A 4-5X expansion microscopy protocol for high‑resolution imaging of olfactory sensory neuron cilia in mouse olfactory epithelium

Here, we present a protocol for 4-5× expansion of mouse olfactory epithelium (OE), optimized for high-resolution imaging of olfactory sensory neuron (OSN) cilia. The method combines rapid dissection of the OE on a cold platform to preserve delicate ciliary structures with hydrogel embedding and expansion microscopy (ExM) to achieve isotropic 4-5 fold expansion (based on references 2-4). This approach enables detailed visualization of OSN ciliary architecture using conventional confocal microscopy, without the need for specialized super-resolution instrumentation. OSNs were labeled using anti-Olfactory Marker Protein (OMP) and their cilia were labeled using anti-acetyl-tubulin antibodies.