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An optimized tissue processing pipeline for multiscale imaging of the intact peripheral auditory pathway.

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Τίτλος: An optimized tissue processing pipeline for multiscale imaging of the intact peripheral auditory pathway.
Συγγραφείς: Ahrend JF; Auditory Neuroscience Group, Institute of Physiology, Medical University Innsbruck, Innsbruck, Austria., Kuenkel LS; Auditory Neuroscience Group, Institute of Physiology, Medical University Innsbruck, Innsbruck, Austria., Degenhart G; Core facility MicroCT, University Clinic for Radiology, Medical University Innsbruck, Innsbruck, Austria., Vogl C; Auditory Neuroscience Group, Institute of Physiology, Medical University Innsbruck, Innsbruck, Austria. Electronic address: christian.vogl@i-med.ac.at.
Πηγή: Hearing research [Hear Res] 2026 Sep; Vol. 479, pp. 109748. Date of Electronic Publication: 2026 Jul 14.
Τύπος έκδοσης: Journal Article; Research Support, Non-U.S. Gov't
Γλώσσα: English
Στοιχεία περιοδικού: Publisher: Elsevier/North-Holland Biomedical Press Country of Publication: Netherlands NLM ID: 7900445 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1878-5891 (Electronic) Linking ISSN: 03785955 NLM ISO Abbreviation: Hear Res Subsets: MEDLINE
Imprint Name(s): Original Publication: Amsterdam, Elsevier/North-Holland Biomedical Press.
Ιατρικοί όροι (MeSH): Auditory Pathways*/diagnostic imaging , Auditory Pathways*/physiology , Cochlea*/diagnostic imaging , Cochlea*/physiology , Cochlear Nucleus*/diagnostic imaging , X-Ray Microtomography*, Spiral Ganglion/diagnostic imaging ; Temporal Bone/diagnostic imaging ; Animals ; Microscopy, Fluorescence ; Microscopy, Confocal ; Mice, Inbred C57BL
Περίληψη: We present a comprehensive tissue processing and imaging pipeline tailored for multiscale analyses of the intact peripheral auditory pathway, encompassing the cochlea, cochlear root, and cochlear nucleus (CN) within a single specimen. This workflow integrates advanced imaging modalities - including micro-computed tomography (µCT), light-sheet fluorescence microscopy, confocal microscopy, and super-resolution STED microscopy - to enable a holistic assessment of fixed temporal bones. This methodological pipeline incorporates a unique and novel dissection technique that maintains the delicate connection between the cochlea and CN, facilitating volumetric imaging of neuronal populations and precise quantification of spiral and vestibular ganglion neurons without sectioning-related loss. The presented workflow further supports vascular network segmentation and nanoscale protein distribution analyses, thus offering versatility for diverse cochlear research applications. To promote widespread adoption, an adapted ethyl cinnamate-based clearing protocol, a novel slice preparation procedure as well as a validated antibody library are provided that enable robust labeling of cochlear structures ranging from whole-organ visualization to synaptic-scale analyses. By maximizing information extraction from individual specimens, this accessible pipeline aligns with 3R principles and provides a unified framework for investigating auditory function and pathology, advancing the study of hearing impairment and related genetic models.
(Copyright © 2026 The Authors. Published by Elsevier B.V. All rights reserved.)
Competing Interests: Declaration of competing interest The authors declare no competing interests.
Contributed Indexing: Keywords: Auditory system; Hair cells; Immunofluorescence analysis; Light-sheet fluorescence microscopy (LSFM); Spiral ganglion neurons; Vestibular ganglion neurons
Entry Date(s): Date Created: 20260721 Date Completed: 20260729 Latest Revision: 20260807
Update Code: 20260808
DOI: 10.1016/j.heares.2026.109748
PMID: 42480389
Βάση Δεδομένων: MEDLINE
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  Data: An optimized tissue processing pipeline for multiscale imaging of the intact peripheral auditory pathway.
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  Data: We present a comprehensive tissue processing and imaging pipeline tailored for multiscale analyses of the intact peripheral auditory pathway, encompassing the cochlea, cochlear root, and cochlear nucleus (CN) within a single specimen. This workflow integrates advanced imaging modalities - including micro-computed tomography (µCT), light-sheet fluorescence microscopy, confocal microscopy, and super-resolution STED microscopy - to enable a holistic assessment of fixed temporal bones. This methodological pipeline incorporates a unique and novel dissection technique that maintains the delicate connection between the cochlea and CN, facilitating volumetric imaging of neuronal populations and precise quantification of spiral and vestibular ganglion neurons without sectioning-related loss. The presented workflow further supports vascular network segmentation and nanoscale protein distribution analyses, thus offering versatility for diverse cochlear research applications. To promote widespread adoption, an adapted ethyl cinnamate-based clearing protocol, a novel slice preparation procedure as well as a validated antibody library are provided that enable robust labeling of cochlear structures ranging from whole-organ visualization to synaptic-scale analyses. By maximizing information extraction from individual specimens, this accessible pipeline aligns with 3R principles and provides a unified framework for investigating auditory function and pathology, advancing the study of hearing impairment and related genetic models.<br /> (Copyright © 2026 The Authors. Published by Elsevier B.V. All rights reserved.)
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  Data: Declaration of competing interest The authors declare no competing interests.
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  Data: <i>Keywords: </i>Auditory system; Hair cells; Immunofluorescence analysis; Light-sheet fluorescence microscopy (LSFM); Spiral ganglion neurons; Vestibular ganglion neurons
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