Loike J D, Silverstein S C
J Immunol Methods. 1983 Feb 25;57(1-3):373-9. doi: 10.1016/0022-1759(83)90097-2.
A fluorescence quenching method, using trypan blue, is described for quantifying the ingestion of either glutaraldehyde-fixed sheep red blood cells or antibody coated glutaraldehyde-fixed sheep red blood cells by murine peritoneal macrophages. This method is based on the observations that glutaraldehyde-fixed red blood cells fluoresce at about 585 nm when excited at 490 nm and that when trypan blue is in intimate contact with fixed red blood cells, the fluorescence is converted from a chartreuse to a red color. Thus, the ingestion of fixed erythrocytes by murine macrophages can be monitored by fluorescence microscopy after the addition of 1 mg/ml of trypan blue at the end of the assay. Extracellular glutaraldehyde-fixed red blood cells fluoresce a red color whereas the intracellular particles continue to fluoresce a chartreuse color. This method offers a simple and convenient technique for rapidly distinguishing between intracellular and extracellular glutaraldehyde-fixed red blood cells in macrophages.
描述了一种使用台盼蓝的荧光猝灭方法，用于定量小鼠腹腔巨噬细胞对戊二醛固定的绵羊红细胞或抗体包被的戊二醛固定的绵羊红细胞的摄取。该方法基于以下观察结果：戊二醛固定的红细胞在490nm激发时在约585nm处发出荧光，并且当台盼蓝与固定的红细胞紧密接触时，荧光从黄绿色转变为红色。因此，在测定结束时加入1mg/ml台盼蓝后，可通过荧光显微镜监测小鼠巨噬细胞对固定红细胞的摄取。细胞外戊二醛固定的红细胞发出红色荧光，而细胞内颗粒继续发出黄绿色荧光。该方法提供了一种简单方便的技术，用于快速区分巨噬细胞内和细胞外戊二醛固定的红细胞。
