Deformability based cell margination-A simple microfluidic design for malaria-infected erythrocyte separation
Deformability based cell margination-A simple microfluidic design for malaria-infected erythrocyte separation
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DOI：
10.1039/c003873c
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发表时间：
2010-01-01
期刊：
影响因子：
6.1
通讯作者：
Lim, Chwee Teck
中科院分区：
文献类型：
--
作者：
Hou, Han Wei;Bhagat, Ali Asgar S.;Lim, Chwee Teck
在管腔直径小于300 μ m的血管中，由于血管内的流速梯度，尺寸比白细胞小且更易变形的红细胞（RBC）迁移到血管的轴向中心。这种现象将白细胞转移到血管壁，并恰当地称为边集。在这里，我们使用微流体技术证明，较硬的疟疾感染的红细胞（iRBC）的行为与白细胞相似，并向侧壁边缘化。这提供了对微循环中iRBC的血流动力学效应及其对与细胞粘附至内皮相关的病理生理学结果的贡献的更好理解。在这项工作中，基于iRBC降低的变形性，模拟细胞边缘化以从全血中分离iRBC。在由聚二甲基硅氧烷制成的简单的长直通道微流体装置中测试疟疾感染的样品。在该微通道中，细胞边集沿着通道宽度被引导，iRBC在每个侧壁附近对齐，然后随后使用3出口系统去除，从而实现分离。使用环期和晚期滋养体/滋养体期iRBC进行测试。通过分析这些iRBC在出口处的微通道宽度上的分布以及进行流式细胞术分析来量化装置性能。结果表明，在侧出口处，早期iRBC的回收率接近75%，晚期iRBC的回收率>90%。简单和被动的系统操作使得该技术非常适合在资源有限的环境中进行现场iRBC富集，并且可以应用于以细胞硬度变化为特征的其他血细胞疾病，例如镰状细胞性贫血和白血病。
In blood vessels with luminal diameter less than 300 mu m, red blood cells (RBCs) which are smaller in size and more deformable than leukocytes, migrate to the axial centre of the vessel due to flow velocity gradient within the vessels. This phenomenon displaces the leukocytes to the vessel wall and is aptly termed as margination. Here, we demonstrate using microfluidics that stiffer malaria-infected RBCs (iRBCs) behave similar to leukocytes and undergo margination towards the sidewalls. This provides better understanding of the hemodynamic effects of iRBCs in microcirculation and its contribution to pathophysiological outcome relating to cytoadherence to endothelium. In this work, cell margination is mimicked for the separation of iRBCs from whole blood based on their reduced deformability. The malaria infected sample was tested in a simple long straight channel microfluidic device fabricated in polydimethylsiloxane. In this microchannel, cell margination was directed along the channel width with the iRBCs aligning near each sidewall and then subsequently removed using a 3-outlet system, thus achieving separation. Tests were conducted using ring stage and late trophozoite/schizont stage iRBCs. Device performance was quantified by analyzing the distribution of these iRBCs across the microchannel width at the outlet and also conducting flow cytometry analysis. Results indicate recovery of similar to 75% for early stage iRBCs and >90% for late stage iRBCs at the side outlets. The simple and passive system operation makes this technique ideal for on-site iRBCs enrichment in resource-limited settings, and can be applied to other blood cell diseases, e.g. sickle cell anemia and leukemia, characterized by changes in cell stiffness.