title: Whole-slide, quadruple immunofluorescence labeling of routinely processed paraffin sections.
abstract: Whole-slide images (WSI) have acquired a stable place in diagnostic histopathology and immunohistochemistry. Immunofluorescence (IF) techniques hold a limited and selective role in diagnostics (eg, renal and cutaneous pathology) and so far remain excluded from the digital pathology evolution, with notable exceptions, such as quantitative immunopathology. We explored the ability of a commercial fluorescent slide scanner to provide 4-color IF WSI from routinely processed tissues. With minor modifications and a careful match between filters and fluorochromes, we show that 4-color IF WSI can be obtained from routine material with negligible autofluorescence, good sensitivity, and diagnostic power.
authors.affiliation: *Department of Pathology, Universitá degli Studi di Milano-Bicocca †Azienda Ospedaliera San Gerardo, Monza, Milano, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Buscone
authors.forename: Maria C
authors.initials: MC
authors.lastname: Argentieri
authors.forename: Daniela
authors.initials: D
authors.lastname: Pilla
authors.forename: Giorgio
authors.initials: G
authors.lastname: Cattoretti
mesh_terms: D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000473:pathology; D005404:Fixatives; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes / Q000737:chemistry*; D005557:Formaldehyde; D006801:Humans; D008867:Microtomy; D009506:Nevus / Q000175:diagnosis* / Q000473:pathology; D010232:Paraffin; D013194:Staining and Labeling / Q000379:methods*; D013601:T-Lymphocytes / Q000473:pathology; D016610:Tissue Embedding
chemical_list: D000911:Antibodies, Monoclonal; D005404:Fixatives; D005456:Fluorescent Dyes; D005557:Formaldehyde; D010232:Paraffin