Bio-Electrical Engineering Innovation Hub, University of Warwick, Coventry CV4 7AL, U.K.
Department of Chemistry, University of Warwick, Coventry CV4 7AL, U.K.
Anal Chem. 2020 Dec 15;92(24):16024-16032. doi: 10.1021/acs.analchem.0c03653. Epub 2020 Nov 26.
This paper reports on the use of scanning ion conductance microscopy (SICM) to locally map the ionic properties and charge environment of two live bacterial strains: the Gram-negative and the Gram-positive . SICM results find heterogeneities across the bacterial surface and significant differences among the Gram-positive and Gram-negative bacteria. The bioelectrical environment of the was found to be considerably more negatively charged compared to . SICM measurements, fitted to a simplified finite element method (FEM) model, revealed surface charge values of -80 to -140 mC m for the Gram-negative . The Gram-positive show a much higher conductivity around the cell wall, and surface charge values between -350 and -450 mC m were found using the same simplified model. SICM was also able to detect regions of high negative charge near , not detected in the topographical SICM response and attributed to the extracellular polymeric substance. To further explore how the cell wall structure can influence the SICM current response, a more comprehensive FEM model, accounting for the physical properties of the Gram-positive cell wall, was developed. The new model provides a more realistic description of the cell wall and allows investigation of the relation between its key properties and SICM currents, building foundations to further investigate and improve understanding of the Gram-positive cellular microenvironment.
本文报告了扫描离子电导显微镜(SICM)在两种活细菌菌株(革兰氏阴性菌和革兰氏阳性菌)的局部离子特性和电荷环境映射中的应用。SICM 结果发现细菌表面存在异质性,革兰氏阳性菌和革兰氏阴性菌之间存在显著差异。与 相比, 被发现带负电荷的程度要大得多。拟合简化有限元方法(FEM)模型的 SICM 测量结果表明,革兰氏阴性菌的表面电荷值为-80 至-140 mC m。革兰氏阳性菌在细胞壁周围显示出更高的电导率,使用相同的简化模型发现表面电荷值在-350 至-450 mC m 之间。SICM 还能够检测到 附近带高负电荷的区域,这在形貌 SICM 响应中未检测到,归因于细胞外聚合物。为了进一步探索 细胞壁结构如何影响 SICM 电流响应,开发了一个更全面的 FEM 模型,该模型考虑了革兰氏阳性菌细胞壁的物理特性。新模型提供了细胞壁的更真实描述,并允许研究其关键特性与 SICM 电流之间的关系,为进一步研究和提高对革兰氏阳性细胞微环境的理解奠定了基础。