a-c Amplitude of the subthreshold membrane potential (Vm) and corresponding quantifications. a, b Population-averaged Vm during 40 Hz DBS (a, n = 22 neurons) and 140 Hz DBS (b, n = 26 neurons). Vm is calculated as the fluorescence at each time point divided by the averaged amplitude of all spikes detected in a recording session for a given neuron. DBS-induced Vm changes were computed as normalized Vm by subtracting the mean of the Vm during the pre-stimulation baseline period. Shaded gray area is the standard error of the mean (SEM). c Quantification of Vm change from baseline during the transient (0–0.15 s) and sustained (0.15–1 s) periods of 40 Hz and 140 Hz DBS. Data are visualized as violin plots with the outer shape representing the data kernel density and box plots showing interquartile range (1x, 1.5x). The white lines in the boxes are the mean. For 40 Hz DBS, paired t-test, df = 21 neurons, transient: p = 0.002; sustained: p = 0.003, transient vs. sustained: p = 0.0156. For 140 Hz DBS, paired t-test, transient: p = 3.1 × 10−4, df = 25; sustained: p = 0.001, transient vs. sustained: p = 0.78. Comparison between 40 Hz and 140 Hz DBS, independent t-test, df = 46, transient: p = 0.045; sustained: p = 0.79. d-f Spike rate and corresponding quantifications. d, e Population-averaged spike rate during 40 Hz DBS (d, n = 22) and 140 Hz DBS (e, n = 26). Shaded gray area is SEM. f Violin plots of spike rate change from baseline during the transient and sustained periods of 40 Hz and 140 Hz DBS. For 40 Hz DBS, paired t-test, df = 21 neurons, transient: p = 0.0013; sustained: p = 0.002, transient vs. sustained: p = 0.19. For 140 Hz DBS, paired t-test, transient: p = 0.0071, df = 25; sustained: p = 0.11, transient vs. sustained: p = 0.01. Comparison between 40 Hz and 140 Hz DBS, independent t-test, df = 46, transient: p = 0.97; sustained: p = 0.089. g-i Time-frequency spectrum power of Vm and corresponding quantifications. Population-averaged Vm power during 40 Hz DBS (g, n = 22) and 140 Hz DBS (h, n = 26). i Violin plots of Vm 40 Hz or 140 Hz power change relative to the baseline during the transient and sustained periods of 40 Hz and 140 Hz DBS. For 40 Hz DBS, paired t-test, df = 21 neurons, transient: p = 1.05 × 10−4; sustained: p = 1.23 × 10−4, transient vs. sustained: p = 0.94. For 140 Hz DBS, paired t-test, transient: p = 9.77 × 10−6, df = 25; sustained: p = 0.0017, transient vs. sustained: p = 5.76 × 10−5. Comparison between 40 Hz and 140 Hz DBS, independent t-test, df = 46, transient: p = 4.87 × 10−4; sustained: p = 4.05 × 10−4. ns = non-significant, *<0.05, **<0.01, and ***<0.001. All paired or independent t-tests are two-sided. Source data are provided as a Source Data file.