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Related Experiment Video

Updated: Jun 28, 2026

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging

Published on: April 8, 2016

Multiplex, Quantitative, High-Resolution Imaging of Protein:Protein Complexes via Hybridization Chain Reaction.

Samuel J Schulte¹, Boyoung Shin¹, Ellen V Rothenberg¹

¹Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, California 91125, United States.

ACS Chemical Biology

|January 17, 2024

Summary

This summary is machine-generated.

Hybridization chain reaction (HCR) imaging now visualizes protein:protein complexes with high signal and low background. This method enables simultaneous multiplex imaging of RNA, protein, and protein complexes for spatial exploration.

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Area of Science:

Molecular Biology
Biotechnology
Cell Biology

Background:

Hybridization chain reaction (HCR) is a powerful technique for amplifying signals in biological imaging.
Current HCR applications primarily focus on imaging RNA and protein targets.
There is a need for methods to visualize protein:protein interactions with high resolution and specificity.

Purpose of the Study:

To extend HCR capabilities for imaging protein:protein complexes.
To develop a method for simultaneous, multiplex, quantitative, and high-resolution imaging of RNA, protein, and protein:protein targets.
To establish a unified framework for multi-target spatial biology.

Main Methods:

Utilized proximity-dependent cooperative probes for conditional signal amplification upon protein colocalization.
Employed HCR probes and amplifiers for automatic background suppression.
Demonstrated the technique in human cells, mouse proT cells, and FFPE human breast tissue sections.

Main Results:

Achieved high signal-to-background ratio for protein:protein complex imaging.
Successfully demonstrated multiplex imaging of three different protein:protein complexes simultaneously.
Validated accurate and precise relative quantitation of protein:protein complexes with subcellular resolution.
Established a unified framework for simultaneous multiplex imaging of RNA, protein, and protein:protein targets using one-step, isothermal, enzyme-free HCR amplification.

Conclusions:

HCR technology can be effectively adapted for the visualization and quantitation of protein:protein complexes.
The developed unified framework enables simultaneous multiplex imaging across RNA, protein, and protein complex targets.
This advancement offers a powerful tool for spatial exploration of complex biological networks.