After cryosectioning, muscle sections were stained with haematoxylin & eosin for assessment of muscle architecture, CD31 to visualize capillaries, Van Gieson’s stain to identify collagen and fibrosis, SDH activity as a general marker of mitochondrial (oxidative) activity, and periodic acid–Schiff stain for glycogen content75 (link). The muscle fibre cross-sectional area was measured after immunolabelling of laminin; fibre type was identified by staining myosin heavy chain I and myosin heavy chain IIa. Digital images of stained sections (four images per muscle section) were obtained using an upright microscope (20× objective) with a camera (Axio Imager D1, ZEISS) and AxioVision AC software (AxioVision AC, release 4.7.1, ZEISS) for acquisition. Myofibre cross-sectional area was quantified as described previously76 (link),77 (link).
Other organizations :
Nestlé (Switzerland), University of Toyama, University of Melbourne, USA Mitchell Cancer Institute, University of South Alabama, University of Southampton, Novo Nordisk Foundation, University of Copenhagen, Tehran University of Medical Sciences, Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research, National University Hospital, National University of Singapore, École Polytechnique Fédérale de Lausanne
Collagen and fibrosis identification (Van Gieson's stain)
Mitochondrial (oxidative) activity (SDH activity)
Glycogen content (periodic acid–Schiff stain)
Muscle fibre cross-sectional area (after immunolabelling of laminin)
Muscle fibre type (staining myosin heavy chain I and myosin heavy chain IIa)
control variables
Microscope objective (20×)
Imaging software (AxioVision AC, release 4.7.1)
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