对疟原虫感染的红细胞进行示踪染色以观察免疫吞噬效应

Henrique Borges da Silva; Carlos Eduardo Tadokoro; Maria Regina D’Império Lima

doi:10.21769/BioProtoc.1884

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

Cell Tracer Violet and CellTracker Red CMTPX Staining of Purified Mature Plasmodium-infected Red Blood Cells

对疟原虫感染的红细胞进行示踪染色以观察免疫吞噬效应

HS Henrique Borges da Silva email

CT Carlos Eduardo Tadokoro

ML Maria Regina D’Império Lima email

发布: 2016年08月05日第6卷第15期 DOI: 10.21769/BioProtoc.1884 浏览次数: 11260

评审: Pinchas TsukermanAnonymous reviewer(s)

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 Pathogens, featuring study using the protocol.

Feb 2015

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

相关实验方案

饮食诱导性肥胖模型中脂肪组织炎症性与代谢激活型巨噬细胞的鉴定与分选

Dan Wu [...] Weidong Wang

2025年10月20日 2094 阅读

基于 PKD2L1 启动子驱动的慢病毒系统在体外选择性富集与鉴定脑脊液接触神经元

Wei Tan [...] Qing Li

2025年11月20日 1244 阅读

重新优化原代小胶质细胞分离流程：改进的小胶质细胞提取方法

Jianwei Li [...] Guohui Lu

2025年12月05日 1121 阅读

Abstract

Efficient staining methods to identify Plasmodium-infected red blood cells (iRBCs) are crucial to discriminate precisely the immune cells responsible for their elimination from circulation. Here, we describe the protocol for the purification of iRBCs and their subsequent staining with the vital dyes Cell Tracer Violet (CTV) or CellTracker Red CMTPX (CMTPX), both of which readily diffuse into cells and bind covalently to intracellular amines. The iRBCs stained by using this protocol were used in ex vivo phagocytosis assays, to determine the ability of splenic dendritic cells of phagocytizing these parasites (Borges da Silva et al., 2015).

Materials and Reagents

15 ml tubes (TPP, catalog number: 91015 )
1 ml syringes (BD Biosciences, catalog number: 309659 )
B6.Rag2^-/- (RAGKO) mice (Jackson Laboratories, C57BL/6)
Plasmodium chabaudi (AS strain)
RPMI 1640 (Thermo Fisher Scientific, catalog number: 11875093 )
Fetal bovine serum heat inactivated (Thermo Fisher Scientific, catalog number: 10437028 )
Penicillin-Streptomycin (Thermo Fisher Scientific, catalog number: 15140122 )
L-glutamine (Thermo Fisher Scientific, catalog number: 25030081 )
Sodium pyruvate (Thermo Fisher Scientific, catalog number: 11360070 )
2-mercaptoethanol (Thermo Fisher Scientific, catalog number: 21985023 )
Cell Tracer Violet (CTV) (Thermo Fisher Scientific, catalog number: C34557 )
CellTracker Red CMTPX (CMTPX) (Thermo Fisher Scientific, catalog number: C34552 )
Halothane (Sigma-Aldrich, catalog number: H0150000 )
NaCl
KCl
Na₂HPO₄
KH₂PO₄
Percoll (GE Healthcare Dharmacon, catalog number: 17089101 )
1x phosphate buffered saline (PBS) (see Recipes)
74% Percoll (see Recipes)

Equipment

Centrifuge (Eppendorf, model: 5804 )
1-ml micropipettes
25 gauge needle (BD Biosciences, catalog number: 305122 )
Laminar flow hood (AirClean Systems, catalog number: AC8000HLF )
Epi-Fluorescent microscope (Leica Microsystems, model: DM6000B )

Procedure

English

中文翻译

文章信息

版权信息

如何引用

Readers should cite both the Bio-protocol article and the original research article where this protocol was used:

Silva, H. B. D., Tadokoro, C. E. and D’Império-Lima, M. R. (2016). Cell Tracer Violet and CellTracker Red CMTPX Staining of Purified Mature Plasmodium-infected Red Blood Cells. Bio-protocol 6(15): e1884. DOI: 10.21769/BioProtoc.1884.
Borges da Silva, H., Fonseca, R., Cassado Ados, A., Machado de Salles, E., de Menezes, M. N., Langhorne, J., Perez, K. R., Cuccovia, I. M., Ryffel, B., Barreto, V. M., Marinho, C. R., Boscardin, S. B., Alvarez, J. M., D'Imperio-Lima, M. R. and Tadokoro, C. E. (2015). In vivo approaches reveal a key role for DCs in CD4+ T cell activation and parasite clearance during the acute phase of experimental blood-stage malaria. PLoS Pathog 11(2): e1004598.

Download Citation in RIS Format

分类

您对这篇实验方法有问题吗？

在此处发布您的问题，我们将邀请本文作者来回答。同时，我们会将您的问题发布到Bio-protocol Exchange，以便寻求社区成员的帮助。

发布问题

0 Q&A

提交稿件