Cryosectioning and Microscopic Imaging of Skeletal Muscles

Ten micrometer thick cross-sections of TA and GA were performed at −25 °C using a cryostat (HM500M Microm International, Fisher Scientific, Illkirch, France) and stained [22 (link),23 (link)]. Observations and image acquisitions were performed using a photonic microscope in bright field mode (Olympus BX-51, Tokyo, Japan), coupled to a high-resolution cooled digital camera (Olympus DP72) and Cell-D software (Olympus Soft Imaging Solutions, Münster, Germany) [22 (link),23 (link)]. After the image acquisition for each muscle section, the image analysis was performed using the Visilog 6.9 software (Noesis, Crolles, France).