利用粘附式细胞仪测定培养巨噬细胞内pH值

Intracellular pH measurement in living cells utilizing adherent cell analysis and sorting cytometer

用ｐＨ值敏感荧光探针Ｓｎａｆｌｃａｌｃｅｉｎ－ＡＭ标记体外培养４８小时的小鼠腹腔巨噬细胞，５７０型粘附式细胞仪观察巨噬细胞在对刀豆素Ａ进行受体介导内吞过程中细胞内ｐＨ值的变化。结果显示，开始内吞后细胞内ｐＨ值即降低，内吞进行到第５分钟细胞内ｐＨ值降到最低，在此后的６分钟内均维持于这一较低水平。本法灵敏度高，简便快捷，适用范围广，可无损伤观察活细胞或细胞器ｐＨ值的变化。

By using fluorescence staining method, the effect of receptor-mediated endocytosis on intracellular pH (pHi) of mouse macrophages was studied with adherent cell analysis and sorting cytometer 570 (ACAS 570). The results showed that the pHi decreased immediately when macrophages started endocytosis, and reached a low level 5 min later. After that the pHi maintained in the low level for at least 6 min. It is suggested that the method is sensitive, quick and simple, and can be adopted widely in studing pH of living cells and organelles without injury. The method may be one of the most effective methods in measurement pHi.