Cellular enrichment through microfluidic fractionation based on cell biomechanical properties.
Cellular enrichment through microfluidic fractionation based on cell biomechanical properties.
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DOI：
10.1007/s10404-015-1608-y
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发表时间：
2015-10
影响因子：
2.8
通讯作者：
Sulchek T
中科院分区：
文献类型：
--
作者：
Wang G;Turbyfield C;Crawford K;Alexeev A;Sulchek T
患病细胞群的生物力学特性显示出与健康细胞群的差异，然而这些生物力学特性的重叠限制了它们在疾病细胞富集和检测中的应用。我们报道了一种新的微流体细胞富集技术，该技术通过生物力学特性的差异连续分离细胞，从而产生高度纯净的细胞亚群。细胞分离是在微流体通道中实现的，微流体通道具有一系列对角脊，旨在将生物力学上不同的细胞分离到通道中的不同位置。由于在细胞压缩过程中施加弹性和粘性力，这是细胞生物力学特性（包括尺寸和粘弹性）的功能，更大、更硬和更少粘性的细胞平行于对角脊移动，并表现出正向的横向位移。另一方面，更小、更软、更粘的细胞由于脊引起的循环流动而垂直于对角脊移动，导致负侧向位移。然后利用多个出口来收集细胞，这些细胞具有更细的细胞生物力学特性差异。结果是，与二元输出相比，细胞分离显著提高了细胞分离效率，并能够测量单个细胞类型内细微的生物力学差异。作为概念验证演示，我们混合了两种不同的白血病细胞系（K562和HL60），并利用细胞分离实现了超过45倍的细胞群增强，每种细胞系的高纯度细胞富集（90%至99%）。此外，我们展示了单个细胞类型（K562细胞）的细胞分离成亚群，并用原子力显微镜表征分离细胞的生物力学特性的变化。这些结果将有利于获得细胞混合物的无标记分离，或更好地研究单个细胞类型的生物力学差异的起源。
The biomechanical properties of populations of diseased cells are shown to have differences from healthy populations of cells, yet the overlap of these biomechanical properties can limit their use in disease cell enrichment and detection. We report a new microfluidic cell enrichment technology that continuously fractionates cells through differences in biomechanical properties, resulting in highly pure cellular subpopulations. Cell fractionation is achieved in a microfluidic channel with an array of diagonal ridges that are designed to segregate biomechanically distinct cells to different locations in the channel. Due to the imposition of elastic and viscous forces during cellular compression, which are a function of cell biomechanical properties including size and viscoelasticity, larger, stiffer and less viscos cells migrate parallel to the diagonal ridges and exhibit positive lateral displacement. On the other hand, smaller, softer and more viscous cells migrate perpendicular to the diagonal ridges due to circulatory flow induced by the ridges and result in negative lateral displacement. Multiple outlets are then utilized to collect cells with finer gradation of differences in cell biomechanical properties. The result is that cell fractionation dramatically improves cell separation efficiency compared to binary outputs and enables the measurement of subtle biomechanical differences within a single cell type. As a proof-of-concept demonstration, we mix two different leukemia cell lines (K562 and HL60) and utilize cell fractionation to achieve over 45-fold enhancement of cell populations, with high purity cellular enrichment (90% to 99%) of each cell line. In addition, we demonstrate cell fractionation of a single cell type (K562 cells) into subpopulations and characterize the variations of biomechanical properties of the separated cells with atomic force microscopy. These results will be beneficial to obtaining label-free separation of cellular mixtures, or to better investigate the origins of biomechanical differences in a single cell type.