Breast Cancer Diagnostics Using Microfluidic Multiplexed Immunohistochemistry

  • Minseok S. Kim
  • Seyong Kwon
  • Je-Kyun ParkEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 949)


A quantitative, reproducible, fast and inexpensive multiplexed immunohistochemistry (IHC) system might play a locomotive role in drug screening and personalized medicine. Currently, fully automated IHC machines and sequential multiplexed IHC methods based upon multiple color reagents have been developed, with the evolution of such methods having revealed novel biological findings over the conventional IHC method, which is time consuming and labor intensive. We describe a novel parallel multiplexed IHC method using a microfluidic multiplexed immunohistochemistry (MMIHC) device for quantitative pathological diagnosis of breast cancer. The key factors for success of parallel multiplexed IHC are the fabrication of a robust microfluidic device, the interface between the device and a tissue slide, and an accurate fluidic control for multiple IHC reagents. In order to apply conventional thin-section tissues into on-chip systems without any additional modification process, a tissue slide-compatible assembler was developed for optimal compatibility of conventional IHC methods. With this approach, a perfect fluid control for various solutions was demonstrated without any leakage, bubble formation or cross-contamination. The results presented in this chapter indicate that the microfluidic IHC protocol developed can provide the possibility of tailored cancer treatments as well as precise histopathological diagnosis using numerous specific biomarkers.

Key words

Breast cancer Immunohistochemistry Microfluidics Multiplexing Pathology Quantification Tissue 



This research was supported by a National Leading Research Laboratory program (grant no. 2011-0018607). We acknowledge Eun Sook Lee, Chul Hwan Kim, Soim Kwon, and Bumi Kwon for clinical application, and Taemin Kim for image analysis.


  1. 1.
    Coons AH, Creech HJ, Jones RN, Berliner E (1941) The demonstration of pneumococcal antigen in tissues by the use of fluorescent antibody. J Immunol 45:159Google Scholar
  2. 2.
    Graham RC Jr, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302CrossRefGoogle Scholar
  3. 3.
    Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG (1970) The unlabeled antibody enzyme method of immunohistochemistry: preparation and properties of soluble antigen-antibody complex (horseradish peroxidase-antihorseradish peroxidase) and its use in identification of spirochetes. J Histochem Cytochem 18:315–333CrossRefGoogle Scholar
  4. 4.
    Taylor CR, Levenson RM (2006) Quantification of immunohistochemistry–issues concerning methods, utility and semiquantitative assessment II. Histopathology 49:411–424CrossRefGoogle Scholar
  5. 5.
    Lakhani SR, Ashworth A (2001) Microarray and histopathological analysis of tumours: the future and the past? Nat Rev Cancer 1:151–157CrossRefGoogle Scholar
  6. 6.
    Ludwig JA, Weinstein JN (2005) Biomarkers in cancer staging, prognosis and treatment selection. Nat Rev Cancer 5:845–856CrossRefGoogle Scholar
  7. 7.
    Kim MS, Kim T, Kong S-Y, Kwon S, Bae CY, Choi J et al (2010) Breast cancer diagnosis using a microfluidic multiplexed immunohistochemistry platform. PLoS One 5:e10441. doi: 10.1371/journal.pone.0010441 CrossRefGoogle Scholar
  8. 8.
    Kim MS, Kwon S, Kim T, Lee ES, Park J-K (2011) Quantitative proteomic profiling of breast cancers using a multiplexed microfluidic platform for immunohistochemistry and immunocytochemistry. Biomaterials 32:1396–1403CrossRefGoogle Scholar
  9. 9.
    Vestad T, Marr DWM (2004) Flow resistance for microfluidic logic operations. Appl Phys Lett 84:5074–5075CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media,LLC 2013

Authors and Affiliations

  1. 1.Department of Bio and Brain EngineeringKorea Advanced Institute of Science and Technology (KAIST)DaejeonRepublic of Korea
  2. 2.KAIST Institute for the NanoCenturyDaejeonRepublic of Korea

Personalised recommendations