Chazotte Brad
CSH Protoc. 2008 Apr 1;2008:pdb.prot4950. doi: 10.1101/pdb.prot4950.
INTRODUCTIONThe eukaryotic cell has evolved to compartmentalize its functions and transport various metabolites among cellular compartments. Therefore, in cell biology, the study of organization and structure/function relationships is of great importance. The nucleus contains almost all of the cell's DNA and is bounded by a double membrane. Inside and adjacent to the inner membrane of the nuclear envelope is the nuclear lamina. It is composed of a fibrous meshwork comprising one or more of three major intermediate filament-like polypeptides: lamins A, B, and C. The outer nuclear membrane is contiguous with the endoplasmic reticulum. Several fluorescent stains are available that label DNA and allow easy visualization of the nucleus in interphase cells and chromosomes in mitotic cells. These stains include Hoechst and 4',6-diamidino-2-phenylindole (DAPI), which are used in this article. Although not as bright as the vital Hoechst stains for DNA, DAPI has greater photostability. One advantage of Hoechst 33342 over DAPI is that the former is membrane-permeant and is, therefore, useful for imaging living cells, because it does not require cell fixation or permeabilization.
引言
真核细胞已进化出将其功能进行区室化并在细胞区室之间运输各种代谢物的能力。因此，在细胞生物学中，对细胞组织以及结构/功能关系的研究至关重要。细胞核几乎包含细胞的所有DNA，并由双层膜界定。核纤层位于核被膜内膜的内部及与之相邻处。它由一个纤维状网络组成，该网络包含三种主要的中间丝样多肽中的一种或多种：核纤层蛋白A、B和C。核外膜与内质网相连。有几种荧光染料可用于标记DNA，从而便于观察间期细胞中的细胞核和有丝分裂细胞中的染色体。这些染料包括本文中使用的Hoechst染料和4',6-二脒基-2-苯基吲哚（DAPI）。尽管DAPI对DNA的染色不如活性Hoechst染料明亮，但它具有更高的光稳定性。Hoechst 33342相对于DAPI的一个优点是，前者可透过细胞膜，因此可用于活细胞成像，因为它不需要细胞固定或通透处理。
