Nagata T
Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, Japan.
Histol Histopathol. 1997 Oct;12(4):1091-124.
The techniques for light and electron microscopic radioautography developed in our laboratory are described. The outlines of the applications to various tissues and organs are also briefly summarized. First, the techniques are explained in detail. These techniques can be divided into two categories, i.e., chemical fixation followed by wet-mounting radioautography and cryo-fixation followed by dry-mounting radioautography. The former is limited in application to only insoluble compounds, while the latter is universally applicable to any kind of compounds including soluble compounds. Both procedures, the chemical fixation wet-mounting procedure and the cryo-fixation dry-mounting procedure at light and electron microscopic levels are described. Next, the applications of both the procedures to various compounds are briefly mentioned. Chemical fixation with glutaraldehyde and osmium tetroxide followed by wet-mounting radioautography demonstrates insoluble macromolecular synthesis such as nucleic acids (both DNA and RNA), proteins, glucides and lipids. On the other hand, cyrofixation followed by dry-mounting radioautography demonstrates soluble small molecular compounds such as macromolecular precursors, hormones, neurotransmitters, vitamins, inorganic compounds and drugs or toxins. These procedures are expected to be applied for various inorganic and organic compounds in living organisms to elucidate the sites of their incorporation, synthesis and discharge.
本文描述了我们实验室开发的光镜和电镜放射自显影技术。同时也简要总结了这些技术在各种组织和器官中的应用概况。首先,详细解释这些技术。这些技术可分为两类,即化学固定后湿装放射自显影和冷冻固定后干装放射自显影。前者仅适用于不溶性化合物,而后者普遍适用于任何类型的化合物,包括可溶性化合物。本文描述了光镜和电镜水平下化学固定湿装法和冷冻固定干装法这两种方法。接下来,简要提及这两种方法在各种化合物中的应用。用戊二醛和四氧化锇进行化学固定,然后进行湿装放射自显影,可显示不溶性大分子的合成,如核酸(DNA和RNA)、蛋白质、糖类和脂质。另一方面,冷冻固定后干装放射自显影可显示可溶性小分子化合物,如大分子前体、激素、神经递质、维生素、无机化合物以及药物或毒素。预计这些方法可应用于生物体中的各种无机和有机化合物,以阐明它们的掺入、合成和释放位点。