结缔组织是器官发育、修复和再生的重要组成部分。然而，我们对结缔组织动力学的理解还处于初级阶段。在本文中，我们详细介绍了一种方法，利用胺基和N-羟基琥珀酰亚胺酯之间的交联化学反应，在体内追踪器官细胞外基质（ECM）的命运。该方法能够对ECM蛋白进行可靠的标记，补充了先前基于亲和力的单蛋白方法。该方案适用于初级科学家，标记步骤需花费5至10分钟。使用N-羟基琥珀酰亚胺酯对ECM进行荧光标记，能够可视化ECM的空间变化，特别适用于研究器官纤维化、肿瘤基质形成、伤口愈合和再生中的结缔组织动力学。这种体内化学命运追踪方法在任何组织/器官系统上都具有高度的适应性，可以补充细胞命运追踪技术。此外，由于蛋白质的基本化学性质在物种间高度保守，这种方法也适用于跨物种比较ECM动力学的研究。© 2023。Springer Nature Limited.

Connective tissues are essential building blocks for organ development, repair and regeneration. However, we are at the early stages of understanding connective tissue dynamics. Here, we detail a method that enables in vivo fate mapping of organ extracellular matrix (ECM) by taking advantage of a crosslinking chemical reaction between amine groups and N-hydroxysuccinimide esters. This methodology enables robust labeling of ECM proteins, which complement previous affinity-based single-protein methods. This protocol is intended for entry-level scientists and the labeling step takes between 5 and 10 min. ECM 'tagging' with fluorophores using N-hydroxysuccinimide esters enables visualization of ECM spatial modifications and is particularly useful to study connective tissue dynamics in organ fibrosis, tumor stroma formation, wound healing and regeneration. This in vivo chemical fate mapping methodology is highly versatile, regardless of the tissue/organ system, and complements cellular fate-mapping techniques. Furthermore, as the basic chemistry of proteins is highly conserved between species, this method is also suitable for cross-species comparative studies of ECM dynamics.© 2023. Springer Nature Limited.