Skin TRITC Painting to Track Dendritic Cells Migrating to the Lymph Nodes
通过皮肤TRITC染色追踪树突细胞向淋巴结的迁移
发布: 2016年09月20日第6卷第18期 DOI: 10.21769/BioProtoc.1939 浏览次数: 9012
评审: Ivan ZanoniAnonymous reviewer(s)
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参见作者原研究论文
The authors used this protocol in:
Sep 2015
Abstract
Our protocol describes a simple method that allows tracking of dendritic cells (DC) migration from the flank skin to draining lymph nodes (LN) using a red fluorescent dye tetramethylrhodamine-5-isothiocyanate (TRITC). TRITC is a photostable dye that readily labels cells including DC in the skin and can survive repeated exposure of two-photon laser excitation for prolonged imaging duration. This method can be combined with various fluorescent antibody labels or transgenic mouse strains (such as CD11c-EYFP) to visualize distinct DC populations simultaneously.
Keywords: Dendritic cells (树突状细胞)
Skin (皮肤)
Lymph nodes (淋巴结)
Intravital imaging (活体成像)
Materials and Reagents
- 10-20 μl pipette tips
- Paraffin film (Cole-Parmer, catalog number: PM996 )
- Eppendorf tubes (Eppendorf, catalog number: 022364111 )
- Felt-tip marker (Sharpie, fine point permanent marker)
- Mouse (C57BL/6)
- Tetramethylrhodamine-5-(and-6)-isothiocyanate (TRITC) (Thermo Fisher Scientific, catalog number: T490 )
- Acetone (Chem Supply, catalog number: AL008-2.5L-P )
- DMSO (Dimethyl sulfoxide) (Sigma-Aldrich, catalog number: D2650 )
- Veet (sensitive skin hair removal cream) (Reckitt Benckiser)
- Eye gel (e.g., Poly Visc, Alcon Laboratories)
- Ilium Ketamil (Ketamine) (Troy Laboratories)
- Ilium Xylazil-20 (Xylazine) (Troy Laboratories)
- Ketamine/Xylazine solution (see Recipes)
Equipment
- 10-20 μl pipette
- Spatula
- Fine balance for weighing 0.1 μg
- Wahl super trimmer (Wahl, catalog number: 5996415011081 )
- Heat pad (Stoelting, model: 50300 )
- Clippers
- Tissue paper/gauze
- Ruler
Procedure
文章信息
版权信息
© 2016 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Hor, J. L. and Mueller, S. N. (2016). Skin TRITC Painting to Track Dendritic Cells Migrating to the Lymph Nodes. Bio-protocol 6(18): e1939. DOI: 10.21769/BioProtoc.1939.
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分类
免疫学 > 动物模型 > 小鼠
免疫学 > 免疫细胞功能 > 树突细胞
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