An effective manual deboning method to prepare intact mouse nasal tissue with preserved anatomical organization.
An effective manual deboning method to prepare intact mouse nasal tissue with preserved anatomical organization.
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DOI：
10.3791/50538
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发表时间：
2013-08-10
期刊：
影响因子：
--
通讯作者：
Lin W
中科院分区：
文献类型：
--
作者：
Dunston D;Ashby S;Krosnowski K;Ogura T;Lin W
哺乳动物的鼻子是一个具有复杂内部结构的多功能器官。鼻腔内有各种上皮，如嗅觉上皮、呼吸上皮和鳞状上皮，它们在解剖位置、形态和功能上都有明显的差异。在成年小鼠中，鼻子被各种头骨覆盖，限制了对内部结构的实验访问，特别是那些在后部的结构，如主要的嗅觉上皮(MOE)。在这里，我们描述了一种有效的方法，以获得几乎完整和完整的鼻部组织保存解剖组织。使用解剖显微镜下的手术工具，我们依次取出鼻部组织周围的头盖骨。这一过程既可以在多聚甲醛固定的小鼠头上进行，也可以在肉质解剖的小鼠头上进行。整个脱钙过程大约需要20-30分钟，比传统的化学脱钙所需的实验时间要短得多。此外，我们提出了一种简单的方法来去除鼻甲之间的气泡，这对于从鼻部组织制备中获得完整的薄的水平或冠状或矢状面是至关重要的。用我们的方法制备的鼻组织可用于整体上皮细胞的整体观察，以及形态、免疫细胞化学、RNA原位杂交和生理学研究，特别是在需要进行区域特异性检查和比较的研究中。
The mammalian nose is a multi-functional organ with intricate internal structures. The nasal cavity is lined with various epithelia, such as olfactory, respiratory and squamous epithelia which differ markedly in anatomical locations, morphology and functions. In adult mice, the nose is covered with various skull bones, limiting experimental access to internal structures, especially those in the posterior, such as the main olfactory epithelium (MOE). Here we describe an effective method for obtaining almost entire and intact nasal tissues with preserved anatomical organization. Using surgical tools under a dissecting microscope, we sequentially remove the skull bones surrounding the nasal tissue. This procedure can be performed on both paraformaldehyde-fixed and fleshly dissected, skinned mouse heads. The entire deboning procedure takes about 20–30 min, which is significantly shorter than the experimental time required for conventional chemical-based decalcification. In addition, we present an easy method to remove air bubbles trapped between turbinates, which is critical for obtaining intact thin horizontal or coronal or sagittal sections from the nasal tissue preparation. Nasal tissue prepared using our method can be used for whole mount observation of the entire epithelia, as well as morphological, immunocytochemical, RNA in situ hybridization and physiological studies, especially in studies where region-specific examination and comparison are of interest.
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DOI：
10.3791/1850
发表时间：
2010-03-02
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Cygnar, Katherine D;Stephan, Aaron B;Zhao, Haiqing
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影响因子：
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DOI：
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发表时间：
2011-06-15
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
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通讯作者：
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DOI：
10.1073/pnas.94.7.3388
发表时间：
1997-04-01
影响因子：
11.1
作者：
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通讯作者：
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DOI：
10.3791/3564
发表时间：
2012-07-30
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
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通讯作者：
Shain, William