title: Immunohistochemical techniques for the human inner ear.
abstract: In this review, we provide a description of the recent methods used for immunohistochemical staining of the human inner ear using formalin-fixed frozen, paraffin and celloidin-embedded sections. We also show the application of these immunohistochemical methods in auditory and vestibular endorgans microdissected from the human temporal bone. We compare the advantages and disadvantages of immunohistochemistry (IHC) in the different types of embedding media. IHC in frozen and paraffin-embedded sections yields a robust immunoreactive signal. Both frozen and paraffin sections would be the best alternative in the case where celloidin-embedding technique is not available. IHC in whole endorgans yields excellent results and can be used when desiring to detect regional variations of protein expression in the sensory epithelia. One advantage of microdissection is that the tissue is processed immediately and IHC can be made within 1 week of temporal bone collection. A second advantage of microdissection is the excellent preservation of both morphology and antigenicity. Using celloidin-embedded inner ear sections, we were able to detect several antigens by IHC and immunofluorescence using antigen retrieval methods. These techniques, previously applied only in animal models, allow for the study of numerous important proteins expressed in the human temporal bone potentially opening up a new field for future human inner ear research.
authors.affiliation: Head and Neck Surgery Department, David Geffen School of Medicine at UCLA, 951794, 35-64 Rehab, 1000 Veteran Avenue, Los Angeles, CA, 90095, USA.
authors.forename: Ivan A
authors.initials: IA
authors.lastname: Lopez
authors.affiliation: Neurology Department, David Geffen School of Medicine at UCLA, 3-250 RNRC, Los Angeles, CA, 90095, USA.
authors.forename: Gail
authors.initials: G
authors.lastname: Ishiyama
authors.affiliation: Head and Neck Surgery Department, David Geffen School of Medicine at UCLA, 951794, 35-64 Rehab, 1000 Veteran Avenue, Los Angeles, CA, 90095, USA.|Otorhinolaryngology/Head and Neck Surgery Department, School of Medicine, Hamamatsu University, 1-20-1 Handayama, Hamamatsu, 431-3192, Shizuoka, Japan.
authors.forename: Seiji
authors.initials: S
authors.lastname: Hosokawa
authors.affiliation: Head and Neck Surgery Department, David Geffen School of Medicine at UCLA, 951794, 35-64 Rehab, 1000 Veteran Avenue, Los Angeles, CA, 90095, USA.|Otorhinolaryngology/Head and Neck Surgery Department, School of Medicine, Hamamatsu University, 1-20-1 Handayama, Hamamatsu, 431-3192, Shizuoka, Japan.
authors.forename: Kumiko
authors.initials: K
authors.lastname: Hosokawa
authors.affiliation: Head and Neck Surgery Department, David Geffen School of Medicine at UCLA, 951794, 35-64 Rehab, 1000 Veteran Avenue, Los Angeles, CA, 90095, USA.
authors.forename: Dora
authors.initials: D
authors.lastname: Acuna
authors.affiliation: Head and Neck Surgery Department, David Geffen School of Medicine at UCLA, 951794, 35-64 Rehab, 1000 Veteran Avenue, Los Angeles, CA, 90095, USA.
authors.forename: Fred H
authors.initials: FH
authors.lastname: Linthicum
authors.affiliation: Head and Neck Surgery Department, David Geffen School of Medicine at UCLA, 951794, 35-64 Rehab, 1000 Veteran Avenue, Los Angeles, CA, 90095, USA. ishiyama@ucla.edu.
authors.forename: Akira
authors.initials: A
authors.lastname: Ishiyama
mesh_terms: D007758:Ear, Inner / Q000166:cytology*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D016707:Tissue Fixation
keywords: Antigen retrieval; Archival human temporal bone; Celloidin-embedded sections; Cochlea; Cryosections; Immunohistochemistry; Microdissection; Paraffin; Vestibule