Improved method for transmission electron microscopy of ciliated cell monolayers maintained on gas‐permeable membranes
Improved method for transmission electron microscopy of ciliated cell monolayers maintained on gas‐permeable membranes
复制标题
透气膜上维持的纤毛细胞单层的透射电子显微镜改进方法
作者：
M. G. Gabridge;E. Dougherty
本文介绍了一种制备纤毛呼吸道上皮细胞透射电镜薄片的新方法。用胶原酶处理气管外植培养物以促进上皮迁移。当外植体在腔室/培养皿中的FEP蚀刻Teflon膜上孵育时，在外植体附近出现单层生长斑块。单层细胞保持高度分化，如纤毛运动活跃所证明。将单层固定并原位处理。在包埋过程中，将腔室支撑在固体特氟隆圆柱体上，以在加热膜以使树脂固化时保持膜拉紧。去除膜，重新包埋单层细胞并切片。培养物的电子显微镜检查显示在平坦、均匀的基面上的透明细胞轮廓。这种方法提供了一个正常的空间配置的细胞在一个单层的格式，并应证明有用的电子显微镜研究的任何细胞培养的柔性薄膜。
A new method is described for preparing thin sections for transmission electron microscopy of ciliated respiratory epithelial cells cultivated in vitro on Teflon membranes. Hamster tracheal explant cultures were treated with collagenase to promote epithelial migration. Patches of monolayer growth occurred in the vicinity of the explants when the latter were incubated on FEP‐etched Teflon membranes in Chamber/Dishes. Cells in the monolayer maintained a high degree of differentiation as evidenced by active ciliary motion. The monolayer was fixed and processed in situ. During the embedding process the chamber was supported on a solid Teflon cylinder to keep the membrane taut as it was heated to polymerize the resin. The membrane was removed and the monolayered cells were re‐embedded and sectioned. Electron microscopic examination of cultures revealed clear cell profiles on a flat, even basal plane. This method provides for a normal spatial configuration of cells in a monolayer format, and should prove useful for electron microscopy studies of any cells cultured on flexible films.