title: Multiparameter immunofluorescence on paraffin-embedded tissue sections.
abstract: Immunohistochemical techniques have gained increasing importance in diagnostics and research. While formalin-fixed, paraffin-embedded human tissue retains excellent morphology, the detection of antigens by immunofluorescence in its sections and especially the demonstration of multiple simultaneous antibodies have limitations. Double immunofluorescence labeling of routinely processed paraffin sections has been described previously. The signal intensity observed after triple labeling has been reported to be significantly inferior to that obtained by application of double fluorochromes. The authors show multicolor labeling of three and four primary antibodies in routinely processed paraffin-embedded tissue sections using a standardized immunofluorescence technique. In addition, procedures to reduce background staining and to avoid nonspecific double staining are described.
authors.affiliation: Institute of Pathology, University of Regensburg, Germany. frauke.bataille@klinik.uni-regensburg.de
authors.forename: Frauke
authors.initials: F
authors.lastname: Bataille
authors.forename: Sabine
authors.initials: S
authors.lastname: Troppmann
authors.forename: Frank
authors.initials: F
authors.lastname: Klebl
authors.forename: Gerhard
authors.initials: G
authors.lastname: Rogler
authors.forename: Benjamin
authors.initials: B
authors.lastname: Stoelcker
authors.forename: Ferdinand
authors.initials: F
authors.lastname: Hofstadter
authors.forename: Anja-Katrin
authors.initials: AK
authors.lastname: Bosserhoff
authors.forename: Petra
authors.initials: P
authors.lastname: Rümmele
mesh_terms: D000906:Antibodies / Q000032:analysis* / Q000737:chemistry; D001808:Blood Vessels / Q000166:cytology / Q000473:pathology; D005455:Fluorescent Antibody Technique / Q000379:methods*; D005456:Fluorescent Dyes; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods*; D007413:Intestinal Mucosa / Q000473:pathology*; D016612:Paraffin Embedding; D013194:Staining and Labeling / Q000379:methods*; D013601:T-Lymphocytes / Q000276:immunology
chemical_list: D000906:Antibodies; D005456:Fluorescent Dyes