Theranostics 2017; 7(19):4710-4721. doi:10.7150/thno.20440

Research Paper

Precisely Enumerating Circulating Tumor Cells Utilizing a Multi-Functional Microfluidic Chip and Unique Image Interpretation Algorithm

Mingxing Zhou1, 2*, Hui Zheng1*, Zhaoba Wang2*, Ren Li1, Xiaoran Liu3, Weikai Zhang1, Zihua Wang1, Huiping Li3✉, Zewen Wei1✉, Zhiyuan Hu1, 4, 5✉

1. CAS Key Laboratory of Standardization and Measurement for Nanotechnology, CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology of China, Beijing 100190, China;
2. School of Information and Communication Engineering, North University of China, Taiyuan 030051, China;
3. Key Laboratory of Carcinogenesis and Translational Research (Ministry of Education), Department of Breast Oncology, Peking University Cancer Hospital & Institute, Beijing 100142, China;
4. Sino-Danish College, University of Chinese Academy of Sciences, Beijing 100049, China;
5. Yangtze River Delta Academy of Nanotechnology and Industry Development Research, Jiaxing 314000, China.
* These authors contributed equally to this work.

Abstract

Enumerating circulating tumor cells (CTCs) has been demonstrably useful in cancer treatment. Although there are several approaches that have proved effective in isolating CTC-like cells, the crucial identification of CTCs continues to rely on the manual interpretation of immunofluorescence images of all cells that have been isolated. This procedure is time consuming and more importantly, CTC identification relies on subjective criteria that may differ between examiners. In this study, we describe the design, testing, and verification of a microfluidic platform that provides accurate and automated CTC enumeration using a common objective criterion.

Methods: The platform consists of a multi-functional microfluidic chip and a unique image processing algorithm. The microfluidic chip integrates blood filtering, cell isolation, and single cell positioning to ensure minimal cell loss, efficient cell isolation, and fixed arraying of single cells to facilitate downstream image processing. By taking advantage of the microfluidic chip design to reduce calculation loads and eliminate measurement errors, our specially designed algorithm has the capability of rapidly interpreting hundreds of images to provide accurate CTC counts.

Results: Following intensive optimization of the microfluidic chip, the image processing algorithm, and their collaboration, we verified the complete platform by enumerating CTCs from six clinical blood samples of patients with breast cancer. Compared to tube-based CTC isolation and manual CTC identification, our platform had better accuracy and reduced the time needed from sample loading to result review by 50%.

Conclusion: This automated CTC enumeration platform demonstrates not only a sound strategy in integrating a specially designed multi-functional microfluidic chip with a unique image processing algorithm for robust, accurate, and “hands-free” CTC enumeration, but may also lead to its use as a novel in vitro diagnostic device used in clinics and laboratories as readily as a routine blood test.

Keywords: circulating tumor cells, microfluidic chip, breast cancer, automated cell enumeration.

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How to cite this article:
Zhou M, Zheng H, Wang Z, Li R, Liu X, Zhang W, Wang Z, Li H, Wei Z, Hu Z. Precisely Enumerating Circulating Tumor Cells Utilizing a Multi-Functional Microfluidic Chip and Unique Image Interpretation Algorithm. Theranostics 2017; 7(19):4710-4721. doi:10.7150/thno.20440. Available from http://www.thno.org/v07p4710.htm