Overview of multiplex immunohistochemistry/immunofluorescence techniques in the era of cancer immunotherapy

Abstract

Conventional immunohistochemistry (IHC) is a widely used diagnostic technique in tissue pathology. However, this technique is associated with a number of limitations, including high inter-observer variability and the capacity to label only one marker per tissue section. This review details various highly multiplexed techniques that have emerged to circumvent these constraints, allowing simultaneous detection of multiple markers on a single tissue section and the comprehensive study of cell composition, cellular functional and cell-cell interactions. Among these techniques, multiplex Immunohistochemistry/Immunofluorescence (mIHC/IF) has emerged to be particularly promising. mIHC/IF provides high-throughput multiplex staining and standardized quantitative analysis for highly reproducible, efficient and cost-effective tissue studies. This technique has immediate potential for translational research and clinical practice, particularly in the era of cancer immunotherapy.

A robust multiplex immunofluorescence and digital pathology workflow for the characterisation of the tumour immune microenvironment

“…Equally, there are several dia software that are compatible with most mIF imaging modalities, including Indica Labs' HALO and Visiopharm's Oncotopix. However, unlike qupath , these proprietary software do not offer scripting functionalities, a feature considered essential to our study [35]. We also demonstrate the importance of minimising pre‐analytical variables for the assessment of the resultant digital image, including biomarker combinations of interest and tissue thickness.…”

Prognostic Significance of CD4+ and CD8+ Tumor-Infiltrating Lymphocytes in Head and Neck Squamous Cell Carcinoma: A Meta-Analysis

“…Novel development of multi-parameter assessment methods to view several cell types and corresponding marker expression may facilitate these goals. Such as multiplex fluorescence immunohistochemistry, which is a promising technique that has the ability to simultaneously assess multiple cell subsets in situ, maximize data harvesting per tissue section, improve the quality and detail of pathological analysis and thus efficient tissue use [ 58 ]. Finally, at present it is unclear whether conventional treatment protocols for HNSCC inhibit or promote tumor immune response mechanisms.…”

Transcriptional Spatial Profiling of Cancer Tissues in the Era of Immunotherapy: The Potential and Promise

“…Multiplex immunohistochemistry/immunofluorescence (mIHC/IF) is one commonly used tool that enables the simultaneous detection of up to 40 markers of interest in a single tissue sample [ 15 , 16 ]. This approach was better able to predict the cellular response to PD-1/PD-L1 therapy compared to analyses of the tumour mutational burden (TMB) or gene expression profile (GEP) alone [ 17 ].…”