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<p>Multiplexed fluorescence imaging provides valuable biological insights from the cellular to the tissue level but remains limited in live-mammal studies by the lack of a fluorescent palette capable of overcoming photon scattering and autofluorescence noise for real-time, multiplexed in vivo imaging. Here we present a fluorophore palette engineered from erbium(III)-phthalocyanine complexes, termed the lanthanide rainbow (Lanbow), which offers tunable near-infrared absorption and a unified 1,530 nm emission with brightness surpassing that of existing molecular dyes. Lanbow uses excitation-encoded and efficient single-band detection in the 1,500–1,900 nm shortwave infrared subregion, where tissue scattering and autofluorescence are minimized, enabling up to nine-colour imaging in deep tissues. We also demonstrate fluorescence-guided surgery featuring multiparametric anatomical identification and functional assessment, with deep-learning networks automating real-time analysis for intraoperative guidance. This study establishes a transformative platform for real-time, highly multiplexed imaging in live mammals.</p>

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Excitation-encoded single-emission shortwave infrared lanthanide fluorophore palette for real-time in vivo multispectral imaging

  • 期刊论文

  • 发表日期：

    2025年8月27日

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  • Lu Zhang,
  • Ri Cheng,
  • Zuyang He,
  • Mei Mei,
  • Bin Wu,
  • Weimin Tan,
  • Bo Yan,
  • Shangfeng Wang,
  • Fan Zhang

摘要

Multiplexed fluorescence imaging provides valuable biological insights from the cellular to the tissue level but remains limited in live-mammal studies by the lack of a fluorescent palette capable of overcoming photon scattering and autofluorescence noise for real-time, multiplexed in vivo imaging. Here we present a fluorophore palette engineered from erbium(III)-phthalocyanine complexes, termed the lanthanide rainbow (Lanbow), which offers tunable near-infrared absorption and a unified 1,530 nm emission with brightness surpassing that of existing molecular dyes. Lanbow uses excitation-encoded and efficient single-band detection in the 1,500–1,900 nm shortwave infrared subregion, where tissue scattering and autofluorescence are minimized, enabling up to nine-colour imaging in deep tissues. We also demonstrate fluorescence-guided surgery featuring multiparametric anatomical identification and functional assessment, with deep-learning networks automating real-time analysis for intraoperative guidance. This study establishes a transformative platform for real-time, highly multiplexed imaging in live mammals.