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

在研究单元（桶皮质功能）的更新提案中，基本上所有子项目的中心主题仍然是由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 et al.，2012; Progress in Neurobiology，in revision）。近年来，由于许多新的和令人兴奋的遗传/分子生物学技术（cre-driver和-reporter lines，brainbow，channelrhodopsins，genetically encoded calcium indicators，etc.）的出现，啮齿动物桶状皮质结构与功能的成功关系显示出了很大的进展。已经变得可用，其允许在同一组实验中用荧光标记物在结构上可视化功能上操纵和分析的单个神经元或神经元群体。此外，只有在桶皮层中存在一个容易可视化的模板，可以将不同小组和实验的结果映射到该模板上：桶作为功能柱的结构相关物。为了精确地分类神经元，并建立基于计算机的神经元模型，以模拟柱状系综的功能，形态学特性必须以定量的方式获得。然而，这种定量重建是一个非常耗时的过程。因此，完全或至少半自动化的程序是非常期望的。该中心项目（i）提供了一个技术平台和能力中心，将支持所有处理单个或少量标记神经元分析的子项目，这些神经元在生理实验过程中将被染料标记，（ii）提出了组织学程序，成像方法和软件可用性，这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.