title: Breast cancer diagnosis using a microfluidic multiplexed immunohistochemistry platform.
abstract: BACKGROUND
  Biomarkers play a key role in risk assessment, assessing treatment response, and detecting recurrence and the investigation of multiple biomarkers may also prove useful in accurate prediction and prognosis of cancers. Immunohistochemistry (IHC) has been a major diagnostic tool to identify therapeutic biomarkers and to subclassify breast cancer patients. However, there is no suitable IHC platform for multiplex assay toward personalized cancer therapy. Here, we report a microfluidics-based multiplexed IHC (MMIHC) platform that significantly improves IHC performance in reduction of time and tissue consumption, quantification, consistency, sensitivity, specificity and cost-effectiveness.
  
  
  RESULTS
  By creating a simple and robust interface between the device and human breast tissue samples, we not only applied conventional thin-section tissues into on-chip without any additional modification process, but also attained perfect fluid control for various solutions, without any leakage, bubble formation, or cross-contamination. Four biomarkers, estrogen receptor (ER), human epidermal growth factor receptor 2 (HER2), progesterone receptor (PR) and Ki-67, were examined simultaneously on breast cancer cells and human breast cancer tissues. The MMIHC method improved immunoreaction, reducing time and reagent consumption. Moreover, it showed the availability of semi-quantitative analysis by comparing Western blot. Concordance study proved strong consensus between conventional whole-section analysis and MMIHC (n = 105, lowest Kendall's coefficient of concordance, 0.90). To demonstrate the suitability of MMIHC for scarce samples, it was also applied successfully to tissues from needle biopsies.
  
  
  CONCLUSIONS
  The microfluidic system, for the first time, was successfully applied to human clinical tissue samples and histopathological diagnosis was realized for breast cancers. Our results showing substantial agreement indicate that several cancer-related proteins can be simultaneously investigated on a single tumor section, giving clear advantages and technical advances over standard immunohistochemical method. This novel concept will enable histopathological diagnosis using numerous specific biomarkers at a time even for small-sized specimens, thus facilitating the individualization of cancer therapy.
authors.affiliation: Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea.
authors.forename: Minseok S
authors.initials: MS
authors.lastname: Kim
authors.forename: Taemin
authors.initials: T
authors.lastname: Kim
authors.forename: Sun-Young
authors.initials: SY
authors.lastname: Kong
authors.forename: Soim
authors.initials: S
authors.lastname: Kwon
authors.forename: Chae Yun
authors.initials: CY
authors.lastname: Bae
authors.forename: Jaekyu
authors.initials: J
authors.lastname: Choi
authors.forename: Chul Hwan
authors.initials: CH
authors.lastname: Kim
authors.forename: Eun Sook
authors.initials: ES
authors.lastname: Lee
authors.forename: Je-Kyun
authors.initials: JK
authors.lastname: Park
mesh_terms: D001706:Biopsy; D001940:Breast / Q000473:pathology; D001943:Breast Neoplasms / Q000175:diagnosis* / Q000473:pathology*; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D044085:Microfluidics / Q000379:methods*; D008954:Models, Biological; D009367:Neoplasm Staging; D015203:Reproducibility of Results; D013194:Staining and Labeling
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