# Multiplex immunofluorescence

Source: https://onco.cc/technologies/multiplex-immunofluorescence/  
OnCo record `multiplex-immunofluorescence` (Technology). Data CC BY-NC 4.0, attribute "Data from OnCo (onco.cc)"; commercial use needs a licence.

## TL;DR

Staining one tumour slide for several proteins in different colours, then using software to count immune and cancer cells and measure how close they are.

## Summary

Multiplex immunofluorescence extends standard immunohistochemistry to six to eight markers per slide using tyramide signal amplification and multispectral imaging (Akoya Phenoptics), or to dozens with cyclic staining (CODEX/PhenoCycler). Combined with image analysis it quantifies immune-cell composition and spatial arrangement, which large meta-analyses have found to predict response to checkpoint inhibitors better than PD-L1 staining alone.

## Fields

- Kind: Technology
- Status: established
- Last checked: 2026-09-04
- Principle: Sequential antibody staining with fluorescent tyramide deposition, antibody stripping between rounds, and spectral unmixing to separate channels.
- Strengths: Uses standard pathology sections; Captures cell-to-cell proximity; Better predictor of immunotherapy response than single-marker IHC in pooled studies
- Limitations: Panel design and validation are laborious; Inter-laboratory standardisation incomplete; Not yet a routine clinical test

## Sources

- Wikipedia: https://en.wikipedia.org/wiki/Immunofluorescence
- Wikipedia: https://en.wikipedia.org/wiki/Immunofluorescence

## Connected records

- fronts: [Diagnostics & Biomarkers](https://onco.cc/fronts/diagnostics/)
- technologies: [Digital pathology & AI](https://onco.cc/technologies/digital-pathology-ai/)

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