PDF(Pubmed)
Abstract：
Imaging neuronal architecture has been a recurrent challenge over the years, and the localization of synaptic proteins is a frequent challenge in neuroscience. To quantitatively detect and analyze the structure of synapses, we recently developed free SODA software to detect the association of pre and postsynaptic proteins. To fully take advantage of spatial distribution analysis in complex cells, such as neurons, we also selected some new dyes for plasma membrane labeling. Using Icy SODA plugin, we could detect and analyze synaptic association in both conventional and single molecule localization microscopy, giving access to a molecular map at the nanoscale level. To replace those molecular distributions within the neuronal three-dimensional (3D) shape, we used MemBright probes and 3D STORM analysis to decipher the entire 3D shape of various dendritic spine types at the single-molecule resolution level. We report here the example of synaptic proteins within neuronal mask, but these tools have a broader spectrum of interest since they can be used whatever the proteins or the cellular type. Altogether with SODA plugin, MemBright probes thus provide the perfect toolkit to decipher a nanometric molecular map of proteins within a 3D cellular context.
摘要：
多年来，成像神经元结构一直是一个反复出现的挑战，突触蛋白的定位是神经科学中的一个常见挑战。为了定量检测和分析突触的结构，我们最近开发了免费的SODA软件来检测突触前和突触后蛋白的关联。为了充分利用复杂细胞中的空间分布分析，比如神经元，我们还选择了一些新的染料用于质膜标记。使用冰冷的SODA插件，我们可以在常规和单分子定位显微镜中检测和分析突触关联，获得纳米级分子图谱。为了取代神经元三维(3D)形状内的分子分布，我们使用MemBright探针和3DSTORM分析以单分子分辨率水平破译各种树突状脊柱类型的整个3D形状.我们在这里报告了神经元掩模中的突触蛋白的例子，但是这些工具具有更广泛的兴趣，因为它们可以用于任何蛋白质或细胞类型。连同SODA插件，因此，MemBright探针提供了完美的工具包来破译3D细胞环境中蛋白质的纳米分子图。
