Department of Nano Science and Engineering Center for Nano Manufacturing, Inje University , Gimhae 621-749, Republic of Korea.
Anal Chem. 2016 May 3;88(9):4857-63. doi: 10.1021/acs.analchem.6b00570. Epub 2016 Apr 19.
This paper introduces a single-cell isolation technology for circulating tumor cells (CTCs) using a microfluidic device (the "SIM-Chip"). The SIM-Chip comprises a lateral magnetophoretic microseparator and a microdispenser as a two-step cascade platform. First, CTCs were enriched from whole blood by the lateral magnetophoretic microseparator based on immunomagnetic nanobeads. Next, the enriched CTCs were electrically identified by single-cell impedance cytometer and isolated as single cells using the microshooter. Using 200 μL of whole blood spiked with 50 MCF7 breast cancer cells, the analysis demonstrated that the single-cell isolation efficiency of the SIM-Chip was 82.4%, and the purity of the isolated MCF7 cells with respect to WBCs was 92.45%. The data also showed that the WBC depletion rate of the SIM-Chip was 2.5 × 10(5) (5.4-log). The recovery rates were around 99.78% for spiked MCF7 cells ranging in number from 10 to 90. The isolated single MCF7 cells were intact and could be used for subsequent downstream genetic assays, such as RT-PCR. Single-cell culture evaluation of the proliferation of MCF7 cells isolated by the SIM-Chip showed that 84.1% of cells at least doubled in 5 days. Consequently, the SIM-Chip could be used for single-cell isolation of rare target cells from whole blood with high purity and recovery without cell damage.
本文介绍了一种使用微流控装置（“SIM-Chip”）分离循环肿瘤细胞（CTCs）的单细胞分离技术。SIM-Chip 由侧向磁分离微分离器和微分配器组成，作为两步级联平台。首先，通过基于免疫磁珠的侧向磁分离微分离器从全血中富集 CTCs。接下来，通过单细胞阻抗细胞仪对富集的 CTCs 进行电识别，并使用微喷射器将其分离为单细胞。使用 200μL 全血加 50 个 MCF7 乳腺癌细胞进行分析，结果表明 SIM-Chip 的单细胞分离效率为 82.4%，分离出的 MCF7 细胞相对于白细胞（WBCs）的纯度为 92.45%。数据还显示，SIM-Chip 的 WBC 清除率为 2.5×10(5)（5.4-log）。当 MCF7 细胞的数量从 10 到 90 时，回收率约为 99.78%。分离出的单个 MCF7 细胞完整无损，可用于随后的下游遗传分析，如 RT-PCR。SIM-Chip 分离出的 MCF7 细胞的单细胞培养评估显示，84.1%的细胞在 5 天内至少倍增一次。因此，SIM-Chip 可用于从全血中以高纯度和回收率分离稀有靶细胞，而不会造成细胞损伤。
