美国霍华德休斯医学研究所Kayvon Pedram团队使用糖基结合荧光染料对细胞外基质进行活体成像。2025年2月6日，《自然—方法学》杂志在线发表了这项成果。

研究人员展示了一种细胞不可渗透的小分子荧光染料，称为Rhobo6，它在可逆结合糖基后会激活并发生红移。鉴于大多数细胞外基质（ECM）成分都密集地糖基化，该染料使得可以在从体外底物到体内小鼠乳腺肿瘤等系统中实现免洗可视化。

与现有技术相比，Rhobo6提供了广泛的底物谱、优越的组织渗透性、非干扰性标记和极小的光漂白。这项工作建立了一种简单的方法，用于成像活组织和生物体中ECM的分布，并降低了研究细胞外生物学的门槛。

据悉，所有多细胞系统都产生并动态调节ECM，这些基质在生物化学和机械信号传递中发挥着重要作用。

尽管这些细胞外组装的空间排列对其生物学功能至关重要，但由于组成ECM的生物分子难以单独和共同地进行荧光标记，因此可视化ECM结构具有挑战性。

附：英文原文

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

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

Issue&Volume: 2025-02-06

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.

DOI: 10.1038/s41592-024-02590-2

Source: https://www.nature.com/articles/s41592-024-02590-2