title: Immunodetection of NETs in Paraffin-Embedded Tissue.
abstract: The pathogenic potential of neutrophil extracellular traps (NETs) was recently described, and their detection in tissue could serve as a prognostic marker. NETs are delicate and filigree structures; hence good tissue preservation is essential for their detection. Indeed, analysis of paraffin-embedded tissue has proven superior to the study of cryo sections. Though, under favorable conditions, the presence of NETs can be detected in tissue sections stained with histological dyes, definitive identification of NETs needs the colocalization of immunofluorescent signals for both nuclear and granular (or cytoplasmic) NET components. We tested diverse antigen retrieval methods and various combinations of commercially available antibodies and present here staining protocols to detect NETs in human and murine tissue sections.
authors.affiliation: Microscopy Core Facility, Max Planck Institute for Infection Biology , Berlin , Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Brinkmann
authors.affiliation: Microscopy Core Facility, Max Planck Institute for Infection Biology, Berlin, Germany; Department of Cellular Microbiology, Max Planck Institute for Infection Biology, Berlin, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Abu Abed
authors.affiliation: Microscopy Core Facility, Max Planck Institute for Infection Biology , Berlin , Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Goosmann
authors.affiliation: Department of Cellular Microbiology, Max Planck Institute for Infection Biology , Berlin , Germany.
authors.forename: Arturo
authors.initials: A
authors.lastname: Zychlinsky
keywords: NETs; antigen retrieval; image analysis; immunodetection; paraffin-embedded tissue