Central Research Laboratory, Sree Balaji Medical College & Hospital (SBMCH), Bharath Institute for Higher Education & Research (BIHER), Bharath University, Chennai, Tamil Nadu, 600073, India.
Laboratory of Integrative Multiscale Engineering Materials and Systems, Department of Aerospace Engineering, Indian Institute of Science, Bangalore, 560012, India.
Biotechnol Lett. 2020 May;42(5):853-863. doi: 10.1007/s10529-020-02835-y. Epub 2020 Feb 10.
This study aims to detect pathogenic Escherichia coli (E. coli) bacteria using non-destructive fluorescence microscopy and micro-Raman spectroscopy.
Raman vibrational spectroscopy provides additional information regarding biochemical changes at the cellular level. We have used two nanomaterials zinc oxide nanoparticles (ZnO-NPs) and gold nanoparticles (Au-NPs) to detect pathogenic E. coli. The scanning electron microscope (SEM) with energy dispersive X-ray (EDAX) spectroscopy exhibit surface morphology and the elemental composition of the synthesized NPs. The metal NPs are useful contrast agents due to the surface plasmon resonance (SPR) to detect the signal intensity and hence the bacterial cells. The changes due to the interaction between cells and NPs are further correlated to the change in the surface charge and stiffness of the cell surface with the help of the fluorescence microscopic assay.
We conclude that when two E. coli strains (MTCC723 and MTCC443) and NPs are respectively mixed and kept overnight, the growth of bacteria are inhibited by ZnO-NPs due to changes in cell membrane permeability and intracellular metabolic system under fluorescence microscopy. However, SPR possessed Au-NPs result in enhanced fluorescence of both pathogens. In addition, with the help of Raman microscopy and element analysis, significant changes are observed when Au-NPs are added with the two strains as compared to ZnO-NPs due to protein, lipid and DNA/RNA induced conformational changes.
本研究旨在利用非破坏性荧光显微镜和微拉曼光谱检测致病性大肠杆菌(E. coli)细菌。
拉曼振动光谱提供了有关细胞水平生化变化的附加信息。我们使用了两种纳米材料氧化锌纳米粒子(ZnO-NPs)和金纳米粒子(Au-NPs)来检测致病性大肠杆菌。扫描电子显微镜(SEM)和能谱(EDAX)显示了合成纳米粒子的表面形态和元素组成。金属纳米粒子是有用的对比剂,因为表面等离子体共振(SPR)可以检测信号强度,从而检测细菌细胞。通过荧光显微镜测定,进一步将细胞与纳米粒子相互作用引起的变化与细胞表面电荷和表面硬度的变化相关联。
我们得出结论,当两种大肠杆菌菌株(MTCC723 和 MTCC443)和 NPs 分别混合并放置过夜时,由于细胞膜通透性和细胞内代谢系统的变化,ZnO-NPs 抑制了细菌的生长,在荧光显微镜下观察到。然而,SPR 赋予 Au-NPs 的特性导致两种病原体的荧光增强。此外,借助拉曼显微镜和元素分析,当与两种菌株一起添加 Au-NPs 时,与 ZnO-NPs 相比,观察到蛋白质、脂质和 DNA/RNA 诱导的构象变化导致了明显的变化。