Related Experiment Video
Updated: Feb 25, 2026
High-Throughput Automated Multiplex Immunofluorescence Assays for Translational Research
Published on: June 10, 2025
Immunohistochemistry Microarrays
Huiyan Li1, Gabrielle Brewer1, Grant Ongo1
1Biomedical Engineering Department, ‡McGill University and Genome Quebec Innovation Centre, §Department of Pathology, McGill University Health Centre, and ∥Department of Neurology and Neurosurgery, McGill University , Montreal, Quebec H3A0G1, Canada.
Insights
We developed IHC microarrays (IHCμA) for multiplexed staining of intact tissues, enabling simultaneous analysis of multiple proteins. This technique preserves tissue architecture and spatial information, advancing biomarker discovery in cancer research.
Area of Science:
- Biomedical Engineering
- Molecular Pathology
- Cancer Research
Background:
- Conventional immunohistochemistry (IHC) is crucial for cancer diagnosis but limited to single-protein analysis per tissue section.
- Existing high-throughput methods like tissue microarrays and microfluidics have limitations in multiplexing, complexity, or analyzing intact tissues.
- There is a need for methods that allow multiplexed protein analysis on intact tissues while preserving histological and spatial context.
Purpose of the Study:
- To introduce and validate IHC microarrays (IHCμA) as a novel method for multiplexed protein staining on intact tissue sections.
- To demonstrate the ability of IHCμA to preserve histological and spatial information during multiplexed staining.
- To evaluate the potential of IHCμA for high-throughput biomarker discovery and validation in cancer.
Main Methods:
- Development of IHC microarrays (IHCμA) by prespotting antibody-containing dextran solution droplets onto slides.
- Application of IHCμA to formalin-fixed, paraffin-embedded (FFPE) tissue sections by snapping prespotted slides onto tissue slices.
- Evaluation of sampling considerations and demonstration of multiplexed staining of 8 cancer proteins using 180 spots on a breast cancer tissue section.
Main Results:
- IHCμA enables multiplexed staining of intact tissues with preserved histological and spatial information, similar to conventional IHC.
- The method allows prespotting and storage of antibody microarrays, reducing the need for complex instrumentation and immediate preparation.
- Demonstrated successful multiplexed staining of 8 cancer proteins on a breast cancer tissue, showcasing the technique's potential.
Conclusions:
- IHC microarrays (IHCμA) offer a powerful new approach for multiplexed immunohistochemistry on intact tissue slices.
- This technique overcomes limitations of current methods, facilitating simultaneous analysis of multiple proteins with preserved spatial context.
- IHCμA holds significant potential for accelerating the discovery and validation of tissue biomarkers in human tumors.
Abstract:
Immunohistochemistry (IHC) on tissue sections is widely used for quantifying the expression patterns of proteins and is part of the standard of care for cancer diagnosis and prognosis, but is limited to staining a single protein per tissue. Tissue microarray and microfluidics staining methods have emerged as powerful high throughput techniques, but they either only permit the analysis of a single protein per slide or require complex instrumentation and expertise while only staining isolated areas. Here, we introduce IHC microarrays (IHCμA) for multiplexed staining of intact tissues with preserved histological and spatial information. Droplets of a dextran solution containing antibodies were prespotted on a slide and snapped onto a preprocessed formalin-fixed, paraffin-embedded (FFPE) tissue section soaked in a polyethylene glycol solution. The antibodies are confined within the dextran droplets and locally stain the tissue below with a contrast similar to the one obtained by conventional IHC. The microarray of antibody droplets can be prespotted on a slide and stored, thus neither the preparation of the antibody solutions nor a sophisticated microarray spotter is needed. Sampling considerations with IHCμA were evaluated by taking three tissues with varying levels of cancer cells. A multiplex IHCμA with 180 spots targeting 8 cancer proteins was performed on a breast cancer tissue section to illustrate the potential of this method. This work opens the avenue of applying microarray technologies for conducting IHC on intact tissue slices and has great potential to be used in the discovery and validation of tissue biomarkers in human tumors.
Related Concept Videos
02:34DNA Microarrays
01:22Immunocytochemistry and Immunohistochemistry
These...