Kawamoto T
Department of Biochemistry, School of Dental Medicine, Tsurumi University, Yokohama, Japan.
J Histochem Cytochem. 1990 Dec;38(12):1805-14. doi: 10.1177/38.12.2254645.
An approach using autoradiography for the study of early changes in the distribution of water-soluble materials and the chemography involved was investigated. Radioactive calcium chloride (45Ca) was injected into the iliac vein of a rat. Ten seconds after the injection the rat was frozen in hexane (-90 degrees C). The frozen rat was embedded in 5% sodium carboxymethyl cellulose and blocked in the coolant. A sheet of plastic tape coated with a synthetic rubber glue was fastened to the trimmed block surface, and whole-body sections 2-10 microns thick were cut with a disposable microtome knife. Selected sections were freeze-dried and then covered with a dried autoradiographic emulsion film about 1 microns thick. The autoradiograph clearly showed the distribution of radioactive calcium in the calcification zone of long bones. The samples chosen to assess chemographic artifacts showed positive and negative chemographies on most of the tissues when these were kept at 23 degrees C, and although both chemographic effects were significantly reduced when the samples were kept at -20 degrees C, cells in several tissues still exhibited positive and negative chemographies. The technique can be used for the study of any animal whose size is suitable for whole-body freeze-sectioning.
研究了一种利用放射自显影术来研究水溶性物质分布早期变化及相关化学成像的方法。将放射性氯化钙（45Ca）注入大鼠的髂静脉。注射后十秒，将大鼠置于己烷（-90摄氏度）中冷冻。将冷冻的大鼠嵌入5%的羧甲基纤维素钠中，并在冷却剂中进行包埋。将一片涂有合成橡胶胶水的塑料胶带固定在修整后的包埋块表面，用一次性切片刀切成2 - 10微米厚的全身切片。选取的切片进行冷冻干燥，然后覆盖上一层约1微米厚的干燥放射自显影乳剂膜。放射自显影片清晰显示了放射性钙在长骨钙化区的分布。用于评估化学成像伪影的样本在23摄氏度保存时，大多数组织呈现出正性和负性化学成像，尽管当样本在-20摄氏度保存时，两种化学成像效应均显著降低，但几个组织中的细胞仍表现出正性和负性化学成像。该技术可用于研究任何大小适合全身冷冻切片的动物。
