Abstract
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.
Highlights
The total size of a high-speed capillary electrophoresis (CE) bioanalyzer could be minimized to 90 × 75 × 77 mm (Fig. 1) and the instrument cost could be reduced to ca. $500, which integrated multiple modules including picoliter-scale translational spontaneous sample injection, short capillary CE, high-voltage power supply, orthogonal laser induced fluorescence (LIF) detection (Fig. 2), battery, system control, on-line data acquisition, processing, storage, and display modules (Fig. 1)
The performance of the instrument was demonstrated in multi-mode separations for amino acids, DNA fragments, amino acid enantiomers and proteins with fast speed and high efficiency, as well as in analysis of PCR products and restriction fragment length polymorphism (RFLP) digestion products of KRAS proto-oncogene for actual colorectal cancer diagnosis
To our knowledge, it is the smallest high-speed CE apparatus with LIF detection reported in the literatures
Summary
In 2003, Jackson et al.[23] developed a miniaturized CE instrument based on amperometric detection It integrated a glass CE chip, battery, dual-source high-voltage power supply, interface circuit and modules, with a total size of ca. The device integrated glass microchip, high-voltage power supply, confocal LIF detector with 532/633 nm laser and photomultiplier (PMT), and electronic control, processing and display modules with a total size of approximately 230 × 200 × 130 mm. We developed a compact high-speed CE bioanalyzer integrated modules of sample injection and changing, short capillary-based CE separation, LIF detection, as well as a custom designed tablet computer for data processing, instrument controlling, and result displaying with a desktop instrument size of 230 × 170 × 190 mm[33]. The performance of the instrument was demonstrated in multi-mode separations for amino acids, DNA fragments, amino acid enantiomers and proteins with fast speed and high efficiency, as well as in analysis of PCR products and restriction fragment length polymorphism (RFLP) digestion products of KRAS proto-oncogene for actual colorectal cancer diagnosis
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