Development of microscopic time‐domain dual lifetime referencing luminescence detection for pH monitoring in microfluidic free‐flow isoelectric focusing

Development of microscopic time‐domain dual lifetime referencing luminescence detection for pH monitoring in microfluidic free‐flow isoelectric focusing
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开发用于微流体自由流等电聚焦中 pH 监测的微观时域双寿命参考发光检测

DOI:
10.1002/elsc.201400081
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发表时间:
2015
影响因子:
2.7
通讯作者:
S. Nagl
S. Nagl
中科院分区:
工程技术3区
文献类型:
--
作者:
E. Poehler;C. Herzog;M. Suendermann;S.A. Pfeiffer;S. Nagl

文献摘要

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基于时间域双寿命参考(t-DLR)原理,研制了一种基于寿命的比率式微尺度pH传感器系统和荧光显微镜装置,用于在线观测自由流动等电聚焦过程中的pH。T-DLR方法已被开发用于pH监测和环境监测和生物技术领域的各种其他应用。在这里,我们介绍了微流控通道中t-DLR的pH传感器微结构的集成,以及t-DLR方案在小型化电泳过程中的应用。该pH传感器被喷墨打印在玻璃上,长度为10 mm,高度为404±18 nm,宽度为371±28μm,并集成到由激光切割和层压技术产生的微流控芯片中。其工作范围为pH 4~8,pKa值为6.10±0.01,快速响应时间小于500ms。该传感器用于在线观察β-乳球蛋白A、刀白蛋白和肌红蛋白等电聚焦过程中的pH梯度,并通过等电点对它们进行识别。所得等电点与文献数据吻合较好,证明了该pH传感器在微流控连续分离中的适用性。
A lifetime‐based ratiometric microscale pH sensor system and a fluorescence microscopic setup was developed for the in‐line observation of pH in free‐flow isoelectric focusing based on the principle of time‐domain dual lifetime referencing (t‐DLR). The t‐DLR method has been developed for pH monitoring and various other applications in the fields of environmental monitoring and biotechnology. Here, we introduce the integration of pH sensor microstructures for t‐DLR in microfluidic channels and the application of the t‐DLR scheme for pH sensing in miniaturized electrophoretic procedures. The pH sensor was inkjet‐printed on glass in rows with a length of 10 mm, a height of 404 ± 18 nm, and a width of 371 ± 28 μm and integrated into a microfluidic chip generated by a laser cutting and lamination technique. It had a working range from pH 4 to 8 with a pKaof 6.10 ± 0.01 and fast response times under 500 ms. The sensor was used for the in‐line observation of the pH gradient during isoelectric focusing of the proteins β‐lactoglobulin A, conalbumin, and myoglobin and their identification by their pI. The obtained pIs were in good agreement with literature data demonstrating the applicability of the pH sensor in microfluidic continuous separations.
具有集成微点阵列的微流体排列,用于表征 pH 值和溶剂极性
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期刊:
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