Tissue clearing for confocal imaging of native and bio-artificial skeletal muscle
Tissue clearing for confocal imaging of native and bio-artificial skeletal muscle
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DOI：
10.3109/10520295.2015.1019564
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发表时间：
2015-01-01
影响因子：
1.6
通讯作者：
Thorrez, L.
中科院分区：
文献类型：
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作者：
Decroix, L.;Van Muylder, V.;Thorrez, L.
新颖的清除技术已经彻底改变了大脑的三维共聚焦成像，而不需要物理组织切片。我们评估了三种清除方法，ScaleA2, Clear（T2）和3DISCO，用于共聚焦显微镜下观察天然肌肉和组织工程肌肉。我们发现，Clear（T2）处理略微提高了免疫组化染色的显示深度，但没有提高内源性绿色荧光蛋白（GFP）的显示深度。ScaleA2更好地保存了内源性GFP信号，并允许更深层的GFP成像，但与原肌球蛋白免疫组化染色不相容。disco处理保留了内源性GFP和免疫组织化学染色，并允许明显更深的成像。与ScaleA2和Clear（T2）相比，3DISCO过程的清除时间较短。我们认为3DISCO是天然骨骼肌组织和组织工程骨骼肌组织较好的清除方法。
Novel clearing techniques have revolutionized three-dimensional confocal imaging of the brain without the need for physical tissue sectioning. We evaluated three clearing methods, ScaleA2, Clear(T2), and 3DISCO for visualizing native and tissue engineered muscle by confocal microscopy. We found that Clear(T2) treatment improved the depth of visualization of immunohistochemical staining slightly, but did not improve depth of visualization of endogenous green fluorescent protein (GFP). ScaleA2 preserved endogenous GFP signal better and permitted significantly deeper GFP imaging, but it was incompatible with tropomyosin immunohistochemical staining. 3DISCO treatment preserved both endogenous GFP and immunohistochemical staining, and permitted significantly deeper imaging. Clearing time for the 3DISCO procedure is short compared to ScaleA2 and Clear(T2). We suggest that 3DISCO is the preferable clearing method for native and tissue engineered skeletal muscle tissue.