title: Capture and Identification of Heterogeneous Circulating Tumor Cells Using Transparent Nanomaterials and Quantum Dots-Based Multiplexed Imaging.
abstract: BACKGROUND
  Capture and identification of circulating tumor cells (CTCs) in the blood system can help guide therapy and predict the prognosis of cancer patients. However, simultaneous capture and identification of CTCs with both epithelial and mesenchymal phenotypes remains a formidable technical challenge for cancer research. This study aimed at developing a system to efficiently capture and identify these CTCs with heterogeneous phenotypes using transparent nanomaterials and quantum dots (QDs)-based multiplexed imaging.
  
  
  METHODS
  Hydroxyapatite-chitosan (HA-CTS) nanofilm-coated substrates were modified based on our previous work to increase the capture efficiency of cancer cell lines by extending baking and incubating time. QDs-based imaging was applied to detect cytokeratin, epithelial cell adhesion molecule (EpCAM), and vimentin of cancer cells to demonstrate the feasibility of multiplexed imaging. And QDs-based multiplexed imaging of CD45, cytokeratin and vimentin was applied to detect CTCs from different cancer patients that were captured using HA-CTS nanofilm substrates.
  
  
  RESULTS
  Comparisons of the capture efficiencies of cancer cells at different baking time of film formation and incubating time of cell capture revealed the optimal baking and incubating time. Optimal time was chosen to develop a modified CTCs capture system that could capture EpCAM-positive cancer cells at an efficiency > 80%, and EpCAM-negative cancer cells at an efficiency > 50%. QDs-based imaging exhibited comparable detection ability but higher photostability compared to organic dyes imaging in staining cells. In addition, QDs-based multiplexed imaging also showed the molecular profiles of cancer cell lines with different phenotypes well. The integrated CTCs capture and identification system successfully captured and imaged CTCs with different sub-phenotypes in blood samples from cancer patients.
  
  
  CONCLUSIONS
  This study demonstrated a reliable capture and detection system for heterogeneous CTCs that combined enrichment equipment based on HA-CTS nanofilm substrates with QDs-based multiplexed imaging.
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Yuan-Yuan
authors.initials: YY
authors.lastname: Chen
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Bo-Ran
authors.initials: BR
authors.lastname: Cheng
authors.affiliation: 2. Key Laboratory of Artificial Micro- and Nano-Structures of Ministry of Education, School of Physics and Technology, Wuhan University, Wuhan, Hubei, China.
authors.forename: Zhao-Bo
authors.initials: ZB
authors.lastname: He
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Shu-Yi
authors.initials: SY
authors.lastname: Wang
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Zhen-Meng
authors.initials: ZM
authors.lastname: Wang
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Min
authors.initials: M
authors.lastname: Sun
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Hai-Bin
authors.initials: HB
authors.lastname: Song
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Yuan
authors.initials: Y
authors.lastname: Fang
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Chen
authors.affiliation: 1. Department of Oncology, Zhongnan Hospital of Wuhan University; Hubei Key Laboratory of Tumor Biological Behaviors & Hubei Cancer Clinical Study Center; Wuhan, Hubei, 430071, P. R. China;
authors.forename: Bin
authors.initials: B
authors.lastname: Xiong
keywords: Circulating tumor cells; epithelial; hydroxyapatite-chitosan; mesenchymal; quantum dots.