Live-cell imaging of dendritic spines by STED microscopy
Live-cell imaging of dendritic spines by STED microscopy
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DOI：
10.1073/pnas.0810028105
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发表时间：
2008-12-02
影响因子：
11.1
通讯作者：
Bonhoeffer, Tobias
中科院分区：
文献类型：
--
作者：
Naegerl, U. Valentin;Willig, Katrin I.;Bonhoeffer, Tobias
时间推移荧光成像已成为神经生物学研究的重要手段之一。特别是，共聚焦和双光子显微镜已被用来研究突触形态的活动依赖性变化。然而，光学显微镜的衍射极限分辨率往往不足，迫使研究人员通过EM补充活细胞成像策略。在这里，我们报告的第一次使用远场光学技术与亚衍射分辨率的非侵入性图像活动依赖的形态可塑性的树突棘。具体来说，我们表明，时间推移刺激发射耗尽成像的树突棘的YFP阳性海马神经元的器官切片优于共聚焦显微镜揭示重要的结构细节。该技术大大提高了量化的形态参数，如颈部宽度和弯曲的头部的棘，这被认为是发挥关键作用的功能和可塑性的突触连接。
Time lapse fluorescence imaging has become one of the most important approaches in neurobiological research. In particular, both confocal and two-photon microscopy have been used to study activity-dependent changes in synaptic morphology. However, the diffraction-limited resolution of light microscopy is often inadequate, forcing researchers to complement the live cell imaging strategy by EM. Here, we report on the first use of a far-field optical technique with subdiffraction resolution to noninvasively image activity-dependent morphological plasticity of dendritic spines. Specifically we show that time lapse stimulated emission depletion imaging of dendritic spines of YFP-positive hippocampal neurons in organotypic slices outperforms confocal microscopy in revealing important structural details. The technique substantially improves the quantification of morphological parameters, such as the neck width and the curvature of the heads of spines, which are thought to play critical roles for the function and plasticity of synaptic connections.