Perforated Patch-Clamp Recording of Olfactory Sensory Neurons in Mice

Take the severed head of a genetically engineered mouse. The olfactory epithelium in its nose contains olfactory sensory neurons, or OSNs, with fluorophore-tagged odor receptors.

Remove the skin and dissect the head to isolate the septum with attached olfactory epithelium.

Separate and secure the epithelium in a recording chamber.

Using a fluorescence microscope, detect the labeled receptors to target the OSNs' dendritic knob.

Bring a recording pipette with an intracellular solution containing nystatin, a membrane-perforating antibiotic.

Apply positive pressure to avoid pipette clogging. Insert the pipette into the epithelium and approach an OSN.

Switch to negative pressure, forming a seal with the cell membrane. Apply current to maintain the membrane at the resting potential.

Nystatin perforates the membrane, allowing measurement of intracellular ionic currents.

Position a pipette containing an odor stimulant near the OSN. Release the stimulant, which binds to the odor receptors, triggering ion channel opening to allow ion influx. Record the resulting electrophysiological signals.