Single-cell bacteria growth monitoring by automated DEP-facilitated image analysis
Single-cell bacteria growth monitoring by automated DEP-facilitated image analysis
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DOI：
10.1039/c004691d
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
van Leeuwen, Rien
中科院分区：
文献类型：
--
作者：
Peitz, Ingmar;van Leeuwen, Rien
生长监测是许多测定病原体的存在或性质的选择方法，例如。G.在诊断和食品质量方面。依赖于培养大量细菌的既定方法相当耗时，而在医疗保健中，时间往往至关重要。已经发表了几种新的方法，主要是针对测定少数细菌的生长或其他特性。然而，到目前为止，还没有一种方法可以方便且容易地实现单细胞分辨率，从而提供自动化和高通量的可能性。在这项研究中，我们通过采用微流体电极结构中的细菌介电电泳捕获，光学检测和自动细菌识别以及图像分析算法计数来证明这些好处。对于原理验证实验，我们选择了大肠杆菌和多粘菌素B的抗生素敏感性试验。在单细胞上证明了生长监测，并显示了抗生素对生长速率的影响。作为标准诊断参数的最小抑制浓度来自剂量-反应曲线。该报告是进一步将图像分析代码集成到设备控制中的基础。最终，可以同时使用光学微扫描仪和许多电极结构来创建自动化和并行化的设置。然后，在一个单一的实验中就可以产生足够的数据来进行合理的统计评估和确认初步发现。
Growth monitoring is the method of choice in many assays measuring the presence or properties of pathogens, e. g. in diagnostics and food quality. Established methods, relying on culturing large numbers of bacteria, are rather time-consuming, while in healthcare time often is crucial. Several new approaches have been published, mostly aiming at assaying growth or other properties of a small number of bacteria. However, no method so far readily achieves single-cell resolution with a convenient and easy to handle setup that offers the possibility for automation and high throughput. We demonstrate these benefits in this study by employing dielectrophoretic capturing of bacteria in microfluidic electrode structures, optical detection and automated bacteria identification and counting with image analysis algorithms. For a proof-of-principle experiment we chose an antibiotic susceptibility test with Escherichia coli and polymyxin B. Growth monitoring is demonstrated on single cells and the impact of the antibiotic on the growth rate is shown. The minimum inhibitory concentration as a standard diagnostic parameter is derived from a dose-response plot. This report is the basis for further integration of image analysis code into device control. Ultimately, an automated and parallelized setup may be created, using an optical microscanner and many of the electrode structures simultaneously. Sufficient data for a sound statistical evaluation and a confirmation of the initial findings can then be generated in a single experiment.