A novel m-xylylene-diamine/glucose based-supramolecular eutectogels with tissue clearing for three dimensional histological imaging

Hydrogel-based tissue clearing technologies have shown significant promise for deep-tissue imaging and subcellular-level optical 3D reconstruction of whole organs. This study proposes a novel approach utilizing a deep eutectic solvent (DES) formulated with glucose and m-xylylene-diamine (MXDA) to create a highly efficient tissue-clearing hydrogel system named the passive hydrogel clearing system (PHCS). PHCS achieved efficient tissue clearing through a single-step tissue gelation process. The resulting hydrogel-tissue complex exhibited thermoreversible properties, transitioning into a sol state upon heating and vice versa upon cooling. Notably, PHCS enabled media embedding, facilitating immunofluorescence histopathology. Additionally, the system demonstrated compatibility with various fluorescent probes, particularly lipophilic dyes. Our study successfully employed PHCS for the reconstruction of vascular structures within the intestine, enabling the generation of a 3D pathology model. These findings suggest that PHCS is a promising novel method for fabricating hydrogels for tissue clearing and holds great potential for application as a mounting medium for morphological imaging. [Display omitted] •A highly efficient tissue-clearing deep eutectic hydrogel system was successfully formulated.•The PHCS hydrogel was revealed as a thermoreversible gel in response to temperature stimulation.•PHCS can form a co-transparent gel with tissues via a self-assembled method.•PHCS has no effect on the fine structure of tissues and is compatible with various fluorescent dyes.•PHCS can be used in resolving 3D reconstruction, also be sliced to use for 2D pathological analysis.