Efficient capture of circulating tumor cells with a novel immunocytochemical microfluidic device
Efficient capture of circulating tumor cells with a novel immunocytochemical microfluidic device
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DOI：
10.1063/1.3623748
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发表时间：
2011-09-01
期刊：
影响因子：
3.2
通讯作者：
Leonard, Edward F.
中科院分区：
文献类型：
--
作者：
Dickson, Mary Nora;Tsinberg, Pavel;Leonard, Edward F.
对癌症患者血液中的循环肿瘤细胞（CTC）进行细胞遗传学检查的能力对于实现靶向个体化治疗至关重要。与正常（旁观者）血细胞相比，CTC是罕见的，发现比例低至1：10（9）。最成功的分离技术是免疫细胞化学技术，其标记CTC以基于将其与正常旁观者细胞区分开的独特表面抗原进行分离。这里讨论的方法利用选择性结合CTC的生物素标记的抗体。将抗体引入血细胞的悬浮液中，旨在仅CTC将展示表面生物素分子。接下来，使细胞悬浮液通过含有约9000个横向的链霉亲和素包被的柱的微流体通道。与柱接触的CTC有机会参与固定细胞的生物素-链霉亲和素反应。旁观者血细胞保持悬浮状态并通过通道。本研究的目的是建立这些通道的技术性能作为抗原密度和操作条件的函数，特别是流速。在18 μ L/min时，超过70%的细胞在大于30000位点/细胞的抗原密度下被捕获，而50%的细胞在大于10000的抗原密度下被捕获。据发现，较低的流速导致细胞捕获概率降低，表明一些流线的发展是从来没有足够接近一个职位，以允许细胞后接触。使用计算流体动力学软件的未来建模和流线研究可以帮助优化捕获稀有细胞的通道性能。(C)2011年美国物理学会。[doi：10.1063/1.3623748]
Ability to perform cytogenetic interrogations on circulating tumor cells (CTCs) from the blood of cancer patients is vital for progressing toward targeted, individualized treatments. CTCs are rare compared to normal (bystander) blood cells, found in ratios as low as 1:10(9). The most successful isolation techniques have been immunocytochemical technologies that label CTCs for separation based on unique surface antigens that distinguish them from normal bystander cells. The method discussed here utilizes biotin-tagged antibodies that bind selectively to CTCs. The antibodies are introduced into a suspension of blood cells intending that only CTCs will display surface biotin molecules. Next, the cell suspension is passed through a microfluidic channel that contains about 9000 transverse, streptavidin coated posts. A CTC making contact with a post has the opportunity to engage in a biotin-streptavidin reaction that immobilizes the cell. Bystander blood cells remain in suspension and pass through the channel. The goal of the present study is to establish the technical performance of these channels as a function of antigen density and operating conditions, especially flow rate. At 18 mu L/min, over 70% of cells are captured at antigen densities greater than 30 000 sites/cell while 50% of cells are captured at antigen densities greater than 10 000. It is found that lower flow rates lead to decreasing cell capture probabilities, indicating that some streamlines develop which are never close enough to a post to allow cell-post contact. Future modeling and streamline studies using computational fluid dynamics software could aid in optimization of channel performance for capture of rare cells. (C) 2011 American Institute of Physics. [doi: 10.1063/1.3623748]