A novel, automated technology for multiplex biomarker imaging and application to breast cancer.
Histopathology
; 64(2): 242-55, 2014 Jan.
Article
en En
| MEDLINE
| ID: mdl-24330149
ABSTRACT
AIMS:
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. METHODS ANDRESULTS:
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.CONCLUSIONS:
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 positional shifts, while the fluorescent dye cycling approach dramatically expands the number of biomarkers which can be visualized and quantified on a single tissue section.Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Mama
/
Neoplasias de la Mama
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Inmunohistoquímica
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Biomarcadores de Tumor
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Técnica del Anticuerpo Fluorescente
Límite:
Female
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Humans
Idioma:
En
Revista:
Histopathology
Año:
2014
Tipo del documento:
Article
País de afiliación:
Canadá