A Bright Ratiometric Dipyrene Probe for Functional Imaging of Condensate Microenvironments in Living Cells
Biomolecular condensates exhibit spatially heterogeneous microenvironments shaped by intertwined physicochemical factors, yet practical small-molecule tools for mapping these states in living cells remain limited. Here, we report PyLUMI, a bright dipyrene ratiometric probe, optimized by linker engineering to improve fluorescence output under aqueous conditions while preserving excimer-to-monomer (E/M) readout capability. Solution-phase characterization identifies a scaffold with enhanced fluorescence efficiency and an E/M response to coupled polarity-viscosity microenvironmental changes. In recombinant protein condensates, PyLUMI provides substantially increased signal and enables pixel-resolved visualization of intra-condensate heterogeneity. In living cells, the probe enables low-exposure ratiometric imaging of centrosome-associated condensate compartments and reveals microenvironmental signatures linked to cell-cycle progression and pathology-associated centrosome amplification beyond morphological size differences. These results establish PyLUMI as a functional chemical reporter for spatially resolved profiling of integrated physicochemical fingerprints in condensate-associated compartments.
Authors
- Shogo Amemori (ORCID: https://orcid.org/0000-0002-0676-9721)
- Dini Kurnia Ikliptikawati (ORCID: https://orcid.org/0000-0001-5194-2266)
- Yoshio Nishiyama (ORCID: https://orcid.org/0000-0002-3147-4634)
- De‐Chen Lin (ORCID: https://orcid.org/0000-0002-1951-367X)
- Shih-Cheng Chen (ORCID: https://orcid.org/0000-0002-4764-0510)
- Motohiro Mizuno (ORCID: https://orcid.org/0000-0002-1712-9308)
- Kei Makiyama
- Koki Matsumoto (ORCID: https://orcid.org/0000-0002-4220-7304)
- Takahiro Soeta (ORCID: https://orcid.org/0000-0001-9883-4772)
- Koichi Ogami (ORCID: https://orcid.org/0000-0001-9380-9666)
- Chien‐Hung Yu (ORCID: https://orcid.org/0000-0003-0337-4473)
- Kenji Takahashi (ORCID: https://orcid.org/0000-0002-6775-9850)
- Masaharu Hazawa (ORCID: https://orcid.org/0000-0003-0342-9016)
- Yuta Kozuka
- Kazuma Ogawa (ORCID: https://orcid.org/0000-0002-1691-7302)
- Wei‐Min Liu (ORCID: https://orcid.org/0000-0003-3607-1911)
- Hiroshi Suzuki (ORCID: https://orcid.org/0000-0003-4682-5086)
- Maho Tobita
- Kentaro Ohira
- Richard W. Wong
- Sora Kitai
Institutions
- Kanazawa University (JP)
- Fu Jen Catholic University (TW)
- University of Southern California (US)
- Kanazawa Institute of Technology (JP)
- National Health Research Institutes (TW)
- Kanazawa Medical University (JP)
- Inamori Foundation (JP)
- Life Science Institute (JP)
- Aichi Cancer Center (JP)
- Nagoya University (JP)
- National Cheng Kung University (TW)
Publication Details
- Journal
- Advanced Science
- Published
- 2026-07-16
- DOI
- https://doi.org/10.1002/advs.76419
- Primary Topic
- Advanced Fluorescence Microscopy Techniques
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Japan Agency for Medical Research and Development