Cellular, subcellular and functional in vivo labeling of the spinal cord using vital dyes
Cellular, subcellular and functional in vivo labeling of the spinal cord using vital dyes
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DOI：
10.1038/nprot.2013.022
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发表时间：
2013-02
期刊：
影响因子：
14.8
通讯作者：
E. Romanelli;C. Sorbara;Ivana Nikić;Athanasios Dagkalis;T. Misgeld;M. Kerschensteiner
中科院分区：
文献类型：
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作者：
E. Romanelli;C. Sorbara;Ivana Nikić;Athanasios Dagkalis;T. Misgeld;M. Kerschensteiner
在这里，我们提供了一个协议，用于快速标记不同的细胞类型，不同的亚细胞区室和关键损伤介质在活小鼠脊髓。这种方法是基于对手术暴露的脊髓应用合成活性染料。以适当浓度应用的合适的活体染料导致活体标记的可靠性，其可以与遗传标签组合，并且在许多情况下被保存用于固定后分析。结合体内成像，这种方法允许在细胞、亚细胞和功能水平上直接观察中枢神经系统生理学和病理生理学。脊髓的手术暴露和准备可以在不到1小时内完成，然后需要应用染料30-60分钟，然后标记的脊髓可以成像数小时。
Here we provide a protocol for rapidly labeling different cell types, distinct subcellular compartments and key injury mediators in the spinal cord of living mice. This method is based on the application of synthetic vital dyes to the surgically exposed spinal cord. Suitable vital dyes applied in appropriate concentrations lead to reliablein vivolabeling, which can be combined with genetic tags and in many cases preserved for postfixation analysis. In combination within vivoimaging, this approach allows the direct observation of central nervous system physiology and pathophysiology at the cellular, subcellular and functional level. Surgical exposure and preparation of the spinal cord can be achieved in less than 1 h, and then dyes need to be applied for 30–60 min before the labeled spinal cord can be imaged for several hours.