利用全载免疫荧光和薄层显微镜观察斑马鱼幼体大脑中prps6阳性细胞的方法。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-01-18 DOI:10.1016/j.xpro.2024.103587

Olga Doszyn, Tomasz Dulski, Justyna Zmorzynska

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引用次数: 0

摘要

由于它们的体积小和透明，幼体斑马鱼是全脑成像的有用模型。在这里，我们提出了一种方案，用于可视化磷酸化Rps6，雷帕霉素复合体1 （mTORC1）活性的机制靶标，在受精后5天（dpf）的斑马鱼大脑中，使用全载免疫荧光和光片显微镜。我们描述了样品制备，储存，染色和成像的步骤。该方案也可以修改为针对其他蛋白质的抗体染色。有关本协议使用和执行的完整细节，请参见Doszyn等人1。

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Protocol for the visualization of pRps6-positive cells in larval zebrafish brains using whole-mount immunofluorescence and light-sheet microscopy.

Due to their small size and transparency, larval zebrafish are a useful model for whole-brain imaging. Here, we present a protocol for the visualization of phosphorylated Rps6, a marker of mechanistic target of rapamycin complex 1 (mTORC1) activity, in the zebrafish brains at 5 days post fertilization (dpf), using whole-mount immunofluorescence and light-sheet microscopy. We describe steps for sample preparation, storage, staining, and imaging. This protocol can also be modified for staining with antibodies against other proteins. For complete details on the use and execution of this protocol, please refer to Doszyn et al.¹.

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