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首页> 外文期刊>Journal of Neuroscience Methods >Fix the problem – A practical guide to whole-mount immunohistochemistry of teased nerve fibres
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Fix the problem – A practical guide to whole-mount immunohistochemistry of teased nerve fibres

机译：解决问题 - 一个实用的柚子神经纤维的免疫组织化学指南

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Highlights ? Biocompatible adhesives withstand antibody retrieval and counteract nerve fibre recoil. ? Prefixative permeabilisation allows for epitope detection throughout teased fibre bundles. ? After fixation, antibody penetration requires enzymatic digestion by Proteinase-K. ? Sealed-chamber incubation reduces the volume of required sera to 100–150 μl per 10–15 teased fibre bundles. Abstract Background Immunohistochemical staining of entire nerve fibres allows for studying the molecular composition of functional fibre subunits and may add to the diagnostic value of nerve fibre teasing. New method In this study, we established a sealed-slide method for reproducible immunostaining of deep axoplasmic proteins in permanently straightened nerve fibres. Results Immunostaining of teased nerve fibres very much is facilitated by tip-fixation with biocompatible glass adhesives. Antibody penetration in fresh nerves can be achieved by thermic and chemical permeabilisation while enzymatic digestion allows for sufficient permeability after aldehyde fixation. Comparison with existing methods The methods recommended herein are easy to perform and represent a reliable and reproducible way to whole mount immunostaining. Conclusions Sealed-slide immunostaining of tip-fixed and permeabilised nerve biopsies will help to validate neurophysiological abnormalities and to screen for target molecules and predictive markers of peripheral nerve disorders such as in inherited neuropathies and Guillain-Barré syndrome. ]]>
机译：强调 ？生物相容性粘合剂耐受抗体检索和抵消神经纤维反冲。还前缀渗透允许在整个尖端的纤维束中检测表位检测。还在固定后，抗体渗透需要蛋白酶-K酶消化。还密封室孵育将所需血清的体积降低至每10-15次取出的纤维束100-150μL。摘要背景全神经纤维的免疫组织化学染色允许研究功能纤维亚基的分子组成，并可以增加神经纤维戏弄的诊断价值。本研究的新方法，我们建立了一种密封的滑动方法，用于永久拉直神经纤维中深轴质蛋白的可再现免疫染色。结果通过用生物相容性玻璃粘合剂的尖端固定促进柚子神经纤维的免疫染色。通过热和化学渗透可以实现新鲜神经中的抗体渗透，而酶消化允许在醛固定后允许足够的渗透性。与现有方法的比较本文建议的方法易于执行，并且代表全部坐式免疫染色的可靠和可重复的方式。结论尖端固定和渗透神经活检的密封滑动免疫染色将有助于验证神经生理异常和筛选靶分子和外周神经疾病的预测标志，如遗传神经病变和Guillain-Barré综合征。 ]]>

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