Sawaguchi Akira, Aoyama Fumiyo, Ide Soyuki, Goto Yoshiteru, Suganuma Tatsuo
Department of Anatomy, Ultrastructural Cell Biology, Faculty of Medicine, University of Miyazaki, Japan.
J Electron Microsc (Tokyo). 2008 Dec;57(6):203-6. doi: 10.1093/jmicro/dfn020. Epub 2008 Oct 21.
High-pressure freezing (HPF) has been generally accepted as the most reliable method for cryofixation of biological samples, yielding a deep vitreous freezing. In recent cell biology, mammalian cultured cells are widely used, but HPF of cultured cell monolayer has not reached its full potential. In this study, we developed a new reliable device for HPF of cultured cell monolayer by using a 10-microm-thin stainless disc both as culture plate and specimen carrier. We describe the practical procedure, and demonstrate fine structures of HeLa cells cultured and cryofixed on the stainless discs as results.
高压冷冻（HPF）已被公认为是对生物样品进行冷冻固定的最可靠方法，能实现深度玻璃化冷冻。在当代细胞生物学中，哺乳动物培养细胞被广泛使用，但培养细胞单层的高压冷冻尚未发挥出其全部潜力。在本研究中，我们开发了一种新型可靠装置，通过使用一个10微米厚的薄不锈钢盘作为培养板和样品载体，来对培养细胞单层进行高压冷冻。我们描述了实际操作步骤，并展示了在不锈钢盘上培养和冷冻固定的HeLa细胞的精细结构。
