Fig. 2: Molecular design and spectra properties of cyanines.

Chemical structures of a CyP7, b bioorthogonal-activated cyanines, and c control cyanines. Bioorthogonal-moiety Tz (red color group) with methyl (shorter) (CyP7T) or PEG3 (longer) (CyP7N) distances from substrate structures. CyP7TT/CyP7NT activated by CyP7T/CyP7N and TCO via IEDDA reaction. d Normalized absorption (black)/emission (red) spectra of dyes. e Fluorescence intensity at the maximum emission spectra with different concentrations of cyanines. f–h Fluorescence loss with concentrations. Dotted lines represent ideal linear relationship between fluorescence intensity and concentration, indicating non-quenching in blue range. i ACQ effect for control group (CyP7NT formed by CyP7N and TCO).