Red Fluorescence from Organic Microdots: Leveraging Foldamer‐Linked Azobenzene for Enhanced Stability and Intensity in Bioimaging Applications

Azobenzene, while relevant, has faced constraints in biological system applications due to its suboptimal quantum yield and short‐wavelength emission. This study presents a pioneering strategy for fabricating organic microdots by coupling foldamer‐linked azobenzene, resulting in robust fluorescence intensity and stability, especially in aggregated states, thereby showing promise for bioimaging applications. Comprehensive experimental and computational examinations elucidate the mechanisms underpinning enhanced photostability and fluorescence efficacy. In vitro and in vivo evaluations disclose that the external layer of cis‐azo‐foldamer microdots performs a self‐sacrificial function during photo‐bleaching. Consequently, these red‐fluorescent microdots demonstrate extraordinary structural and photochemical stabilities over extended periods. The conjugation of a β‐peptide foldamer to the azobenzene chromophore through a glycine linker instigates a blue‐shifted and amplified π*–n transition. Molecular dynamics simulations reveal that the aggregated state of cis‐azo‐foldamers fortifies the stability of cis isomers, thereby augmenting fluorescence efficiency. This investigation furnishes crucial insights into conceptualizing novel, biologically inspired materials, promising stable and enduring imaging applications, and carries implications for diverse arenas such as medical diagnostics, drug delivery, and sensing technologies. This study highlights the enhanced fluorescence and photostability of a specialized azo‐foldamer (cis‐F1) featuring a chromophore/linker/β‐peptide sequence in its aggregated state. Biological assays reveal a self‐sacrificial mechanism in cis‐F1 microdots, preserving fluorescence post‐injection. Computational and molecular dynamics (MD) simulations confirm the stability and prolonged fluorescence of cis‐F1, offering insights for designing bio‐inspired materials for stable imaging applications.