In vivo atomic force microscopy-infrared spectroscopy of bacteria
In vivo atomic force microscopy-infrared spectroscopy of bacteria
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DOI：
10.1098/rsif.2018.0115
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发表时间：
2018-03-01
影响因子：
3.9
通讯作者：
Wood, Bayden R.
中科院分区：
文献类型：
--
作者：
Kochan, Kamila;Perez-Guaita, David;Wood, Bayden R.
介绍了一种利用原子力显微镜-红外光谱(AFM-IR)探测活细菌纳米级分子变化的新实验平台。这种近场技术非常适合单个细菌细胞的研究。在这里，我们报告了它在金黄色葡萄球菌细胞分裂过程中监测细胞壁动态变化的应用，使用AFM来显示细胞的分裂，并使用AFM-IR记录光谱来显示隔膜的增厚。在这项工作之后，对几个细菌物种的单细胞进行了调查，特别强调了细胞壁的特征。具体地说，可以在纳米空间分辨率上识别和绘制包含复杂碳水化合物和磷酸二酯基的金黄色葡萄球菌和大肠杆菌的主要细胞壁成分，包括肽多聚糖和磷壁酸。通过对6种活细菌的AFM-IR谱进行主成分分析，可以根据主要来源于细胞壁成分的谱带区分革兰氏阳性菌和革兰氏阴性菌。在纳米尺度上监测细菌细胞过程中体内分子变化的能力为研究环境影响和其他影响细菌化学的因素打开了一个新的平台。
A new experimental platform for probing nanoscale molecular changes in living bacteria using atomic force microscopy-infrared (AFM-IR) spectroscopy is demonstrated. This near-field technique is eminently suited to the study of single bacterial cells. Here, we report its application to monitor dynamical changes occurring in the cell wall during cell division in Staphylococcus aureus using AFM to demonstrate the division of the cell and AFM-IR to record spectra showing the thickening of the septum. This work was followed by an investigation into single cells, with particular emphasis on cell-wall signatures, in several bacterial species. Specifically, mainly cell wall components from S. aureus and Escherichia coli containing complex carbohydrate and phosphodiester groups, including peptidoglycans and teichoic acid, could be identified and mapped at nanometre spatial resolution. Principal component analysis of AFM-IR spectra of six living bacterial species enabled the discrimination of Gram-positive from Gram-negative bacteria based on spectral bands originating mainly from the cell wall components. The ability to monitor in vivo molecular changes during cellular processes in bacteria at the nanoscale opens a new platform to study environmental influences and other factors that affect bacterial chemistry.