The mice (n = 4/group) were anesthetized and perfused (4% paraformaldehyde, 0.1% glutaraldehyde, and 15% picric acid in phosphate buffer [PB]), and their brains were processed for electron microscopic analysis. Ultrathin sections were cut on a Leica Ultramicrotome, collected on Formvar-coated single-slot grids, and analyzed with a Tecnai 12 BioTWIN electron microscope (FEI). Analysis of mitochondria (area, coverage, density, and contact with the endoplasmic reticulum [ER]) was performed in an unbiased fashion, as previously described (21 (link),22 (link)).
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Ultrastructural Analysis of Mitochondria
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Brains
Buffer
Endoplasmic reticulum
Formvar
Glutaraldehyde
Mice
Microscope electron
Mitochondria
Paraformaldehyde
Phosphate
Picric acid
Ultramicrotome
Corresponding Organization : University of Veterinary Medicine
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Variable analysis
independent variables
- N (number of mice per group)
- Mitochondrial area
- Mitochondrial coverage
- Mitochondrial density
- Contact between mitochondria and endoplasmic reticulum (ER)
- Anesthesia
- Perfusion (4% paraformaldehyde, 0.1% glutaraldehyde, and 15% picric acid in phosphate buffer [PB])
- Brain processing for electron microscopic analysis
- Cutting of ultrathin sections on a Leica Ultramicrotome
- Collection of sections on Formvar-coated single-slot grids
- Analysis with a Tecnai 12 BioTWIN electron microscope (FEI)
- Unbiased analysis of mitochondria, as previously described (21, 22)
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