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Framework Integration Engineered Asymmetric Cyanine Dyes for In Vivo High-Performance NIR-II Imaging and Multiplexed

Yufei Qin1, Jiaming Guo1, Yujie Qin1

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|December 1, 2025
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Researchers developed novel near-infrared-II (NIR-II) dyes called AFlavs for advanced bioimaging. These bright, stable fluorophores enable high-resolution, multicolor NIR-II imaging in vivo.

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Area of Science:

  • Chemical Biology
  • Biomedical Imaging
  • Organic Chemistry

Background:

  • Developing small-molecule near-infrared-II (NIR-II) fluorophores for high-contrast bioimaging presents significant challenges.
  • Existing NIR-II fluorophores often lack the required brightness, photostability, or multiplexing capabilities for advanced applications.

Purpose of the Study:

  • To design and synthesize a new class of small-molecule NIR-II fluorophores with enhanced properties for bioimaging.
  • To evaluate the performance of these novel fluorophores in various in vivo imaging applications, including vascular imaging and multicolor imaging.

Main Methods:

  • Integration of flavylium and polymethine scaffolds to create asymmetric NIR-II dyes, termed AFlavs.
  • Characterization of photophysical properties, including brightness, photostability, and quenching resistance.
  • In vivo evaluation of AFlav-4 for vascular imaging, tumor vasculature assessment, ischemia-reperfusion studies, and bone-targeted delivery.
  • Demonstration of multicolor imaging capabilities through spectral separation analysis.

Main Results:

  • The synthesized AFlavs exhibit high brightness (227-2396 M-1 cm-1) and superior photostability.
  • AFlav-4 demonstrated effective performance in various vascular imaging tasks, including tumor vasculature disruption and ischemia-reperfusion evaluation.
  • AFlavs enabled unprecedented two-, three-, and four-color multiplex imaging with spectral orthogonality (>100 nm absorption separation).

Conclusions:

  • AFlavs represent a promising new class of small-molecule NIR-II fluorophores with excellent photophysical properties.
  • These dyes are suitable for high-resolution, multicolor, and long-term in vivo NIR-II imaging.
  • The developed AFlavs hold significant potential for advancing various biomedical imaging applications.