通过流式细胞术和组织学鉴定鼠类嗜碱性粒细胞。

Identification of Murine Basophils by Flow Cytometry and Histology.

机构信息

Institute of Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen, Erlangen, Germany.

Infektionsbiologische Abteilung, Universitätsklinikum Erlangen, Erlangen, Germany.

出版信息

Methods Mol Biol. 2020;2163:367-375. doi: 10.1007/978-1-0716-0696-4_29.

DOI:10.1007/978-1-0716-0696-4_29

PMID:32766990

Abstract

Here, we describe how murine basophils can be detected in vivo by flow cytometry and immunofluorescence staining. Basophils constitute a homogeneous population of CD4CD19CD49bIgE cells in flow cytometric analysis. When IgE levels are low, one can also use anti-FcεRI or anti-CD200R3 antibodies instead of anti-IgE. For immunofluorescence staining, we use an anti-Mcpt8 antibody since Mcpt8 is a specific marker for murine basophils. We describe how to prepare the tissue to cut cryo-sections and how to perform the staining using a tyramide-based amplification kit.

摘要

在这里，我们描述了如何通过流式细胞术和免疫荧光染色在体内检测鼠类嗜碱性粒细胞。在流式细胞术分析中，嗜碱性粒细胞构成了 CD4CD19CD49bIgE 细胞的同质群体。当 IgE 水平较低时，也可以使用抗 FcεRI 或抗 CD200R3 抗体代替抗 IgE。对于免疫荧光染色，我们使用抗 Mcpt8 抗体，因为 Mcpt8 是鼠类嗜碱性粒细胞的特异性标志物。我们描述了如何准备组织以切割冷冻切片，以及如何使用基于酪胺的扩增试剂盒进行染色。