Light-Sheet Microscopy Enables Three-Dimensional Fluorescence Imaging and Live Imaging of Satellite Cells on Skeletal Muscle Fibers.

Q4 Biochemistry, Genetics and Molecular Biology
Hira Asif Khan, Nick Van Hateren, Anne-Gaëlle Borycki
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引用次数: 0

Abstract

The ex vivo myofiber culture system has proven to be a useful methodology to explore the biology and behavior of satellite cells within their niche environment. However, a limitation of this system is that myofibers and their associated satellite cells are commonly examined using conventional fluorescence microscopy, which renders a three-dimensional system into two-dimensional imaging, leading to the loss of precious information or misleading interpretation of observations. Here, we report on the use of light-sheet fluorescence microscopy to generate three-dimensional and live imaging of satellite cells on myofibers. Light-sheet microscopy offers high imaging speed and good spatial resolution with minimal photo-bleaching, allowing live imaging and three-dimensional acquisition of skeletal muscle fiber specimen. The potentials of this technology are wide, ranging from the visualization of satellite cell behavior such as cell division and cell migration to imaging the sub-cellular localization of proteins or organelles.

光片显微镜可对骨骼肌纤维上的卫星细胞进行三维荧光成像和实时成像。
体内外肌纤维培养系统已被证明是探索卫星细胞在其生态环境中的生物学和行为的有用方法。然而,该系统的一个局限是,肌纤维及其相关卫星细胞通常使用传统的荧光显微镜进行检查,这使得三维系统变成了二维成像,导致珍贵信息的丢失或对观察结果的误导性解释。在此,我们报告了利用光片荧光显微镜对肌纤维上的卫星细胞进行三维和活体成像的情况。光片显微镜具有成像速度快、空间分辨率高、光漂白少等特点,可对骨骼肌纤维标本进行活体成像和三维采集。这项技术具有广泛的潜力,从卫星细胞行为(如细胞分裂和细胞迁移)的可视化,到蛋白质或细胞器的亚细胞定位成像,不一而足。
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来源期刊
Methods in molecular biology
Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
2.00
自引率
0.00%
发文量
3536
期刊介绍: For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.
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