Department of Biotechnology, MES Abasaheb Garware College, Pune 411004, India.
Serum Institute of India Pvt. Ltd., 212/2, Hadapsar, Pune 411028, India.
J Microbiol Methods. 2020 Jun;173:105920. doi: 10.1016/j.mimet.2020.105920. Epub 2020 Apr 15.
Many methods are developed to assess antimicrobial action of ZnO nanoparticles (NPs). A large number of methods associated with the use of fluorescent probes are developed, including Spectrofluorometry, fluorescence microscopy, and cytometry. In this study, flowcytometry, Spectrofluorometry and fluorescent microscopy was used to measure membrane potential variation of E. coli and S. aureus cells treated with two different sizes of zinc oxide (ZnO) NPs and were compared with conventional methods. In order to estimate change in membrane potential, E. coli and S. aureus cells were treated with iopnophore agent carbonyl cyanide m-chlorophenylhydrazone (CCCP) and membrane potential was evaluated using fluorescent probe 3,3'-Diethyloxacarbocyanine, iodide (DIOC(3)). All the three methods showed similar results and among these Spectrofluorometry was easy to use and inexpensive to assess the viability of bacterial cells via their membrane potential.
许多方法被开发出来以评估氧化锌纳米粒子(NPs)的抗菌作用。大量与荧光探针使用相关的方法被开发出来,包括分光荧光法、荧光显微镜和细胞计数法。在这项研究中,使用流式细胞术、分光荧光法和荧光显微镜来测量用两种不同尺寸的氧化锌(ZnO)纳米粒子处理的大肠杆菌和金黄色葡萄球菌细胞的膜电位变化,并与传统方法进行了比较。为了估计膜电位的变化,用离子载体羰基氰化物 m-氯代苯腙(CCCP)处理大肠杆菌和金黄色葡萄球菌细胞,并使用荧光探针 3,3'-二乙基氧杂羰花青碘化物(DIOC(3))评估膜电位。这三种方法都显示出相似的结果,其中分光荧光法易于使用且价格低廉,可通过细菌细胞的膜电位来评估其活力。