HIGH-RESOLUTION AUTORADIOGRAPHY .1. METHODS
HIGH-RESOLUTION AUTORADIOGRAPHY .1. METHODS
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DOI：
10.1083/jcb.15.2.173
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发表时间：
1962-01-01
影响因子：
7.8
通讯作者：
VANTUBERGEN, RP
中科院分区：
文献类型：
--
作者：
CARO, LG;KOLB, JA;VANTUBERGEN, RP
在获得高分辨率的放射自显影，特别强调电子显微镜放射自显影技术，所使用的方法，连同控制实验设计，以建立最佳的条件或程序。基于这些实验，选择的乳液是IIford L-4，其晶体尺寸略大于0.1微米。它以由单层银晶体组成的凝胶膜的形式施加到样品上。背景，当存在时，可以通过简单的方法根除。制剂可以在干燥剂存在下、在室温下或在冰箱中储存。照相显影是在Microdol或一种特殊的细粒“物理”显影剂中进行的。为了在电子显微镜中进行检查，将切片用铀酰或铅染色剂染色。这些方法在亚细胞水平上给出了标记的良好定位，并且在相对颗粒计数中具有良好的再现性。
Methods used in obtaining high resolution in autoradiography, with special emphasis on the technique of electron microscopic autoradiography, are described, together with control experiments designed to establish the optimum conditions or procedures. On the basis of these experiments the emulsion selected was IIford L-4, with a crystal size slightly larger than 0.1 micron. It is applied to the specimen in the form of a gelled film consisting of a monolayer of silver crystals. Background, when present, can be eradicated by a simple method. The preparations can be stored, in presence of a drying agent, at room temperature or in a refrigerator. Photographic development is done in Microdol, or in a special fine grain "physical" developer. For examination in the electron microscope the sections are stained with uranyl or lead stains. These methods give a good localization of the label, at the subcellular level, and good reproducibility in relative grain counts.