2.3. Skeletal Muscle Histology

Embedded tibialis anterior muscle blocks from four groups were cut into 5 μm sections and stained with hematoxylin-eosin. Images were observed under a microscope and captured with a digital camera (Olympus, Tokyo, Japan). Tibialis anterior muscle was embedded in optimal cutting temperature compound (VWR, Radnor, PA, USA) and was cut into 20 μm sections. The sections were stained with 0.4% oil red O (ORO) solution (Sigma, St. Louis, MO, USA). Quantification of neutral lipids by ORO analysis was done using ImageJ (National Institutes of Health, Bethesda, MA, USA).