在片上实验室评估脉冲介电泳-场流分馏分离和光纤检测技术,作为一种分离相似细菌细胞的技术

IF 2.5 4区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Mohd Firdaus Kamuri, Zurina Zainal Abidin, Mohd Hanif Yaacob, Mohd Nizar Hamidon
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引用次数: 0

摘要

这项研究解决了分离大肠杆菌和嗜水气单胞菌等结构相似细菌的难题。这种方法采用脉冲场介电泳,并在集成光学检测功能的片上实验室系统中以层流分馏法辅助。在脉冲模式下,细菌细胞通过 30 微米的微电极,微电极的频率为 1 兆赫,峰-峰值电压为 14 伏特,同时流体流动将细菌带向腔室的末端。特定脉冲间隔的通断电场使细菌细胞受到各种力的作用,包括动力学力、介电泳力、重力、阻力和扩散力,从而产生促进其运动的净力。对脉冲时间、流速和电压的变化进行了研究,以确定高效分离的最佳组合。接下来,使用光纤根据细菌的吸光度对其进行检测。结果表明,在流速为 9.6 μL min-1、脉冲时间为 4 秒、介质电导率为 40 μS cm-1 的条件下,90 分钟内的分离效率为 30%。与大肠杆菌相比,在施加电场的过程中,纤毛虫聚集体受到了更大的去污力,并保留在微电极上,而大肠杆菌则更快地向光学检测方向移动。有电场和无电场时的分离机制是不同的,因此在电场关闭期间精确控制细胞的移动对减少不受控制的扩散非常重要。虽然光学检测部分取得了成功,但为了更好地进行分离,建议延长时间和分离长度。精心调整脉冲时间、流速、电压和微电极设计的组合对于这个集成片上实验室系统高效分离和检测密切相关的微生物至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Assessment of pulsed dielectrophoretic-field flow fractionation separation coupled with fibre-optic detection on a lab-on-chip as a technique to separate similar bacteria cells

Assessment of pulsed dielectrophoretic-field flow fractionation separation coupled with fibre-optic detection on a lab-on-chip as a technique to separate similar bacteria cells

This study addresses the challenge of separating bacteria with similar structures such as Escherichia coli and Aeromonas hydrophila. This approach employs pulsed field dielectrophoresis assisted by laminar flow fractionation in a lab-on-a-chip system with integrated optical detection. Bacterial cells passed through 30-µm microelectrodes subjected at 1 MHz and 14 V peak-to-peak in pulsed mode, while fluid flow carried bacteria towards the chamber’s end. The on-and-off electric field at specific pulse intervals expose bacterial cells to diverse forces, including kinetics, dielectrophoresis, gravity, drag, and diffusion, resulting in a net force facilitating their movement. Variations of pulsing time, flow rates, and voltage were investigated to identify the optimal combination for efficient separation. Next, the bacteria were detected using an optical fibre based on their absorbance. Results demonstrated a 30% separation efficiency in 90 min at 9.6 μL min−1 flow rates, 4 s pulsing time, and 40 μS cm−1 medium conductivity. A. hydrophila aggregates experienced greater DEP force and retained at microelectrodes during electric field application compared to E. coli, which moved faster towards optical detection. The separation mechanism with and without electric field was different, and precise control of cell movement during field-off periods is important to minimize uncontrolled diffusion. While the optical detection part has been successful, longer time and separation length are recommended for better separation. A carefully tuned combination of pulsing time, flow rates, voltage, and microelectrode design is crucial for this integrated lab-on-chip system to be efficient for separating and detecting closely related microorganisms.

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来源期刊
Biotechnology and Bioprocess Engineering
Biotechnology and Bioprocess Engineering 工程技术-生物工程与应用微生物
CiteScore
5.00
自引率
12.50%
发文量
79
审稿时长
3 months
期刊介绍: Biotechnology and Bioprocess Engineering is an international bimonthly journal published by the Korean Society for Biotechnology and Bioengineering. BBE is devoted to the advancement in science and technology in the wide area of biotechnology, bioengineering, and (bio)medical engineering. This includes but is not limited to applied molecular and cell biology, engineered biocatalysis and biotransformation, metabolic engineering and systems biology, bioseparation and bioprocess engineering, cell culture technology, environmental and food biotechnology, pharmaceutics and biopharmaceutics, biomaterials engineering, nanobiotechnology, and biosensor and bioelectronics.
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