de Grooth B G, Terstappen L W, Puppels G J, Greve J
Department of Applied Physics, University of Twente, Enschede, The Netherlands.
Cytometry. 1987 Nov;8(6):539-44. doi: 10.1002/cyto.990080602.
Polarization measurement of orthogonal light scattering is introduced as a new optical parameter in flow cytometry. In the experimental setup, the electrical field of the incident laser beam is polarized in the direction of the sample flow. The intensity of the orthogonal light scattering polarized along the direction of the incoming laser beam is called depolarized orthogonal light scattering. Theoretical analysis shows that for small values of the detection aperture, the measured depolarization is caused by anisotropic cell structures and multiple scattering processes inside the cell. Measurements of the orthogonal depolarized light scattering in combination with the normal orthogonal light scattering of human leucocytes revealed two populations of granulocytes. By means of cell sorting it was shown that the granulocytes with a relatively high depolarization are eosinophilic granulocytes. Similar experiments with human lymphocytes revealed a minor subpopulation of yet-unidentified lymphocytes with a relative large orthogonal light-scattering depolarization. The results were obtained with an argon ion laser tuned at different wavelengths as well as with a 630-nm helium neon laser. These results show that measurement of depolarized orthogonal light scattering is a useful new parameter for flow-cytometric cell differentiation.
正交光散射的偏振测量作为流式细胞术中一种新的光学参数被引入。在实验装置中，入射激光束的电场沿样品流方向偏振。沿入射激光束方向偏振的正交光散射强度称为去偏振正交光散射。理论分析表明，对于小检测孔径，测得的去偏振是由细胞内各向异性的细胞结构和多次散射过程引起的。对人白细胞的正交去偏振光散射与正常正交光散射进行联合测量，揭示了粒细胞的两个群体。通过细胞分选表明，去偏振相对较高的粒细胞是嗜酸性粒细胞。对人淋巴细胞进行的类似实验显示，有一小部分尚未鉴定的淋巴细胞亚群具有相对较大的正交光散射去偏振。这些结果是使用调谐到不同波长的氩离子激光器以及630纳米的氦氖激光器获得的。这些结果表明，去偏振正交光散射测量是流式细胞术细胞分化的一个有用的新参数。
