Establishing a Whole-Cell Patch Clamp for Electrophysiological Recording from Vomeronasal Sensory Neurons

Take a coronal slice of the chemosensory vomeronasal organ from a transgenic mouse expressing fluorescently labeled sensory neurons.

The sensory epithelium contains these labeled neurons, which are exposed to the lumen connected to the nasal cavity for sensing chemical cues.

Secure the slice in an imaging chamber using a slice anchor.

Transfer the chamber to a recording setup and flow oxygenated extracellular solution to maintain tissue viability.

Position a perfusion pipette near the slice to introduce chemical stimuli and a recording pipette over the sensory neurons.

Using a fluorescence microscope, identify the labeled neurons.

Apply positive pressure to prevent pipette clogging, and approach a neuron until a dimple forms on the membrane.

Switch to negative pressure, drawing the membrane into the pipette.

Apply suction to rupture the membrane and establish continuity with the cytoplasm, called whole-cell patch clamp, allowing the introduction of chemical stimuli and recording the resulting electrophysiological signals.

Transfer a VNO slice to the imaging chamber, and fix the slice position using stainless steel anchor wired with 0.1-millimeter thick synthetic fibers. Do not cove