Hu Tianbao, Han Xue, Wu Lei, Sun Bangyong, Li Gang
Key Laboratory of Optoelectronic Technology and Systems, Defense Key Disciplines Lab of Novel Micro-Nano Devices and System Technology, Ministry of Education, Chongqing University, Chongqing, 400044, China.
College of Electrical Engineering, Henan University of Technology, Zhengzhou, 450001, China.
Sci Rep. 2025 Jul 13;15(1):25301. doi: 10.1038/s41598-025-11525-6.
Traditional plate culturing remains the "gold standard" in microbiology labs, but it is hindered by labor-intensive workflows, prolonged incubation times, and limited single-cell resolution. labor-intensive, time-consuming, and resource-heavy. Herein, we propose a digital plating (DP) platform that integrates the principles of traditional plate culturing with cutting-edge digital bioassay technology, enabling rapid isolation, quantification, and phenotypic characterization of microorganisms. The DP consists of a high-density picoliter microwell array chip covered with a replaceable agar sheet. In the DP platform, bacterial suspension is partitioned into a high-density picoliter microwell array chip via a self-pumping mechanism and incubated after covering with a specific nutrient- or/and chemical-laden agar sheet. Thanks to the replaceability of the agar cover "plate", the DP allows flexible change of the microenvironment in picowells for culturing or screening microbes, which significantly extends its application range. Meanwhile, the DP platform enables precise bacterial quantification within hours, significantly faster than conventional plate culturing (e.g., in the case of Escherichia coli, 6-7 h for the DP vs. 16-24 h for traditional methods). The platform's versatility is demonstrated through (i) single-cell isolation from mixed microbial communities, (ii) selective enrichment using differential media, (iii) rapid antibiotic susceptibility testing (< 6 h), and (iv) quantitative assessment of microbial interactions. By combining digital quantification with agar-based workflows, the DP platform bridges the gap between high-throughput microfluidics and practical laboratory routines, offering a scalable, cost-effective solution for clinical diagnostics, environmental microbiology, and synthetic biology.
传统的平板培养仍然是微生物实验室中的“金标准”，但它受到劳动强度大的工作流程、较长的培养时间和有限的单细胞分辨率的阻碍，既耗费人力、时间，又消耗资源。在此，我们提出了一种数字平板（DP）平台，该平台将传统平板培养的原理与前沿的数字生物测定技术相结合，能够对微生物进行快速分离、定量和表型表征。DP由一个覆盖有可更换琼脂片的高密度皮升微孔阵列芯片组成。在DP平台中，细菌悬液通过自泵机制被分配到高密度皮升微孔阵列芯片中，并用特定的富含营养物或/和化学物质的琼脂片覆盖后进行培养。由于琼脂覆盖“平板”的可更换性，DP允许灵活改变皮升孔中的微环境以培养或筛选微生物，这显著扩展了其应用范围。同时，DP平台能够在数小时内实现精确的细菌定量，比传统平板培养快得多（例如，对于大肠杆菌，DP需要6 - 7小时，而传统方法需要16 - 24小时）。该平台的多功能性通过以下方式得到证明：（i）从混合微生物群落中进行单细胞分离，（ii）使用差异培养基进行选择性富集，（iii）快速抗生素敏感性测试（< 6小时），以及（iv）对微生物相互作用进行定量评估。通过将数字定量与基于琼脂的工作流程相结合，DP平台弥合了高通量微流控与实际实验室操作之间的差距，为临床诊断、环境微生物学和合成生物学提供了一种可扩展、经济高效的解决方案。
