Live imaging of the extracellular matrix with a glycan-binding fluorophore

All multicellular systems produce and dynamically regulate extracellular matrices (ECMs) that play essential roles in both biochemical and mechanical signaling. Though the spatial arrangement of these extracellular assemblies is critical to their biological functions, visualization of ECM structure is challenging, in part because the biomolecules that compose the ECM are difficult to fluorescently label individually and collectively. Here, we present a cell-impermeable small-molecule fluorophore, termed Rhobo6, that turns on and red shifts upon reversible binding to glycans. Given that most ECM components are densely glycosylated, the dye enables wash-free visualization of ECM, in systems ranging from in vitro substrates to in vivo mouse mammary tumors. Relative to existing techniques, Rhobo6 provides a broad substrate profile, superior tissue penetration, non-perturbative labeling, and negligible photobleaching. This work establishes a straightforward method for imaging the distribution of ECM in live tissues and organisms, lowering barriers for investigation of extracellular biology.

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Publication Details

Journal
Nature Methods
Published
2025-02-06
DOI
https://doi.org/10.1038/s41592-024-02590-2
Citations
20
Primary Topic
Cell Adhesion Molecules Research
Type
article
Field-Weighted Citation Impact
32.44

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article

Live imaging of the extracellular matrix with a glycan-binding fluorophore

Thomas A. Ravenscroft, Valerie M. Weaver, Kayvon Pedram, Tanya L. Dilan et al.
20 citations
Nature Methods
Cell Adhesion Molecules Research
32.44
article

Live imaging of the extracellular matrix with a glycan-binding fluorophore

Thomas A. Ravenscroft, Valerie M. Weaver, Kayvon Pedram, Tanya L. Dilan, Antonio Fiore, Misha B. Ahrens, Ronak Patel, Hari Shroff, Pratik Kumar, Jason J. Northey, G. Yu, Shaohe Wang, Wiert Kolkman, Virginia M. S. Ruetten, Richard Ikegami
article en
20 citations

Abstract

All multicellular systems produce and dynamically regulate extracellular matrices (ECMs) that play essential roles in both biochemical and mechanical signaling. Though the spatial arrangement of these extracellular assemblies is critical to their biological functions, visualization of ECM structure is challenging, in part because the biomolecules that compose the ECM are difficult to fluorescently label individually and collectively. Here, we present a cell-impermeable small-molecule fluorophore, termed Rhobo6, that turns on and red shifts upon reversible binding to glycans. Given that most ECM components are densely glycosylated, the dye enables wash-free visualization of ECM, in systems ranging from in vitro substrates to in vivo mouse mammary tumors. Relative to existing techniques, Rhobo6 provides a broad substrate profile, superior tissue penetration, non-perturbative labeling, and negligible photobleaching. This work establishes a straightforward method for imaging the distribution of ECM in live tissues and organisms, lowering barriers for investigation of extracellular biology.

Nature MethodsVol. 22(5)
Howard Hughes Medical Institute (US), University of California, San Francisco (US), Janelia Research Campus (US)
Howard Hughes Medical Institute, National Institutes of Health
Openalex Percentile: Top 0%
Cell Adhesion Molecules Research
32.44
DOIOpen Access PDFOpenAlex
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