A robust vessel-labeling pipeline with high tissue clearing compatibility for 3D mapping of vascular networks

The combination of vessel-labeling, tissue-clearing, and light-sheet imaging techniques provides a potent tool for accurately mapping vascular networks, enabling the assessment of vascular remodeling in vascular-related disorders. However, most vascular labeling methods face challenges such as inadequate labeling efficiency or poor compatibility with current tissue clearing technology, which significantly undermines the image quality. To address this limitation, we introduce a vessel-labeling pipeline, termed Ultralabel, which relies on a specially designed dye hydrogel containing lysine-fixable dextran and gelatins for double enhancement. Ultralabel demonstrates not only excellent vessel-labeling capability but also strong compatibility with all tissue clearing methods tested, which outperforms other vessel-labeling methods. Consequently, Ultralabel enables fine mapping of vascular networks in diverse organs, as well as multi-color labeling alongside other labeling techniques. Ultralabel should provide a robust and user-friendly method for obtaining 3D vascular networks in different biomedical applications. [Display omitted] •Ultralabel, a vessel casting method based on lysine-fixable dextran hydrogels•Ultralabel enables efficient vascular labeling for both large vessels and capillaries•Ultralabel is well compatible with diverse tissue clearing methods•Ultralabel enables 3D mapping and profiling of vascular networks in health and disease Biological sciences; Biological sciences tools; Optical imaging