Center for Neural Informatics, Structures, & Plasticity, Krasnow Institute for Advanced Study, George Mason University, Fairfax, VA 22030, USA.
Allen Institute for Brain Science, Seattle, WA 98109, USA.
Sci Data. 2018 Jan 23;5:170207. doi: 10.1038/sdata.2017.207.
Several efficient procedures exist to digitally trace neuronal structure from light microscopy, and mature community resources have emerged to store, share, and analyze these datasets. In contrast, the quantification of intracellular distributions and morphological dynamics is not yet standardized. Current widespread descriptions of neuron morphology are static and inadequate for subcellular characterizations. We introduce a new file format to represent multichannel information as well as an open-source Vaa3D plugin to acquire this type of data. Next we define a novel data structure to capture morphological dynamics, and demonstrate its application to different time-lapse experiments. Importantly, we designed both innovations as judicious extensions of the classic SWC format, thus ensuring full back-compatibility with popular visualization and modeling tools. We then deploy the combined multichannel/time-varying reconstruction system on developing neurons in live Drosophila larvae by digitally tracing fluorescently labeled cytoskeletal components along with overall dendritic morphology as they changed over time. This same design is also suitable for quantifying dendritic calcium dynamics and tracking arbor-wide movement of any subcellular substrate of interest.
现已有几种有效的方法可用于从光学显微镜中数字化追踪神经元结构,并且已经出现了成熟的社区资源来存储、共享和分析这些数据集。相比之下,细胞内分布和形态动态的量化尚未标准化。目前对神经元形态的广泛描述是静态的,不足以进行亚细胞特征描述。我们引入了一种新的文件格式来表示多通道信息,并提供了一个开源的 Vaa3D 插件来获取这种类型的数据。接下来,我们定义了一种新的数据结构来捕获形态动态,并演示了其在不同时间推移实验中的应用。重要的是,我们将这两项创新设计为经典 SWC 格式的明智扩展,从而确保与流行的可视化和建模工具完全向后兼容。然后,我们通过数字化追踪荧光标记的细胞骨架成分以及整体树突形态,在活体果蝇幼虫的发育神经元上部署了组合的多通道/时变重建系统,随着时间的推移,这些结构会发生变化。这种设计同样适用于量化树突钙动态,并跟踪任何感兴趣的亚细胞基质的整个树突范围的运动。