Neuronal Morphology Visualization and Analysis

Dendrites and cell body of fluorescent dentate neurons were traced using a computer-controlled microscope-based system (Axio Imager A2 Zeiss microscope, 100X oil-immersion objective) with a software (Neurolucida® software; MicroBrigthField Bioscience) that provides neuron tracing tools to trace from a live camera image (QImaging).
For spine analysis, confocal stacks of images were obtained with a Leica SP5 confocal microscope (63X oil-immersion objective; XY dimensions: 41.0 μm; z-axis interval: 0.13 μm). The dendritic length of each segment was measured on Z projections, and the number of dendritic spines was counted using NeuronStudio software ^{35 (link)}. Before spine analysis, images were deconvoluted using AutoQuantX3 software (Media Cybernetics). A minimum of 30 dendritic segments per experimental group and time point were examined for spine analysis. Morphometric analysis of the AIS was done as previously described ^{36 (link)} using a Leica SP5 microscope (63X water-immersion objective; XY dimensions: 82.0 μm; z-axis interval: 0.21 μm).

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Kerloch T., Farrugia F., Bouit L., Maître M., Terral G., Koehl M., Mortessagne P., Heng J.I., Blanchard M., Doat H., Leste-Lasserre T., Goron A., Gonzales D., Perrais D., Guillemot F., Abrous D.N, & Pacary E. (2021). The atypical Rho GTPase Rnd2 is critical for dentate granule neuron development and anxiety-like behavior during adult but not neonatal neurogenesis. Molecular Psychiatry, 26(12), 7280-7295.

Publication 2021

SP5 confocal microscope AutoQuantX3 software

Axis Cell body Confocal microscope Dendritic Dendritic spines Immersion Microscope Neuron Spine

Corresponding Organization :

Other organizations : Université de Bordeaux, Neurocentre Magendie, Inserm, Centre National de la Recherche Scientifique, Institut Interdisciplinaire de Neuroscience, Curtin University, The Francis Crick Institute

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Variable analysis

independent variables

Microscope-based system (Axio Imager A2 Zeiss microscope, 100X oil-immersion objective) with a software (Neurolucida® software; MicroBrigthField Bioscience) that provides neuron tracing tools
Confocal microscope (Leica SP5, 63X oil-immersion objective)
Deconvolution software (AutoQuantX3, Media Cybernetics)

dependent variables

Dendritic length of each segment
Number of dendritic spines
Morphometric analysis of the AIS

control variables

XY dimensions of confocal stacks (41.0 μm)
Z-axis interval of confocal stacks (0.13 μm)
XY dimensions of AIS analysis (82.0 μm)
Z-axis interval of AIS analysis (0.21 μm)

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