Optical validation of in vitro extra-cellular neuronal recordings

Simultaneous calcium imaging and extra-cellular recordings from cultured cortical rat neurons were performed to directly map the efficacy of extra-cellular recordings with microelectrodes. For the first time, we can associate extra-cellular recordings with neuronal activity of specific neurons in the vicinity of the electrode. We demonstrate that recorded cells can be identified by correlating the electrical signals and the calcium response. Our data demonstrate that in sparse cultures, microelectrodes record exclusively from cells which reside at very close proximity to the recording electrode. Moreover, we show that recording appears to be limited to only a partial subset of the cells residing in this range. We further show that even in cases of strong neuron–electrode coupling, extra-cellular signals recorded from single, well-identified neurons vary in shape over time rendering spike sorting and network activity rate analysis incongruous. As multi-electrode array technology is becoming increasingly widespread, the visualization technique we report here will help users better understand the limits of this versatile and useful method.