Wideband ratiometric measurement of tonic and phasic dopamine release in the striatum

Reward learning, cognition, and motivation are supported by changes in neurotransmitter levels across multiple timescales. Current measurement technologies for various neuromodulators (such as dopamine and serotonin) do not bridge timescales of fluctuations, limiting the ability to define the behavioral significance, regulation, and relationship between fast (phasic) and slow (tonic) dynamics. To help resolve longstanding debates about the behavioral significance of dopamine across timescales, we developed a novel quantification strategy, augmenting extensively used carbon-fiber Fast Scan Cyclic Voltammetry (FSCV). We iteratively engineered the FSCV scan sequence to rapidly modify electrode sensitivity within a sampling window and applied ratiometric analysis for wideband dopamine measurement. This allowed us to selectively eliminate artifacts unrelated to electrochemical detection (i.e., baseline drift), overcoming previous limitations that precluded wideband dopamine detection from milliseconds to hours. We extensively characterize this approach in vitro, validate performance in vivo with simultaneous microdialysis, and deploy this technique to measure wideband dopamine changes across striatal regions under pharmacological, optogenetic, and behavioral manipulations. We demonstrate that our approach can extend to additional analytes, including serotonin and pH, providing a robust platform to assess the contributions of multi-timescale neuromodulator fluctuations to cognition, learning, and motivation.