Imaging Membrane Repair in Single CellsUsing Correlative Light and ElectronMicroscopy

Coralie Croissant; Flora Bouvet; Sisareuth Tan; Anthony Bouter

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Imaging Membrane Repair in Single CellsUsing Correlative Light and ElectronMicroscopy

机译：使用相关光和电子显微镜进行成像膜修复单细胞

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Many cells possess the ability to repair plasma membrane disruption in physiologicalconditions. Growing evidence indicates a correlation between membranerepair and many human diseases. For example, a negative correlationis observed in muscle where failure to reseal sarcolemma may contribute tothe development of muscular dystrophies. Instead, a positive correlation isobserved in cancer cells where membrane repair may be exacerbated duringmetastasis. Here we describe a protocol that combines laser technology formembrane damage, immunostaining with gold nanoparticles and imaging byfluorescence microscopy and transmission electron microscopy (TEM), whichallows the characterization of the molecular machinery involved in membranerepair. Fluorescence microscopy enables to determine the subcellular localizationof candidate proteins in damaged cells while TEM offers high-resolutionultrastructural analysis of the μm2-disruption site, which enables to decipherthe membrane repair mechanism. Here we focus on the study of human skeletalmuscle cells, for obvious clinical interest, but this protocol is also suitable forother cell types.

机译：许多细胞具有修复生理学条件中的血浆膜破坏的能力。越来越多的证据表明膜线和许多人类疾病之间的相关性。例如，在肌肉中观察到的阴性相关性，在其中未能重新抗炎的Sarcolemma可能有助于肌营养不良的发育。相反，在癌细胞中呈阳性相关性，其中膜修复在Metaastasis期间可能会加剧。在这里，我们描述了一种结合激光技术型型碎片损伤，用金纳米颗粒的免疫染色和通过荧光显微镜的成像和透射电子显微镜（TEM）进行免疫染色的方案，这使得膜的分子机械的表征。荧光显微镜使得能够在TEM提供受损电池中候选蛋白质的亚细胞定位，而TEM提供μm2-破坏部位的高分辨率恒定的分析，这使得能够破译膜修复机制。在这里，我们专注于对人体骨骼细胞的研究，以实现明显的临床兴趣，但该方案也适合其他细胞类型。

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来源
《Current Protocols in Cell Biology》 |2018年第1期|共19页
作者
Coralie Croissant; Flora Bouvet; Sisareuth Tan; Anthony Bouter;
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作者单位

Institute of Chemistry and Biology of Membranes and Nano-objects UMR 5248 CNRS University of Bordeaux Pessac France;

Institute of Chemistry and Biology of Membranes and Nano-objects UMR 5248 CNRS University of Bordeaux Pessac France;

Institute of Chemistry and Biology of Membranes and Nano-objects UMR 5248 CNRS University of Bordeaux Pessac France;

Institute of Chemistry and Biology of Membranes and Nano-objects UMR 5248 CNRS University of Bordeaux Pessac Franc;

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原文格式 PDF
正文语种 eng
中图分类细胞生物学;
关键词
correlative light and electron microscopy; gold nanoparticles; laser; membrane repair; human skeletal muscle;

机译：相关光和电子显微镜;金纳米颗粒;激光;膜修复;人骨骼肌;

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专利

1. Imaging Membrane Repair in Single CellsUsing Correlative Light and ElectronMicroscopy [J] . Coralie Croissant, Flora Bouvet, Sisareuth Tan, Current Protocols in Cell Biology . 2018,第1期

机译：使用相关光和电子显微镜进行成像膜修复单细胞
2. Correlative mass spectrometry imaging, applying time-of-flight secondary ion mass spectrometry and atmospheric pressure matrix-assisted laser desorption/ionization to a single tissue section [J] . Desbenoit N., Walch A., Spengler B., Rapid Communications in Mass Spectrometry: RCM . 2018,第2期

机译：相关质谱成像，施加飞行时间二次离子质谱和大气压矩阵辅助激光解吸/电离到单个组织部分
3. Spatially Resolved Membrane Transport in a Single Cell Imaged by Second Harmonic Light Scattering [J] . Gh Mohammad Sharifian, Wilhelm Michael J., Moore Michael, Biochemistry . 2019,第14期

机译：通过二次谐波光散射成像的单个细胞中的空间分离的膜运输
4. Feature-based registration for correlative light and electron microscopy images [C] . Nam D., Mantell J., Hodgson L., IEEE International Conference on Image Processing . 2014

机译：基于特征的配准，用于相关的光学和电子显微镜图像
5. Investigation of Membrane Protein Dynamics using Correlative Single-particle Tracking and Super-resolution Microscopy Combined with Bayesian Inference of Diusion in Arbitrary Landscapes [D] . Mazloom-Farsibaf, Hanieh. 2019

机译：使用相关单颗粒跟踪和超分辨率显微镜的膜蛋白动力学的研究与贝叶斯推动在任意景观中
6. Correlative mass spectrometry imaging applying time‐of‐flight secondary ion mass spectrometry and atmospheric pressure matrix‐assisted laser desorption/ionization to a single tissue section [O] . N. Desbenoit, A. Walch, B. Spengler, -1

机译：相关质谱成像将飞行时间二次离子质谱和大气压基质辅助的激光解吸/电离应用于单个组织切片
7. Correlative mass spectrometry imaging, applying time-of-flight secondary ion mass spectrometry and atmospheric pressure matrix-assisted laser desorption/ionization to a single tissue section [O] . N. Desbenoit, A. Walch, B. Spengler, 2017

机译：相关质谱成像，施加飞行时间二次离子质谱和大气压矩阵辅助激光解吸/电离到单个组织部分

Imaging Membrane Repair in Single CellsUsing Correlative Light and ElectronMicroscopy

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