荧光显微镜法显示体外中性粒细胞胞外杀菌网

Koji Tanaka; Tadanobu Shimura; Yuji Toiyama; Yoshinaga Okugawa; Yasuhiro Inoue; Toshimitsu Araki; Yasuhiko Mohri; Akira Mizoguchi; Masato Kusunoki

doi:10.21769/BioProtoc.1550

Improve Research Reproducibility A Bio-protocol resource

提交稿件
订阅
登录
/
注册
- 个人主页
- 编辑个人信息
- 修改密码
- 退出
CN
- EN - English
- CN - 中文

Peer-reviewed

Visualization of ex vivo Neutrophil Extracellular Traps by Fluorescence Microscopy

荧光显微镜法显示体外中性粒细胞胞外杀菌网

Koji Tanaka email

发布: 2015年08月05日第5卷第15期 DOI: 10.21769/BioProtoc.1550 浏览次数: 11554

评审: Ivan ZanoniAchille BroggiMarco Di Gioia

PDF

Q&A

引用

Cited by

参见作者原研究论文

The authors used this protocol in:

Cover of PLOS ONE, featuring study using the protocol.

Nov 2014

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

利用CRISPRainbow在人体细胞中实时成像特定位点基因组

Thomas J. Versosky [...] Li-Chun Tu

2025年09月05日 2419 阅读

快速且均一的 NHS 酯基膜蛋白标记方法用于活体哺乳动物细胞

Alyssa Burgess [...] Ying S. Hu

2025年10月05日 433 阅读

基于荧光寿命的 FAST 标记细胞区室分离

Aidar R. Gilvanov [...] Yulia A. Bogdanova

2025年10月05日 209 阅读

Abstract

Neutrophil extracellular traps (NETs) are extracellular DNAs decorated with nuclear and granular proteins such as histones, neutrophil elastase or myeloperoxidase. They exhibit fibrous mesh-like, web-like, or string-like structures. Here, we describe our protocol regarding visualization of ex vivo NETs released from neutrophils activated by lipopolysaccharide (LPS) using fluorescence microscopy.

Materials and Reagents

Whole blood from wild-type C57/BL6 mice (Japan SLC, Inc.)
Whole blood from human volunteers
LPS (Escherichia coli, serotype 0111:B4) (Sigma-Aldrich, catalog number: L4391 )
Polymorphprep^TM (Axis Shield PoC AS, catalog number: 1114683 )
RPMI 1640 medium (no phenol red) (Life Technologies, catalog number: 32404-014 )
Fetal bovine serum (Life Technologies, catalog number: 12483-020 )
ACK (Ammonium-chloride-potassium) lysing buffer (Lonza, catalog number: 10-548E )
SYTOX Green (Life Technologies, Invitrogen^TM, catalog number: S7020 )

Equipment

Glass Pasteur pipets (Iwaki brand, Asahi Techno Glass Corporation)
96-well plates (TPP Techno Plastic Products AG)
CO₂ Incubator (SANYO)
Fluorescence microscopy (Olympus, model: IX71 )

Procedure

English

中文翻译

文章信息

版权信息

如何引用

Tanaka, K., Shimura, T., Toiyama, Y., Okugawa, Y., Inoue, Y., Araki, T., Mohri, Y., Mizoguchi, A. and Kusunoki, M. (2015). Visualization of ex vivo Neutrophil Extracellular Traps by Fluorescence Microscopy. Bio-protocol 5(15): e1550. DOI: 10.21769/BioProtoc.1550.