Abstract

AbstractControlling the growth of bacterial biofilms in a specific pattern greatly enhances the study of cell‐to‐cell interactions and paves the way for expanding their biological applications. However, the development of simple, cost‐effective, and highly resolved biopatterning approaches remains a persistent challenge. Herein, a pioneering photodynamic biopatterning technique for the creation of living bacterial biofilms with customized geometries at high resolutions is presented. First of all, an outstanding aggregation‐induced emission photosensitizer is synthesized to enable efficient photodynamic bacterial killing at a low concentration. By combining with custom‐designed photomasks featuring both opaque and transparent patterns, the viability of photosensitizer‐coated bacteria is successfully manipulated by controlling the degree of light transmittance. This process leads to the formation of living bacterial biofilms with specific patterns replicated from the photomask. Such an innovative strategy can be employed to generate living bacterial biofilms composed of either mono‐ or multispecies, with a spatial resolution of approximately 24 µm. Furthermore, its potential applications in information storage/encryption and antibiotic screening are explored. This study provides an alternative way to understand and investigate the intricate interactions among bacteria within 3D biofilms, holding great promise in the controlled fabrication of dynamic biological systems for advanced applications.

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