Preparation of Cochlear Samples for SEM Imaging
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Corresponding Organization : University of Sussex
Variable analysis
- Fixing the cochleae by perfusing with 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.2)
- Decalcifying the cochleae in 0.5 M EDTA pH 8.0 for 2-3 days at 4°C
- Morphological changes in the organ of Corti observed using scanning electron microscopy (SEM)
- PH of the sodium cacodylate buffer (7.2)
- Temperature of the EDTA decalcification (4°C)
- Duration of the EDTA decalcification (2-3 days)
- Osmium tetroxide post-fixation (1% for 3 h at room temperature)
- Dehydration through a series of ascending concentrations of ethanol
- Critical-point drying
- Platinum sputter coating
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