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通过光诱导电动学从红细胞中快速、无标记地分离伯基特淋巴瘤细胞。

Rapid and label-free separation of Burkitt's lymphoma cells from red blood cells by optically-induced electrokinetics.

作者信息

Liang Wenfeng, Zhao Yuliang, Liu Lianqing, Wang Yuechao, Dong Zaili, Li Wen Jung, Lee Gwo-Bin, Xiao Xiubin, Zhang Weijing

机构信息

State Key Laboratory of Robotics, Shenyang Institute of Automation, Chinese Academy of Sciences, Shenyang, China; University of Chinese Academy of Sciences, Beijing, China; Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Kowloon, Hong Kong.

Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Kowloon, Hong Kong.

出版信息

PLoS One. 2014 Mar 7;9(6):e90827. doi: 10.1371/journal.pone.0090827. eCollection 2014.

Abstract

Early stage detection of lymphoma cells is invaluable for providing reliable prognosis to patients. However, the purity of lymphoma cells in extracted samples from human patients' marrow is typically low. To address this issue, we report here our work on using optically-induced dielectrophoresis (ODEP) force to rapidly purify Raji cells' (a type of Burkitt's lymphoma cell) sample from red blood cells (RBCs) with a label-free process. This method utilizes dynamically moving virtual electrodes to induce negative ODEP force of varying magnitudes on the Raji cells and RBCs in an optically-induced electrokinetics (OEK) chip. Polarization models for the two types of cells that reflect their discriminate electrical properties were established. Then, the cells' differential velocities caused by a specific ODEP force field were obtained by a finite element simulation model, thereby established the theoretical basis that the two types of cells could be separated using an ODEP force field. To ensure that the ODEP force dominated the separation process, a comparison of the ODEP force with other significant electrokinetics forces was conducted using numerical results. Furthermore, the performance of the ODEP-based approach for separating Raji cells from RBCs was experimentally investigated. The results showed that these two types of cells, with different concentration ratios, could be separated rapidly using externally-applied electrical field at a driven frequency of 50 kHz at 20 Vpp. In addition, we have found that in order to facilitate ODEP-based cell separation, Raji cells' adhesion to the OEK chip's substrate should be minimized. This paper also presents our experimental results of finding the appropriate bovine serum albumin concentration in an isotonic solution to reduce cell adhesion, while maintaining suitable medium conductivity for electrokinetics-based cell separation. In short, we have demonstrated that OEK technology could be a promising tool for efficient and effective purification of Raji cells from RBCs.

摘要

淋巴瘤细胞的早期检测对于为患者提供可靠的预后至关重要。然而,从人类患者骨髓中提取的样本中淋巴瘤细胞的纯度通常较低。为了解决这个问题,我们在此报告我们使用光诱导介电泳(ODEP)力从红细胞(RBC)中快速纯化Raji细胞(一种伯基特淋巴瘤细胞)样本的无标记过程的工作。该方法利用动态移动的虚拟电极在光诱导电动(OEK)芯片中对Raji细胞和红细胞施加不同大小的负ODEP力。建立了反映两种细胞不同电学性质的极化模型。然后,通过有限元模拟模型获得特定ODEP力场引起的细胞差异速度,从而建立了使用ODEP力场分离两种细胞的理论基础。为确保ODEP力主导分离过程,利用数值结果对ODEP力与其他重要的电动学力进行了比较。此外,还对基于ODEP的从红细胞中分离Raji细胞的方法进行了实验研究。结果表明,在20Vpp、50kHz的驱动频率下,通过施加外部电场,可以快速分离这两种不同浓度比的细胞。此外,我们发现为了便于基于ODEP的细胞分离,应尽量减少Raji细胞对OEK芯片基底的粘附。本文还展示了我们在等渗溶液中寻找合适的牛血清白蛋白浓度以减少细胞粘附,同时保持适合基于电动学的细胞分离的介质电导率的实验结果。简而言之,我们已经证明OEK技术可能是一种从红细胞中高效纯化Raji细胞的有前途的工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a3bb/3946566/3dbcd5432db8/pone.0090827.g001.jpg

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