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Enhancing the Resolution of Micro Free Flow Electrophoresis through Spatially Controlled Sample Injection
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-06-25 00:00:00 , DOI: 10.1021/acs.analchem.8b01205 Kadi L. Saar 1 , Thomas Müller 2 , Jérôme Charmet 3 , Pavan Kumar Challa 1 , Tuomas P. J. Knowles 1, 4
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-06-25 00:00:00 , DOI: 10.1021/acs.analchem.8b01205 Kadi L. Saar 1 , Thomas Müller 2 , Jérôme Charmet 3 , Pavan Kumar Challa 1 , Tuomas P. J. Knowles 1, 4
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Free flow electrophoresis is a versatile technique for the continuous separation of mixtures with both preparative and analytical applications. Microscale versions of free flow electrophoresis are particularly attractive strategies because of their fast separation times, ability to work with small sample volumes, and large surface area to volume ratios facilitating rapid heat transfer, thus minimizing the detrimental effects of Joule heating even at high voltages. The resolution of microscale free flow electrophoresis, however, is limited by the broadening of the analyte beam in the microfluidic channel, an effect that becomes especially pronounced when the analyte is deflected significantly away from its original position. Here, we describe and demonstrate how restricting spatially the sample injection and collection to the regions where the gradients in the velocity distribution of the carrier medium are the smallest allows this broadening effect to be substantially suppressed and hence the resolution of microscale free flow electrophoresis devices to be increased. To demonstrate this concept, we fabricated microfluidic free flow electrophoresis devices with spatially restricted injection nozzles implemented through the use of multilayer soft-photolithography and further integrated quartz based observation areas for fluorescent detection and imaging. With these devices, we demonstrated a 5-fold reduction in the extent of beam broadening relative to conventional free flow electrophoresis approaches with nonrestricted sample introduction. The manifold enhancement in the achievable resolution of microscale free flow electrophoresis devices opens up the possibility of rapid separation and analysis of complex mixtures.
中文翻译：
通过空间控制的样品注入提高微自由流电泳的分辨率
自由流动电泳是一种用于制备和分析应用中连续分离混合物的通用技术。微型版本的自由流电泳技术特别有吸引力，因为它们具有快速的分离时间，处理小样品量的能力以及大的表面积与体积之比，从而促进了快速的热传递，因此即使在高电压下，也可以将焦耳加热的不利影响降至最低。然而，微尺度自由流电泳的分辨率受到微流体通道中分析物束的展宽的限制，当分析物明显偏离其原始位置时，这种作用尤为明显。这里，我们描述并证明了如何在空间上将样品的注入和收集限制在载体介质速度分布的梯度最小的区域，从而可以基本上抑制这种加宽效应，从而提高微尺度自由流电泳设备的分辨率。为了证明这一概念，我们制造了具有有限空间的喷嘴的微流体自由流动电泳装置，该装置通过使用多层软光刻技术和进一步集成的石英基观察区进行荧光检测和成像。使用这些设备，我们证明了与不加限制样品导入的常规自由流电泳方法相比，束增宽程度降低了5倍。
更新日期：2018-06-25
中文翻译：
通过空间控制的样品注入提高微自由流电泳的分辨率
自由流动电泳是一种用于制备和分析应用中连续分离混合物的通用技术。微型版本的自由流电泳技术特别有吸引力，因为它们具有快速的分离时间，处理小样品量的能力以及大的表面积与体积之比，从而促进了快速的热传递，因此即使在高电压下，也可以将焦耳加热的不利影响降至最低。然而，微尺度自由流电泳的分辨率受到微流体通道中分析物束的展宽的限制，当分析物明显偏离其原始位置时，这种作用尤为明显。这里，我们描述并证明了如何在空间上将样品的注入和收集限制在载体介质速度分布的梯度最小的区域，从而可以基本上抑制这种加宽效应，从而提高微尺度自由流电泳设备的分辨率。为了证明这一概念，我们制造了具有有限空间的喷嘴的微流体自由流动电泳装置，该装置通过使用多层软光刻技术和进一步集成的石英基观察区进行荧光检测和成像。使用这些设备，我们证明了与不加限制样品导入的常规自由流电泳方法相比，束增宽程度降低了5倍。
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