Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor MicroenvironmentNicolas Huyghe 1, Elena Benidovskaya 1, Simon Beyaert 1, Aurélie Daumerie 4, Finoula Maestre Osorio 1, Frank Aboubakar Nana 2,3, Caroline Bouzin 4, Marc Van den Eynde 1,5
1Institut de Recherche Expérimentale et Clinique (IREC), Pôle MIRO, Université Catholique de Louvain (UCLouvain), 2Institut de Recherche Expérimentale et Clinique (IREC), Pôle de Pneumologie, ORL et Dermatologie (PNEU), Université Catholique de Louvain (UCLouvain), 3Division of Pneumology, Cliniques Universitaires St-Luc, Université Catholique de Louvain (UCLouvain), 4IREC Imaging Platform, Université Catholique de Louvain (UCLouvain), 5Institut Roi Albert II, Department of Medical Oncology and Gastroenterology, Cliniques Universitaires St-Luc
In this article, a protocol for manual tyramide signal amplification (TSA) multiplex immunofluorescence (mIF) combined with image analysis and spatial analysis is described. This protocol can be used with formalin-fixed paraffin-embedded (FFPE) sections for the staining of two to six antigens per slide depending on the slide scanner available in the laboratory.