Protocol for the quantitative analysis of images retrieved by multiplex immunofluorescence staining to allow cell type-specific spatial phenotyping of markers of interest in the human placenta.

In contrast to other tissues, the placenta consists of numerous functionally different cell types, distributed in a markedly dissimilar manner within one placenta and between different cases. To evaluate pathology-specific changes in cell phenotype and expression of molecular markers it is important to establish a multi staining method combining immunohistological identification of the cell type with staining of proteins of interest. We successfully established a protocol for a 6-plex antibody panel for multiplex immunofluorescence. Here, we report the staining protocol and the establishment of the quantification algorithm we developed.