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. 2017 Aug 25;7:9542. doi: 10.1038/s41598-017-09284-0

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Next-generation sequencing analysis in captured CTC. (A–D) Cells isolated with the CellCollector were analyzed microscopically by means of immunofluorescence staining according to the staining procedure in the clinical setup (A,B). Cell Collector was fragmented and subjected to WGA (C). After WGA, quality of WGA DAN was analyzed with PCR assay. Four specific fragments were amplificated with WGA DNA, PBMC and ultrapure water. S, indicated WGA DNA; P indicated positive control; NTC indicated ultrapure water. (E) Gene mutations were analyzed with Hotspot panel v2 in five patients’ CTCs. Different colors indicated specific mutation sites in patients. (F) Cancer-related mutation genes were shown with form and pie chart. Different colors indicated different proportion. Numbers one to five indicated gene mutation exist in patient number.