Goodell Margaret A
Baylor College of Medicine, Houston, Texas, USA.
Curr Protoc Cytom. 2005 Nov;Chapter 9:Unit9.18. doi: 10.1002/0471142956.cy0918s34.
This unit describes the use of Hoechst 33342 to identify and purify murine hematopoietic stem cells, the so-called side population. Three properties of the dye contribute to the ability to distinguish stem cells in this way. Hoechst is a DNA-binding dye. It has at least two binding modes that result in different spectral properties, allowing resolution of multiple populations by viewing fluorescence at two wavelengths simultaneously. The ability to discriminate SP cells is based on the differential efflux of the dye by a multi-drug-like transporter. Particular attention is given to the critical aspects of Hoechst concentration, cell concentration, staining time, and staining temperature.
本单元描述了使用Hoechst 33342来鉴定和纯化小鼠造血干细胞，即所谓的侧群细胞。这种染料的三个特性有助于以这种方式区分干细胞。Hoechst是一种DNA结合染料。它至少有两种结合模式，会导致不同的光谱特性，从而通过同时观察两个波长的荧光来分辨多个细胞群。区分侧群细胞的能力基于染料通过一种多药样转运蛋白的差异外排。文中特别关注了Hoechst浓度、细胞浓度、染色时间和染色温度等关键方面。
