Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli.
Leveraging microfluidic dielectrophoresis to distinguish compositional variations of lipopolysaccharide in E. coli.
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DOI：
10.3389/fbioe.2023.991784
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发表时间：
2023
影响因子：
5.7
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中科院分区：
文献类型：
--
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脂多糖（LPS）是构成革兰氏阴性细菌细胞包膜外层的独特特征。 LPS结构的变化影响许多生理过程，包括外膜通透性、抗菌素耐药性、宿主免疫系统的识别、生物膜形成和细菌间竞争。 LPS 特性的快速表征对于研究这些 LPS 结构变化与细菌生理学之间的关系至关重要。然而，目前对 LPS 结构的评估需要 LPS 提取和纯化，然后进行繁琐的蛋白质组分析。本文展示了第一个直接区分具有不同 LPS 结构的大肠杆菌的高通量和非侵入性策略。在线性电动分析中结合使用基于三维绝缘体的介电电泳 (3DiDEP) 和细胞追踪，我们阐明了大肠杆菌 LPS 寡糖的结构变化对电动迁移率和极化率的影响。我们证明我们的平台足够灵敏，可以在分子水平上检测 LPS 结构变化。为了将 LPS 的电动特性与外膜渗透性联系起来，我们进一步研究了 LPS 结构变化对细菌对粘菌素敏感性的影响，粘菌素是一种已知通过靶向 LPS 破坏外膜的抗生素。我们的结果表明，采用 3DiDEP 的微流体电动平台可以成为根据 LPS 糖型分离和选择细菌的有用工具。这些平台的未来迭代可用于根据病原体表面 LPS 结构特征快速分析病原体。
Lipopolysaccharide (LPS) is the unique feature that composes the outer leaflet of the Gram-negative bacterial cell envelope. Variations in LPS structures affect a number of physiological processes, including outer membrane permeability, antimicrobial resistance, recognition by the host immune system, biofilm formation, and interbacterial competition. Rapid characterization of LPS properties is crucial for studying the relationship between these LPS structural changes and bacterial physiology. However, current assessments of LPS structures require LPS extraction and purification followed by cumbersome proteomic analysis. This paper demonstrates one of the first high-throughput and non-invasive strategies to directly distinguish Escherichia coli with different LPS structures. Using a combination of three-dimensional insulator-based dielectrophoresis (3DiDEP) and cell tracking in a linear electrokinetics assay, we elucidate the effect of structural changes in E. coli LPS oligosaccharides on electrokinetic mobility and polarizability. We show that our platform is sufficiently sensitive to detect LPS structural variations at the molecular level. To correlate electrokinetic properties of LPS with the outer membrane permeability, we further examined effects of LPS structural variations on bacterial susceptibility to colistin, an antibiotic known to disrupt the outer membrane by targeting LPS. Our results suggest that microfluidic electrokinetic platforms employing 3DiDEP can be a useful tool for isolating and selecting bacteria based on their LPS glycoforms. Future iterations of these platforms could be leveraged for rapid profiling of pathogens based on their surface LPS structural identity.
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