Institute of Microanalytical Systems, Department of Chemistry and Innovation Center for Cell Signaling Network, Zhejiang University, Hangzhou, 310058, China.
Department of Cell Biology, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang, 110001, China.
Sci Rep. 2018 Jan 29;8(1):1791. doi: 10.1038/s41598-018-20058-0.
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.
在这项工作中,我们开发了一种集成了所有模块的微型手掌高速毛细管电泳(CE)系统,包括皮升级别的样品进样、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。我们采用了一种最小化的微型化策略,结合最小系统设计和低成本系统构建,实现了仪器的微型化,成本极低,这与当前大多数报道的微型 CE 系统中使用的微加工策略不同。采用这种策略,生物分析仪的总尺寸最小化到 90×75×77mm(长×宽×高),仪器成本降低到约 500 美元,这表明该系统是迄今为止报道的具有 LIF 检测的最小和最低成本的 CE 仪器。本生物分析仪还表现出与以前报道的高速 CE 系统相当的分析性能。获得了 1.02nM 荧光素钠的检测限。该仪器能够高效快速分离多种类型的样品,如氨基酸、氨基酸对映体、DNA 片段和蛋白质。我们应用该仪器通过聚合酶链反应-限制性片段长度多态性(PCR-RFLP)方法检测结直肠癌中的 KRAS 突变状态。