Kabakov Alexander E, Kudryavtsev Vladimir A, Gabai Vladimir L
Department of Radiation Biochemistry, Medical Radiology Research Center, Obninsk, Russia.
Methods Mol Biol. 2011;787:231-44. doi: 10.1007/978-1-61779-295-3_17.
Cell death (in particular, apoptosis and necrosis) is accompanied by appearance of certain hallmarks that are manifested as specific alterations in cellular membranes, cytoplasm, nucleus and mitochondria. Some of those hallmarks are easily detectable in situ and, therefore, they can be applied for the assessment of dying or dead cells. In turn, there are also signs of viable cells that include a set of features, such as normal functioning of their membranes and organelles, ability to proliferate, etc. The present chapter provides descriptions of several convenient methods for quantitative determination of dead (apoptotic and necrotic) cells and also methods for determination of survived and viable cells. Here, we describe in details the methods of annexin V/propidium iodide (PI) staining, TUNEL assay, Hoechst/PI staining, MTS tetrazolium assay, and colony formation assay, with the principles, advantages, and drawbacks of each technique.
细胞死亡（尤其是凋亡和坏死）伴随着某些特征的出现，这些特征表现为细胞膜、细胞质、细胞核和线粒体的特定改变。其中一些特征很容易在原位检测到，因此可用于评估濒死或死亡细胞。反过来，活细胞也有一些特征，包括一系列特性，如细胞膜和细胞器的正常功能、增殖能力等。本章介绍了几种定量测定死亡（凋亡和坏死）细胞的简便方法以及测定存活和活细胞的方法。在此，我们详细描述膜联蛋白V/碘化丙啶（PI）染色、TUNEL检测、Hoechst/PI染色、MTS四唑盐检测和集落形成检测方法，以及每种技术的原理、优点和缺点。
