Granick S, Levere R D
J Cell Biol. 1965 Jul;26(1):167-76. doi: 10.1083/jcb.26.1.167.
A new technique for the intracellular localization of minute amounts of heme and hemoproteins is described. The specimen is treated with 1.5 M perchloric acid in the presence of SH groups, followed by ultraviolet light irradiation in a fluorescence microscope. This fixes the proteins in situ and converts the heme to a porphyrin which fluoresces and is readily visualized. With this technique, hemoglobin has been demonstrated in the nuclei of avian erythrocytes, and in the nuclei of human normoblasts at an earlier stage than previously described. In addition, hemoproteins, presumably cytochromes, have been detected in the cytoplasm and nuclei of myelocytes, in thymus lymphocyte nuclei, in chick embryo liver cytoplasm, and in chick embryo somites.
本文描述了一种用于微量血红素和血红蛋白细胞内定位的新技术。将标本在存在巯基的情况下用1.5 M高氯酸处理,然后在荧光显微镜下进行紫外线照射。这可将蛋白质原位固定,并将血红素转化为发出荧光且易于观察的卟啉。运用该技术,已在禽类红细胞核以及人类早幼红细胞核中证实了血红蛋白的存在,且其出现阶段比先前描述的更早。此外,在髓细胞的细胞质和细胞核、胸腺淋巴细胞核、鸡胚肝细胞质以及鸡胚体节中均检测到了血红素蛋白,推测为细胞色素。