Node-pore sensing enables label-free surface-marker profiling of single cells.
Node-pore sensing enables label-free surface-marker profiling of single cells.
复制标题
节点 - 孔隙传感能够对单细胞进行无标记的表面标志物分析。
DOI：
10.1021/ac504613b
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发表时间：
2015-03-03
影响因子：
7.4
通讯作者：
Sohn, Lydia L.
中科院分区：
文献类型：
--
作者：
Balakrishnan, Karthik R.;Whang, Jeremy C.;Hwang, Richard;Hack, James H.;Godley, Lucy A.;Sohn, Lydia L.
流式细胞术是表征细胞群体的普遍存在的多参数方法。然而，随着需要同时筛选的蛋白质数量的增加，这种方法会变得越来越复杂：荧光团的光谱发射重叠以及随后的补偿需求，冗长的样品制备以及需要单独进行的多个控制测试都必须考虑在内。这些因素导致成本增加，因此，流式细胞术在核心设施中进行，由专门的技术人员操作仪器。在这里，我们描述了一种低成本，无标记的微流控方法，可以确定单细胞的表型。我们的方法采用节点孔传感来测量细胞的通过时间，因为它们与一系列不同的抗体相互作用，每个抗体对应于特定的细胞表面抗原，这些抗体已经在单个微流体通道中功能化。我们证明了我们的方法的能力，不仅通过筛选两个急性早幼粒细胞白血病人类细胞系，（NB 4和AP-1060）的骨髓抗原、CD 13、CD 14、CD 15和CD 33的检测，而且还通过基于表面标志物CD 13和HLA-DR区分相似大小的细胞的混合物-AP-1060和NALM-1。我们表明，我们的方法可以筛选临床样本中的复杂亚群：我们成功地鉴定了来自急性髓性白血病患者的原代人骨髓样本中的原始细胞群，并筛选了这些细胞的CD 13、CD 34和HLA-DR。我们表明，我们的无标记方法是一种经济实惠、高度敏感，和用户友好的技术，有可能改变细胞筛选在实验室。
Flow cytometry is a ubiquitous, multiparametric method for characterizing cellular populations. However, this method can grow increasingly complex with the number of proteins that need to be screened simultaneously: spectral emission overlap of fluorophores and the subsequent need for compensation, lengthy sample preparation, and multiple control tests that need to be performed separately must all be considered. These factors lead to increased costs, and consequently, flow cytometry is performed in core facilities with a dedicated technician operating the instrument. Here, we describe a low-cost, label-free microfluidic method that can determine the phenotypic profiles of single cells. Our method employs Node-Pore Sensing to measure the transit times of cells as they interact with a series of different antibodies, each corresponding to a specific cell-surface antigen, that have been functionalized in a single microfluidic channel. We demonstrate the capabilities of our method not only by screening two acute promyelocytic leukemia human cells lines (NB4 and AP-1060) for myeloid antigens, CD13, CD14, CD15, and CD33, simultaneously, but also by distinguishing a mixture of cells of similar size—AP-1060 and NALM-1—based on surface markers CD13 and HLA-DR. Furthermore, we show that our method can screen complex subpopulations in clinical samples: we successfully identified the blast population in primary human bone marrow samples from patients with acute myeloid leukemia and screened these cells for CD13, CD34, and HLA-DR. We show that our label-free method is an affordable, highly sensitive, and user-friendly technology that has the potential to transform cellular screening at the benchside.
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影响因子：
16.8
作者：
Bendall SC;Nolan GP;Roederer M;Chattopadhyay PK
通讯作者：
Chattopadhyay PK
影响因子：
3.7
作者：
Normolle, Daniel P.;Donnenberg, Vera S.;Donnenberg, Albert D.
通讯作者：
Donnenberg, Albert D.
影响因子：
3.7
作者：
Autissier, Patrick;Soulas, Caroline;Williams, Kenneth C.
通讯作者：
Williams, Kenneth C.
DOI：
10.1126/science.1198704
发表时间：
2011-05-06
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Bendall SC;Simonds EF;Qiu P;Amir el-AD;Krutzik PO;Finck R;Bruggner RV;Melamed R;Trejo A;Ornatsky OI;Balderas RS;Plevritis SK;Sachs K;Pe'er D;Tanner SD;Nolan GP
通讯作者：
Nolan GP
影响因子：
1.6
作者：
DEBLOIS, RW;BEAN, CP
通讯作者：
BEAN, CP