Surface-micromachined microfiltration membranes for efficient isolation and functional immunophenotyping of subpopulations of immune cells.
Surface-micromachined microfiltration membranes for efficient isolation and functional immunophenotyping of subpopulations of immune cells.
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DOI：
10.1002/adhm.201200378
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发表时间：
2013-07
影响因子：
10
通讯作者：
Fu, Jianping
中科院分区：
文献类型：
--
作者：
Chen, Weiqiang;Huang, Nien-Tsu;Oh, Boram;Lam, Raymond H. W.;Fan, Rong;Cornell, Timothy T.;Shanley, Thomas P.;Kurabayashi, Katsuo;Fu, Jianping
准确测量免疫系统疾病患者的免疫状态对于评估疾病阶段和调整药物治疗至关重要。功能性细胞免疫测试是诊断免疫功能障碍的一种有前途的方法。传统的功能性细胞免疫测试涉及测量外周血单核细胞在离体刺激时产生促炎细胞因子的能力。然而，这种“批量”测定测量淋巴细胞和单核细胞群体的整体反应性，因此很难查明所涉及的反应性免疫细胞的表型或真实身份。在这项研究中，我们开发了一种具有高孔隙率的大表面微加工聚二甲基硅氧烷（PDMS）微滤膜（PMM），并将其集成在微流体微滤平台中。使用具有与特定细胞表面蛋白抗体缀合的功能化微珠的 PMM，我们展示了对血液样本中免疫细胞亚群的快速、高效和高通量的片上分离、富集和刺激。此外，PMM 集成微滤平台与免洗均质化学发光测定（“AlphaLISA”）相结合，使我们能够对直接从微量血液样本中分离的免疫细胞亚群进行快速、灵敏的片上免疫表型分析。
An accurate measurement of the immune status in patients with immune system disorders is critical in evaluating the stage of diseases and tailoring drug treatments. The functional cellular immunity test is a promising method to establish the diagnosis of immune dysfunctions. The conventional functional cellular immunity test involves measurements of the capacity of peripheral blood mononuclear cells to produce pro-inflammatory cytokines when stimulated ex vivo. However, this “bulk” assay measures the overall reactivity of a population of lymphocytes and monocytes, making it difficult to pinpoint the phenotype or real identity of the reactive immune cells involved. In this research, we develop a large surface micromachined polydimethylsiloxane (PDMS) microfiltration membrane (PMM) with high porosity, which is integrated in a microfluidic microfiltration platform. Using the PMM with functionalized microbeads conjugated with antibodies against specific cell surface proteins, we demonstrated rapid, efficient and high-throughput on-chip isolation, enrichment, and stimulation of subpopulations of immune cells from blood specimens. Furthermore, the PMM-integrated microfiltration platform, coupled with a no-wash homogeneous chemiluminescence assay (“AlphaLISA”), enables us to demonstrate rapid and sensitive on-chip immunophenotyping assays for subpopulations of immune cells isolated directly from minute quantities of blood samples.
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影响因子：
120.7
作者：
Boomer, Jonathan S.;To, Kathleen;Chang, Kathy C.;Takasu, Osamu;Osborne, Dale F.;Walton, Andrew H.;Bricker, Traci L.;Jarman, Stephen D., II;Kreisel, Daniel;Krupnick, Alexander S.;Srivastava, Anil;Swanson, Paul E.;Green, Jonathan M.;Hotchkiss, Richard S.
通讯作者：
Hotchkiss, Richard S.
影响因子：
2.2
作者：
Guerkov, REM;Targoni, OS;Schwander, SK
通讯作者：
Schwander, SK
影响因子：
3.2
作者：
Revzin, Alexander;Maverakis, Emanual;Chang, H. -C.
通讯作者：
Chang, H. -C.
影响因子：
56.9
作者：
SALGAME, P;ABRAMS, JS;BLOOM, BR
通讯作者：
BLOOM, BR
影响因子：
6
作者：
Remick, Daniel G.
通讯作者：
Remick, Daniel G.