Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps
Summary: 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 la...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166723003398 |
| _version_ | 1797797170166890496 |
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| author | Valentina Poli Marco Di Gioia Ivan Zanoni |
| author_facet | Valentina Poli Marco Di Gioia Ivan Zanoni |
| author_sort | Valentina Poli |
| collection | DOAJ |
| description | Summary: 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)1 and Poli et al. (2022).2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-03-13T03:44:08Z |
| format | Article |
| id | doaj.art-f8c7dd2695a34cc990390696f2f1b5c5 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-03-13T03:44:08Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-f8c7dd2695a34cc990390696f2f1b5c52023-06-23T04:44:33ZengElsevierSTAR Protocols2666-16672023-09-0143102372Quantitative cytofluorimetric analysis of mouse neutrophil extracellular trapsValentina Poli0Marco Di Gioia1Ivan Zanoni2Harvard Medical School, Boston, MA 02115, USA; Boston Children’s Hospital, Division of Immunology, Boston, MA 02115, USAHarvard Medical School, Boston, MA 02115, USA; Boston Children’s Hospital, Division of Immunology, Boston, MA 02115, USAHarvard 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; Corresponding authorSummary: 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)1 and Poli et al. (2022).2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166723003398ImmunologyMicrobiology |
| spellingShingle | Valentina Poli Marco Di Gioia Ivan Zanoni Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps STAR Protocols Immunology Microbiology |
| title | Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps |
| title_full | Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps |
| title_fullStr | Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps |
| title_full_unstemmed | Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps |
| title_short | Quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps |
| title_sort | quantitative cytofluorimetric analysis of mouse neutrophil extracellular traps |
| topic | Immunology Microbiology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166723003398 |
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