Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas 79409, USA.
Anal Chem. 2011 Feb 1;83(3):774-81. doi: 10.1021/ac102975g. Epub 2011 Jan 5.
Cell separation based on microfluidic affinity chromatography is a widely used methodology in cell analysis research when rapid separations with high purity are needed. Several successful examples have been reported with high separation efficiency and purity; however, cell capture at the inlet area and inlet design have not been extensively described or studied. The most common inlets-used to connect the microfluidic chip to pumps, tubing, etc.-are vertical (top-loading) inlets and parallel (in-line) inlets. In this work, we investigated the cell capture behavior near the affinity chip inlet area and compared the different performances of vertical inlet devices and parallel inlet devices. Vertical inlet devices showed significant cell capture capability near the inlet area, which led to the formation of cell blockages as the separation progressed. Cell density near the inlet area was much higher than that in the remaining channel, whereas for parallel inlet chips cell density at the inlet area was similar to that in the rest of the channel. In this paper, we discuss the effects of inlet type on chip fabrication, nonspecific binding, cell capture efficiency, and separation purity. We also discuss the possibility of using vertical inlets in negative-selection separations. Our findings show that inlet design is critical and must be considered when fabricating cell affinity microfluidic devices.
基于微流控亲和层析的细胞分离是细胞分析研究中广泛使用的方法，当需要快速、高纯度分离时尤其如此。已经有许多成功的例子报道了这种方法具有高分离效率和纯度；然而，细胞在入口区域的捕获和入口设计并没有得到广泛的描述或研究。最常用的入口——用于将微流控芯片与泵、管道等连接——是垂直（顶部加载）入口和平行（在线）入口。在这项工作中，我们研究了亲和芯片入口区域附近的细胞捕获行为，并比较了垂直入口装置和平行入口装置的不同性能。垂直入口装置在入口区域附近显示出显著的细胞捕获能力，这导致随着分离的进行形成细胞堵塞。入口区域附近的细胞密度远高于其余通道中的细胞密度，而对于平行入口芯片，入口区域的细胞密度与通道其余部分的细胞密度相似。在本文中，我们讨论了入口类型对芯片制造、非特异性结合、细胞捕获效率和分离纯度的影响。我们还讨论了在负选择分离中使用垂直入口的可能性。我们的研究结果表明，入口设计至关重要，在制造细胞亲和微流控器件时必须考虑。
