Department of Biomedical Engineering, University of Wisconsin Madison, Madison, Wisconsin, USA.
Anal Chem. 2010 Oct 1;82(19):8320-6. doi: 10.1021/ac101866p.
Cell concentration via centrifugation is a ubiquitous step in many cell culture procedures. At the macroscale, centrifugation suffers from a number of limitations, particularly when dealing with small numbers of cells (e.g., less than 50,000). On the other hand, typical microscale methods for cell concentration can affect cell physiology and bias readouts of cell behavior and function. In this paper, we present a microfluidic concentrator device that utilizes the effects of gravity to allow cells to gently settle out of a suspension into a collection region without the use of specific adhesion ligands. Dimensional analysis was performed to compare different device designs and was verified with flow modeling to optimize operational parameters. We are able to concentrate low-density cell suspensions in a microfluidic chamber, achieving a cell loss of only 1.1 ± 0.6% (SD, n = 7) with no observed loss during a subsequent cell staining protocol which incorporates ∼36 complete device volume replacements. This method provides a much needed interface between rare cell samples and microfluidic culture assays.
通过离心进行细胞浓度分离是许多细胞培养过程中的常见步骤。在宏观层面上，离心存在许多限制，尤其是在处理少量细胞（例如，少于 50000 个）时。另一方面，典型的微尺度细胞浓缩方法可能会影响细胞生理学，并影响细胞行为和功能的读出结果。在本文中，我们提出了一种微流控浓缩器装置，该装置利用重力的作用，使细胞能够从悬浮液中缓慢沉降到收集区域，而无需使用特定的粘附配体。通过尺寸分析比较了不同的器件设计，并通过流动建模进行了验证，以优化操作参数。我们能够在微流控腔室中浓缩低密度细胞悬浮液，仅损失 1.1±0.6%（SD，n=7）的细胞，并且在随后的细胞染色方案中没有观察到损失，该方案包含约 36 次完整的器件体积替换。该方法为稀有细胞样本与微流控培养分析之间提供了一个非常需要的接口。
