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培养的人成纤维细胞的多角度光散射流动光度测定法:正常细胞与含有细胞质内含物的突变株的比较。

Multiangle light scattering flow photometry of cultured human fibroblasts: comparison of normal cells with a mutant line containing cytoplasmic inclusions.

作者信息

Schafer I A, Jamieson A M, Petrelli M, Price B J, Salzman G C

出版信息

J Histochem Cytochem. 1979 Jan;27(1):359-65. doi: 10.1177/27.1.220324.

Abstract

Multi-angle light scattering flow photometry was used to study the light scattering properties of normal cultured fibroblasts and a mutant fibroblast line containing cytoplasmic lysosomal inclusions. The effect of glutaraldehyde fixation on the light scattering properties of the cells was also examined and correlated with their ultrastructure. Normal fibroblasts showed uniform organelle distribution with few vacuoles or dense bodies in the cytoplasm while the mutant line showed abnormal cytoplasmic inclusions of varying morphology, density and lucency. As predicted by light scattering theory, the mutant cells containing the cytoplasmic inclusions scattered more light at large angles (greater than theta = 1.85 degrees) than did the normal cells. Glutaraldehyde fixation decreased light scattering at small angles (less than theta = 1.85 degrees), increased light scattering at larger angles (greater than theta = 1.85 degrees) in both normal and mutant cells and enhanced resolution of the light scattering signatures. The mutant line scattered 2-3 times more light at a wide angle (greater than theta = 12.74 degrees) than did the normal cells. These data suggest that abnormal lysosomal storage inclusion bodies in the cytoplasm of the cells can be detected by differential light scattering methods.

摘要

多角度光散射流动光度法用于研究正常培养的成纤维细胞和含有细胞质溶酶体包涵体的突变成纤维细胞系的光散射特性。还检测了戊二醛固定对细胞光散射特性的影响,并将其与细胞超微结构相关联。正常成纤维细胞的细胞器分布均匀,细胞质中几乎没有空泡或致密体,而突变细胞系则显示出形态、密度和透明度各异的异常细胞质包涵体。正如光散射理论所预测的,含有细胞质包涵体的突变细胞在大角度(大于θ = 1.85度)比正常细胞散射更多的光。戊二醛固定降低了小角度(小于θ = 1.85度)的光散射,增加了正常细胞和突变细胞在较大角度(大于θ = 1.85度)的光散射,并提高了光散射特征的分辨率。突变细胞系在广角(大于θ = 12.74度)比正常细胞多散射2 - 3倍的光。这些数据表明,细胞内异常的溶酶体储存包涵体可以通过差分光散射方法检测到。

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