Levitt D, King M
J Immunol Methods. 1987 Feb 11;96(2):233-7. doi: 10.1016/0022-1759(87)90319-x.
Fixation and immunofluorescent staining methods were developed for analyzing intracellular antigens with the cell flow cytometer. Fixing cell suspensions with 100% methanol provided best preservation of morphology, lowest fluorescent background staining and most intense specific immunofluorescence. Immunoglobulins present in B cell lines that were representative of different developmental stages could be distinguished quantitatively. Fluorescence histograms were compared with fluorescence microscope presentation of stained cells. Intracellular antigens that reacted with monoclonal antibodies could also be evaluated by flow cytometry. This method was utilized to assess plasmacyte development in mouse spleen cell cultures after stimulation with lipopolysaccharide.
为了使用细胞流式细胞仪分析细胞内抗原，开发了固定和免疫荧光染色方法。用100%甲醇固定细胞悬液可实现最佳的形态保存、最低的荧光背景染色以及最强的特异性免疫荧光。可以定量区分代表不同发育阶段的B细胞系中存在的免疫球蛋白。将荧光直方图与染色细胞的荧光显微镜图像进行比较。与单克隆抗体反应的细胞内抗原也可以通过流式细胞术进行评估。该方法用于评估脂多糖刺激后小鼠脾细胞培养物中浆细胞的发育。
