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Three‐dimensional reconstructions from optical sections of thick mouse inner ears using confocal microscopy
Author(s) -
KOPECKY B.J.,
DUNCAN J.S.,
ELLIOTT K.L.,
FRITZSCH B.
Publication year - 2012
Publication title -
journal of microscopy
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.569
H-Index - 111
eISSN - 1365-2818
pISSN - 0022-2720
DOI - 10.1111/j.1365-2818.2012.03673.x
Subject(s) - troubleshooting , optical sectioning , confocal microscopy , microscopy , computer science , confocal , 3d reconstruction , light sheet fluorescence microscopy , computer vision , biomedical engineering , artificial intelligence , optics , scanning confocal electron microscopy , physics , engineering , operating system
Summary 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.