Bidulock Allison C E, van den Berg Albert, Eijkel Jan C T
BIOS/The Lab-on-a-Chip Group, MESA + Institute for Nanotechnology and MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente Twente, The Netherlands.
Electrophoresis. 2015 Mar;36(6):875-83. doi: 10.1002/elps.201400399. Epub 2015 Feb 20.
To realize portable systems for routine measurements in point-of-care settings, MCE methods are required to be robust across many single-use chips. While it is well-known internal standards (ISTDs) improve run-to-run precision, a systematic investigation is necessary to determine the significance of chip-to-chip imprecision in MCE and how ISTDs account for it. This paper addresses this question by exploring the reproducibility of Na quantification across six basic, in-house fabricated microchips. A dataset of 900 electrophoerograms was collected from analyzing five concentrations of NaCl with two ISTDs (CsCl and LiCl). While both improved the peak area reproducibility, the Na/Cs ratio was superior to the Na/Li ratio (improving the RSD by a factor of 2-4, depending on the Na concentration). We attribute this to the significant variation in microchannel surface properties, which was accounted for by cesium but not lithium. Microchip dimension and detector variations were only a few percent, and could be improved through commercial fabrication over in-house made microchips. These results demonstrate that ISTDs not only correct for intrachip imprecision, but are also a viable means to correct for chip-to-chip imprecision inherent in disposable, point-of-care MCE devices. However, as expected, the internal standard must be carefully chosen.
为实现用于即时护理环境中常规测量的便携式系统,需要微芯片电泳(MCE)方法在许多一次性芯片上具有稳健性。虽然众所周知内标(ISTD)可提高批间精度,但有必要进行系统研究,以确定MCE中芯片间不精确性的重要性以及内标如何对此进行解释。本文通过探索六种自制的基本微芯片上钠定量的重现性来解决这个问题。通过分析五种浓度的氯化钠和两种内标(氯化铯和氯化锂)收集了900个电泳图谱的数据集。虽然两者都提高了峰面积重现性,但钠/铯比优于钠/锂比(根据钠浓度,相对标准偏差(RSD)提高了2至4倍)。我们将此归因于微通道表面性质的显著变化,铯可以解释这种变化,而锂则不能。微芯片尺寸和检测器的变化仅为百分之几,并且通过商业制造而非自制微芯片可以得到改善。这些结果表明,内标不仅可以校正芯片内的不精确性,而且还是校正一次性即时护理MCE设备中固有的芯片间不精确性的可行方法。然而,正如预期的那样,必须谨慎选择内标。