Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama University, Saitama, Japan.
Laboratory of Engineering NanoBiotechnology, Department of Engineering Geoecology, University of Mining and Geology "St. Ivan Rilski", Sofia, Bulgaria.
Microsc Res Tech. 2021 May;84(5):967-975. doi: 10.1002/jemt.23657. Epub 2020 Nov 27.
The rapid photothermal lysis of Escherichia coli O157:H7 treated with light-triggered Janus nanomotors was visualized by Hilbert differential contrast transmission electron microscopy (HDC-TEM). The extraordinary advantage of this high-resolution microscopic technique was that it revealed the detailed ultrastructure alterations of the treated cells at a state close to their native one. The micrographs demonstrated that Janus nanomotors (mesoporous silica nanoparticles with gold hemisphere and half-capped with cysteamine) were able to target and bind to the pathogenic E. coli. The biorecognition reaction proceeded at slightly acid pH thankful to the formed electrostatic adhesion between positively charged amino groups on nanoparticles surface and the negatively charged cell envelope. The exposure of labeled cells to near infrared laser irradiation leaded to occurrence of effective photothermal damage of their plasma membranes, which was enough strong to lyse bacteria. It was because of the overheating obtained by the photon-to-thermal conversion reaction generated by the surface plasmon resonance response of Janus nanomotors. The good efficiency of photothermal lysis to inactivate E. coli O157:H7 was confirmed by staining with LIVE/DEAD viability kit and quantification of the few survived cells in epifluorescence microscope. Furthermore, HDC-TEM images of ice-embedded inhibited bacteria documented the labeling, membrane disruptions and lysis due to the designed operation of Janus nanomotors. The reported microscopic technique provides a novel strategy for developing of Janus nanomachines as promising platform for nondrug treatment and defeating of antibiotic-resistant pathogenic microorganisms.
希尔伯特微分衬度透射电子显微镜(HDC-TEM)可视化了受光触发的 Janus 纳米马达处理的大肠杆菌 O157:H7 的快速光热裂解。这种高分辨率显微镜技术的非凡优势在于,它揭示了处理细胞的详细超微结构变化,使其接近其自然状态。显微照片表明,Janus 纳米马达(具有金半球和半胱氨酸封端的介孔硅纳米粒子)能够靶向并结合致病性大肠杆菌。由于纳米粒子表面上带正电荷的氨基与细胞包膜之间形成的静电吸附,在稍酸性 pH 值下进行了生物识别反应。将标记的细胞暴露于近红外激光照射下,导致其细胞膜发生有效的光热损伤,足以裂解细菌。这是因为 Janus 纳米马达的表面等离子体共振响应产生的光到热转换反应导致过热。通过 LIVE/DEAD 活力试剂盒对大肠杆菌 O157:H7 的失活进行染色,并在荧光显微镜下定量检测少数存活细胞,证实了光热裂解对灭活大肠杆菌 O157:H7 的良好效率。此外,冰嵌入抑制细菌的 HDC-TEM 图像记录了由于 Janus 纳米马达的设计操作而导致的标记、膜破裂和裂解。所报道的显微镜技术为开发 Janus 纳米机器提供了一种新策略,作为非药物治疗和击败抗生素耐药性致病微生物的有前途的平台。