Research Topic

Multiplex Immunohistochemistry/Immunofluorescence Technique: The Potential and Promise for Clinical Application

About this Research Topic

Cancer immunotherapy (such as anti-PD-L1/PD-1 therapy) has advanced exponentially in recent years. However, only a limited number of patients benefit from this treatment, and there is a huge drop off from discoveries in research laboratories making it to the clinical practice. This is largely due to the limitation of suitable testing technique and technology. Conventional immunohistochemistry (IHC) has long been a “gold standard” diagnostic technique in tissue pathology. However, this technique suffers from limitations including high inter-observer variability and the small labeling capacity for one to two biomarkers per tissue section. This represents a major drawback with missed opportunities to harvest additional information of diagnostic-prognostic-predictive importance from patient samples.

On the other hand, Multiplex Immunohistochemistry/Immunofluorescence (mIHC/IF) has emerged to complement the conventional IHC as it can detect multiple biomarkers on a single tissue section, providing high-throughput multiplex staining and standardized quantitative analysis for highly reproducible, efficient, and cost-effective tissue testing. The ability to label multiple markers on a single section is of particular significance when studying samples taken from rare donors, where tissues may be of low availability. Having insufficient tissue for testing is a serious concern in the day-to-day clinical practice, especially during clinical trials and when using biopsy samples. For instance, the majority of biopsy samples have to be spared for molecular testing, which leaves limited samples for diagnosis by using conventional IHC. Cutting more consecutive sections may be impossible in this scenario, and this patient would lose the opportunity to benefit from the era of personalized medicine for immunotherapy.

In summary there is an urgent need for mIHC/IF for routine clinical practice in the era of cancer immunotherapy. Multiple biomarkers are needed to stratify patients, identifying responsive patients who can benefit, and sparing non-responders from unnecessary side-effects and cost. This Research Topic seeks to coordinate and discuss the promise and potential barriers for the clinical application of mIHC/IF from both scientific and technical perspectives in immuno-oncology and immuno-pathology. The topic will include both original articles and extensive reviews highlighting influential discoveries and applications of mIHC/IF in translational research. Topics may cover, but are not limited to, the following:

• Biomarkers Discovery by using mIHC/IF
• Spatial information of Tumor immune micro-environment by using mIHC/IF
• Immuno-phenotyping by using mIHC/IF
• Technical Reproducibility of mIHC/IF
• Comparison of mIHC/IF with other modality such as conventional IHC, RNAseq, NanoString
• High-dimensional analysis of mIHC/IF
• Analytic pipeline and algorithm of mIHC/IF
• Automation, standardization and upscaling the throughput of of mIHC/IF
• Comparison of two or more mIHC/IF techniques (e.g. DNA barcoding vs fluorescence vs metal-conjugation)
• Clinical application and translation of Biomarkers with mIHC/IF


Professor Bernard A Fox is the CEO of Ubivac, and Dr. Jaime Rodriguez is a full time employee of AstraZeneca. All other Topic Editors report no competing interests.


Keywords: multiplex IHC/IF, immunohistochemistry, clinical translation, pathology, immunotherapy


Important Note: All contributions to this Research Topic must be within the scope of the section and journal to which they are submitted, as defined in their mission statements. Frontiers reserves the right to guide an out-of-scope manuscript to a more suitable section or journal at any stage of peer review.

Cancer immunotherapy (such as anti-PD-L1/PD-1 therapy) has advanced exponentially in recent years. However, only a limited number of patients benefit from this treatment, and there is a huge drop off from discoveries in research laboratories making it to the clinical practice. This is largely due to the limitation of suitable testing technique and technology. Conventional immunohistochemistry (IHC) has long been a “gold standard” diagnostic technique in tissue pathology. However, this technique suffers from limitations including high inter-observer variability and the small labeling capacity for one to two biomarkers per tissue section. This represents a major drawback with missed opportunities to harvest additional information of diagnostic-prognostic-predictive importance from patient samples.

On the other hand, Multiplex Immunohistochemistry/Immunofluorescence (mIHC/IF) has emerged to complement the conventional IHC as it can detect multiple biomarkers on a single tissue section, providing high-throughput multiplex staining and standardized quantitative analysis for highly reproducible, efficient, and cost-effective tissue testing. The ability to label multiple markers on a single section is of particular significance when studying samples taken from rare donors, where tissues may be of low availability. Having insufficient tissue for testing is a serious concern in the day-to-day clinical practice, especially during clinical trials and when using biopsy samples. For instance, the majority of biopsy samples have to be spared for molecular testing, which leaves limited samples for diagnosis by using conventional IHC. Cutting more consecutive sections may be impossible in this scenario, and this patient would lose the opportunity to benefit from the era of personalized medicine for immunotherapy.

In summary there is an urgent need for mIHC/IF for routine clinical practice in the era of cancer immunotherapy. Multiple biomarkers are needed to stratify patients, identifying responsive patients who can benefit, and sparing non-responders from unnecessary side-effects and cost. This Research Topic seeks to coordinate and discuss the promise and potential barriers for the clinical application of mIHC/IF from both scientific and technical perspectives in immuno-oncology and immuno-pathology. The topic will include both original articles and extensive reviews highlighting influential discoveries and applications of mIHC/IF in translational research. Topics may cover, but are not limited to, the following:

• Biomarkers Discovery by using mIHC/IF
• Spatial information of Tumor immune micro-environment by using mIHC/IF
• Immuno-phenotyping by using mIHC/IF
• Technical Reproducibility of mIHC/IF
• Comparison of mIHC/IF with other modality such as conventional IHC, RNAseq, NanoString
• High-dimensional analysis of mIHC/IF
• Analytic pipeline and algorithm of mIHC/IF
• Automation, standardization and upscaling the throughput of of mIHC/IF
• Comparison of two or more mIHC/IF techniques (e.g. DNA barcoding vs fluorescence vs metal-conjugation)
• Clinical application and translation of Biomarkers with mIHC/IF


Professor Bernard A Fox is the CEO of Ubivac, and Dr. Jaime Rodriguez is a full time employee of AstraZeneca. All other Topic Editors report no competing interests.


Keywords: multiplex IHC/IF, immunohistochemistry, clinical translation, pathology, immunotherapy


Important Note: All contributions to this Research Topic must be within the scope of the section and journal to which they are submitted, as defined in their mission statements. Frontiers reserves the right to guide an out-of-scope manuscript to a more suitable section or journal at any stage of peer review.

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Submission Deadlines

29 January 2021 Manuscript
26 February 2021 Manuscript Extension

Participating Journals

Manuscripts can be submitted to this Research Topic via the following journals:

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Topic Editors

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Submission Deadlines

29 January 2021 Manuscript
26 February 2021 Manuscript Extension

Participating Journals

Manuscripts can be submitted to this Research Topic via the following journals:

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