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Studying molecular changes occurring in individual epithelial cell types requires a reliable method for removing these cells from the larynx. Our objective was to develop a method to harvest individual epithelial cells from the mouse larynx while minimizing contamination from non\u2010laryngeal sites and non\u2010epithelial laryngeal cells.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Mice were euthanized, and the larynx was carefully exposed and separated from non\u2010laryngeal sites. A small dental brush was inserted into the laryngeal inlet and rotated to obtain epithelial cells. Cells were transferred to collection media, counted, and cytospin preparations stained for laryngeal epithelial (i.e., Pan\u2010Keratin, EpCAM, NGFR, p63, K5, \u03b2\u2010tubulin, MUC5AC) and non\u2010epithelial (i.e., vimentin) cell markers. Histopathology was completed on brushed laryngeal tissue sections to evaluate the depth of cell collection. Preliminary Single\u2010cell RNA sequencing (scRNA\u2010seq) was performed to confirm this method can capture diverse laryngeal cell types.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We collected 6000\u20138000 cells from a single larynx and 35000\u201340000 cells from combining brushings from three tissues. Histopathology demonstrated brushing removed the epithelial layer of the larynx and some underlying tissue. Immunofluorescence staining demonstrated the phenotype of harvested cells was primarily epithelial. Preliminary scRNA\u2010seq was successfully conducted and displayed nine unique cell clusters.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>We developed a reliable method of harvesting individual epithelial cells from the mouse larynx. This method will be useful for collection of laryngeal cells for a variety of downstream cellular and molecular assays, including scRNA\u2010seq, protein analyses, and cell\u2010culture\u2010based experiments, following laryngeal injury.<\/jats:p><\/jats:sec><jats:sec><jats:title>Level of Evidence<\/jats:title><jats:p>NA <jats:italic>Laryngoscope<\/jats:italic>, 134:786\u2013794, 2024<\/jats:p><\/jats:sec>","DOI":"10.1002\/lary.30970","type":"journal-article","created":{"date-parts":[[2023,8,22]],"date-time":"2023-08-22T02:46:47Z","timestamp":1692672407000},"page":"786-794","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Method for Collecting Single Epithelial Cells from the Mouse Larynx"],"prefix":"10.1002","volume":"134","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3020-1324","authenticated-orcid":false,"given":"Mohammed Imran","family":"Khan","sequence":"first","affiliation":[{"name":"Department of 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