Department of Biomedical Engineering , Hanyang University , Seoul 04763 , Republic of Korea.
Graduate School of Medical Science and Engineering, Korea Advanced Institute of Science and Technology (KAIST) , Daejeon 34141 , Republic of Korea.
Anal Chem. 2019 Oct 15;91(20):13230-13236. doi: 10.1021/acs.analchem.9b03673. Epub 2019 Sep 26.
One-step purification of white blood cells (WBCs) is essential to automate blood sample preparation steps for WBC analysis, but conventional methods such as red blood cell (RBC) lysis and density-gradient centrifugation typically require harsh chemical or physical treatment, followed by repeated manual washing steps. Alternative microfluidic separation methods show limited separation performances due to the trade-off between purity and throughput. Herein, an integrated microfluidic device is developed to decouple the trade-off by synergistically combining a slant array ridge-based WBC enrichment unit as a throughput enhancer and a slant, asymmetric lattice-based WBC washing unit as a purity enhancer. The enrichment unit can maintain a high sample-infusion throughput while lowering the flow rate into the washing unit, thus enabling WBC-selective washing without significant influence by the overwhelming number of RBCs and inertial forces. The device delivers efficient separation performances by rejecting 99.9% of RBCs as well as 99.9% of blood plasma from canine and human whole blood in a single round of purification at a high throughput of 60 μL/min. The purified WBC population well preserves the composition of lymphocyte subpopulations, the major components of the adaptive immune system, thus providing the potential for the integrated device to be used as an essential sample-preparation tool for immunologic investigations.
一步法纯化白细胞（WBC）对于自动化血液样本制备步骤进行 WBC 分析至关重要，但传统方法，如红细胞（RBC）裂解和密度梯度离心，通常需要苛刻的化学或物理处理，然后进行多次手动洗涤步骤。替代的微流分离方法由于纯度和通量之间的权衡而显示出有限的分离性能。本文开发了一种集成微流控装置，通过协同结合基于倾斜阵列脊的白细胞富集单元作为高通量增强器和基于倾斜、不对称格子的白细胞洗涤单元作为纯度增强器，来解耦这种权衡。富集单元可以在降低进入洗涤单元的流速的同时保持高样品注入通量，从而能够进行白细胞选择性洗涤，而不受压倒性数量的 RBC 和惯性力的显著影响。该装置在 60 μL/min 的高通量下，在单个净化循环中，从犬和人全血中有效分离，排斥 99.9%的 RBC 和 99.9%的血浆，分离性能高效。经纯化的白细胞群体很好地保留了淋巴细胞亚群的组成，这是适应性免疫系统的主要成分，从而为集成设备提供了作为免疫研究基本样本制备工具的潜力。
