title: High-throughput microfluidic system for long-term bacterial colony monitoring and antibiotic testing in zero-flow environments.
abstract: In this study, a high-throughput microfluidic system is presented. The system is comprised of seven parallel channels. Each channel contains 32 square-shaped microchambers. After simulation studies on samples loaded into the microchambers, and the solute exchange between the microchambers and channels, the long-term culture of Escherichia coli (E. coli) HB101 in the microchambers is realized. Using the principle that L-arabinose (L-Ara) can induce recombinant E. coli HB101 pGLO to synthesize green fluorescent protein (GFP), the real-time analysis of GFP expression in different initial bacterial densities is performed. The results demonstrate that higher initial loading densities of the bacterial colony cause bacterial cell to enter log-phase proliferation sooner. High or low initial loading densities of the bacterial cell suspension induce the same maximum growth rates during the log-phase. Quantitative on-chip analysis of tetracycline and erythromycin inhibition on bacterial cell growth is also conducted. Bacterial morphology changes during antibiotic treatment are observed. The results show that tetracycline and erythromycin exhibit different inhibition activities in E. coli cells. Concentrations of 3 μg/mL tetracycline can facilitate the formation of long filamentous bacteria with the average length of more than 50 μm. This study provides an on-chip framework for bacteriological research in a high-throughput manner and the development of recombinant bacteria-based biosensors for the detection of specific substances.
authors.affiliation: College of Science, Northwest A&F University, Yangling, Shaanxi 712100, PR China.
authors.forename: Peng
authors.initials: P
authors.lastname: Sun
authors.forename: Yang
authors.initials: Y
authors.lastname: Liu
authors.forename: Jun
authors.initials: J
authors.lastname: Sha
authors.forename: Zhiyun
authors.initials: Z
authors.lastname: Zhang
authors.forename: Qin
authors.initials: Q
authors.lastname: Tu
authors.forename: Peng
authors.initials: P
authors.lastname: Chen
authors.forename: Jinyi
authors.initials: J
authors.lastname: Wang
mesh_terms: D000900:Anti-Bacterial Agents / Q000008:administration & dosage*; D001681:Biological Assay / Q000295:instrumentation*; D019149:Bioreactors / Q000382:microbiology; D015374:Biosensing Techniques / Q000295:instrumentation; D002470:Cell Survival / Q000187:drug effects; D015169:Colony Count, Microbial / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000166:cytology* / Q000187:drug effects*; D005434:Flow Cytometry / Q000295:instrumentation*; D017022:Flow Injection Analysis / Q000295:instrumentation; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008991:Monitoring, Physiologic / Q000295:instrumentation
chemical_list: D000900:Anti-Bacterial Agents