title: The application of direct immunofluorescence to intraoperative neurosurgical diagnosis.
abstract: A diagnostic problem can occur at the time of intraoperative consultation of neurosurgical tumors as to whether the tumor is of neuroectodermal origin or whether it represents an epithelial metastasis from another site. Intraoperative diagnoses based on hematoxylin and eosin stained frozen sections are often later confirmed by immunocytochemical analysis of formalin-fixed, paraffin-embedded tissue sections that are not available at the time of surgery. The objective of the current study was to demonstrate that the application of direct immunofluorescence to the intraoperative diagnosis of neurosurgical tumors would provide unequivocal, and nearly immediate results. This report describes a new application of an existing technique for an optimized, rapid procedure utilizing direct immunocytochemistry with fluorescence-labeled primary antibodies to analyze surgical biopsies intraoperatively. The examination of five neurosurgical biopsies established a neuroectodermal origin of three tumors via immunolabeling for glial fibrillary acidic protein (GFAP) and lack of labeling with keratin markers, whereas several metastatic lung carcinomas were identified by immunostaining for keratin, but not GFAP, markers. The results of the direct immunolabeling method were unequivocal and required only minutes. The same diagnoses were confirmed by standard immunocytochemical labeling of formalin-fixed, paraffin-embedded sections, though it required several days to obtain the results. Direct immunofluorescence using fluorescently conjugated primary antibodies is a practical and rapid method for deciding whether a neurosurgical tumor is a primary glial or an epithelial metastatic tumor in origin. It is the first reported application of the technique for this aspect of rapid neurosurgical diagnosis.
authors.affiliation: Department of Physiology and Biophysics, University of Washington, Seattle 98195, USA. siwamoto@u.washington.edu
authors.forename: S
authors.initials: S
authors.lastname: Iwamoto
authors.forename: R C
authors.initials: RC
authors.lastname: Burrows
authors.forename: D E
authors.initials: DE
authors.lastname: Born
authors.forename: M
authors.initials: M
authors.lastname: Piepkorn
authors.forename: M
authors.initials: M
authors.lastname: Bothwell
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D001254:Astrocytoma / Q000378:metabolism* / Q000473:pathology / Q000648:ultrastructure; D001932:Brain Neoplasms / Q000175:diagnosis* / Q000378:metabolism* / Q000473:pathology / Q000601:surgery; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D007432:Intraoperative Period; D007633:Keratins / Q000378:metabolism*; D009493:Neurosurgery / Q000379:methods*; D016612:Paraffin Embedding
chemical_list: D000911:Antibodies, Monoclonal; D005904:Glial Fibrillary Acidic Protein; D007633:Keratins