Mary-Keara Boss, Braden Burdekin, Lauren Harrison, Madison Rivera, Adam Harris, Emily Rout, Erin Trageser, Patricia Gualtieri, Ber-In Lee, Steven Dow
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Saline Lavage for Sampling of the Canine Nasal Immune Microenvironment.
Evaluating the local immune microenvironment of the canine nasal cavity can be important for investigating normal tissue health and disease conditions, particularly those associated with local inflammation. We have optimized a technique to evaluate the local nasal immune microenvironment of dogs via serial nasal lavage. Briefly, with dogs under anesthesia and positioned in sternal recumbency, prewarmed sterile saline is flushed into the affected nostril using a flexible soft rubber catheter. The fluid backflow is collected into conical tubes, and this process is repeated. The fluids containing dislodged cells and proteins are pooled, and the pooled nasal lavage samples are filtered through a cell strainer to remove large debris and mucus. Samples are centrifuged and the cell pellets are isolated for analysis. Once the samples have been processed, analyses that may follow nasal lavage include flow cytometry, transcriptomic analysis of cells via bulk or single-cell RNA seq, and/or quantification of cytokines present in the lavage fluid.
期刊介绍:
JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.