Quantitative Immunofluorescent Imaging of Immune Cells in Mucosal Tissues.
Quantitative Immunofluorescent Imaging of Immune Cells in Mucosal Tissues.
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DOI：
10.1007/978-1-0716-2051-9_9
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发表时间：
2022-01-01
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影响因子：
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通讯作者：
Prodger, Jessica L
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文献类型：
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作者：
Buchanan, Lane B;Shao, Zhongtian;Prodger, Jessica L
了解皮肤和粘膜组织中细菌、病原体和免疫细胞之间的相互作用对于改善无数疾病的预防和治疗至关重要。虽然高参数流式细胞术是目前血液中免疫细胞表征的金标准，但它不太适用于粘膜组织，其中结构和空间信息在组织解聚期间丢失。免疫荧光克服了这一限制，作为一个很好的替代研究粘膜组织中的免疫细胞。然而，使用免疫荧光显微镜分析临床样品受到缺乏高通量定量分析技术的阻碍。在这一章中，我们描述了人类包皮组织切片，染色和成像的方法。我们还描述了从免疫荧光图像中自动定量免疫细胞的方法，包括图像预处理和使用开源软件定量圆形和不规则形状免疫细胞的方法。
Understanding the interplay between commensals, pathogens, and immune cells in the skin and mucosal tissues is critical to improve prevention and treatment of a myriad of diseases. While high-parameter flow cytometry is the current gold standard for immune cell characterization in blood, it is less suitable for mucosal tissues, where structural and spatial information is lost during tissue disaggregation. Immunofluorescence overcomes this limitation, serving as an excellent alternative for studying immune cells in mucosal tissues. However, the use of immunofluorescent microscopy for analyzing clinical samples is hampered by a lack of high-throughput quantitative analysis techniques. In this chapter, we describe methods for sectioning, staining, and imaging whole sections of human foreskin tissue. We also describe methods to automate immune cell quantification from immunofluorescent images, including image preprocessing and methods to quantify both circular and irregularly shaped immune cells using open-source software.