Open AccessCancer immunophenotyping by seven‐colour multispectral imaging without tyramide signal amplification
Author(s) -
Ijsselsteijn Marieke E,
Brouwer Thomas P,
Abdulrahman Ziena,
Reidy Eileen,
Ramalheiro Ana,
Heeren A Marijne,
Vahrmeijer Alexander,
Jordanova Ekaterina S,
de Miranda Noel FCC
Publication year - 2019
Publication title -
the journal of pathology: clinical research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.849
H-Index - 21
ISSN - 2056-4538
DOI - 10.1002/cjp2.113
Subject(s) - immunophenotyping , multiplex , context (archaeology) , multispectral image , immunotherapy , computational biology , biology , immune system , computer science , immunology , bioinformatics , flow cytometry , artificial intelligence , paleontology
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‐free method for the simultaneous detection of seven 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.