A study of fixation for electron microscopy.
A study of fixation for electron microscopy.
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DOI：
10.1084/jem.95.3.285
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发表时间：
1952-03
期刊：
影响因子：
--
通讯作者：
PALADE GE
中科院分区：
文献类型：
--
作者：
PALADE GE
通常，在组织块的四氧化锇固定之前先对组织进行酸化。这种酸化可能是造成形态变化的原因，这种变化在电子显微镜下尤其令人不安。直接在 OsO4 溶液中匀浆组织或在固定剂中添加酶抑制剂（氟化物、碘西酰胺）无法防止酸化和由此产生的形态变化。使用 OsO4 缓冲溶液进行的固定实验表明，固定细胞的外观取决于固定剂的 pH 值。通过将 OsO4 溶液缓冲在 pH 7.3-7.5 下，可以显着提高固定质量。醋酸盐-维隆缓冲液似乎是测试的缓冲液中最有利的。由于这些发现，建议使用醋酸盐-维隆缓冲液在 pH 7.3-7.5 缓冲的 1% OsO4 作为电子显微镜的合适固定剂。
Osmium tetroxide fixation of tissue blocks, as usually effected, is preceded by an acidification of the tissue. This acidification is probably responsible for morphological alterations which are notably disturbing in electron microscopy. The acidification and the resulting morphological alterations cannot be prevented by homogenizing the tissue directly in OsO4 solutions or by adding enzyme inhibitors (fluoride, iodoscetamide) to the fixative. Fixation experiments with buffered OsO4 solutions have shown that the appearance of the fixed cells is conditioned by the pH of the fixative. The quality of fixation can be materially improved by buffering the OsO4 solutions at pH 7.3-7.5, The acetate-veronal buffer appeared to be the most favorable of the buffers tested, Because of these findings, 1 per cent OsO4 buffered at pH 7.3-7.5 with acetate-veronal buffer is recommended as an appropriate fixative for electron microscopy.