通过皮肤TRITC染色追踪树突细胞向淋巴结的迁移

Jyh Liang Hor; Scott N. Mueller

doi:10.21769/BioProtoc.1939

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Peer-reviewed

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

Jyh Liang Hor

Scott N. Mueller email

发布: 2016年09月20日第6卷第18期 DOI: 10.21769/BioProtoc.1939 浏览次数: 9011

评审: Ivan ZanoniAnonymous reviewer(s)

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The authors used this protocol in:

Cover of Immunity, featuring study using the protocol.

Sep 2015

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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

English

中文翻译

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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.