Harvard Medical School, Boston, MA 02115, USA; Boston Children's Hospital, Division of Immunology, Boston, MA 02115, USA.
Harvard Medical School, Boston, MA 02115, USA; Boston Children's Hospital, Division of Immunology, Boston, MA 02115, USA; Boston Children's Hospital, Division of Gastroenterology, Boston, MA 02115, USA.
STAR Protoc. 2023 Sep 15;4(3):102372. doi: 10.1016/j.xpro.2023.102372. Epub 2023 Jun 22.
The release of neutrophil extracellular traps (NETs) has been involved in numerous infectious and non-infectious diseases. Nevertheless, quantitative analysis of NETs in vivo has been challenging. Here, we present a protocol for NET quantification by flow cytometry in the bronchoalveolar lavage fluid (BALF) of mice upon pulmonary infection with S. aureus. We describe steps for bacteria growth and instillation and BALF recovery. We then detail staining to quantify the release of NETs and neutrophils recruited to the site of infection. For complete information on the generation and use of this protocol, please refer to Poli et al. (2021) and Poli et al. (2022)..
中性粒细胞胞外陷阱（NETs）的释放与许多感染性和非感染性疾病有关。然而，体内 NETs 的定量分析一直具有挑战性。在这里，我们提出了一种通过流式细胞术在金黄色葡萄球菌肺部感染的小鼠支气管肺泡灌洗液（BALF）中定量 NETs 的方案。我们描述了细菌生长和注入以及 BALF 回收的步骤。然后，我们详细介绍了染色方法，以定量评估 NETs 的释放以及招募到感染部位的中性粒细胞。有关该方案的生成和使用的完整信息，请参见 Poli 等人（2021 年）和 Poli 等人（2022 年）。
