Trusal L R, Baker C J, Guzman A W
Stain Technol. 1979 Mar;54(2):77-83. doi: 10.3109/10520297909112638.
Plastic coverslips made of polymethylpentene serve as excellent substrates for growth of bovine endothelial cells, and are easily processed for both transmission (TEM) and scanning (SEM) electron microscopy. Portions of the same coverslip (monolayer) are used for both SEM and TEM examination and are fixed, postfixed, and dehydrated as a single entity. The portion of the coverslip for SEM is then excised, critical point dried, and mounted for sputter coating prior to viewing. The remaining piece of coverslip used for TEM is Epon-Araldite embedded, polymerized, separated from the coverslip by liquid nitrogen immersion, and sectioned either "en face" or in cross section for viewing. Coated glass coverslips are not required and organic solvents such as propylene oxide, acetone, and amyl acetate can be used for dehydration and infiltration. Furthermore, specimens do not require re-embedding or blocks to be glued onto blank capsules before sectioning. The number of cells needed to achieve a monolayer is significantly reduced compared to the usual culture flasks, but are abundant enough to assess ultrastructural changes accurately. Support films may be required to prevent folding of the ultrathin section which can obstruct viewing of cells located on the edge of the section.
由聚甲基戊烯制成的塑料盖玻片是牛内皮细胞生长的优良基质，并且易于处理用于透射电子显微镜（TEM）和扫描电子显微镜（SEM）观察。同一块盖玻片（单层）的部分用于SEM和TEM检查，并作为一个整体进行固定、后固定和脱水。然后将用于SEM的盖玻片部分切除，临界点干燥，并在观察前安装用于溅射镀膜。用于TEM的盖玻片剩余部分用Epon-Araldite包埋、聚合，通过液氮浸泡与盖玻片分离，并切成“正面”或横截面用于观察。不需要涂覆玻璃盖玻片，并且可以使用诸如环氧丙烷、丙酮和乙酸戊酯等有机溶剂进行脱水和渗透。此外，标本在切片前不需要重新包埋或把块粘到空白包埋盒上。与通常的培养瓶相比，形成单层所需的细胞数量显著减少，但数量足够多以准确评估超微结构变化。可能需要支撑膜以防止超薄切片折叠，这可能会妨碍观察位于切片边缘的细胞。
