Related Experiment Video
Updated: Aug 8, 2026
Quantitation of Protein Expression and Co-localization Using Multiplexed Immuno-histochemical Staining and Multispectral Imaging
Published on: April 8, 2016
Interpretation and quantification of immunostains
T Seidal1, A J Balaton, H Battifora
1Department of Pathology, Central Hospital, Karlstad, Sweden. tomas.seidal@liv.se
Insights
Standardizing immunohistochemistry (IHC) requires clear interpretation guidelines and reproducible methods. Implementing interlaboratory controls and automated analysis can enhance IHC precision and communication in pathology.
Area of Science:
- Pathology and laboratory medicine
- Biomedical research
- Diagnostic techniques
Background:
- Immunohistochemistry (IHC) is a cornerstone of diagnostic pathology and cytology.
- Current IHC practices lack sufficient standardization, impacting diagnostic reliability.
- Reproducibility and clear interpretation criteria are essential for IHC.
Purpose of the Study:
- To highlight the need for standardization in immunohistochemistry.
- To propose methods for improving the reproducibility and precision of IHC.
- To facilitate better communication within the field of IHC.
Main Methods:
- Defining interpretation parameters: microanatomic distribution, cell proportion, staining intensity, and cutoff levels.
- Emphasizing the need for reproducible parameters in publications.
- Suggesting interlaboratory control materials with defined antigen amounts.
- Recommending automated computer-assisted microspectrophotometry for quantitative IHC.
Main Results:
- Standardization of IHC is currently insufficient despite its widespread use.
- Clear, reproducible interpretation parameters are crucial for reliable immunostaining.
- Interlaboratory controls and advanced analytical methods can improve IHC accuracy.
Conclusions:
- Adoption of standardized interpretation and quantitative methods is vital for IHC.
- Improved standardization will enhance diagnostic accuracy and inter-laboratory communication.
- Implementing control materials and automated analysis promises more reliable IHC results.
Abstract:
Despite the fact that immunohistochemistry is widely used in routine diagnostic work and is a very common part of scientific reports in pathology and cytology, its standardization still lags behind. Interpretation of immunostains should be based on microanatomic distribution of the staining, proportion of positively stained cells, staining intensity, if relevant, and cutoff levels. These parameters should be shown to be reasonably reproducible and should be clearly defined in publications. Uniformity in the setting of thresholds could probably benefit from interlaboratory control materials containing defined amounts of the target antigen. Reliable and precise quantitative immunohistochemistry requires the use of control materials containing defined amounts of the target antigen and processed alongside the specimen combined with automated computer-assisted microspectrophotometry. Application of these suggestions is hoped to improve standardization and to facilitate communication in the field of immunohistochemistry.
