Improvement of Opal Multiplex Immunofluorescence Workflow for Human Tissue Sections.

ABSTRACT: The Opal multiplex technique is an established methodology for the detection of multiple biomarkers in one section. The protocol encompasses iterative single stainings and heating-mediated removal of the primary and secondary antibodies after each staining round, leaving untouched the Opal fluorophores which are deposited onto the antigen of interest. According to our experience, repetitive heating of skin sections often results in tissue damage, indicating an urgent need for milder alternatives to strip immunoglobulins. In this study, we demonstrate that considerable heating-related damage was found not only in skin but also in tissues of different origin, mostly characterized by low cell density. Importantly, the morphology remained fully intact when sections were repetitively exposed to

SUBMITTER: Willemsen M 

PROVIDER: S-EPMC8091416 | biostudies-literature | 2021 May

REPOSITORIES: biostudies-literature