A fundamental understanding of extracellular microenvironments of O 2 and reactive oxygen species (ROS) such as H 2 O 2 , ubiquitous in microbiology, demands high-throughput methods of mimicking, controlling, and perturbing gradients of O 2 and H 2 O 2 at microscopic scale with high spatiotemporal precision. However, there is a paucity of high-throughput strategies of microenvironment design, and it remains challenging to achieve O 2 and H 2 O 2 heterogeneities with microbiologically desirable spatiotemporal resolutions. Here, we report the inverse design, based on machine learning (ML), of electrochemically generated microscopic O 2 and H 2 O 2 profiles relevant for microbiology. Microwire arrays with suitably designed electrochemical catalysts enable the independent control of O 2 and H 2 O 2 profiles with spatial resolution of ∼10 1 μm and temporal resolution of ∼10° s. Neural networks aided by data augmentation inversely design the experimental conditions needed for targeted O 2 and H 2 O 2 microenvironments while being two orders of magnitude faster than experimental explorations. Interfacing ML-based inverse design with electrochemically controlled concentration heterogeneity creates a viable fast-response platform toward better understanding the extracellular space with desirable spatiotemporal control.
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