Multi - neuron intracellular recording 1 in vivo via interacting autopatching 2 robots . 3

The activities of groups of neurons in a circuit or brain region are important for 21 neuronal computations that contribute to behaviors and disease states. Traditional extracellular 22 recordings have been powerful and scalable, but much less is known about the intracellular 23 processes that lead to spiking activity. We present a robotic system, the multipatcher, capable of 24 automatically obtaining blind whole-cell patch clamp recordings from multiple neurons 25 simultaneously. The multipatcher significantly extends automated patch clamping, or 26 ”autopatching”, to guide four interacting electrodes in a coordinated fashion, avoiding mechanical 27 coupling in the brain. We demonstrate its performance in the cortex of anesthetized and awake 28 mice. A multipatcher with four electrodes took an average of 10 min to obtain dual or triple 29 recordings in 29% of trials in anesthetized mice, and in 18% of the trials in awake mice, thus 30 illustrating practical yield and throughput to obtain multiple, simultaneous whole-cell recordings 31 in vivo. 32