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JoVE Journal
Immunology and Infection
单细胞悬液的细胞荧光分析从不同的鼠标皮肤区域准备
单细胞悬液的细胞荧光分析从不同的鼠标皮肤区域准备
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

单细胞悬液的细胞荧光分析从不同的鼠标皮肤区域准备

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.

皮肤是复杂免疫细胞网络的家园。我们描述了一种从动物身体不同部位消化小鼠皮肤的有效方法,以获得单细胞悬液并通过流式细胞术分析驻留在皮肤中的不同白细胞群。

该程序的总体目标是从小鼠皮肤制备单细胞悬液,以通过细胞荧光分析鉴定不同的常驻免疫细胞群。识别和分离不同的皮肤常驻免疫细胞群。使用这种技术,研究人员可以轻松消化来自小鼠皮肤不同区域的样本,而不会损坏感兴趣的免疫细胞表面制造者。

为了从小鼠耳朵中获取皮肤样本,切掉耳朵的无毛部分,并使用镊子拉开器官的背侧和腹侧。然后,轻轻刮擦内耳以去除任何残留的软骨。要获得大块皮肤样本,请用一把剪刀剃掉动物的所有毛发,然后涂抹脱毛化妆水。

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