Olfactory mucosa sampling and processing
Mary Xylaki
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Abstract
This protocol describes the olfactory mucosa sampling and processing for RT-QuIC assays and was adapted from A. Stefani et al. (doi:10.1093/brain/awab005).
Steps
Olfactory mucosa sampling
Two olfactory mucosa samples are collected per person, one nasal swabbing from each nostril.
A sterile, disposable swab is inserted in the nasal cavity, (similar to a covid test but more towards the top) to reach the olfactory and not the epithelial mucosa.
The swab is gently rolled on the mucosal surface, removed, and the tip is cut and placed in a 15-ml conical centrifuge tube with 0.9% NaCl solution.
Olfactory mucosa processing
Olfactory mucosa cells are extracted from the swab by vortexing the tube for 1 min at room temperature.
Swab is removed from the tube and the cell suspension is centrifuged at 2000 x g for 20 mins at 4 °C.
Supernatant is removed and the pellet is directly processed or frozen at -80 °C for processing later.
An inoculation loop is used to collect equal amounts of pellet (∼ 2 μl) which are then dipped into a tube containing 120 μl PBS.
Pellets are disolved by sonicating the tube in a bath sonicator at 120 W for at least 1 min.
Samples can be directly used for RT-QuIC assay.
