The Use of Low Vacuum Scanning Electron Microscopy (LVSEM) to Analyze Peripheral Nerve Samples.
The Use of Low Vacuum Scanning Electron Microscopy (LVSEM) to Analyze Peripheral Nerve Samples.
复制标题
使用低真空扫描电子显微镜 (LVSEM) 分析周围神经样本。
DOI：
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发表时间：
2018
影响因子：
--
通讯作者：
D. Parkinson
中科院分区：
文献类型：
--
作者：
P. Bond;D. Parkinson
电子显微镜的使用可以分析发育期间以及神经再生和修复过程中的周围神经的微结构。我们描述了一种使用低真空扫描电子显微镜方案快速分析和定量周围神经中髓磷脂的新方法。对于这种方法，为传统的透射电子显微镜 (TEM) 成像准备切除的神经，但在拍摄半薄切片的阶段，树脂块是在扫描电子显微镜 (SEM) 中使用背散射电子信号在低真空下成像。固定过程中含有高浓度锇的组织中的任何特征（例如髓磷脂）都在图像中显示为明亮区域。因此，髓磷脂在图像中很容易识别，并且由于它与周围组织之间存在高对比度差异，因此使用标准和免费提供的图像分析软件（例如 ImageJ）自动测量髓磷脂厚度（例如 G 比）很容易实现、一致且可重复。 This method therefore greatly speeds up the analysis of nerve samples and, in many cases, will obviate the need for ultrathin sections and TEM for analyzing nerve morphology.Low vacuum (LV) SEM imaging has benefits compared with light microscopy;放大倍数连续，分辨率更高，对比度和亮度可控。由于树脂块不需要金属涂层即可在 LV 模式下成像，因此一旦收集了图像，该树脂块仍可用于光学显微镜 (LM) 和 TEM 切片。
The use of electron microscopy allows analysis of the microarchitecture of peripheral nerves both during development and in the processes of nerve regeneration and repair.We describe a novel method for the rapid analysis and quantification of myelin in peripheral nerve using a low vacuum scanning electron microscopy protocol. For this methodology, excised nerves are prepared for traditional transmission electron microscopy (TEM) imaging, but at the stage where semi-thin sections would be taken, the resin block is instead imaged at low vacuum in the scanning electron microscope (SEM) using the backscattered electron signal. Any features in the tissue which have incorporated high concentrations of osmium from the fixation process (e.g., myelin) appear as bright regions in the image.Myelin therefore is easily identifiable in the images, and since there is a high contrast difference between it and the surrounding tissue, automated measurements for myelin thickness (e.g., G ratio) using standard and freely available image analysis software (e.g., ImageJ) are easily achievable, consistent, and repeatable. This method therefore greatly speeds up the analysis of nerve samples and, in many cases, will obviate the need for ultrathin sections and TEM for analyzing nerve morphology.Low vacuum (LV) SEM imaging has benefits compared with light microscopy; magnification is continuous, resolution is higher, and contrast and brightness are controllable. Since the resin block does not need a metal coating for imaging in LV mode, once the images have been collected, the block is still ready to section for both light microscopy (LM) and TEM.
DOI：
10.1083/jcb.201606052
发表时间：
2017-02
期刊：
The Journal of cell biology
影响因子：
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作者：
Mindos T;Dun XP;North K;Doddrell RD;Schulz A;Edwards P;Russell J;Gray B;Roberts SL;Shivane A;Mortimer G;Pirie M;Zhang N;Pan D;Morrison H;Parkinson DB
通讯作者：
Parkinson DB