Pancreatic Islet Morphometry and Insulin Pathway Protein Expression

After being paraffin embedding, the sections (4 μm thickness) from each paraffin block were stained with Eosin and Hematoxylin. The size of pancreatic islets was calculated using ImageJ software Immunohistochemistry was performed with specific antibodies against TNF-α, p-IRS1, p-Akt, and Glut-4 (Santa Cruz, CA, USA), in order to detect the inflammation and the insulin pathway proteins expression in target tissues (white adipose and liver). The antibodies reactivity was detected using a streptavidin-peroxidase Histostaining-SP kit. Positive immunohistochemistry (IHC) stains were defined as yellow-brown color. The score was determined and expressed according two-variable factors as described by Fki et al. [13 (link)].