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Quantitative chemical biosensing by bacterial chemotaxis in microfluidic chips: Chemotaxis-microfluidic biosensor

机译:在微流控芯片中通过细菌趋化性进行定量化学生物传感:趋化性-微流控生物传感器

摘要

Whole-cell bacterial bioreporters are proposed as alternatives to chemical analysis of, for example, pollutants in environmental compartments. Commonly based on reporter gene induction, bioreporters produce a detectable signal within 30 min to a few hours after exposure to the chemical target, which is impractical for applications aiming at a fast response. In an attempt to attain faster readout but maintain flexibility of chemical targeting, we explored the concept for quantitative chemical sensing by bacterial chemotaxis. Chemotaxis was quantified from enrichment of cells across a 600 µm- wide chemical gradient stabilized by parallel flow in a microfluidic chip, further supported by transport and chemotaxis steady state and kinetic modelling. As proof- of-concept, we quantified Escherichia coli chemotaxis towards serine, aspartate and methylaspartate as a function of attractant concentration and exposure time. E. coli chemotaxis enrichment increased sharply between 0 and 10 µM serine, before saturating at 100 µM. The chemotaxis accumulation rate was maximal at 10 µM serine, leading to observable cell enrichment within 5 min. The potential application for biosensing of environmental toxicants was investigated by quantifying chemotaxis of Cupriavidus pinatubonensis JMP134 towards the herbicide 2,4- dichlorophenoxyacetate. Our results show that bacterial chemotaxis can be quantified on a scale of minutes and may be used for developing faster bioreporter assays.
机译:提出了全细胞细菌生物报告者作为化学分析的替代方法,例如对环境隔室中的污染物进行化学分析。通常,基于报告基因的诱导,生物报告者会在暴露于化学靶标后30分钟到几小时内产生可检测的信号,这对于旨在快速响应的应用是不切实际的。为了获得更快的读数但保持化学靶向的灵活性,我们探索了通过细菌趋化性进行定量化学传感的概念。通过在微流控芯片中通过平行流动稳定的600 µm宽化学梯度中的细胞富集,对趋化作用进行定量,并进一步得到运输和趋化作用的稳态及动力学建模的支持。作为概念验证,我们定量分析了大肠杆菌对丝氨酸,天冬氨酸和天冬氨酸甲酯的趋化性与引诱剂浓度和暴露时间的关系。在100 µM饱和之前,大肠杆菌的趋化性富集在0至10 µM丝氨酸之间急剧增加。趋化性累积速率在10 µM丝氨酸最大,导致5分钟内可观察到细胞富集。通过定量测定Cupriavidus pinatubonensis JMP134对除草剂2,4-二氯苯氧乙酸酯的趋化性,研究了在生物传感环境有毒物质方面的潜在应用。我们的结果表明,细菌趋化性可以在几分钟内量化,并且可以用于开发更快的生物报告试剂。

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