title: Technical Aspects and Applications for Developing in situ Hybridization Procedures for Formalin-Fixed and Paraffin-Embedded (FFPE) Tissues for Diagnosis of Fungal Infections.
abstract: Although histopathology is required for definitive diagnosis of fungal infections, conclusive identification and discrimination of fungi in tissue sections and cytological preparations remain technically difficult. Therefore, new diagnostic tools are needed for the routine diagnosis of pathogenic fungi. In situ hybridization (ISH) is a non-culture based procedure that has many advantages over traditional diagnostics for identification of pathogenic fungi in histological specimens. This review highlights the basic ISH technique, with particular emphasis on using pretreatment of tissue sections prior to hybridization to solve problems associated with formalin fixation. With this modification, ISH has become a valuable tool that complements conventional histopathological diagnoses in formalin-fixed and paraffinembedded (FFPE) tissues. However, understanding the limitations imposed by formalin fixation is essential in developing suitable ISH protocols for fungal identification.
authors.affiliation: Department of Surgical Pathology, Toho University School of Medicine.
authors.forename: Minoru
authors.initials: M
authors.lastname: Shinozaki
authors.forename: Naobumi
authors.initials: N
authors.lastname: Tochigi
authors.forename: Sota
authors.initials: S
authors.lastname: Sadamoto
authors.forename: Somay
authors.initials: S
authors.lastname: Yamagata Murayama
authors.forename: Megumi
authors.initials: M
authors.lastname: Wakayama
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Nemoto
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Shibuya
mesh_terms: D005557:Formaldehyde; D005658:Fungi / Q000302:isolation & purification* / Q000472:pathogenicity*; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008828:Microbiological Techniques / Q000379:methods*; D009181:Mycoses / Q000175:diagnosis* / Q000382:microbiology; D016612:Paraffin Embedding / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods*
chemical_list: D005557:Formaldehyde