次测量。这种方法大大增加了可用于流式细胞术的染料组合的数量，从而消除了单照明仪器的显著限制。
Flow cytometers based on optical sensing utilize external light sources and fluorescent dyes to measure 1 or more specific components or properties of individual cells or subcellular particles in liquid suspension. To provide for independent excitation of 2 dyes used in double staining experiments a high resolution flow cytometer that uses 2 laser beams to provide 2 wavelengths of excitation was constructed. These beams were separated spatially so that cells flow through them sequentially, with a time separation of about 20 .mu.s. Since the dyes were excited sequentially their emission occurred at different times and their emission spectra may overlap without causing any difficulty in analysis. New light collection optics were developed that permit up to 4 measurements on each cell. This approach greatly increases the number of dye combinations that can be used in flow cytometry, thus removing a significant limitation of single illumination instruments.