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

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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.