Liu Po C, Lee Yi T, Wang Chun Y, Yang Ya-Tang
Electrical Engineering, National Tsing Hua University.
Electrical Engineering, National Tsing Hua University;
J Vis Exp. 2016 Sep 27(115):54426. doi: 10.3791/54426.
We describe a low cost, configurable morbidostat for characterizing the evolutionary pathway of antibiotic resistance. The morbidostat is a bacterial culture device that continuously monitors bacterial growth and dynamically adjusts the drug concentration to constantly challenge the bacteria as they evolve to acquire drug resistance. The device features a working volume of ~10 ml and is fully automated and equipped with optical density measurement and micro-pumps for medium and drug delivery. To validate the platform, we measured the stepwise acquisition of trimethoprim resistance in Escherichia coli MG 1655, and integrated the device with a multiplexed microfluidic platform to investigate cell morphology and antibiotic susceptibility. The approach can be up-scaled to laboratory studies of antibiotic drug resistance, and is extendible to adaptive evolution for strain improvements in metabolic engineering and other bacterial culture experiments.
我们描述了一种用于表征抗生素耐药性进化途径的低成本、可配置的动态恒温器。该动态恒温器是一种细菌培养装置，可连续监测细菌生长并动态调整药物浓度，以便在细菌进化获得耐药性时持续对其进行挑战。该装置的工作体积约为10毫升，完全自动化，并配备用于测量光密度以及输送培养基和药物的微型泵。为了验证该平台，我们测定了大肠杆菌MG 1655对甲氧苄啶耐药性的逐步获得情况，并将该装置与一个多重微流控平台整合，以研究细胞形态和抗生素敏感性。该方法可扩大规模用于抗生素耐药性的实验室研究，并且可扩展至代谢工程中的菌株改良及其他细菌培养实验的适应性进化研究。
