Yu Zeta Tak For, Joseph Jophin George, Liu Shirley Xiaosu, Cheung Mei Ki, Haffey Parker James, Kurabayashi Katsuo, Fu Jianping
Department of Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109, USA.
Department of Chemical Engineering, Indian Institute of Technology Hyderabad, Kandi, Sangareddy, Telangana, India 502285.
Sens Actuators B Chem. 2017 Jun;245:1050-1061. doi: 10.1016/j.snb.2017.01.113. Epub 2017 Jan 23.
Sorting and enumeration of immune cells from blood are critical operations involved in many clinical applications. Conventional methods for sorting and counting immune cells from blood, such as flow cytometry and hemocytometers, are tedious, inaccurate, and difficult for implementation for point-of-care (POC) testing. Herein we developed a microscale centrifugal technology termed Centrifugal Microfluidic Chip (CMC) capable of sorting immune cells from blood and cellular analysis in a laboratory setting. Operation of the CMC entailed a blood specimen layered on a density gradient medium and centrifuged in microfluidic channels where immune cell subpopulations could rapidly be sorted into distinct layers according to their density differentials. We systematically studied effects of different blocking molecules for surface passivation of the CMC. We further demonstrated the applicability of CMCs for rapid separation of minimally processed human whole blood without affecting immune cell viability. Multi-color imaging and analysis of immune cell distributions and enrichment such as recovery and purity rates of peripheral blood mononuclear cells (PBMCs) were demonstrated using CMCs. Given its design and operation simplicity, portability, blood cell sorting efficiency, and cellular analysis capability, the CMC holds promise for blood-based diagnosis and disease monitoring in POC applications.
从血液中对免疫细胞进行分选和计数是许多临床应用中涉及的关键操作。用于从血液中分选和计数免疫细胞的传统方法,如流式细胞术和血细胞计数器,既繁琐、不准确,又难以用于即时检测(POC)。在此,我们开发了一种称为离心微流控芯片(CMC)的微尺度离心技术,它能够在实验室环境中从血液中分选免疫细胞并进行细胞分析。CMC的操作包括将血液样本铺在密度梯度介质上,并在微流控通道中进行离心,在该通道中免疫细胞亚群可根据其密度差异迅速被分选为不同的层。我们系统地研究了不同封闭分子对CMC表面钝化的影响。我们进一步证明了CMC在不影响免疫细胞活力的情况下,可用于快速分离经过最少处理的人全血。使用CMC展示了对免疫细胞分布以及富集情况(如外周血单核细胞(PBMC)的回收率和纯度)的多色成像和分析。鉴于其设计和操作的简便性、便携性、血细胞分选效率以及细胞分析能力,CMC在POC应用中的基于血液的诊断和疾病监测方面具有广阔前景。