Document Detail


Three-dimensional reconstructions from optical sections of thick mouse inner ears using confocal microscopy.
MedLine Citation:
PMID:  23140378     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
B J Kopecky; J S Duncan; K L Elliott; B Fritzsch
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of microscopy     Volume:  248     ISSN:  1365-2818     ISO Abbreviation:  J Microsc     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-11-12     Completed Date:  2013-04-15     Revised Date:  2014-07-29    
Medline Journal Info:
Nlm Unique ID:  0204522     Medline TA:  J Microsc     Country:  England    
Other Details:
Languages:  eng     Pagination:  292-8     Citation Subset:  IM    
Copyright Information:
© 2012 The Authors Journal of Microscopy © 2012 Royal Microscopical Society.
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MeSH Terms
Descriptor/Qualifier:
Animals
Ear, Inner / anatomy & histology*
Imaging, Three-Dimensional / methods*
Mice
Microscopy, Confocal / methods*
Grant Support
ID/Acronym/Agency:
DC010362/DC/NIDCD NIH HHS; R01 DC005590/DC/NIDCD NIH HHS; R01-DC055095590/DC/NIDCD NIH HHS; TL1 RR024981/RR/NCRR NIH HHS; UL1RR024979/RR/NCRR NIH HHS
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