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组织化学研究。LVII. 用干涉显微镜和X射线微放射照相法测定人红细胞的总干质量。

Studies in histochemistry. LVII. Determination of the total dry mass of human erythrocytes by interference microscopy and x-ray microradiography.

作者信息

GAMBLE C N, GLICK D

出版信息

J Biophys Biochem Cytol. 1960 Sep;8(1):53-60. doi: 10.1083/jcb.8.1.53.

Abstract

The total dry mass of human erythrocytes was determined by both interference microscopy and x-ray microradiography. The determination of mass per unit area, and calculation of total dry mass per cell were simplified by changing the shape of the cells to spheres which were then flattened to discs of constant thickness when smeared on glass slides for measurement of fixed cells by interferometry, and to oblate spheroids when smeared on parlodion-coated slides for measurement of fixed cells by x-ray absorption. From x-ray measurements of 100 smeared and alcohol-fixed cells a mean dry mass per cell of 33.7 x 10(-12) g was obtained. Interference measurements of 100 fresh cells suspended in isotonic saline gave a mean value of 32.4 x 10(-12)g while interference measurement of 100 smeared and alcohol-fixed cells gave a mean value of 30.8 x 10(-12)g. The first two values compare well with a mean corpuscular hemoglobin of 31.2 x 10(-12)g, obtained from determinations of erythrocyte count and hemoglobin, since 95 per cent of the dry mass of the cell is hemoglobin. The difference in interference values between the fixed and fresh cells is possibly due to a difference between the specific refractive increment of alcohol-denatured hemoglobin and that of the unmodified substance. The value for the latter was used since that of the former is unknown.

摘要

采用干涉显微镜和X射线显微放射照相法测定了人红细胞的总干质量。通过将细胞形状转变为球体,简化了单位面积质量的测定以及每个细胞总干质量的计算。当将细胞涂片在载玻片上用于干涉测量固定细胞时,球体被压平成厚度恒定的圆盘;当涂片在火棉胶包被的载玻片上用于通过X射线吸收测量固定细胞时,球体变成扁球体。通过对100个涂片并用酒精固定的细胞进行X射线测量,得出每个细胞的平均干质量为33.7×10⁻¹² g。对100个悬浮在等渗盐溶液中的新鲜细胞进行干涉测量,得到的平均值为32.4×10⁻¹² g,而对100个涂片并用酒精固定的细胞进行干涉测量,得到的平均值为30.8×10⁻¹² g。前两个值与通过红细胞计数和血红蛋白测定得到的平均红细胞血红蛋白31.2×10⁻¹² g相比吻合良好,因为细胞干质量的95%是血红蛋白。固定细胞和新鲜细胞干涉值的差异可能是由于酒精变性血红蛋白与未修饰物质的比折射增量不同所致。由于前者的值未知,所以使用了后者的值。

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