Ryu Hyunryul, Choi Kyungyong, Qu Yanyan, Kwon Taehong, Lee Janet S, Han Jongyoon
Research Laboratory of Electronics, Massachusetts Institute of Technology.
Research Laboratory of Electronics, Massachusetts Institute of Technology; Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology.
J Vis Exp. 2018 Jun 7(136):57673. doi: 10.3791/57673.
Airway secretions contain a large number of immune-related cells, e.g., neutrophils, macrophages, and lymphocytes, which can be used as a major resource to evaluate a variety of pulmonary diseases, both for research and clinical purposes. However, due to the heterogeneous and viscous nature of patient mucus, there is currently no reliable dissociation method that does not damage the host immune cells in the patient airway secretion. In this research, we introduce a sample preparation method that uses inertial microfluidics for the patient's immune assessment. Regardless of the heterogeneous fluidic properties of the clinical samples, the proposed method recovers more than 95% of neutrophils from airway secretion samples that are diluted 1,000-fold with milliliters of clean saline. By recirculating the concentrated output stream to the initial sample reservoir, a high concentration, recovery, and purity of the immune cells are provided; recirculation is considered a trade-off to the single-run syringe-based operation of inertial microfluidics. The closed-loop operation of spiral microfluidics provides leukocytes without physical or chemical disturbance, as demonstrated by the phorbol 12-myristate 13-acetate (PMA)-induced elastase release of sorted neutrophils.
气道分泌物包含大量免疫相关细胞，例如中性粒细胞、巨噬细胞和淋巴细胞，它们可作为评估各种肺部疾病的主要资源，用于研究和临床目的。然而，由于患者黏液的异质性和黏性，目前尚无可靠的解离方法能在不损害患者气道分泌物中宿主免疫细胞的情况下进行解离。在本研究中，我们引入了一种使用惯性微流控技术进行患者免疫评估的样本制备方法。无论临床样本的流体性质如何异质，该方法都能从用数毫升生理盐水稀释1000倍的气道分泌物样本中回收超过95%的中性粒细胞。通过将浓缩的输出流再循环到初始样本储存器，可提供高浓度、高回收率和高纯度的免疫细胞；再循环被认为是对基于注射器的惯性微流控单次操作的一种权衡。螺旋微流控的闭环操作可在无物理或化学干扰的情况下提供白细胞，佛波醇12 -肉豆蔻酸酯13 -乙酸酯（PMA）诱导分选的中性粒细胞释放弹性蛋白酶的实验证明了这一点。
