Infection and Immunity, University College London, Cruciform Building, Gower Street, London, WC1E 6BT, United Kingdom.
The Bateson Centre, University of Sheffield, Firth Court, Western Bank, Sheffield, S10 2TN, United Kingdom.
Sci Rep. 2020 Feb 21;10(1):3149. doi: 10.1038/s41598-020-59932-1.
We describe new open source software called QuantiFish for rapid quantitation of fluorescent foci in zebrafish larvae, to support infection research in this animal model. QuantiFish extends the conventional measurements of bacterial load and number of bacterial foci to include measures for dissemination of infection. These are represented by the proportions of bacteria between foci and their spatial distribution. We showcase these measures by comparison of intravenous and hindbrain routes of Mycobacterium marinum infection, which are indistinguishable by measurement of bacterial load and not consistently differentiated by the number of bacterial foci. The intravenous route showed dose dependent dissemination of infection, reflected by increased spatial dispersion of bacteria and lower proportions of bacteria distributed across many foci. In contrast, hindbrain infection resulted in localised disease, limited to a smaller area and higher proportions of bacteria distributed across fewer foci. The application of QuantiFish may extend beyond models of infection, to study other pathologies such as metastatic cancer.
我们描述了一种名为 QuantiFish 的新开源软件，用于快速定量斑马鱼幼虫中的荧光焦点，以支持该动物模型中的感染研究。QuantiFish 将细菌负荷和细菌焦点数量的常规测量扩展到包括感染传播的测量。这些由焦点之间的细菌比例及其空间分布来表示。我们通过比较分枝杆菌感染的静脉内和后脑途径来展示这些措施，这些措施通过细菌负荷的测量无法区分，并且通过细菌焦点的数量也不一致。静脉内途径显示感染的扩散呈剂量依赖性，反映在细菌的空间分散增加，以及分布在许多焦点中的细菌比例降低。相比之下，后脑感染导致局部疾病，仅限于较小的区域，并且分布在较少焦点中的细菌比例较高。QuantiFish 的应用可能不仅限于感染模型，还可以用于研究其他病理学，如转移性癌症。
