Guinea pig temporal bones were harvested to access the cochleae and to measure the pressure generated by the laser pulses. Either scala tympani (PST) or scala vestibuli pressure (PSV) was measured in different cochleae. To measure the PST, a 400 μm diameter cochleostomy was created in scala tympani (ST) with a motorized drill (World Precision Instruments, Sarasota, FL) approximately 0.5 mm from the bony rim of the round window (RW). Using 3D micromanipulators (MHW103, Narishige, Tokyo, Japan), the pressure sensor was inserted approximately 200 μm through the cochleostomy into ST. A 200 μm optical fiber (P200-5-VIS-NIR, Ocean Optics, Dunedin, FL) was placed in front of the RW with its orientation towards the spiral ganglion neurons. The tip of the optical fiber was 200 μm away from the modiolus. To measure PSV, a 400 μm diameter cochleostomy was made in SV close to the oval window in addition to the cochleostomy in ST. The pressure sensor was inserted approximately 200 μm into SV through SV cochleostomy, and the optical fiber was inserted into ST cochleostomy such that its tip was 200 μm away from the modiolus.
Mhw103
Manufactured by Narishige
Sourced in Japan
The MHW103 is a manual hydraulic micromanipulator manufactured by Narishige. It is designed for precise positioning and control of microelectrodes or other microtools during microscopic procedures. The device features a hydraulic system that allows for smooth and accurate movements along three axes.
Automatically generated - may contain errors
2 protocols using mhw103
1
Measuring Cochlear Pressure in Guinea Pigs
2
Microinjection and Electroporation of Tardigrades
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Glass capillaries (GD-1, NARISHIGE) were pulled by a puller (PC-100, NARISHIGE); the temperatures were set at 66.2 and 62.0 °C. Our microinjection system consists of an inverted microscope (AXIO Vert. A1, Zeiss) equipped with an injector (IM-31, NARISHIGE) and manipulators (MMN-1 and MHW-103, NARISHIGE). Tardigrades were mounted as described previously by Tenlen et al. for RNAi experiments without anesthesia (Fig. 1B ) (54 (link), 55 (link)). After the microinjections, individuals were collected and transferred to a cuvette (CUY505P5, NEPA GENE) for electroporation using a super electroporator (NEPA21 type 2, NEPA GENE). The poring pulse was emitted twice at 250 V for 5 ms with a 50-ms interval, and the transfer pulse was emitted five times at 30 V for 50 ms with a 50-ms interval. Tardigrades were maintained with algal food on an agar plate until observation at 17 °C.
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