Dec 18, 2013

A novel, automated technology for multiplex biomarker imaging and application to breast cancer

Gina M ClarkeMartin J Yaffe


Multiplexed immunofluorescence is a powerful tool for validating multigene assays and understanding the complex interplay of proteins implicated in breast cancer within a morphological context. We describe a novel technology for imaging an extended panel of biomarkers on a single, formalin-fixed paraffin-embedded breast sample and evaluating biomarker interaction at a single-cell level, and demonstrate proof-of-concept on a small set of breast tumours, including those which co-express hormone receptors with Her2/neu and Ki-67. Using a microfluidic flow cell, reagent exchange was automated and consisted of serial rounds of staining with dye-conjugated antibodies, imaging and chemical deactivation. A two-step antigen retrieval process was developed to satisfy all epitopes simultaneously, and key parameters were optimized. The imaging sequence was applied to seven breast tumours, and compared with conventional immunohistochemistry. Single-cell correlation analysis was performed with automated image processing. We have described a novel platform for evaluating biomarker co-localization. Expression in multiplexed images is consistent with conventional immunohistochemistry. Automation reduces inconsistencies in staining and positiona...Continue Reading

  • References37
  • Citations12


  • References37
  • Citations12


Mentioned in this Paper

Establishment and Maintenance of Localization
ErbB-2 Receptor
Immunofluorescence Assay
Hormone Receptor
Tumor Lysis Syndrome
Tumor Markers
Fluorescent stain

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