Kingsley R E, Cole N L
Indiana University School of Medicine, South Bend Center, Notre Dame 46556.
J Electron Microsc Tech. 1988 Sep;10(1):77-85. doi: 10.1002/jemt.1060100110.
Common methods for the preparation of cultured cells for concurrent light microscopy (LM), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) are not completely satisfactory. This article describes how we grow mammalian cells on plastic disks made from Aclar film. Aclar is a transparent fluorinated-chlorinated thermoplastic that contains no volatile components and is, for all practical purposes, chemically inert. Cells adhere to it readily and remain attached after fixation, dehydration, and critical-point drying or embedding. The film also accepts heavy metal coating by ionic bombardment and is extremely stable in the vacuum of the SEM. LM observations are unhindered by Aclar, since the film is as transparent as glass. Fluorescence microscopy is possible with this film, since it exhibits no detectable autofluorescence. During SEM observation, the film has great dimensional stability, and the cells and heavy metal coating remain attached to the Aclar even under high-resolution operating conditions. TEM processing of specimens grown on Aclar is simplified by the fact that Aclar does not stick to the epoxy resins used in EM. Furthermore, Aclar is easily sectioned and does not damage knives used in ultramicrotomy. The use of Aclar film considerably simplifies the preparation of cultured cells for all types of microscopy. This method is particularly useful in correlating surface features between SEM and TEM observations.
用于同时进行光学显微镜（LM）、扫描电子显微镜（SEM）和透射电子显微镜（TEM）观察的培养细胞制备的常用方法并不完全令人满意。本文描述了我们如何在由阿克莱尔薄膜制成的塑料圆盘上培养哺乳动物细胞。阿克莱尔是一种透明的氟氯化热塑性塑料，不含挥发性成分，实际上在化学上是惰性的。细胞很容易附着在上面，并且在固定、脱水、临界点干燥或包埋后仍能保持附着。该薄膜还能通过离子轰击接受重金属涂层，并且在扫描电子显微镜的真空中极其稳定。由于该薄膜像玻璃一样透明，因此光学显微镜观察不受其影响。这种薄膜可以进行荧光显微镜观察，因为它没有可检测到的自发荧光。在扫描电子显微镜观察期间，该薄膜具有很高的尺寸稳定性，即使在高分辨率操作条件下，细胞和重金属涂层也能保持附着在阿克莱尔薄膜上。在阿克莱尔薄膜上生长的标本进行透射电子显微镜处理时得到简化，因为阿克莱尔不粘用于电子显微镜的环氧树脂。此外，阿克莱尔很容易切片，并且不会损坏超薄切片机中使用的刀具。使用阿克莱尔薄膜大大简化了用于所有类型显微镜观察的培养细胞的制备。这种方法在关联扫描电子显微镜和透射电子显微镜观察之间的表面特征方面特别有用。
