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Microscreening toxicity system based on living magnetic yeast and gradient chips

机译:基于活酵母和梯度芯片的微筛毒系统

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摘要

There is an increasing demand for easy and cost-effective methods to screen the toxicological impact of the growing number of chemical mixtures being generated by industry. Such a screening method has been developed using viable, genetically modified green fluorescent protein (GFP) reporter yeast that was magnetically functionalised and held within a microfluidic device. The GFP reporter yeast was used to detect genotoxicity by monitoring the exposure of the cells to a well-known genotoxic chemical (methyl methane sulfonate, MMS). The cells were magnetised using biocompatible positively charged PAH-stabilised magnetic nanoparticles with diameters around 15 nm. Gradient mixing was utilised to simultaneously expose yeast to a range of concentrations of toxins, and the effective fluorescence emitted from the produced GFP was measured. The magnetically enhanced retention of the yeast cells, with their facile subsequent removal and reloading, allowed for very convenient and rapid toxicity screening of a wide range of chemicals. This is the first report showing magnetic yeast within microfluidic devices in a simple bioassay, with potential applications to other types of fluorescent reporter yeast in toxicological and biomedical research. The microfluidic chip offers a simple and low-cost screening test that can be automated to allow multiple uses (adapted to different cell types) of the device on a wide range of chemicals and concentrations.
机译:人们越来越需要一种简便且具有成本效益的方法来筛选工业产生的化学混合物数量不断增加的毒理学影响。已经使用磁性功能化并保持在微流体装置中的可行的,遗传修饰的绿色荧光蛋白(GFP)报告酵母开发了这种筛选方法。 GFP记者酵母菌通过监测细胞暴露于众所周知的遗传毒性化学物质(甲磺酸甲酯,MMS)来检测遗传毒性。使用直径约15 nm的生物相容的带正电荷的PAH稳定的磁性纳米粒子对细胞进行磁化。利用梯度混合使酵母同时暴露于一定范围的毒素浓度,并测量了从产生的GFP发出的有效荧光。酵母细胞的磁性增强保留,以及随后轻松的去除和重新装载,使得对各种化学品进行非常方便,快速的毒性筛选成为可能。这是第一个报告,显示了通过简单的生物测定法在微流控装置中的磁性酵母,并在毒理学和生物医学研究中可能应用于其他类型的荧光报告酵母。微流控芯片提供了一种简单且低成本的筛选测试,该测试可以自动化,以允许该设备在多种化学物质和浓度范围内多次使用(适应不同的细胞类型)。

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