研究 SNARE 功能和定位的细胞器和囊泡电子断层扫描。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4314-3_17

Nicholas A Bright, J Paul Luzio

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引用次数: 0

摘要

电子断层扫描可以提供额外的形态学信息不容易获得通过传统的透射电子显微镜薄切片。它使用电子显微镜中的测角仪来倾斜标本，并从不同方向收集一系列2D图像，这些图像组合在一起，提供3D体层析图和感兴趣的形态特征的彩色重建。在这里，我们描述了其用于观察SNARE蛋白VAMP7和VAMP8耗尽后细胞器形态变化的方案，并研究VAMP7在内溶酶体/溶酶体上的定位。

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Electron Tomography of Organelles and Vesicles in the Investigation of SNARE Function and Localization.

Electron tomography can provide additional morphological information not easily obtained by conventional transmission electron microscopy of thin sections. It uses a goniometer stage in the electron microscope to tilt the specimen and collect a series of 2D images from different orientations, which are combined to provide a 3D volume tomogram and a colored reconstruction of the morphological feature(s) of interest. Here we describe the protocols for its use in visualizing changes in organelle morphology after depletion of the SNARE proteins VAMP7 and VAMP8 and to study VAMP7 localization on endolysosomes/lysosomes.

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来源期刊

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.