Manogaran P S, Kausalya S, Pande G
Centre for Cellular and Molecular Biology, Hyderabad, India.
Cytometry. 1995 Apr 1;19(4):320-5. doi: 10.1002/cyto.990190406.
Pulse width analysis, in flow cytometry, has been widely used for optimal cell size resolution in cell kinetics analysis. Pulses, generated by scattered light or fluorescence of cells, are electronically analyzed for their height and width. The information generated from these two properties of the pulses is utilized to distinguish signals from single cells vs. signals from cell clumps or aggregates. Pulse width, unlike pulse height, is more sensitive to differences in cell diameter, and therefore can discriminate very small differences in it, which pulse height cannot. We have exploited this property of pulse widths to measure immunoconjugates between NK cells and their targets. Discrimination of the free target cells from the conjugated ones is possible by the pulse widths of only light scatter signals, both forward and/or orthogonal. This resolution was not obtained if pulse height of the same signals was visualized. Using this resolution it was possible to distinguish single cells from the aggregates between target and effector cells. We propose that this is a better method for distinguishing conjugates than the method in which prior vital staining of cells is used.
在流式细胞术中，脉冲宽度分析已广泛用于细胞动力学分析中的最佳细胞大小分辨率。由细胞的散射光或荧光产生的脉冲会对其高度和宽度进行电子分析。从脉冲的这两个特性产生的信息用于区分来自单细胞的信号与来自细胞团块或聚集体的信号。与脉冲高度不同，脉冲宽度对细胞直径的差异更敏感，因此可以区分其中非常小的差异，而脉冲高度则无法做到。我们利用脉冲宽度的这一特性来测量自然杀伤（NK）细胞与其靶标之间的免疫缀合物。仅通过前向和/或正交光散射信号的脉冲宽度就可以区分游离靶细胞和缀合靶细胞。如果观察相同信号的脉冲高度，则无法获得这种分辨率。利用这种分辨率可以区分单细胞与靶细胞和效应细胞之间的聚集体。我们认为，与使用细胞预先活体染色的方法相比，这是一种区分缀合物的更好方法。
