Clinical and research applications of multiplexed immunohistochemistry and in situ hybridization

J Pathol. 2021 Jul;254(4):405-417. doi: 10.1002/path.5663. Epub 2021 May 6.


Over the past decade, invention and adoption of novel multiplexing technologies for tissues have made increasing impacts in basic and translational research and, to a lesser degree, clinical medicine. Platforms capable of highly multiplexed immunohistochemistry or in situ RNA measurements promise evaluation of protein or RNA targets at levels of plex and sensitivity logs above traditional methods - all with preservation of spatial context. These methods promise objective biomarker quantification, markedly increased sensitivity, and single-cell resolution. Increasingly, development of novel technologies is enabling multi-omic interrogations with spatial correlation of RNA and protein expression profiles in the same sample. Such sophisticated methods will provide unprecedented insights into tissue biology, biomarker science, and, ultimately, patient health. However, this sophistication comes at significant cost, requiring extensive time, practical knowledge, and resources to implement. This review will describe the technical features, advantages, and limitations of currently available multiplexed immunohistochemistry and spatial transcriptomic platforms. © 2021 The Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

Keywords: multiplex immunohistochemistry; multiplex in situ hybridization; single-cell transcriptomes; spatial transcriptomics; surrogate end-points.

Publication types

  • Review

MeSH terms

  • Animals
  • Humans
  • Immunohistochemistry / methods*
  • In Situ Hybridization / methods*