Three-dimensional analysis and quantification of dye-labeled cortical neurons by Neurolucida

在这份关于研究单位(桶状皮质功能)的更新提案中，几乎所有子项目的一个中心主题仍然是兴奋性或抑制性新皮质神经元形成的回路及其与感觉信息的皮质处理的关系(Feldmeyer等人，2012年；神经生物学进展，修订中)。近年来，由于许多新的和令人兴奋的遗传/分子生物学技术(克隆驱动和报告基因、脑弓、通道视紫红质、遗传编码的钙指示物等)的出现，啮齿动物桶状皮质结构与功能的成功关系显示出许多进展。已经出现，允许在同一组实验中使用荧光标记对单个神经元或从功能上进行操作和分析的神经元群体进行结构可视化。此外，只有在桶皮质中存在一个容易可视化的模板，不同小组和实验的结果可以映射到该模板上：桶作为功能柱的结构关联。为了准确地对神经元进行分类，并建立基于计算机的神经元模型来模拟柱状系综的功能，必须以定量的方式获得神经元的形态属性。然而，这种量化重建是一个非常耗时的过程。因此，全自动或至少半自动的程序是非常可取的。这个中央项目(I)提供了一个技术平台和能力中心，它将支持处理单个或小群标记神经元的分析的所有子项目，这些标记神经元在生理实验过程中将被染色；(Ii)提出组织学程序、成像方法和软件可用性，将导致更有效的数据采集。

In this renewal proposal for the Research Unit (Barrel Cortex Function), a central topic of essentially all subprojects still is the circuitry formed by excitatory or inhibitory neocortical neurons and its relationship with the cortical processing of sensory information (Feldmeyer et al., 2012; Progress in Neurobiology, in revision). The successful relation of structure to function in the barrel cortex of rodents recently showed a lot of progress since many new and exciting genetic/molecular biological techniques (cre-driver and –reporter lines, brainbow, channelrhodopsins, genetically-encoded calcium indicators, etc.) have become available which allow in the same set of experiments to structurally visualize, with fluorescent markers, the single neuron or the population of neurons that are functionally manipulated and analyzed. Furthermore, only in the barrel cortex an easily visualizable template exists onto which the results from different groups and experiments can be mapped: barrels as a structural correlate of a functional column. In order to precisely classify neurons and to build computer-based neuronal models with which to simulate the function of columnar ensembles, the morphological properties have to be acquired in a quantitative manner. However, this quantitative reconstruction is a very time-consuming process. Thus, fully- or at least semi-automated procedures are highly desirable. This Central Project (i) provides a technology platform and also competence center which will support all subprojects dealing with the analysis of single, or small populations of labeled neurons which during the course of physiological experiments will be dye-labeled and (ii) brings forward the histological procedures, imaging methods and software usability which will lead to a more efficient data acquisition.