Light-scattering polarization measurements as a new parameter in flow cytometry
Article first published online: 8 MAR 2005
Copyright © 1987 Wiley-Liss, Inc.
Volume 8, Issue 6, pages 539–544, November 1987
How to Cite
de Grooth, B. G., Terstappen, L. W. M. M., Pupples, G. J. and Greve, J. (1987), Light-scattering polarization measurements as a new parameter in flow cytometry. Cytometry, 8: 539–544. doi: 10.1002/cyto.990080602
- Issue published online: 16 JUN 2005
- Article first published online: 8 MAR 2005
- Manuscript Accepted: 8 JUN 1987
- Manuscript Received: 8 AUG 1986
- Eosinophilic granulocytes;
- neutrophilic granulocytes;
- blood cell differentiation
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 argonion 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.