Subcellular resolution 3D light field imaging with genetically encoded voltage indicators
Subcellular resolution 3D light field imaging with genetically encoded voltage indicators
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DOI：
10.1101/2020.05.22.108191
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发表时间：
2020-05
期刊：
影响因子：
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通讯作者：
Peter Quicke;Carmel L. Howe;P. Song;H. V. Jadan;Chenchen Song;T. Knöpfel;M. Neil;P. Dragotti;
中科院分区：
文献类型：
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作者：
Peter Quicke;Carmel L. Howe;P. Song;H. V. Jadan;Chenchen Song;T. Knöpfel;M. Neil;P. Dragotti;
光场显微镜（LFM）能够实现高信噪比（SNR）、快速帧率下的光效率体积成像，并已成功应用于单细胞分辨率功能性神经元钙成像。具有遗传编码电压指示器（GEVI）的电压成像特别受益于光场显微镜的体积成像能力，这是由于所需的高采样率以及有限的探头亮度和功能灵敏度。以前的LFM研究成像GEVI跟踪人口水平的相互作用，只有在无脊椎动物的准备工作，没有单细胞分辨率。在这里，我们展示了亚细胞分辨率GEVI光场成像在急性小鼠脑切片解决树突状电压信号定位在三维。我们表征了不同光场重建技术对信噪比和信号定位的影响，并将信噪比与宽场成像的荧光瞬态进行了比较。我们的研究结果表明，光场电压成像的潜力，研究树突的整合和动作电位的传播和反向传播在3个空间维度。
Light field microscopy (LFM) enables high signal-to-noise ratio (SNR), light efficient volume imaging at fast frame rates, and has been successfully applied to single-cell resolution functional neuronal calcium imaging. Voltage imaging with genetically encoded voltage indicators (GEVIs) stands to particularly benefit from light field microscopy’s volumetric imaging capability due to high required sampling rates, and limited probe brightness and functional sensitivity. Previous LFM studies have imaged GEVIs to track population-level interactions only in invertebrate preparations and without single cell resolution. Here we demonstrate sub-cellular resolution GEVI light field imaging in acute mouse brain slices resolving dendritic voltage signals localized in three dimensions. We characterize the effects of different light field reconstruction techniques on the SNR and signal localization and compare the SNR to fluorescence transients imaged in wide field. Our results demonstrate the potential of light field voltage imaging for studying dendritic integration and action potential propagation and backpropagation in 3 spatial dimensions.