Related Experiment Video
Updated: Jun 6, 2026
Dual-modality Molecular Cartography: Integrating Multiplex mRNA Detection with Protein Imaging Mass Cytometry
Published on: November 14, 2025
Quantitative proteomic profiling of breast cancers using a multiplexed microfluidic platform for immunohistochemistry
Minseok S Kim1, Seyong Kwon, Taemin Kim
1Department of Bio and Brain Engineering, Korea Advanced Institute of Science and Technology (KAIST), Yuseong-gu, Daejeon, Republic of Korea.
Insights
A novel microfluidic platform enables multiplexed immunohistochemistry (IHC) and immunocytochemistry (ICC) for rapid, quantitative proteomic profiling in breast cancer. This technology aids in accurate diagnosis and personalized cancer therapy by analyzing multiple biomarkers simultaneously.
Area of Science:
- Biomedical Engineering
- Cancer Research
- Proteomics
Background:
- Accurate proteomic profiling is crucial for breast cancer subtyping and targeted therapy.
- Conventional immunohistochemistry (IHC) and immunocytochemistry (ICC) methods can be time-consuming and require large sample volumes.
- Multiplexed analysis of biomarkers is essential for comprehensive understanding of cancer heterogeneity.
Purpose of the Study:
- To develop and validate a multiplexed microfluidic platform for high-throughput IHC/ICC.
- To enable quantitative proteomic profiling of breast cancer cell lines and clinical samples.
- To assess the platform's efficiency, accuracy, and applicability in diagnosing breast cancer.
Main Methods:
- A microfluidic device was designed for simultaneous execution of 20 ICC assays.
- Four breast cancer cell lines (AU-565, HCC70, MCF-7, SK-BR-3) were analyzed for proteomic expression.
- The platform's performance was compared to conventional IHC methods using human breast cancer tissue.
Main Results:
- The microfluidic system reduced assay time by 16-fold and enabled quantitative biomarker comparison.
- Protein localization and staining quality were comparable to conventional IHC.
- Distinct proteomic profiles were observed in different cell lines, including triple-negative and basal/myoepithelial markers.
- Successful application to clinical breast cancer tissue demonstrated diagnostic utility.
Conclusions:
- The multiplexed microfluidic IHC/ICC platform offers a rapid and quantitative approach for proteomic profiling.
- This technology facilitates accurate histopathological diagnosis by analyzing numerous biomarkers simultaneously.
- The platform supports personalized cancer therapy by providing detailed insights into individual tumor characteristics.
Abstract:
This paper describes a multiplexed microfluidic immunohistochemistry (IHC)/immunocytochemistry (ICC) platform for quantitative proteomic profiling in breast cancer samples. Proteomic profiling via ICC was examined for four breast cancer cell lines (AU-565, HCC70, MCF-7, and SK-BR-3). The microfluidic device enabled 20 ICC assays on a biological specimen at the same time and a 16-fold decrease in time consumption, and could be used to quantitatively compare the expression level of each biomarker. The immunohistochemical staining from the microfluidic system showed an accurate localization of protein and comparable quality to that of the conventional IHC method. Although AU-565 and SK-BR-3 cell lines were classified by luminal subtype and adenocarcinomas and were derived from the same patient, weak p63 expression was seen only in SK-BR-3. The HCC70 cell line showed a triple-negative (estrogen receptor-negative/progesterone receptor-negative/human epidermal growth factor receptor 2-negative) phenotype and showed only cytokeratin 5 expression, a representative basal/myoepithelial cell marker. To demonstrate the applicability of the system to clinical samples for proteomic profiling, we were also able to apply this platform to human breast cancer tissue. This result indicates that the microfluidic IHC/ICC platform is useful for accurate histopathological diagnoses using numerous specific biomarkers simultaneously, facilitating the individualization of cancer therapy.
