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Updated: Jun 23, 2026
Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
Published on: December 20, 2014
Quantitative, fluorescence-based in-situ assessment of protein expression
Christopher B Moeder1, Jennifer M Giltnane, Sharon Pozner Moulis
1Department of Pathology, Yale University School of Medicine, New Haven, CT, USA.
Insights
A new immunofluorescence-based method, AQUA technology, offers accurate in situ protein quantification for companion diagnostics. This automated approach provides reproducible, continuous data, surpassing traditional immunohistochemistry limitations.
Area of Science:
- Biomedical Engineering
- Molecular Pathology
- Cancer Diagnostics
Background:
- Companion diagnostics require precise in situ protein quantification.
- Traditional immunohistochemistry (IHC) lacks rigorous quantitative capabilities.
- Existing methods struggle with accurate protein measurement in tissues.
Purpose of the Study:
- To develop and validate an accurate, reproducible method for in situ protein quantification.
- To overcome the limitations of traditional IHC for quantitative analysis.
- To introduce the AQUA technology for precise protein expression measurement.
Main Methods:
- Development of an immunofluorescence-based assay.
- Commercialization of the AQUA technology for image analysis.
- Automated staining, image acquisition, and AQUA software analysis of tissue samples.
Main Results:
- AQUA technology enables rapid, efficient, and accurate measurement of protein expression levels.
- High reproducibility and accuracy comparable to ELISA assays.
- Automated process minimizes error, providing continuous scale scores.
Conclusions:
- AQUA technology provides a robust solution for quantitative in situ protein analysis.
- This method enhances the utility of IHC for biomarker discovery and diagnostics.
- The technology facilitates novel observations not possible with traditional methods.
Abstract:
As companion diagnostics grow in prevalence and importance, the need for accurate assessment of in situ protein concentrations has increased. Traditional immunohistochemistry (IHC), while valuable for assessment of context of expression, is less valuable for quantification. The lack of rigorous quantitative potential of traditional IHC led to our development of an immunofluorescence-based method now commercialized as the AQUA technology. Immunostaining of tissue samples, image acquisition, and use of AQUA software allow investigators to quickly, efficiently, and accurately measure levels of expression within user-defined subcellular or architectural compartments. IHC analyzed by AQUA shows high reproducibility and demonstrates protein measurement accuracy similar to ELISA assays. The process is largely automated, eliminating potential error, and the resultant scores are exported on a continuous scale. There are now numerous published examples where observations made with this technology are not seen by traditional methods.
