Visser J W, Jongeling A A, Tanke H J
J Histochem Cytochem. 1979 Jan;27(1):32-5. doi: 10.1177/27.1.35567.
A method was developed to determine the intracellular pH (pHi) of individual cells by use of fluorescence measurements. The method is based on the observation that the fluorescence excitation spectrum of fluorescein is pH-dependent. Fluorescence excitation spectra from individual rat bone marrow cells treated with fluorescein diacetate (FDA) were compared with those of fluorescein solutions of known pH values. Cells which were suspended in media of pH between 4.0 and 8.1 with high to normal buffering capacities had pHi values equal to those of the media. Cells suspended in media with low buffering capacities maintained a pH,i of 6.7 +/- 0.2. Preliminary results indicated that the pHi of individual cells may also be determined by using flow cytometry.
开发了一种通过荧光测量来测定单个细胞内pH值（pHi）的方法。该方法基于以下观察结果：荧光素的荧光激发光谱依赖于pH值。将用二乙酸荧光素（FDA）处理的单个大鼠骨髓细胞的荧光激发光谱与已知pH值的荧光素溶液的光谱进行比较。悬浮在pH值在4.0至8.1之间且具有高至正常缓冲能力的培养基中的细胞，其pHi值与培养基的pHi值相等。悬浮在低缓冲能力培养基中的细胞维持的pHi为6.7±0.2。初步结果表明，也可以使用流式细胞术来测定单个细胞的pHi。
