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

被引：29

作者：

Clarke, Gina M. ^{[1
]}

Zubovits, Judit T. ^{[2
,3
]}

Shaikh, Kashan A. ^{[4
]}

Wang, Dan ^{[1
]}

Dinn, Sean R. ^{[5
]}

Corwin, Alex D. ^{[4
]}

Santamaria-Pang, Alberto ^{[5
]}

Li, Qing ^{[5
]}

Nofech-Mozes, Sharon ^{[2
,3
]}

Liu, Kela ^{[1
]}

Pang, Zhengyu ^{[5
]}

Filkins, Robert J. ^{[5
]}

Yaffe, Martin J. ^{[1
,6
,7
]}

机构：

[1] Sunnybrook Res Inst, Toronto, ON, Canada

[2] Sunnybrook Hlth Sci Ctr, Dept Anat Pathol, Toronto, ON M4N 3M5, Canada

[3] Univ Toronto, Fac Med, Dept Lab Med & Pathobiol, Toronto, ON, Canada

[4] GE Co, Global Res Ctr, Elect Technol & Syst, Niskayuna, NY 12309 USA

[5] GE Co, Global Res Ctr, Diagnost & Biomed Technol, Niskayuna, NY 12309 USA

[6] Univ Toronto, Dept Med Biophys, Fac Med, Toronto, ON, Canada

[7] Univ Toronto, Dept Med Imaging, Fac Med, Toronto, ON, Canada

来源：

HISTOPATHOLOGY | 2014年 / 64卷 / 02期

关键词：

biomarker multiplex imaging; breast cancer; computer-assisted detection; immunofluorescence; microfluidics; ESTROGEN-RECEPTOR; TUMOR HETEROGENEITY; PROGNOSTIC VALUE; KI-67; HER2; PROLIFERATION; SECTIONS; ANTIBODY; PROTEIN; INDEX;

D O I：

10.1111/his.12240

中图分类号：

Q2 [细胞生物学];

学科分类号：

071013 [干细胞生物学];

摘要：

AimsMultiplexed 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 and resultsUsing 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. ConclusionsWe 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.

引用

页码：242 / 255

页数：14

共 43 条

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