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使用氯膦酸脂质体化学清除冈比亚按蚊吞噬血细胞的实验方案。

Protocol to chemically deplete phagocytic hemocytes in Anopheles gambiae using clodronate liposomes.

作者信息

Kwon Hyeogsun, Smith Ryan C

机构信息

Department of Plant Pathology, Entomology, and Microbiology, Iowa State University, Ames, IA 50011, USA.

出版信息

STAR Protoc. 2025 May 15;6(2):103819. doi: 10.1016/j.xpro.2025.103819.

Abstract

Understanding the roles of phagocytic hemocytes in mosquito innate immunity has been significantly limited due to the lack of genetic tools. Here, we present a protocol for depleting phagocytic hemocytes in Anopheles gambiae mosquitoes using clodronate liposomes. We describe steps for mosquito injection, as well as validation by microscopy, quantitative real-time PCR (real-time qPCR), and flow cytometry analysis. This protocol allows for the delineation of phagocytic hemocyte function in mosquito immunity, which can be more broadly applied to other arthropod systems. For complete details on the use and execution of this protocol, please refer to Kwon et al..

摘要

由于缺乏遗传工具,对吞噬血细胞在蚊子先天免疫中的作用的理解一直受到很大限制。在这里,我们提出了一种使用氯膦酸脂质体耗尽冈比亚按蚊中吞噬血细胞的方案。我们描述了蚊子注射步骤,以及通过显微镜、定量实时PCR(实时qPCR)和流式细胞术分析进行验证的方法。该方案有助于阐明吞噬血细胞在蚊子免疫中的功能,并且可以更广泛地应用于其他节肢动物系统。有关该方案使用和执行的完整详细信息,请参考权等人的研究。

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