烧烤方法:通过共聚焦显微镜对受感染细胞的细菌负荷量化进行简化工作流程.
Jacques Augenstreich1, Michael Shuster1, Yongqiang Fan1
1Department of Cell Biology and Molecular Genetics, University of Maryland, College Park, MD 20742, USA.
Biology open
|December 29, 2023
概括
我们开发了两种自动化方法来量化巨细胞 (BBQ) 内的细菌负担. 这些基于PyImageJ和Cellpose的工作流改善了宿主-病原体相互作用研究的准确性和速度.
科学领域:
- 微生物学 微生物学
- 细胞生物学 细胞生物学
- 免疫学 免疫学 免疫学
背景情况:
- 准确量化巨细胞内的细菌负担 (BBQ) 对于理解宿主-病原体动态至关重要.
- 现有的烧烤方法有局限性,影响结果的可靠性.
- 聚焦显微镜是可视化细胞内细菌的一个关键技术.
研究的目的:
- 引入两种新的,用于细菌负荷量化 (BBQ) 的自动化方法.
- 用先进的图像分析来克服当前烧烤技术的局限性.
- 为研究宿主-病原体相互作用提供可靠,公正和快速的工具.
主要方法:
- 使用PyImageJ和Cellpose软件开发了两种用于烧烤的自动化工作流程.
- 方法1:精制的全光量化与过,以排除未感染的细胞.
- 方法2:计算每个细胞的总细菌体积,以减少基于光的偏差.
主要成果:
- 使用Salmonella enterica serovar Typhimurium和Mycobacterium结核病模型验证的工作流.
- 证明了对巨细胞内的细菌负荷的准确和可靠量化.
- 实现了细菌负担的公正和快速评估.
结论:
- 新的自动化工作流程为BBQ提供了显著的改进.
- 这些工具增强了对宿主细胞内的细菌感染的研究.
- 这些方法适用于研究宿主-病原体相互作用的各种研究.
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