Nguyen Hang Thi, O'Donovan Lisa A, Venter Henrietta, Russell Cecilia C, McCluskey Adam, Page Stephen W, Trott Darren J, Ogunniyi Abiodun D
Australia Centre for Antimicrobial Resistance Ecology, School of Animal and Veterinary Sciences, Roseworthy Campus, The University of Adelaide, Roseworthy, SA 5371, Australia.
Department of Pharmacology, Toxicology, Internal Medicine and Diagnostics, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi 100000, Vietnam.
Antibiotics (Basel). 2021 Mar 17;10(3):307. doi: 10.3390/antibiotics10030307.
In this study, we optimized and compared different transmission electron microscopy (TEM) methods to visualize changes to Gram-negative bacterial morphology induced by treatment with a robenidine analogue (NCL195) and colistin combination. Aldehyde-fixed bacterial cells (untreated, treated with colistin or NCL195 + colistin) were prepared using conventional TEM methods and compared with ultrathin Tokuyasu cryo-sections. The results of this study indicate superiority of ultrathin cryo-sections in visualizing the membrane ultrastructure of and , with a clear delineation of the outer and inner membrane as well as the peptidoglycan layer. We suggest that the use of ultrathin cryo-sectioning can be used to better visualize and understand drug interaction mechanisms on the bacterial cell membrane.
在本研究中，我们优化并比较了不同的透射电子显微镜（TEM）方法，以观察用罗苯尼丁类似物（NCL195）和黏菌素联合处理诱导的革兰氏阴性菌形态变化。使用传统TEM方法制备醛固定的细菌细胞（未处理、用黏菌素或NCL195 + 黏菌素处理），并与超薄德永冷冻切片进行比较。本研究结果表明，超薄冷冻切片在观察[细菌名称1]和[细菌名称2]的膜超微结构方面具有优势，能清晰勾勒出外膜、内膜以及肽聚糖层。我们建议使用超薄冷冻切片法可更好地观察和理解药物在细菌细胞膜上的相互作用机制。
