Nagata T
Department of Anatomy and Cell Biology, Shinshu University School of Medicine, Matsumoto, Japan.
Braz J Med Biol Res. 1998 Feb;31(2):185-95. doi: 10.1590/s0100-879x1998000200002.
Standard techniques for radioautography used in biological and medical research can be classified into three categories, i.e., macroscopic radioautography, light microscopic radioautography and electron microscopic radioautography. The routine techniques used in these three procedures are described. With regard to macroscopic radioautography, whole body radioautography is a standard technique which employs freezing and cryosectioning and can demonstrate organ distributions of both soluble and insoluble compounds. In contrast, in light and electron microscopic radioautography, soluble and insoluble techniques are separated. In order to demonstrate insoluble labeled compounds, conventional chemical fixations such as formalin for light microscopy or buffered glutaraldehyde and osmium tetroxide for both light and electron microscopy followed by dehydration, embedding and wet-mounting applications of radioautographic emulsions can be used. For the demonstration of soluble labeled compounds, however, cryotechniques such as cryofixation, cryosectioning, freeze-drying, freeze-substitution followed by dry-sectioning and dry-mounting radioautography should be employed both for light and electron microscopy. The outlines of these techniques, which should be utilized in various fields of biological and medical research, are described in detail.
生物和医学研究中使用的放射自显影标准技术可分为三类，即宏观放射自显影、光学显微镜放射自显影和电子显微镜放射自显影。本文描述了这三种方法中使用的常规技术。关于宏观放射自显影，全身放射自显影是一种标准技术，它采用冷冻和冷冻切片，能够显示可溶性和不溶性化合物在器官中的分布。相比之下，在光学显微镜和电子显微镜放射自显影中，可溶性和不溶性技术是分开的。为了显示不溶性标记化合物，可以使用常规化学固定方法，如用于光学显微镜的福尔马林，或用于光学显微镜和电子显微镜的缓冲戊二醛和四氧化锇，随后进行脱水、包埋以及放射自显影乳剂的湿装片应用。然而，为了显示可溶性标记化合物，对于光学显微镜和电子显微镜，都应采用冷冻技术，如冷冻固定、冷冻切片、冷冻干燥、冷冻置换，随后进行干切片和干装片放射自显影。本文详细描述了这些应在生物和医学研究各个领域中使用的技术要点。
