A System for live sorting of neuronal spiking activity from large-scale recordings

Online monitoring and quantification of neural signals has tremendous value both for neurofeedback experiments and for brain-computer interfaces. Unfortunately, established methods of online monitoring primarily involve the use of thresholded neural activity rather than sorted single-neuron spikes. The recent introduction of large-scale, high-density electrophysiology has enabled the recording of activity from hundreds of neurons simultaneously in both model organisms and human participants. This development highlights the need for a robust and easily implementable system for sorting spikes during data collection for ‘live’ analyses of neuronal signals. Here, we describe a system for live sorting of neuronal activity (LSS) based on the widely used Kilosort platform. The LSS workflow utilizes an initial period of recorded neural data to identify waveform templates using Kilosort 4. LSS then interfaces with the SpikeGLX API to retrieve small batches (e.g. 50 ms) of data and for processing online. We measured the similarity of single-neuron activity sorted live by LSS to that sorted offline in neurophysiological recordings from macaque visual cortex using Neuropixels probes. We show that LSS closely replicates the post-stimulus time histograms and visual response tuning curves of single-neurons obtained using offline sorting. Furthermore, we show that decoding neural signals online with LSS consistently outperforms online decoding of thresholded activity, and that LSS can achieve the same performance as that obtained with offline sorting.