Temporal bone preparation and experimental procedures were similar to methods described previously by our laboratory (29 (link)–31 (link)), as well as other authors (23 (link)), modified accommodate for the preparation and experimental time required for using whole head specimens. Preparation and experimentation were typically completed on separate days, thus in order to minimize degradation to the tissue, the following schedule was followed for hemi-cephalic/whole head specimens. First, specimens were thawed and temporal bones were prepared in one or both ears and refrozen within approximately 12 or 24 hours. Second, specimens were rethawed; one ear was tested within approximately 12 hours in hemicephalic, and both ears were tested during the course of two consecutive days (~48 h) in whole heads. The total duration that each specimen was left at room temperature was < ~24 hours for hemi-cephalic, and < 72 hours in whole head specimens.
Temporal bones were prepared using the following procedure: specimens were thawed in warm water, and the external ear canal and tympanic membrane were inspected for damage. A canal-wall-up mastoidectomy and extended facial recess approach was performed to visualize the incus, stapes, and round window (30 (link)). The cochlear promontory near the oval and round windows was thinned with a small diamond burr in preparation for pressure sensor insertion into the scala vestibuli (SV) and scala tympani (ST).
Cochleostomies into the ST and SV were created under a droplet of water using a fine pick. Pressure sensors (FOP-M260-ENCAP, FISO Inc., Quebec, QC, Canada), were inserted into the SV and ST using rigidly mounted micromanipulators (David Kopf Instruments, Trujunga, CA). Pressure sensor diameter is approximately 310 μm (comprised of a 260 μm glass tube covered in polyimide tubing with ~25 μm wall thickness), and are inserted into the cochleostomy until the sensor tip is just within the bony wall of the cochlea (~100 μm). Cochleostomies were made as small as possible, such that the pressure probes fit snuggly within, but inserted completely into the opening. Pressure sensor sensitivity is rated at ± 1 psi (6895 Pa). The signal is initially processed by a signal conditioner (Veloce 50; FISO Inc., Quebec, QC, Canada), which specifies the precision and resolution of at 0.3% and 0.1% of full scale, or ~20.7 Pa and 6.9 Pa respectively. Sensors were sealed within the cochleostomies with alginate dental impression material (Jeltrate; Dentsply International Inc., York, PA). Location of the cochlostomies with respect to the basilar membrane were verified visually after each experiment by removing the bone between the two cochleostomies.
Out-of-plane velocity of VStap was measured with a single-axis LDV (OFV-534 & OFV-5000; Polytec Inc., Irvine, CA) mounted to a dissecting microscope (Carl Zeiss AG, Oberkochen, Germany). Microscopic retro-reflective glass beads (Polytec Inc., Irvine, CA) were placed on the neck and posterior crus of the stapes to ensure a strong LDV signal since the stapes footplate was typically obscured by the presence of the stapes tendon. In all LDV measurements, the position of the laser was held as constant as possible between experimental conditions (32 (link),33 (link)).
CI electrodes used in these experiments were: Nucleus Hybrid L24 (HL24; Cochlear Ltd, Sydney, Australia), Nucleus CI422 Slim Straight inserted at 20 and 25 mm (SS20 & SS25; Cochlear Ltd, Sydney, Australia), Nucleus CI24RE Contour Advance (NCA; Cochlear Ltd, Sydney, Australia), HiFocus Mid-Scala (MS; Advanced Bionics AG, Stäfa, Switzerland), and HiFocus 1j (1J; Advanced Bionics AG, Stäfa, Switzerland). Electrode dimensions are provided inTable 1 . Electrodes were inserted sequentially, under water, into the ST via a RW approach. Electrodes were typically inserted in order of smallest to largest (i.e. the order listed above) in an attempt to minimize the effects of damage caused by insertion on subsequent recordings. Potential effects of insertion order are expected to be minimal, owing to the similarity in responses across conditions (see Results), and the lack of any observable effect in one experiment in which the electrode insertion order was shuffled. The cochleostomy was sealed following each electrode insertion with alginate dental impression material, and excess water was removed via suction from the middle ear cavity.
Temporal bones were prepared using the following procedure: specimens were thawed in warm water, and the external ear canal and tympanic membrane were inspected for damage. A canal-wall-up mastoidectomy and extended facial recess approach was performed to visualize the incus, stapes, and round window (30 (link)). The cochlear promontory near the oval and round windows was thinned with a small diamond burr in preparation for pressure sensor insertion into the scala vestibuli (SV) and scala tympani (ST).
Cochleostomies into the ST and SV were created under a droplet of water using a fine pick. Pressure sensors (FOP-M260-ENCAP, FISO Inc., Quebec, QC, Canada), were inserted into the SV and ST using rigidly mounted micromanipulators (David Kopf Instruments, Trujunga, CA). Pressure sensor diameter is approximately 310 μm (comprised of a 260 μm glass tube covered in polyimide tubing with ~25 μm wall thickness), and are inserted into the cochleostomy until the sensor tip is just within the bony wall of the cochlea (~100 μm). Cochleostomies were made as small as possible, such that the pressure probes fit snuggly within, but inserted completely into the opening. Pressure sensor sensitivity is rated at ± 1 psi (6895 Pa). The signal is initially processed by a signal conditioner (Veloce 50; FISO Inc., Quebec, QC, Canada), which specifies the precision and resolution of at 0.3% and 0.1% of full scale, or ~20.7 Pa and 6.9 Pa respectively. Sensors were sealed within the cochleostomies with alginate dental impression material (Jeltrate; Dentsply International Inc., York, PA). Location of the cochlostomies with respect to the basilar membrane were verified visually after each experiment by removing the bone between the two cochleostomies.
Out-of-plane velocity of VStap was measured with a single-axis LDV (OFV-534 & OFV-5000; Polytec Inc., Irvine, CA) mounted to a dissecting microscope (Carl Zeiss AG, Oberkochen, Germany). Microscopic retro-reflective glass beads (Polytec Inc., Irvine, CA) were placed on the neck and posterior crus of the stapes to ensure a strong LDV signal since the stapes footplate was typically obscured by the presence of the stapes tendon. In all LDV measurements, the position of the laser was held as constant as possible between experimental conditions (32 (link),33 (link)).
CI electrodes used in these experiments were: Nucleus Hybrid L24 (HL24; Cochlear Ltd, Sydney, Australia), Nucleus CI422 Slim Straight inserted at 20 and 25 mm (SS20 & SS25; Cochlear Ltd, Sydney, Australia), Nucleus CI24RE Contour Advance (NCA; Cochlear Ltd, Sydney, Australia), HiFocus Mid-Scala (MS; Advanced Bionics AG, Stäfa, Switzerland), and HiFocus 1j (1J; Advanced Bionics AG, Stäfa, Switzerland). Electrode dimensions are provided in