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 ] using a Leica SP5 microscope (63X water-immersion objective; XY dimensions: 82.0 µm; z-axis interval: 0.21 µm).
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 ] using a Leica SP5 microscope (63X water-immersion objective; XY dimensions: 82.0 µm; z-axis interval: 0.21 µm).
Free full text: Click here
