Optimizing Visualization of Axonal Transport of Endogenous Cargo by Fluorescence Microscopy in Living <em>Caenorhabditis elegans</em>

Oliver Glomb; Mengya Lyu; Shaul Yogev

doi:10.3791/66236

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Biology

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Optimizing Visualization of Axonal Transport of Endogenous Cargo by Fluorescence Microscopy in Living Caenorhabditis elegans

通过荧光显微镜优化活体秀丽隐杆线虫中内源性货物轴突运输的可视化

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05:47 min

February 16, 2024

DOI: 10.3791/66236-v

Oliver Glomb¹, Mengya Lyu¹, Shaul Yogev¹

¹Department of Neuroscience,Yale School of Medicine

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该论文描述了荧光显微镜采集参数的优化，以可视化活线虫中内源性标记货物在单神经元分辨率下的轴突运输。

经典起始通常用于轴突转运蛋白，以实现轴突转运可视化，但内源性货物行为不同。内源性蛋白质水平低，使可视化具有挑战性。我们提供了优化采集条件的想法，以更好地可视化内源性 GPR3，它标记突触血管前体。

轴突运输区域中胞质蛋白的丰度使得使用常规荧光显微镜难以可视化它们的动力学。该协议建议使用消融步骤以及时间控制的标记方法来克服这些挑战。我们最近能够可视化内源性光谱的轴突运输。

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关键词:轴突运输内源性货物荧光显微镜秀丽隐杆线虫 CRISPR Cas9 突触小泡前体 RAB-3 采集参数漂白优化