Automated whole-cell patch-clamp electrophysiology of neurons in vivo - PubMed (original) (raw)

Figure 2. Autopatcher operation and performance

(a) Representative timecourse of pipette resistance during autopatcher operation, top, with zoomed-in view of the neuron hunting phase, bottom. Roman numerals: i, the first of the series of resistance measurements that indicate neuron detection; ii, the last of the series; iii, when positive pressure is released; iv, when suction is applied; v, when holding potential starts to ramp from −30 mV to −65 mV; vi, when it hits −65 mV; vii, break-in. (b) Raw traces showing patch pipette currents, while a square voltage wave (10 Hz, 10 mV) is applied, at the events flagged by Roman numerals in Fig. 2a. (c–f) Quality of recordings obtained with the autopatcher vs. by manual whole cell patch clamping. (c) left, Plot of access resistances obtained versus pipette depth and right, bar graph summary of access resistances (mean ± s.d.), for the final autopatcher whole cell patch validation test set (closed symbols; n = 23), the test set in which the autopatcher concludes in the gigaseal state (open symbols, n = 24; data acquired after manual break-in), and the test set acquired via manual whole cell patch clamp (grayed symbols; n = 15), for cortical (circles) and hippocampal (triangles) neurons. (d) left, Resting potential versus pipette depth, and right, summary data, plotted as in c. (e) left, Holding current versus pipette depth, and right, summary, plotted as in c. (f) left, Holding times versus pipette depth, and right, summary, plotted as in c (including recordings that were deliberately terminated, as well as recordings terminated spontaneously).