Two-photon calcium imaging of neuronal populations enables optical recording of spiking activity in living animals, but standard laser scanners are too slow to accurately determine spike times. Here we report in vivo imaging in mouse neocortex with greatly improved temporal resolution using random-access scanning with acousto-optic deflectors. We obtained fluorescence measurements from 34–91 layer 2/3 neurons at a 180–490 Hz sampling rate. We detected single action potential–evoked calcium transients with signal-to-noise ratios of 2–5 and determined spike times with near-millisecond precision and 5–15 ms confidence intervals. An automated 'peeling' algorithm enabled reconstruction of complex spike trains from fluorescence traces up to 20–30 Hz frequency, uncovering spatiotemporal trial-to-trial variability of sensory responses in barrel cortex and visual cortex.

By revealing spike sequences in neuronal populations on a fast time scale, high-speed calcium imaging will facilitate optical studies of information processing in brain microcircuits. Sensory-evoked population spiking dynamics in mouse barrel cortex. Spatiotemporal spiking activity of the 56 neurons shown in evoked by the first air puff in each of the eight trials. Responses to all trials are shown in parallel for the time window of 60 ms surrounding each first whisker stimulation.

The movie frame duration was artificially set to 1 ms. Spike times for all neuron were reconstructed with the peeling algorithm. Drag And Drop File Download Html5 Player. The occurrence of a spike is color-coded in red, whereby the color saturation (from light to dark and again to light) follows a Gaussian time course with the appropriate 95% confidence interval (10.4 ms) to indicate the uncertainty in spike detection.

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