Superiority of in vitro over in vivo calibrations of BCECF in vascular smooth muscle cells

摘要: We measured intracellular pH (pHi) in single vascular smooth muscle cells (VSM) cultured from rabbit abdominal aorta, using 2',7'-biscarboxyethyl-5(6)carboxyfluorescein (BCECF) on a microscope-based fluorimetric system. previously found substantial errors introduced by high K+/nigericin to calibrate BCECF (1). also demonstrated that the necessary correction (pHcor) K+/nigericin-calibrated pHi was linearly dependent pHi, increasing with (2). When nigericin calibration data were corrected this pHcor, new similar result of calibrating vitro (higher Rmax, lower Rmin, and pK). Therefore, study possibility is considered calibrations might provide better estimates pHi. Our best estimate for absolute level derives null method bracketing steady-state In VSM cells, only leads t...