Preparation of cells and reagents for flow cytometry.
Preparation of cells and reagents for flow cytometry.
复制标题
DOI：
10.1002/0471142735.im0503s44
复制
发表时间：
2001-11-01
影响因子：
--
通讯作者：
Giorgi, J V
中科院分区：
文献类型：
--
作者：
Holmes, K;Lantz, L M;Giorgi, J V
流式细胞术广泛用于分析细胞表面和细胞内分子的表达（基于每个细胞），表征和定义异质群体中的不同细胞类型，评估分离的亚群的纯度，以及分析细胞大小和体积。该技术主要用于测量荧光标记抗体或配体产生的荧光强度，这些抗体或配体与特定的细胞相关分子结合。本文介绍了一种直接和间接染色淋巴组织或外周血淋巴细胞的单细胞悬液以检测细胞表面膜抗原的方法。此外，支持方案提供了用于纯化抗体的荧光标记的方法。还包括单细胞悬浮液中细胞内抗原的流式细胞术分析的方案。替代方案描述了在存在去污剂的情况下未固定细胞的细胞内染色和非活细胞的染色，以促进在固定或透化细胞制备物中区分死细胞。
Flow cytometry is widely used for analyzing the expression of cell surface and intracellular molecules (on a per cell basis), characterizing and defining different cell types in heterogeneous populations, assessing the purity of isolated subpopulations, and analyzing cell size and volume. This technique is predominantly used to measure fluorescence intensity produced by fluorescent-labeled antibodies or ligands that bind to specific cell-associated molecules. A procedure for direct and indirect staining of single-cell suspensions of lymphoid tissue or peripheral blood lymphocytes to detect cell surface membrane antigens is presented. In addition, support protocols present methods for fluorescence labeling of purified antibodies. A protocol for flow cytometric analysis of intracellular antigens in single-cell suspensions is also included. Alternate protocols describe intracellular staining of unfixed cells in the presence of a detergent and staining of nonviable cells to facilitate discrimination of dead cells in fixed or permeabilized cell preparations.