Ultrafast tissue staining with chemical tags
Ultrafast tissue staining with chemical tags
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DOI：
10.1073/pnas.1411087111
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发表时间：
2014-09-09
影响因子：
11.1
通讯作者：
Jefferis, Gregory S. X. E.
中科院分区：
文献类型：
--
作者：
Kohl, Johannes;Ng, Julian;Jefferis, Gregory S. X. E.
基因编码的荧光蛋白和免疫染色被广泛用于检测固定生物样本中的细胞和亚细胞结构。然而，对于厚的或整体的组织，每种方法都受到限制，包括有限的光谱灵活性和较低的信号或速度、较差的穿透性和较高的背景标记。我们通过使用转基因表达的化学标签对厚厚的生物组织进行快速、均匀、高信号和低背景的标记，克服了这些限制。我们首先构建了一个广泛适用的转基因果蝇报告系平台，证明了化学标记可以将整只苍蝇脑的染色速度提高100倍。使用病毒载体将化学标签传递到小鼠的大脑中，然后我们证明了这种标记策略在小鼠身上效果很好。因此，这种基于标签的方法极大地提高了在完整和/或厚的生物样本中标记基因标记细胞的速度和特异性。
Genetically encoded fluorescent proteins and immunostaining are widely used to detect cellular and subcellular structures in fixed biological samples. However, for thick or whole-mount tissue, each approach suffers from limitations, including limited spectral flexibility and lower signal or slow speed, poor penetration, and high background labeling, respectively. We have overcome these limitations by using transgenically expressed chemical tags for rapid, even, high-signal and low-background labeling of thick biological tissues. We first construct a platform of widely applicable transgenic Drosophila reporter lines, demonstrating that chemical labeling can accelerate staining of whole-mount fly brains by a factor of 100. Using viral vectors to deliver chemical tags into the mouse brain, we then demonstrate that this labeling strategy works well in mice. Thus this tag-based approach drastically improves the speed and specificity of labeling genetically marked cells in intact and/or thick biological samples.