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用于在倒置共聚焦显微镜上对上皮组织动力学进行长期监测的简化活体成像方法。

Streamlined intravital imaging approach for long-term monitoring of epithelial tissue dynamics on an inverted confocal microscope.

作者信息

Hamersky Michael, Tekale Khushi, Winfree L Matthew, Rowan Matthew Jm, Seldin Lindsey

机构信息

Department of Cell Biology, Emory University School of Medicine, Atlanta, GA, USA.

Independent scholar, Snellville, GA.

出版信息

bioRxiv. 2023 May 12:2023.05.10.540242. doi: 10.1101/2023.05.10.540242.

DOI:10.1101/2023.05.10.540242
PMID:37214899
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10197696/
Abstract

UNLABELLED

Understanding normal and aberrant in vivo cell behaviors is necessary to develop clinical interventions to thwart disease initiation and progression. It is therefore critical to optimize imaging approaches that facilitate the observation of cell dynamics in situ, where tissue structure and composition remain unperturbed. The epidermis is the body's outermost barrier as well as the source of the most prevalent human cancers, namely cutaneous skin carcinomas. The accessibility of skin tissue presents a unique opportunity to monitor epithelial and dermal cell behaviors in intact animals using noninvasive intravital microscopy. Nevertheless, this sophisticated imaging approach has primarily been achieved using upright multiphoton microscopes, which represents a significant barrier-for-entry for most investigators. In this study, we present a custom-designed 3D-printed microscope stage insert suitable for use with inverted confocal microscopes that streamlines long-term intravital imaging of ear skin in live transgenic mice. We believe this versatile invention, which may be customized to fit the inverted microscope brand and model of choice, as well as adapted to image additional organ systems, will prove invaluable to the greater scientific research community by significantly enhancing the accessibility of intravital microscopy. This technological advancement is critical to bolster our understanding of live cell dynamics in both normal and disease contexts.

SUMMARY

A new tool to simplify intravital imaging using inverted confocal microscopy.

摘要

未标记

了解正常和异常的体内细胞行为对于开发阻止疾病发生和进展的临床干预措施至关重要。因此,优化成像方法以促进原位观察细胞动态至关重要,因为在原位组织结构和组成保持不受干扰。表皮是人体最外层的屏障,也是最常见的人类癌症即皮肤癌的来源。皮肤组织的可及性提供了一个独特的机会,可使用非侵入性活体显微镜在完整动物中监测上皮和真皮细胞行为。然而,这种复杂的成像方法主要是使用直立式多光子显微镜实现的,这对大多数研究人员来说是一个重大的进入障碍。在本研究中,我们展示了一种定制设计的3D打印显微镜载物台插件,适用于倒置共聚焦显微镜,可简化对活体转基因小鼠耳部皮肤的长期活体成像。我们相信,这项多功能发明可以定制以适合所选的倒置显微镜品牌和型号,也可以适用于对其他器官系统成像,通过显著提高活体显微镜的可及性,将对更广泛的科研界证明是非常宝贵的。这一技术进步对于加强我们对正常和疾病背景下活细胞动态的理解至关重要。

总结

一种使用倒置共聚焦显微镜简化活体成像的新工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/0174de224242/nihpp-2023.05.10.540242v2-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/d56a7444101d/nihpp-2023.05.10.540242v2-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/5c0f1f208567/nihpp-2023.05.10.540242v2-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/615d63a6ec98/nihpp-2023.05.10.540242v2-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/c633b13721cf/nihpp-2023.05.10.540242v2-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/1717c04da8d8/nihpp-2023.05.10.540242v2-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/0174de224242/nihpp-2023.05.10.540242v2-f0006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/d56a7444101d/nihpp-2023.05.10.540242v2-f0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/5c0f1f208567/nihpp-2023.05.10.540242v2-f0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/615d63a6ec98/nihpp-2023.05.10.540242v2-f0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/c633b13721cf/nihpp-2023.05.10.540242v2-f0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/1717c04da8d8/nihpp-2023.05.10.540242v2-f0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f1e/10197696/0174de224242/nihpp-2023.05.10.540242v2-f0006.jpg

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A quantitative 3D intravital look at the juxtaglomerular renin-cell-niche reveals an individual intra/extraglomerular feedback system.对肾小球旁肾素细胞龛进行定量三维活体观察,揭示了一种个体内/肾小球内反馈系统。
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