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bioRxiv · 10.64898/2026.08.21.746318

SPOUT: An open-source hardware and software platform to study decision making while manipulating and recording from neural activity

van den Boom, B. J. G.·Dash, D.·Rutherford, M.·Girasole, A. E.·Gorelik, P.·Mazor, O.·Sabatini, B. L.
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Abstract

Recording and manipulating brain activity during behavior is critical to understanding the underlying mechanisms of decision-making. Linking neural activity to behavior requires behavioral hardware and software tightly integrated with recording and perturbation systems on a shared clock. We built SPOUT (State-machine Platform for Operant Uni/dual-spout Tasks), an open-source, Teensy-driven state-machine platform with a MATLAB interface that runs 10 unique decision-making tasks (with dozens of variations available through user-friendly settings) to study behavior in head-restrained mice. The platform is built on several custom hardware devices: a dual-lick detector, headplate designs for optogenetics and two-photon calcium imaging, a three-axis motorized spout manipulator, and an optogenetics power modulator. The firmware differentiates between one and two lick spout tasks and can be controlled by a user-friendly interface. Task settings can be selected through the interface or by loading predefined settings files. We validated the clock speed and lick detection against an independent, external acquisition system and identified highly precise, sub-millisecond detection of single licks. Using a pseudo-random synchronization pulse generated by SPOUT, we corrected for missing data due to glitches in the acquisition system and clock drift. We showcase the versatility of SPOUT by training mice on an uninstructed lick-left/lick-right task in which the rewarded side switches unexpectedly and found that mice use history-dependent action-outcome associations to guide future behavior. Transiently inhibiting the anterior lateral motor cortex (ALM) during cue presentation induced contralateral deficits, without affecting ipsilateral trials. Finally, two-photon imaging of ALM neurons revealed stronger population responses during contralateral choice licks compared to ipsilateral ones. Together, SPOUT offers an open-source, affordable platform to study decision-making in head-restrained mice while combining neural recordings and manipulations.

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van den Boom, B. J. G., Dash, D., Rutherford, M., Girasole, A. E., Gorelik, P., Mazor, O., Sabatini, B. L.. 2026-08-26. SPOUT: An open-source hardware and software platform to study decision making while manipulating and recording from neural activity. https://doi.org/10.64898/2026.08.21.746318

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