Tinari Antonella, Giammarioli Anna Maria, Manganelli Valeria, Ciarlo Laura, Malorni Walter
Department of Technology and Health, Istituto Superiore di Sanità, Rome, Italy.
Methods Enzymol. 2008;442:1-26. doi: 10.1016/S0076-6879(08)01401-8.
Diverse forms of cell death have initially been described thanks to their observation at the electron microscope. Morphological and ultrastructural features of necrosis, apoptosis, and autophagy, considered here as prototypic cell death processes, allow one to characterize and quantify early and late cytopathological changes occurring in cells undergoing degeneration. Both light microscopy and scanning electron microscopy can provide useful insights, for example, to quantitatively evaluate cell death or to characterize cell surface changes of the cells, respectively. However, transmission electron microscopy preparation allows distinguishing among different forms of cell death. This chapter describes in brief the methods used to characterize cell death forms, including membrane, nucleus, and organelle changes, and shows paradigmatic micrographs. In particular, morphogenetic changes occurring in mitochondria during apoptosis, that is, fission process or, conversely, vacuole formation during autophagy, are shown. Possible artifacts are also described. Ultrastructural analysis seems still to provide essential information for studies on cell death.
多种形式的细胞死亡最初是通过在电子显微镜下的观察得以描述的。坏死、凋亡和自噬的形态学及超微结构特征,在此被视为典型的细胞死亡过程,它们使人们能够对发生退化的细胞中早期和晚期的细胞病理学变化进行表征和量化。光学显微镜和扫描电子显微镜都能提供有用的见解,例如,分别用于定量评估细胞死亡或表征细胞的表面变化。然而,透射电子显微镜制样能够区分不同形式的细胞死亡。本章简要描述了用于表征细胞死亡形式的方法,包括细胞膜、细胞核和细胞器的变化,并展示了具有代表性的显微照片。特别展示了凋亡过程中线粒体发生的形态发生变化,即裂变过程,或者相反,自噬过程中液泡的形成。还描述了可能出现的假象。超微结构分析似乎仍然为细胞死亡研究提供了重要信息。