Aug 25, 2022
Olfactory mucosa sampling and processing
- maria.xylaki 1
- 1University Medical Center Goettingen
External link: https://pubmed.ncbi.nlm.nih.gov/33855335/
Protocol Citation: maria.xylaki 2022. Olfactory mucosa sampling and processing . protocols.io https://dx.doi.org/10.17504/protocols.io.14egn2b86g5d/v1
Manuscript citation:
Stefani A, Iranzo A, Holzknecht E, Perra D, Bongianni M, Gaig C, et al. Alpha-synuclein seeds in olfactory mucosa of patients with isolated REM sleep behaviour disorder. Brain. Oxford University Press; 2021;144:1118–26.
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: August 25, 2022
Last Modified: May 31, 2024
Protocol Integer ID: 69175
Keywords: olfactory, mucosa, nasal, sample, RT-QuIC, SAA, ASAPCRN, olfactory mucosa sampling, quic assay, assay, quic, processing, processing for rt
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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).
Materials
- benchtop centrifuge with cooling function
- waterbath sonicator
- regular lab equipment (pipetts, tips, tubes)
- nylon flocked swabs (like FLOQBrush®; CopanItalia Spa)
- 1μl plastic inoculation loops, disposable, sterile
- NaCl 0.9% (w/v) aqeous solution
- PBS
Troubleshooting
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.