Puvion E, Bernhard W
J Cell Biol. 1975 Oct;67(1):200-14. doi: 10.1083/jcb.67.1.200.
The interphase nucleus of the normal rat hepatocyte has been studied in ultrathin frozen sections after glutaraldehyde fixation and the modification of various staining procedures known to be specific for DNA structures (Moyne's thallium stain, Gautier's osmium-ammine) or preferential for RNP carriers and basic proteins (regressive stains based on the use of EDTA or citrate, negatively charged colloidal iron). The results are comparable to those obtained after classical dehydration and embedding. Particular attention has been paid to the nucleolus and extranucleolar RNP components, such as perichromatin fibrils and granules, as well as interchromatin granules. A striking observation was the uneven size and the strongly increased number of perichromatin granules, and the appearance of a contiguous interchromatin net, containing nucleoproteins. Cryoultramicrotomy without embedding appears to be very useful for the exploration of the nucleus in thick sections which remain sufficiently transparent even with the usual accelerating voltages.
在戊二醛固定后，通过超薄冷冻切片对正常大鼠肝细胞的间期核进行了研究，并对各种已知对DNA结构具有特异性的染色程序（莫因铊染色、 Gautier锇氨染色）或对RNP载体和碱性蛋白具有选择性的染色程序（基于使用EDTA或柠檬酸盐的退行性染色、带负电荷的胶体铁）进行了改进。结果与经典脱水和包埋后获得的结果相当。特别关注了核仁以及核仁外的RNP成分，如染色质周纤维和颗粒，以及染色质间颗粒。一个显著的观察结果是染色质周颗粒大小不均且数量大幅增加，以及出现了一个包含核蛋白的连续染色质间网络。不进行包埋的冷冻超薄切片术对于探索厚切片中的细胞核似乎非常有用，即使在通常的加速电压下，这些厚切片仍保持足够的透明度。
