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JoVE Journal
Immunology and Infection
別のマウス皮膚の領域から細胞蛍光分析のための単一細胞懸濁液の調製
別のマウス皮膚の領域から細胞蛍光分析のための単一細胞懸濁液の調製
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Preparation of Single-cell Suspensions for Cytofluorimetric Analysis from Different Mouse Skin Regions

別のマウス皮膚の領域から細胞蛍光分析のための単一細胞懸濁液の調製

Full Text
19,703 Views
05:58 min
April 20, 2016

DOI: 10.3791/52589-v

Achille Broggi1,2, Clara Cigni1, Ivan Zanoni1,2,3, Francesca Granucci1,3

1Department of Biotechnology and Biosciences,University of Milano-Bicocca, 2Boston Children's Hospital, Division of Gastroenterology,Harvard Medical School, 3Humanitas Clinical and Research Center

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Overview

This article presents a methodology for preparing a single-cell suspension from mouse skin to analyze resident immune cell populations. The technique allows for the digestion of skin samples from various regions without damaging immune cell surface markers.

Key Study Components

Area of Science

  • Immunology
  • Cell Biology
  • Flow Cytometry

Background

  • The skin contains a diverse network of immune cells.
  • Understanding these populations is crucial for studying skin immunity.
  • Flow cytometry is a key technique for analyzing cell populations.
  • Efficient sample preparation is essential for accurate results.

Purpose of Study

  • To develop a reliable method for isolating immune cells from mouse skin.
  • To facilitate the identification of different leukocyte populations.
  • To improve the analysis of immune responses in skin.

Methods Used

  • Sample collection from various regions of mouse skin.
  • Use of forceps and scraping techniques for sample preparation.
  • Application of depilatory lotion for hair removal.
  • Flow cytometric analysis to identify immune cell populations.

Main Results

  • Successful generation of single-cell suspensions from mouse skin.
  • Identification of distinct populations of resident immune cells.
  • Preservation of surface markers during the digestion process.
  • Enhanced understanding of skin immune cell dynamics.

Conclusions

  • The described methodology is effective for studying skin immunity.
  • Flow cytometry provides valuable insights into immune cell populations.
  • This approach can be applied to various research contexts in immunology.

Frequently Asked Questions

What is the main goal of this procedure?
The main goal is to prepare a single-cell suspension from mouse skin to identify different populations of resident immune cells.
How are skin samples collected?
Skin samples are collected from various regions, including the ears, using forceps and scraping techniques.
What technique is used for analyzing the immune cells?
Flow cytometry is used to analyze the different leukocyte populations in the skin.
Why is it important to preserve surface markers?
Preserving surface markers is crucial for accurately identifying and analyzing immune cell populations.
Can this method be applied to other areas of research?
Yes, this methodology can be adapted for various studies in immunology and related fields.

皮膚は複雑な免疫細胞ネットワークに生息しています。私たちは、動物の体のさまざまな部分からマウスの皮膚を消化するための効率的な方法論を説明し、単一細胞懸濁液を取得し、フローサイトメトリーによって皮膚に常在するさまざまな白血球集団を分析します。

この手順の全体的な目標は、マウスの皮膚から単一細胞懸濁液を調製し、細胞蛍光分析を通じて常在免疫細胞の異なる集団を同定し、皮膚常在免疫細胞の異なる集団を同定し、単離することです。この技術を使用すると、研究者は、目的の免疫細胞表面メーカーに損傷を与えることなく、マウスの皮膚のさまざまな領域からのサンプルを簡単に消化できます。

マウスの耳から皮膚サンプルを採取するには、耳の無毛部分を切り取り、鉗子を使用して臓器の背側と腹側を引き離します。次に、内耳をそっとこすり、残っている軟骨を取り除きます。チャンクの皮膚サンプルを取得するには、バリカンで動物のすべての毛を剃り、続いて脱毛ローションを塗布します。

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医学 問題110 白血球 免疫系 皮膚 真皮 フローサイトメトリー 単一細胞調製物 コラゲナーゼ

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