Precision Microfilters as an all in one System for Multiplex Analysis of Circulating Tumor Cells.
Precision Microfilters as an all in one System for Multiplex Analysis of Circulating Tumor Cells.
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DOI：
10.1039/c5ra21524b
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发表时间：
2016
期刊：
影响因子：
3.9
通讯作者：
Stefansson S
中科院分区：
文献类型：
--
作者：
Adams DL;Alpaugh RK;Martin SS;Charpentier M;Chumsri S;Cristofanilli M;Adams DK;Makarova OV;Zhu P;Li S;Tang CM;Stefansson S
从癌症患者血液中计数循环肿瘤细胞（CTC）是一种已建立的诊断测定，用于作为单重测试评估患者状态。然而，在即将到来的个性化医疗时代，患者CTC的多重分析，包括蛋白质组学和基因组学技术，必须与CTC分离平台技术相结合。微制造的进步已经证明，CTC可以使用微流体芯片实验室设备进行分离和分析。然而，到目前为止，大多数微流体装置要么仍处于开发阶段，不适用于所有临床试验，要么尚未商业化。为了克服这些差异，我们描述了一种用于临床适用的CTC测定的分离和多重化的一体化装置。微过滤器提供了一个理想的芯片实验室平台，用于分析CTC，因为无毒和惰性材料允许从细胞生长到临床染色技术的多种测试，所有这些都没有背景干扰。光刻制造的微过滤器，可以制成具有高孔隙率，精确的孔尺寸，排列的孔分布，并优化CTC的大小为基础的分离。在这项研究中，我们描述了使用微过滤器分离和原位分析CTC，使用多种连续技术，包括培养，FISH，组织病理学分析，H&E染色，光漂白和再染色。此外，作为原理证明，我们描述了从微滤器定量释放患者来源的CTCS的能力，以用于下游基因组/蛋白质组学分析。
Enumeration of circulating tumor cells (CTCs) from cancer patient blood is an established diagnostic assay used to evaluate patient status as a singleplex test. However, in the coming age of personalized medicine, multiplex analysis of patient CTCs, including proteomic and genomic techniques, will have to be integrated with CTC isolation platform technologies. Advancements in microfabrication have demonstrated that CTCs can be isolated and analyzed using microfluidic lab-on-a-chip devices. However, to date, most microfluidic devices are either still in the development phase, not applicable to all clinical tests, or are not commercially available. To overcome these discrepancies, we describe an all-in-one device for the isolation and multiplexing of clinically applicable CTC assays. Microfilters present an ideal lab-on-a-chip platform for analysis of CTCs as non-toxic and inert materials allow for a multitude of tests from cell growth through clinical staining techniques, all without background interference. Lithographically fabricated microfilters, can be made with high porosity, precise pore dimensions, arrayed pore distribution, and optimized for CTC size-based isolation. In this study we describe microfilter use in isolation and in situ analysis of CTCs using multiple sequential techniques including culture, FISH, histopathological analysis, H&E staining, photobleaching and re-staining. Further, as a proof of principle, we then describe the ability to quantitatively release patient derived CTCS from the microfilters for potential use in downstream genomic/proteomic analysis.
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影响因子：
158.5
作者：
Cristofanilli, M;Budd, GT;Hayes, DF
通讯作者：
Hayes, DF
影响因子：
3.7
作者：
Coumans FA;van Dalum G;Beck M;Terstappen LW
通讯作者：
Terstappen LW
影响因子：
11.1
作者：
Joosse SA;Gorges TM;Pantel K
通讯作者：
Pantel K
影响因子：
9.3
作者：
Harouaka, Ramdane A.;Zhou, Ming-Da;Zheng, Si-Yang
通讯作者：
Zheng, Si-Yang
DOI：
10.1073/pnas.1320198111
发表时间：
2014-03-04
影响因子：
11.1
作者：
Adams, Daniel L.;Martin, Stuart S.;Cristofanilli, Massimo
通讯作者：
Cristofanilli, Massimo