Laboratory for Soft Materials and Interfaces, Department of Materials, ETH Zürich, Switzerland.
Department of Infectious Diseases and Hospital Epidemiology, University Hospital Zürich, University of Zurich, Switzerland.
Lab Chip. 2023 Nov 21;23(23):5018-5028. doi: 10.1039/d3lc00611e.
In microbiology, accessing single-cell information within large populations is pivotal. Here we introduce bio-sCAPA, a technique for patterning bacterial cells in defined geometric arrangements and monitoring their growth in various nutrient environments. We demonstrate bio-sCAPA with a study of subpopulations of antibiotic-tolerant bacteria, known as persister cells, which can survive exposure to high doses of antibiotics despite lacking any genetic resistance to the drug. Persister cells are associated with chronic and relapsing infections, yet are difficult to study due in part to a lack of scalable, single-cell characterisation methods. As >10 cells can be patterned on each template, and multiple templates can be patterned in parallel, bio-sCAPA allows for very rare population phenotypes to be monitored with single-cell precision across various environmental conditions. Using bio-sCAPA, we analysed the phenotypic characteristics of single cells tolerant to flucloxacillin and rifampicin killing. We find that antibiotic-tolerant cells do not display significant heterogeneity in growth rate and are instead characterised by prolonged lag-time phenotypes alone.
在微生物学中，获取大群体中的单细胞信息至关重要。在这里，我们介绍了 bio-sCAPA 技术，该技术可用于在特定几何排列中对细菌细胞进行图案化，并在各种营养环境中监测它们的生长。我们通过对被称为持留细胞的抗生素耐受细菌亚群的研究来展示 bio-sCAPA 技术，持留细胞尽管对药物没有遗传抗性，但可以在暴露于高剂量抗生素的情况下存活。持留细胞与慢性和复发性感染有关，但由于缺乏可扩展的单细胞特征化方法，因此难以研究。由于每个模板上可以图案化 >10 个细胞，并且可以并行图案化多个模板，因此 bio-sCAPA 允许以单细胞精度在各种环境条件下监测非常罕见的群体表型。使用 bio-sCAPA，我们分析了对氟氯西林和利福平杀伤具有耐受性的单个细胞的表型特征。我们发现，抗生素耐受细胞在生长速率方面没有表现出明显的异质性，而是仅表现出延长的迟滞时间表型。
