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相关实验视频

Updated: Jul 26, 2025

Demonstrating Hairy and Glabrous Skin Innervation in a 3D Pattern Using Multiple Fluorescent Staining and Tissue Clearing Approaches

Demonstrating Hairy and Glabrous Skin Innervation in a 3D Pattern Using Multiple Fluorescent Staining and Tissue Clearing Approaches

Published on: May 20, 2022

皮肤整体安装免疫光染色协议,3D可视化和空间图像分析.

Alfonso J Schmidt^1,2, Graham D Wright³, Franca Ronchese²

¹Hugh Green Cytometry Centre, Wellington, New Zealand.

Current protocols

|June 20, 2023

概括

这项研究介绍了一种全方位的皮肤染色协议,用于对免疫细胞和结构的3D成像. 该方法允许对皮肤进行详细的分析.

关键词:

菲律宾菲律宾菲律宾邻近频率和正常化中位数均度 (NME)空间分布指数 (SDI) 是指空间分布的指数.细胞简介器细胞简介器共聚焦激光扫描显微镜 (CLSM)光结合初级抗体的抗体.图像分析图像分析多色色的免疫光染色整体山皮肤的皮肤.

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Horizontal Whole Mount: A Novel Processing and Imaging Protocol for Thick, Three-dimensional Tissue Cross-sections of Skin

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Last Updated: Jul 26, 2025

Demonstrating Hairy and Glabrous Skin Innervation in a 3D Pattern Using Multiple Fluorescent Staining and Tissue Clearing Approaches

Demonstrating Hairy and Glabrous Skin Innervation in a 3D Pattern Using Multiple Fluorescent Staining and Tissue Clearing Approaches

Published on: May 20, 2022

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Published on: March 27, 2021

Horizontal Whole Mount: A Novel Processing and Imaging Protocol for Thick, Three-dimensional Tissue Cross-sections of Skin

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Published on: August 2, 2017

科学领域:

免疫学免疫学免疫学
细胞生物学细胞生物学
显微镜的使用方法

背景情况:

整体皮肤染色提供了皮肤结构和免疫细胞的3D视图,绕过了传统的切割.
了解皮肤对病原体的免疫防御,需要详细描述居民细胞类型及其空间组织.

研究的目的:

提供一个详细的协议,用于多色色的免疫光染色的整个山鼠皮肤.
为了使免疫细胞群和组织结构在三维中可视化和定量分析.
促进研究皮肤在对抗病原体的免疫战略.

主要方法:

用抗体对抗结构标记物 (CD31,LYVE-1) 和免疫细胞 (MHCII,CD64,CD103,CD326) 的多色色免疫光染色,对整个山鼠皮肤进行染色.
同焦激光扫描显微镜 (CLSM) 用于高分辨率的3D成像.
使用开源软件 (ImageJ/FIJI) 进行图像处理和可视化,包括z投影,直角视图,3D染和动画.
使用CellProfiler进行定量空间分析,以评估细胞类型的关系,使用诸如空间分布指数 (SDI),邻近频率 (NF) 和规范中位数均度 (NME) 等指数.

主要成果:

一个优化的染色面板成功地可视化了血管,淋巴网络,抗原呈现细胞,巨细胞,单细胞,树突性表皮T细胞和朗格汉斯细胞.
建立了用于3D图像可视化和细胞分布和相互作用的定量空间分析的管道.
使用商业可用的试剂和自由可用的软件进行全面的皮肤免疫细胞分析的可行性.

结论:

所介绍的协议使得对整个鼠标皮肤进行了强大的染色,成像和分析.
这种方法提升了对皮肤结构和免疫学格局的理解.
研究人员现在可以有效地使用可访问的工具和技术研究皮肤的防御机制.