Journal article
Three-dimensional reconstructions from optical sections of thick mouse inner ears using confocal microscopy
Journal of microscopy (Oxford), Vol.248(3), pp.292-298
12/2012
DOI: 10.1111/j.1365-2818.2012.03673.x
PMCID: PMC3625616
PMID: 23140378
Abstract
Three-dimensional (3D) reconstructions of the vertebrate inner ear have provided novel insights into the development of this complex organ. 3D reconstructions enable superior analysis of phenotypic differences between wild type and mutant ears but can result in laborious work when reconstructed from physically sectioned material. Although nondestructive optical sectioning light sheet microscopy may ultimately prove the ideal solution, these technologies are not yet commercially available, or in many instances are not monetarily feasible. Here we introduce a simple technique to image a fluorescently labelled ear at different stages throughout development at high resolution enabling 3D reconstruction of any component of the inner ear using confocal microscopy. We provide a step-by-step manual from tissue preparation to imaging to 3D reconstruction and analysis including a rationale and troubleshooting guide at each step for researchers with different equipment, protocols, and access to resources to successfully incorporate the principles of this method and customize them to their laboratory settings.
Details
- Title: Subtitle
- Three-dimensional reconstructions from optical sections of thick mouse inner ears using confocal microscopy
- Creators
- B. J Kopecky - Department of Biology, University of Iowa, Iowa City, Iowa, U.S.AJ. S. Duncan - Department of Biology, University of Iowa, Iowa City, Iowa, U.S.AK.L ELLIOTT - Department of Biology, University of Iowa, Iowa City, Iowa, U.S.AB FRITZSCH - Department of Biology, University of Iowa, Iowa City, Iowa, U.S.A
- Resource Type
- Journal article
- Publication Details
- Journal of microscopy (Oxford), Vol.248(3), pp.292-298
- DOI
- 10.1111/j.1365-2818.2012.03673.x
- PMID
- 23140378
- PMCID
- PMC3625616
- NLM abbreviation
- J Microsc
- ISSN
- 0022-2720
- eISSN
- 1365-2818
- Grant note
- R01 DC005590 || DC / National Institute on Deafness and Other Communication Disorders : NIDCD
- Language
- English
- Date published
- 12/2012
- Academic Unit
- Iowa Neuroscience Institute; Biology; Craniofacial Anomalies Research Center
- Record Identifier
- 9984070000902771
Metrics
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