CDr15 is a deep-red fluorescent probe ( Ex =733 nm) that can selectively intercalate into and label bacterial extracellular DNA (eDNA) . CDr15 exhibits extremely high specificity for biofilm imaging and cannot effectively bind to mammalian nuclear DNA. CDr15 enables real-time visualization of the microcolony structure and developmental process of three-dimensional *Pseudomonas aeruginosa* biofilms, and accurately localizes biofilm-forming regions of microorganisms in a mouse corneal infection model. With low background interference signals, CDr15 serves as an ideal diagnostic tool for research fields including bacterial biofilms and corneal infections.
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