Fig. 3. Recovery from acidosis in isolated granulosa cells. Granulosa shells were prepared by removing the oocyte from preantral follicles recovered from 10-day-old mice, and then loaded with SNARF-AM. (A) Left, a series of three photomicrographs illustrating the procedure by which the oocyte (arrowheads) is removed from a preantral follicle. A higher resolution exemplar photomicrograph of the resulting granulosa shell is also shown (center) and a fluorescence micrograph of SNARF-loaded shells (right). Scale bar: 50 µm. (B) Recovery from acidosis in SNARF-loaded granulosa shells following a NH₄Cl pulse. Note the rapid increase in pH when Na⁺ was returned to the bath (experiments were performed in triplicate, n=23 in total). Note also that amiloride retarded Na⁺-dependent acidosis correction (experiments were performed in triplicate, n=23 in total). Amiloride was added at t=10 minutes. (C) Average rate of recovery from acidosis following Na⁺ replacement. ^*, P<0.05.