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首页> 外文期刊>Journal of Microscopy >Three-dimensional reconstructions from optical sections of thick mouse inner ears using confocal microscopy
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Three-dimensional reconstructions from optical sections of thick mouse inner ears using confocal microscopy

机译：使用共聚焦显微镜的厚鼠内耳光部分的三维重建

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摘要

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.
机译：脊椎动物内耳的三维（3D）重建为该复杂器官的发育提供了新的洞察力。 3D重建能够卓越地分析野生型和突变耳之间的表型差异，但在从物理切片材料重建时可能导致艰苦的工作。尽管非破坏性光学切片灯纸显微镜可以最终证明理想的解决方案，但这些技术尚未商业上可获得，或者在许多情况下，这些技术并非单项可行。在这里，我们介绍一种简单的技术，在整个高分辨率下在不同阶段以不同阶段进行荧光标记的耳朵，从而能够使用共聚焦显微镜来实现内耳的任何部件的3D重建。我们提供从组织准备到3D重​​建和分析的逐步手册，包括用于不同设备，协议和资源的研究人员的每个步骤中的基本原理和故障排除指南，以成功地纳入该方法的原则和定制他们到他们的实验室设置。

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