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A Low-Cost Palmtop High-Speed Capillary Electrophoresis Bioanalyzer with Laser Induced Fluorescence Detection

机译:具有激光诱导荧光检测的低成本掌上型高速毛细管电泳生物分析仪

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摘要

In this work, we developed a miniaturized palmtop high-speed capillary electrophoresis (CE) system integrating whole modules, including picoliter-scale sample injection, short capillary-based fast CE, high-voltage power supply, orthogonal laser induced fluorescence (LIF) detection, battery, system control, on-line data acquisition, processing, storage, and display modules. A strategy of minimalist miniaturization combining minimal system design and low-cost system construction was adopted to achieve the instrument miniaturization with extremely low cost, which is differing from the current microfabrication strategy used in most reported miniaturized CE systems. With such a strategy, the total size of the bioanalyzer was minimized to 90 × 75 × 77 mm (length × width × height) and the instrument cost was reduced to ca. $500, which demonstrated the smallest and lowest-cost CE instrument with LIF detection in so far reported systems. The present bioanalyzer also exhibited comparable analytical performances to previously-reported high-speed CE systems. A limit of detection of 1.02 nM sodium fluorescein was obtained. Fast separations were achieved for multiple types of samples as amino acids, amino acid enantiomers, DNA fragments, and proteins with high efficiency. We applied this instrument in colorectal cancer diagnosis for detecting KRAS mutation status by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method.
机译:在这项工作中,我们开发了一个集成了整个模块的微型掌上高速毛细管电泳(CE)系统,包括皮升级样品注射,基于短毛细管的快速CE,高压电源,正交激光诱导荧光(LIF)检测,电池,系统控制,在线数据采集,处理,存储和显示模块。采用最小化微型化的策略,将最小化的系统设计和低成本的系统构造相结合,以极低的成本实现了仪器的微型化,这与目前在大多数报道的微型CE系统中使用的微细加工策略不同。通过这种策略,生物分析仪的总尺寸被最小化到90××75××77×mm(长×宽×高),并且仪器成本降低到了约50毫米。售价$ 500美元,展示了迄今为止报道的系统中最小且成本最低的带有LIF检测功能的CE仪器。本生物分析仪还表现出与先前报道的高速CE系统相当的分析性能。检出限为1.02 nM荧光素钠。快速分离多种类型的样品,例如氨基酸,氨基酸对映体,DNA片段和蛋白质。我们将该仪器应用于大肠癌的诊断,通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测KRAS突变状态。

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