Multiplex fluorescence immunohistochemistry (mIHC) service



Multiplex immunohistochemistry (mIHC) is an advanced multidimensional tissue detection technique. This technique enables panoramic analysis of the tumor immune microenvironment, labeling multiple biomarkers on a single tissue section. It allows for the insitu detection of multiple target sites in cells or tissue samples. By studying the combination and positional relationships of these targets, the interaction mechanisms can be elucidated, solving the challenge of detecting multiple biomarkers of the same species on a single section. Quantitative pathological analysis provides information on the types, components, and expression levels of various targets in situ within tissue cells, as well as the spatial location, qualitative and quantitative information of each target and its interactions.


Multiplex immunohistochemical staining primarily utilizes tyramide signal amplification (TSA) technology, employing horseradish peroxidase (HRP) for high-density in-situ labeling of target proteins or nucleic acids—an enzymatic detection method. Fluorescently labeled tyramide molecules are activated by HRP labeled with secondary antibody in an H2O2 environment, generating a large number of enzymatic reactions. This causes fluorescein to covalently bind to protein residues (including tryptophan, histidine, and tyrosine residues) around the tissue at the antigen-antibody binding site, forming a large amount of fluorescein deposition and achieving signal amplification.




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mIHC Personalized Technology Service Process

1. Multicolor target + multicolor staining 2. Sample species and type 3. Antibody confirmation
Confirm requirements and deliver samples
1. Multicolor target + multicolor staining
2. Sample species and type
3. Antibody confirmation
1. IHC staining localization 2. Single staining test 3. PDF delivery results
Preliminary experiment (single-standard test)
1. IHC staining localization
2. Single staining test
3. PDF delivery results
1. Multi-standard pre-testing 2. PDF deliverables
Preliminary experiments (multiple tests)
1. Multi-standard pre-testing
2. PDF deliverables
1. Multiple staining markers 2. Data acquisition
Formal Experiment
1. Multiple staining markers
2. Data acquisition
1. Panoramic scan results 2. Data analysis using MIKAIA software
Data Acceptance
1. Panoramic scan results
2. Data analysis using MIKAIA software


Service Description

Three-color and four-color, four-color and five-color, five-color and six-color, six-color and seven-color...
Dyeing types:
Three-color and four-color, four-color and five-color, five-color and six-color, six-color and seven-color...
Fixed tissue samples, tissue blocks, paraffin section
Acceptable sample types:
Fixed tissue samples, tissue blocks, paraffin section
Preliminary experiment: 3-5 images/target/panel/sample
Number of images required:
Preliminary experiment: 3-5 images/target/panel/sample
2 panels/sample (1 panel for backup)
Formal Experiment:
2 panels/sample (1 panel for backup)
Adjusted according to specific needs such as the number of targets, panel combinations, and sample quantity to ensure efficient delivery.
Service cycle:
Adjusted according to specific needs such as the number of targets, panel combinations, and sample quantity to ensure efficient delivery.
Preliminary experiments 2-4 weeks, formal experiments 2-4 weeks (from the date all samples and antibodies arrive).
Typical cycle:
Preliminary experiments 2-4 weeks, formal experiments 2-4 weeks (from the date all samples and antibodies arrive).




Multiplex immunohistochemical staining results


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Figure 1: Human colon cancer tissue (CK-pan, CD4, CD8, CD68, PD-L1 multi-target staining)


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Figure 2: Human lung cancer tissue (multi-target staining of CK-pan, CD3, CD4, CD8, CD68, and Ki67)


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Figure 3: Human colon cancer tissue (CK-pan, CD4, CD8, CD68, PD-L1 multi-target staining)


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Figure 4: Human lung cancer tissue (CK-pan, CD4, CD8, Ki-67 multi-target staining)