An Isolated Semi-intact Preparation of the Mouse Vestibular Sensory Epithelium for Electrophysiology and High-resolution Two-photon Microscopy

The overall goal of this procedure is to obtain a near normal preparation of the mouse vestibular sensory epithelium that can be used to study the fundamental cellular and subcellular processes of vestibular hair cells else. This is accomplished by first isolating the bony vestibular labyrinth from the cranial vault. The second step is to carefully scratch away at the bony labyrinth with fine forceps to expose the underlying membranous labyrinth before removing it completely from its bone casing.

Next, the vestibular sensory epithelium is exposed by cutting away the overlying membrane using fine iris scissors. The final step is to mount the semi intact sensory epithelium preparation on a glass bottomed chamber and stabilize it with a weighted nylon net. Ultimately, patch clamp electrophysiology and two photon microscopy are used to characterize the intrinsic membrane properties and calcium signaling profile respectively of each type of vestibular hair, cell, and or primary arine.