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A comprehensive evaluation of collagen-based historical artifacts is crucial for the preservation and inheritance of cultural relics, necessitating interdisciplinary approaches that integrate scientific knowledge with practical expertise to develop effective conservation strategies. The present work was focused on a piece of historical leather armor housed in Turpan Museum, Xinjiang, China. SEM–EDS, XRD, ATR-FTIR, and NMR were carried out on this historical leather armor from macro and micro perspectives, mainly including morphology observation, composition analysis, structural assessment, which could provide both quantitative and qualitative insights into the deterioration of the historical leather armor. Additionally, the non-destructive sampling methods and the third-generation sequencing technology (TGS) were employed to identify a total of 13 bacterial species and 8 fungal species, and their metabolic pathways were predicted, providing guidance for preventive conservation and restoration strategies. By the present work, necessary fundamental knowledge might be provided for the effective preservation and inheritance of collagen-based historical artifacts. [ABSTRACT FROM AUTHOR]