High-resolution immunofluorescence imaging of mouse cochlear hair bundles.
High-resolution immunofluorescence imaging of mouse cochlear hair bundles.
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小鼠耳蜗毛束的高分辨率免疫荧光成像。
DOI：
10.1016/j.xpro.2022.101431
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发表时间：
2022-06-17
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耳蜗毛束的高分辨率免疫荧光成像面临着许多挑战，由于毛束的小尺寸，脆弱的性质，和复杂的组织。在这里，我们描述了一个优化的协议，毛束蛋白免疫染色和定位。详细介绍了小鼠内耳的分离、固定和Corti器的解剖方法和步骤。我们进一步强调了最佳的透化，封闭，染色和安装条件以及高分辨率显微镜成像的参数。有关本方案使用和执行的完整详细信息，请参阅。解剖小鼠耳蜗和Corti器官的技术解剖、透化、阻断参数以检测毛束蛋白质安装方法以定位毛束中的蛋白质出版商说明：进行任何实验方案都需要遵守当地机构的实验室安全和伦理准则。耳蜗毛束的高分辨率免疫荧光成像可能是困难的，因为毛束的小尺寸，脆弱的性质，和复杂的组织。在这里，我们描述了一个优化的协议，毛束蛋白免疫染色和定位。本文详细介绍了小鼠内耳的提取、固定和Corti器的解剖方法和步骤。我们注意到最佳的透化，封闭，染色，安装条件和高分辨率显微镜成像的参数。
High-resolution immunofluorescence imaging of cochlear hair bundles faces many challenges due to the hair bundle’s small dimensions, fragile nature, and complex organization. Here, we describe an optimized protocol for hair-bundle protein immunostaining and localization. We detail the steps and solutions for extracting and fixing the mouse inner ear and for dissecting the organ of Corti. We further emphasize the optimal permeabilization, blocking, staining, and mounting conditions as well as the parameters for high-resolution microscopy imaging. For complete details on the use and execution of this protocol, please refer to. Techniques for dissecting the mouse cochlea and the organ of Corti Dissection, permeabilization, blocking parameters to detect hair bundle proteins Mounting method to localize protein in the hair bundles Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. High-resolution immunofluorescence imaging of cochlear hair bundles can be difficult because of the hair bundle’s small dimensions, fragile nature, and complex organization. Here, we describe an optimized protocol for hair-bundle protein immunostaining and localization. We detail the steps and solutions for extracting and fixation of the mouse inner ear and the dissection of the organ of Corti. We note the optimal permeabilization, blocking, staining, and mounting conditions and the parameters for high-resolution microscopy imaging.