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Rapid Characterization of Bacterial Electrogenicity Using a Single-Sheet Paper-Based Electrofluidic Array

机译:使用单片纸基电液阵列快速表征细菌的电原性

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摘要

Electrogenicity, or bacterial electron transfer capacity, is an important application which offers environmentally sustainable advances in the fields of biofuels, wastewater treatment, bioremediation, desalination, and biosensing. Significant boosts in this technology can be achieved with the growth of synthetic biology that manipulates microbial electron transfer pathways, thereby potentially significantly improving their electrogenic potential. There is currently a need for a high-throughput, rapid, and highly sensitive test array to evaluate the electrogenic properties of newly discovered and/or genetically engineered bacterial species. In this work, we report a single-sheet, paper-based electrofluidic (incorporating both electronic and fluidic structure) screening platform for rapid, sensitive, and potentially high-throughput characterization of bacterial electrogenicity. This novel screening array uses (i) a commercially available wax printer for hydrophobic wax patterning on a single sheet of paper and (ii) water-dispersed electrically conducting polymer mixture, poly(3,4-ethylenedioxythiophene):polystyrene sulfonate, for full integration of electronic and fluidic components into the paper substrate. The engineered 3-D, microporous, hydrophilic, and conductive paper structure provides a large surface area for efficient electron transfer. This results in rapid and sensitive power assessment of electrogenic bacteria from a microliter sample volume. We validated the effectiveness of the sensor array using hypothesis-driven genetically modified Pseudomonas aeruginosa mutant strains. Within 20 min, we observed that the sensor platform successfully measured the electricity-generating capacities of five isogenic mutants of P. aeruginosa while distinguishing their differences from genetically unmodified bacteria.
机译:电性或细菌电子转移能力是重要的应用,它在生物燃料,废水处理,生物修复,淡化和生物传感领域提供了环境可持续的进步。随着操纵微生物电子转移途径的合成生物学的发展,可以显着提高这项技术,从而潜在地显着提高其电化潜力。当前需要高通量,快速和高度灵敏的测试阵列,以评估新发现和/或基因工程的细菌物种的电学性质。在这项工作中,我们报告了一种单页纸基电流体(兼具电子和流体结构)筛选平台,可快速,灵敏和潜在地高通量表征细菌的电原性。这种新颖的筛选阵列使用(i)商业蜡印机在单张纸上进行疏水性蜡构图,以及(ii)水分散的导电聚合物混合物,聚(3,4-乙撑二氧噻吩):聚苯乙烯磺酸盐,以实现完全集成电子和流体成分进入纸质基材。工程化的3-D,微孔,亲水和导电纸结构为有效的电子转移提供了较大的表面积。这样可以从微升样品量中快速,灵敏地评估电原细菌的功率。我们验证了使用假设驱动的基因改造的铜绿假单胞菌突变株传感器阵列的有效性。在20分钟内,我们观察到传感器平台成功测量了铜绿假单胞菌的五个同基因突变体的发电能力,同时将它们与未经遗传修饰的细菌区分开来。

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