Sato Shigeru, Sasaki Yoshihiro, Adachi Akiko, Dai Wei, Liu Xiao-Lan, Namimatsu Shigeki
Central Institute for Electron Microscopic Research, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan.
Med Electron Microsc. 2003 Sep;36(3):179-82. doi: 10.1007/s00795-003-0216-1.
To assess the usefulness of oolong tea extract (OTE) staining for connective tissue observation, we examined the visceral pleural mesothelium of rat lung by transmission electron microscope. Four kinds of electron microscopic staining methods (routine, tannic acid, OTE in distilled water, and OTE in 0.1 M phosphate buffer) were compared to determine the most suitable method for electron microscopic observation of ultrathin sections. Elastin (elastic laminae) was selectively stained by tannic acid and both water and buffered OTE. Except for elastin, connective tissue and cell ultrastructures were also electron enhanced by tannic acid and both water and buffered OTE staining. However, using water OTE, the electron-dense filaments (10-12 nm in diameter) were obscured. In tannic acid staining, the unit membranes of the visceral pleural mesothelial cells were weaker as compared with routine and buffered OTE stains. Thus, the buffered OTE staining method is a highly useful technique for connective tissue observation and electron-enhanced staining in transmission electron microscopic preparations.
为评估乌龙茶提取物(OTE)染色对结缔组织观察的实用性,我们通过透射电子显微镜检查了大鼠肺的脏层胸膜间皮。比较了四种电子显微镜染色方法(常规染色、单宁酸染色、蒸馏水OTE染色和0.1 M磷酸盐缓冲液OTE染色),以确定用于超薄切片电子显微镜观察的最合适方法。弹性蛋白(弹性膜)可被单宁酸以及水和缓冲OTE选择性染色。除弹性蛋白外,单宁酸以及水和缓冲OTE染色也能增强结缔组织和细胞超微结构的电子密度。然而,使用水OTE时,电子致密丝(直径10 - 12 nm)会变得模糊。在单宁酸染色中,与常规和缓冲OTE染色相比,脏层胸膜间皮细胞的单位膜显示较弱。因此,缓冲OTE染色方法是透射电子显微镜制备中用于结缔组织观察和电子增强染色的非常有用的技术。