Cuchens M A, Buttke T M
Cytometry. 1984 Nov;5(6):601-9. doi: 10.1002/cyto.990050608.
A flow cytometric procedure has been developed for performing kinetic studies on the capping of membrane immunoglobulin (mIg) on B lymphocytes. Mouse B cells were stained with fluorescein-conjugated antimouse-Ig antisera and subjected to pulse-shape (width, peak, and area) analyses prior to, during, and after ligand-induced redistribution of mIg. It was found that ring-stained, patched, and capped cells could be discriminated based on the width of the electronic signal curve generated as the cells passed through the laser beam. Additionally, endocytosis and or shedding of the cap could be correlated with a change in the area under the curve. Using these two parameters (width and area), the effects of temperature, cross-linking, and several pharmacological agents on the capping process were examined. Through the use of flow cytometry, the inhibitory effects of various perturbants could be localized to discrete stages of the capping process.
已开发出一种流式细胞术程序，用于对B淋巴细胞上膜免疫球蛋白（mIg）的封帽进行动力学研究。用荧光素偶联的抗小鼠Ig抗血清对小鼠B细胞进行染色，并在mIg的配体诱导再分布之前、期间和之后进行脉冲形状（宽度、峰值和面积）分析。发现可以根据细胞通过激光束时产生的电子信号曲线的宽度来区分环状染色、斑块状和封帽状细胞。此外，封帽的内吞作用和/或脱落可能与曲线下面积的变化相关。使用这两个参数（宽度和面积），研究了温度、交联和几种药物制剂对封帽过程的影响。通过使用流式细胞术，各种干扰剂的抑制作用可以定位到封帽过程的离散阶段。
