用于犬鼻腔免疫微环境采样的生理盐水洗液

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES
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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引用次数: 0

摘要

评估犬鼻腔的局部免疫微环境对于研究正常组织健康和疾病状况,特别是与局部炎症相关的组织健康和疾病状况非常重要。我们优化了一种通过连续鼻腔灌洗来评价犬局部鼻腔免疫微环境的技术。简单地说,狗在麻醉状态下,以胸骨平躺的姿势,用柔软的橡胶导管将预热过的无菌生理盐水注入患病的鼻孔。回流的流体被收集到锥形管中,并重复此过程。将含有移位细胞和蛋白质的液体汇集在一起,并将汇集的鼻灌洗液样本通过细胞过滤器过滤,以去除大块碎片和粘液。将样品离心,分离细胞微球进行分析。一旦样品处理完毕,鼻灌洗后的分析包括流式细胞术、通过大量或单细胞RNA测序对细胞进行转录组分析和/或对灌洗液中存在的细胞因子进行定量。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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.

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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
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.
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