Increasing the time for recovery between applications increases activation efficiency. Single-channel openings with a delay of 10 s between the start of each consecutive application versus a delay of 4 s. (A and B) Currents with a 10-s delay in our standard external solution at pH 8 with an energy-based internal. Displayed as in Fig. 1. (C) Dwell time histogram of the latency to first opening with a 10-s delay and an energy-based internal. Histogram was best fit by two exponentials (dashed lines). (D) Mean (±SEM) success rate for various conditions. *P < 0.05, **P < 0.01, one-way ANOVA (P = 0.0005) with a post-hoc Tukey’s test. No ATP, 4 s is statistically different from all other conditions: versus ATP, 4 s (P = 0.015), versus ATP, 10 s (P = 0.0013), versus no ATP, 10 s (P = 0.035). All other comparisons were not statistically different: ATP, 4 s versus ATP, 10 s (P > 0.99), ATP, 4 s versus no ATP, 10 s (P = 0.75), ATP, 10 s versus no ATP, 10 s (P = 0.65). (E) Mean (±SEM) efficiency (η) of activation for various conditions. **P < 0.01, ****P < 0.001, one-way ANOVA (P < 0.0001) with a post-hoc Tukey’s test. No ATP, 4 s is statistically different from all other conditions: versus ATP, 4 s (P = 0.0074), versus ATP, 10 s (P < 0.0001), versus no ATP, 10 s (P = 0.0056). All other comparisons were not statistically different: ATP, 4 s versus ATP, 10 s (P = 0.95), ATP, 4 s versus no ATP, 10 s (P = 0.87), ATP, 10 s versus no ATP, 10 s (P = 0.38). Note that “not significant” comparisons are not indicated.