Quantifying the dynamics of bacterial secondary metabolites by spectral multiphoton microscopy.
Quantifying the dynamics of bacterial secondary metabolites by spectral multiphoton microscopy.
复制标题
通过光谱多光子显微镜量化细菌次生代谢物的动态。
作者：
N. Sullivan;D. Tzeranis;Yun Wang;P. So;D. Newman
吩嗪类化合物是由铜绿假单胞菌产生的一组荧光小分子，在维持细胞氧化还原稳态中发挥作用。吩嗪由于其氧化还原活性、细胞内和细胞外的存在以及其多样的化学性质而在体内研究中具有挑战性。在这里，我们描述了一种非侵入性的体内光学技术，监测吩嗪浓度的细菌细胞内使用时间推移光谱多光子荧光显微镜。该技术能够同时监测培养物中细菌表达的多种弱荧光分子（吩嗪、铁载体、NAD（P）H）。这项工作提供了第一次在体内测量减少吩嗪浓度，以及第一次描述的时间动力学的吩嗪-NAD（P）H氧化还原系统在铜绿假单胞菌，照亮了一个意想不到的作用1-羟基吩嗪。类似的方法可以用来研究细菌内各种小分子的丰度和氧化还原动力学，无论是作为单细胞还是在群落中。
Phenazines, a group of fluorescent small molecules produced by the bacterium Pseudomonas aeruginosa, play a role in maintaining cellular redox homeostasis. Phenazines have been challenging to study in vivo due to their redox activity, presence both intra- and extracellularly, and their diverse chemical properties. Here, we describe a noninvasive in vivo optical technique to monitor phenazine concentrations within bacterial cells using time-lapsed spectral multiphoton fluorescence microscopy. This technique enables simultaneous monitoring of multiple weakly fluorescent molecules (phenazines, siderophores, NAD(P)H) expressed by bacteria in culture. This work provides the first in vivo measurements of reduced phenazine concentration as well as the first description of the temporal dynamics of the phenazine-NAD(P)H redox system in Pseudomonas aeruginosa, illuminating an unanticipated role for 1-hydroxyphenazine. Similar approaches could be used to study the abundance and redox dynamics of a wide range of small molecules within bacteria, both as single cells and in communities.
影响因子：
56.9
作者：
DENK, W;STRICKLER, JH;WEBB, WW
通讯作者：
WEBB, WW