Cancer immunophenotyping by 7 colour multispectral imaging without tyramide signal amplification

Marieke Ijselstein, Thomas Brouwer, Ziena Abdulrahman, Eileen Reidy, Ana Ramalheiro, AM Heeren, Alexander Vahrmeijer, ES Jordanova, Noel de Miranda

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Checkpoint blockade immunotherapies have revolutionised cancer treatment in the last decade. Nevertheless, these are only beneficial for a small proportion of cancer patients. Important prognosticators for response to immunotherapy are the mutation burden of tumours as well as the quality and quantity of tumour‐infiltrating immune cells. High‐throughput multiplex immunophenotyping technologies have a central role in deciphering the complexity of anti‐tumour immune responses. Current techniques for the immunophenotyping of solid tumours are held back by the lack of spatial context, limitations in the number of targets that can be visualised simultaneously, and/or cumbersome protocols. We developed a tyramide signal amplification (TSA) – free method for the simultaneous detection of 7 cellular targets by immunofluorescence. This method overcomes limitations posed by most widespread techniques and provides a unique tool for extensive phenotyping by multispectral fluorescence microscopy. Furthermore, it can be easily implemented as a high‐throughput technology for validation of discovery sets generated by RNA sequencing or mass cytometry and may serve in the future as a complementary diagnostic tool.
Original languageEnglish
Pages (from-to)3-11
JournalThe Journal of Pathology: Clinical Research
Volume5
Issue number1
Early online date7 Sep 2018
DOIs
Publication statusPublished - Jan 2019

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