4.3. Masson’s Trichrome Staining Method

Masson’s trichrome staining method was used to determine the extent of collagen deposition and fibrosis in mouse kidney tissues. In the corresponding area, H&E staining of the adjacent paraffin section was performed for comparisons of tissue morphology. The experiments were conducted as follows: sections were first deparaffinized and rehydrated in ethanol/water solutions then post-fixed with Bouin’s solution for 1 h at room temperature. The fixation buffer was removed, and slides were stained with iron hematoxylin, Biebrich scarlet-acid fuchsin, and phosphomolybdic-phosphotungstic acid sequentially for 10 min per stain. Slides were then stained with Aniline blue. Finally, slides were washed in 10% acetate solution for 3–5 min and mounted in the mounting medium for observations to be made. The slices were visualized by an Olympus BX-41 microscope.