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. 2017 Aug 29;7:9789. doi: 10.1038/s41598-017-10763-7

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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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Schematic diagram of SE-iFISH and viability examination of the enriched tumor cells. (a) Illustration of schematic experimental flow and application of SE-iFISH. Diverse subtypes of CRCs in tumor animal models or cancer patients’ biofluid samples, including blood, bone marrow (BM), ascites, malignant pleural effusion (MPE), and cerebrospinal fluid (CSF), etc., are effectively and efficiently enriched, identified and characterized. Obtained viable CRCs including CTCs are available for primary tumor cell culture and a series of indicated downstream analyses. (b) Viability examination of tumor cells and WBCs following subtraction enrichment and immunostaining processing. Necrotic breast cancer cell (7-AAD⁺/HER2⁺/Hoechst⁺) (red arrow), and WBC (7-AAD⁺/HER2⁻/Hoechst⁺) (white arrow) are depicted. The same enlarged necrotic WBC is shown in the top right corner box. Healthy cells are 7-AAD negative. (c) Average of viability of the enriched tumor cells (blue) and WBCs (red) is 95.3 ± 2% and 93 ± 2.1% (mean ± SD), respectively. Results represent average of values obtained in 3 separate experiments performed in triplicate. Cytelligen, the copyright holder of the registered iFISH trademark, ensures Nature Publishing Group (NPG) to publish iFISH logo image in all formats under an Open Access License.