S4500 field emission sem

All electrochemical measurements were performed using a lab-built glass cell.^{9 (link)} A mixture of aluminum chloride, dipropylsulfone (DPSO₂), and toluene (1 : 10 : 5 molar ratio) served as the electrolyte solution. A Mo plate (1.3 cm × 1.3 cm), which was used as the current collector, was immersed in conc. HCl for several seconds before use. An Al plate (10 mm in diameter, 0.2 mm in thickness) was used as the counter and reference electrodes in all electrochemical experiments. A glass fiber filter (Advantec, Ltd.) was used as the separator. A test cell was assembled in an Ar-filled glovebox. Cyclic voltammetry and charge/discharge tests were performed at 30 °C using an SI1287 potentiostat (Solartron) and an HJ1001SM8 charge/discharge system (Hokuto Denko), respectively. X-ray diffraction (XRD) spectra were measured by an X-ray diffractometer (50 kV, 30 mA; XRD-6100, Shimadzu) equipped with a Cu Kα source (λ = 0.1541 nm). Scanning electron microscope (SEM) images and energy-dispersive X-ray analysis (EDX) were performed with an S-4500 field-emission SEM (Hitachi) and EDAX system (Ametec), respectively.

Cochleae of P3 and P18 mice were excised from the temporal bone and fixed by perfusion as described above using 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer containing 2 mM calcium chloride and incubation for 2 h. They were stored in 1/10th fixative until further processing. For postfixation, the samples were washed in sodium cacodylate buffer and then immersed in 1% osmium tetroxide in sodium cacodylate for 1 h.
For SEM, cochleae were dissected and then prepared using the OTOTO technique^{73 (link)}. Briefly, segments were incubated in alternating solutions of osmium tetroxide (3X, 2 h) and saturated thiocarbohydrazide (2X, 20 mins) with six washes in water between each change. They were subsequently dehydrated through an ethanol series and critical point dried using a Polaron drier, mounted on platinum stubs using adhesive carbon pads, and examined in a Hitachi S4500 field emission SEM at 5 kV.
For TEM, samples were fixed and dehydrated as for SEM, but then embedded in Spurr’s resin^{73 (link)}, sectioned at 70–100 nm and the sections collected on copper grids. Grids were stained in uranyl acetate and lead citrate and examined in a JEOL 100 S TEM. Images were recorded on Acros Neopan 35 mm negatives and digitized using a HP Canonscan 9000 negative scanner.