Histological Analysis of Iron Deficiency

A long-term iron-deficient diet causes the imbalance of liver function and iron loss. In this research, hepatic tissues fixed in 4% paraformaldehyde solution were dehydrated, embedded in paraffin, and cross-sectioned into 4-μm thick slices. The slices were then treated with HCl (5%) to release ferric ions, followed by treatment with 5% potassium ferrocyanide (Sigma, MO, USA) to produce insoluble ferric ferrocyanide and counterstained with the neutral red (18 (link), 19 (link)). In addition, the H&E staining of colon tissue was carried out according to the previous method (20 (link)). Briefly, colon tissues were cut into 5 μm-thick sections and stained with the H&E kit (Solarbio Science and Technology, Beijing, China). All the stained samples were observed under the microscope (DM1000, Leica Microsystems, Wetzlar, Germany).