From flies to humans, many components of the innate immune system have been conserved during metazoan evolution. This foundational observation has allowed us to develop Drosophila melanogaster, the fruit fly, into a powerful model to study innate immunity in animals. Thanks to an ever‐growing arsenal of genetic tools, an easily manipulated genome, and its winning disposition, Drosophila is now employed to study not only basic molecular mechanisms of pathogen recognition and immune signaling, but also the nature of physiological responses activated in the host by microbial challenge and how dysregulation of these processes contributes to disease. Here, we present a collection of methods and protocols to challenge the fly with an assortment of microbes, both systemically and orally, and assess its humoral, cellular, and epithelial response to infection. Our review covers techniques for measuring the reaction to microbial infection both qualitatively and quantitatively. Specifically, we describe survival, bacterial load, BLUD (a measure of disease tolerance), phagocytosis, melanization, clotting, and ROS production assays, as well as efficient protocols to collect hemolymph and measure immune gene expression. We also offer an updated catalog of online resources and a collection of popular reporter lines and mutants to facilitate research efforts.

中文翻译：

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研究果蝇先天免疫的方法。

从苍蝇到人类，先天免疫系统的许多组成部分在后生动物进化过程中都得到了保留。这一基础性观察使我们能够将果蝇果蝇（Drosophila melanogaster）发展为研究动物先天免疫力的强大模型。得益于不断增长的遗传工具库，易于操作的基因组及其成功的果蝇（Drosophila）配置现在不仅用于研究病原体识别和免疫信号传导的基本分子机制，而且还用于研究微生物攻击在宿主中激活的生理反应的性质以及这些过程的失调如何导致疾病。在这里，我们提出了一系列的方法和协议，以各种微生物在系统和口腔上挑战果蝇，并评估其对感染的体液，细胞和上皮反应。我们的综述涵盖了定性和定量测量对微生物感染反应的技术。具体来说，我们描述存活率，细菌载量，BLUD（衡量疾病耐受性的指标），吞噬作用，黑色素化，凝血和ROS生成分析，以及收集血淋巴和测量免疫基因表达的有效方案。我们还提供了更新的在线资源目录以及一些流行的记者专栏和突变体，以促进研究工作。