Hyperspectral microscope imaging (HMI) technique was employed to assess the changes in physicochemical parameters and microstructure of 'Golden Delicious' apples flesh during storage. Four regions of interest (ROIs), including whole-cell ROI, intercellular space ROI, cytoplasm ROI, and cell wall ROI were investigated to assess their relationships with physicochemical parameters. Different ROIs presented similar vibrational profiles, but with slight differences in spectral intensity, especially in the range of 800-1000 nm. Spectral angle mapper (SAM) was applied to the HMI of apple tissues at different storage stages to clearly show the structural changes of parenchyma cells, while principal component analysis (PCA) could highlight the distribution of sugars, water and pigments in apple flesh at the cellular scale. Simultaneously with the degradation of acid-soluble pectin (ASP), middle lamella dissolution and increased intercellular space were observed using SEM and TEM. Single feature variables were used to construct linear models based on pearson correlation analysis, with R