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活体多光子显微镜在小鼠耳蜗可视化研究及中阶容积变化研究中的应用价值

Usefulness of Intravital Multiphoton Microscopy in Visualizing Study of Mouse Cochlea and Volume Changes in the Scala Media.

作者信息

Ju Hyun Mi, Lee Sun Hee, Kong Tae Hoon, Kwon Seung-Hae, Choi Jin Sil, Seo Young Joon

机构信息

Laboratory of Smile Snail, Yonsei University Wonju College of Medicine, Wonju, South Korea.

Department of Otorhinolaryngology, Yonsei University Wonju College of Medicine, Wonju, South Korea.

出版信息

Front Neurol. 2017 Jul 31;8:332. doi: 10.3389/fneur.2017.00332. eCollection 2017.

DOI:10.3389/fneur.2017.00332
PMID:28824523
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5535263/
Abstract

Conventional microscopy has limitations in viewing the cochlear microstructures due to three-dimensional spiral structure and the overlying bone. But these issues can be overcome by imaging the cochlea with intravital multiphoton microscopy (MPM). By using near-infrared lasers for multiphoton excitation, intravital MPM can detect endogenous fluorescence and second harmonic generation of tissues. In this study, we used intravital MPM to visualize various cochlear microstructures without any staining and non-invasively analyze the volume changes of the scala media (SM) without removing the overlying cochlear bone. The intravital MPM images revealed various tissue types, ranging from thin membranes to dense bone, as well as the spiral ganglion beneath the cochlear bone. The two-dimensional, cross-sectional, and serial z-stack intravital MPM images also revealed the spatial dilation of the SM in the temporal bone of pendrin-deficient mice. These findings suggest that intravital MPM might serve as a new method for obtaining microanatomical information regarding the cochlea, similar to standard histopathological analyses in the animal study for the cochlea. Given the capability of intravital MPM for detecting an increase in the volume of the SM in pendrin-deficient mice, it might be a promising new tool for assessing the pathophysiology of hearing loss in the future.

摘要

由于三维螺旋结构和覆盖其上的骨骼，传统显微镜在观察耳蜗微观结构方面存在局限性。但通过活体多光子显微镜（MPM）对耳蜗进行成像，可以克服这些问题。通过使用近红外激光进行多光子激发，活体MPM能够检测组织的内源性荧光和二次谐波产生。在本研究中，我们使用活体MPM在不进行任何染色的情况下可视化各种耳蜗微观结构，并在不去除覆盖的耳蜗骨的情况下非侵入性地分析中阶（SM）的体积变化。活体MPM图像显示了从薄膜到致密骨骼等各种组织类型，以及耳蜗骨下方的螺旋神经节。二维、横截面和连续z轴堆叠的活体MPM图像还显示了pendrin缺陷小鼠颞骨中SM的空间扩张。这些发现表明，活体MPM可能成为获取有关耳蜗微观解剖信息的一种新方法，类似于动物耳蜗研究中的标准组织病理学分析。鉴于活体MPM能够检测pendrin缺陷小鼠中SM体积的增加，它可能是未来评估听力损失病理生理学的一种有前景的新工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/8931918e0886/fneur-08-00332-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/dd624a1ee913/fneur-08-00332-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/9b7f8c76ef5c/fneur-08-00332-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/98fd69e11db0/fneur-08-00332-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/3e9a78dd66f3/fneur-08-00332-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/cc0bf14401d0/fneur-08-00332-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/e17da02cd222/fneur-08-00332-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/8931918e0886/fneur-08-00332-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/dd624a1ee913/fneur-08-00332-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/9b7f8c76ef5c/fneur-08-00332-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/98fd69e11db0/fneur-08-00332-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/3e9a78dd66f3/fneur-08-00332-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/cc0bf14401d0/fneur-08-00332-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/e17da02cd222/fneur-08-00332-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/615e/5535263/8931918e0886/fneur-08-00332-g007.jpg

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