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Advancing in situ single-cell microbiological analysis through a microwell droplet array with a gradual open sidewall
Lab on a Chip ( IF 6.1 ) Pub Date : 2023-11-06 , DOI: 10.1039/d3lc00590a Jie Wang 1 , Lin Du 2 , Yuwei Han 3 , Dawei Zhang 1 , Dalei Jing 2
Lab on a Chip ( IF 6.1 ) Pub Date : 2023-11-06 , DOI: 10.1039/d3lc00590a Jie Wang 1 , Lin Du 2 , Yuwei Han 3 , Dawei Zhang 1 , Dalei Jing 2
Affiliation
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The utilization of microfluidic analysis technology has resulted in the advancement of fast pathogenic bacteria detection, which can accurately provide information on biochemical reactions in a single cell and enhance detection efficiency. Nevertheless, the achievement of rapid and effective in situ detection of single-bacteria arrays remains a challenge due to the complexity of bacterial populations and low Reynolds coefficient fluid, resulting in insufficient diffusion. We develop microwell droplet array chips from the lateral hydrodynamic wetting approach to address this issue. The sidewall of the microwell gradually opens which aids in advancing the liquid–air interface and facilitates the impregnation of the solid microwells, preserving the Wenzel state and assisting in resisting the liquid force to separation from the drop. The feasibility of preparing cell arrays and identifying them inside the microwells was demonstrated through the simulated streamlined distribution of gradual and traditional microwells with different sizes. The water-based ink diffusion experiment examined the relationship between diffusion efficiency and flow velocity, as well as the position of the microwell relative to the channel. It showed that the smaller gradual microwell still has a good diffusion efficiency rate at a flow velocity of 2.1 μL min−1 and that the infiltration state is easier to adjust. With this platform, we successfully isolated a mixed population containing E. coli and S. aureus, obtained single-bacteria arrays, and performed Gram assays after in situ propagation. After 20 hours of culture, single bacteria reproduced demonstrating the capability of this platform to isolate, cultivate, and detect pathogenic bacteria.
中文翻译：
通过具有逐渐开放侧壁的微孔液滴阵列推进原位单细胞微生物分析
微流控分析技术的运用促进了病原菌快速检测的进步，可以准确提供单细胞内生化反应的信息，提高检测效率。然而，由于细菌群体的复杂性和低雷诺系数流体导致扩散不足，实现快速有效的单细菌阵列原位检测仍然是一个挑战。我们通过横向流体动力润湿方法开发了微孔液滴阵列芯片来解决这个问题。微孔的侧壁逐渐打开，这有助于推进液-气界面并促进固体微孔的浸渍，保持Wenzel状态并有助于抵抗液体力从液滴中分离。通过模拟不同尺寸的渐变微孔和传统微孔的流线型分布，论证了在微孔内制备细胞阵列并进行识别的可行性。水性墨水扩散实验检查了扩散效率和流速之间的关系，以及微孔相对于通道的位置。结果表明，较小的渐变微孔在流速为2.1 μL min -1时仍具有良好的扩散效率，并且浸润状态更容易调节。利用该平台，我们成功分离了含有大肠杆菌和金黄色葡萄球菌的混合群体，获得了单菌阵列，并在原位繁殖后进行了革兰氏分析。 培养20小时后，单个细菌繁殖，证明了该平台分离、培养和检测病原菌的能力。
更新日期：2023-11-06
中文翻译：
通过具有逐渐开放侧壁的微孔液滴阵列推进原位单细胞微生物分析
微流控分析技术的运用促进了病原菌快速检测的进步，可以准确提供单细胞内生化反应的信息，提高检测效率。然而，由于细菌群体的复杂性和低雷诺系数流体导致扩散不足，实现快速有效的单细菌阵列原位检测仍然是一个挑战。我们通过横向流体动力润湿方法开发了微孔液滴阵列芯片来解决这个问题。微孔的侧壁逐渐打开，这有助于推进液-气界面并促进固体微孔的浸渍，保持Wenzel状态并有助于抵抗液体力从液滴中分离。通过模拟不同尺寸的渐变微孔和传统微孔的流线型分布，论证了在微孔内制备细胞阵列并进行识别的可行性。水性墨水扩散实验检查了扩散效率和流速之间的关系，以及微孔相对于通道的位置。结果表明，较小的渐变微孔在流速为2.1 μL min -1时仍具有良好的扩散效率，并且浸润状态更容易调节。利用该平台，我们成功分离了含有大肠杆菌和金黄色葡萄球菌的混合群体，获得了单菌阵列，并在原位繁殖后进行了革兰氏分析。 培养20小时后，单个细菌繁殖，证明了该平台分离、培养和检测病原菌的能力。
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