Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays
Microfluidic sorting and multimodal typing of cancer cells in self-assembled magnetic arrays
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DOI：
10.1073/pnas.1001515107
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发表时间：
2010-08-17
影响因子：
11.1
通讯作者：
Viovy, Jean-Louis
中科院分区：
文献类型：
--
作者：
Saliba, Antoine-Emmanuel;Saias, Laure;Viovy, Jean-Louis
我们提出了一种独特的细胞分选方法，“以弗所”，使用生物功能化的超顺磁珠自组装在微流体通道上的微接触印刷制备的磁阱阵列列。它结合了微流控细胞分选的优点，特别是在细胞和珠之间应用良好控制的流动激活相互作用，以及免疫磁性分选的优点，特别是使用批量制备的、良好表征的携带抗体的珠。在细胞系混合物上，我们证明了优于94%的捕获产率，以及原位培养捕获细胞的可能性。第二个系列的实验涉及临床样本，血液，胸腔积液，细针抽吸物，来自健康捐赠者和B细胞血液恶性肿瘤（白血病和淋巴瘤）患者。在盲法试验中，直接在微流控室中分析B淋巴细胞的免疫表型和形态，并与常规流式细胞术和视觉细胞学数据进行比较。使用麻黄的免疫表型结果与通过流式细胞术获得的结果完全一致。我们获得了原位高分辨率共聚焦三维图像的细胞核，显示核内的细节与传统的细胞学染色一致。因此，Ephesia提供了一种强大的细胞捕获和分型方法，允许全自动高分辨率和定量免疫分型和形态学分析。它所需的样本体积和细胞数量比细胞计数法小至少10倍，可能会增加适应症范围和基于微活检的诊断成功率，并减少分析时间和成本。
We propose a unique method for cell sorting, "Ephesia," using columns of biofunctionalized superparamagnetic beads self-assembled in a microfluidic channel onto an array of magnetic traps prepared by microcontact printing. It combines the advantages of microfluidic cell sorting, notably the application of a well controlled, flow-activated interaction between cells and beads, and those of immunomagnetic sorting, notably the use of batch-prepared, well characterized antibody-bearing beads. On cell lines mixtures, we demonstrated a capture yield better than 94%, and the possibility to cultivate in situ the captured cells. A second series of experiments involved clinical samples-blood, pleural effusion, and fine needle aspirates-issued from healthy donors and patients with B-cell hematological malignant tumors (leukemia and lymphoma). The immunophenotype and morphology of B-lymphocytes were analyzed directly in the microfluidic chamber, and compared with conventional flow cytometry and visual cytology data, in a blind test. Immunophenotyping results using Ephesia were fully consistent with those obtained by flow cytometry. We obtained in situ high resolution confocal three-dimensional images of the cell nuclei, showing intranuclear details consistent with conventional cytological staining. Ephesia thus provides a powerful approach to cell capture and typing allowing fully automated high resolution and quantitative immunophenotyping and morphological analysis. It requires at least 10 times smaller sample volume and cell numbers than cytometry, potentially increasing the range of indications and the success rate of microbiopsy-based diagnosis, and reducing analysis time and cost.