Neurons were manually traced and reconstructed in 3-dimensions with a 63×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axio Imager Vario microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience, Williston, VT, USA). To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma [33 (link),34 (link),75 (link)].Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30-μm increments [34 (link)]. were analyzed in order to assess morphological cellular diversity and potential differences between the animal groups. A total of 50 cells were reconstructed for controls (~8 cells per animal) and a total of 49 cells were reconstructed for neutron irradiated mice (~8 cells per animal).
Neurolucida morphometry software
Manufactured by MBF Biosciences
Sourced in United States
Neurolucida is a comprehensive software solution for neuroanatomical reconstruction and quantitative analysis. It enables detailed mapping and measurement of neuronal structures, including dendritic and axonal arbors, as well as cell bodies and synapses. The software provides a suite of tools for tracing, visualizing, and analyzing complex neuronal networks.
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6 protocols using neurolucida morphometry software
1
Pyramidal Neuron Morphometrics in Irradiated Mice
2
Reconstruction of Hippocampal Pyramidal Neurons
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To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma42 (link)43 (link)44 (link). Neurons meeting these criteria were reconstructed in three-dimensions (3D) with a 40×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience). Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30 μm increments were analyzed in order to assess morphological cellular diversity and potential differences among animals45 (link).
3
Dendritic Morphology Analysis of CA1 Pyramidal Neurons
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To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma
[75 (link), 76 (link), 78 (link)]. Neurons meeting these criteria were reconstructed in 3-dimensions (3D) with a 40×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience). Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30-μm increments
[79 (link)] were analyzed in order to assess morphological cellular diversity and potential differences between the animal groups.
[75 (link), 76 (link), 78 (link)]. Neurons meeting these criteria were reconstructed in 3-dimensions (3D) with a 40×/1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience). Using NeuroExplorer software (MBF Bioscience) total dendritic length, number of intersections, and the amount of dendritic material per radial distance from the soma, in 30-μm increments
[79 (link)] were analyzed in order to assess morphological cellular diversity and potential differences between the animal groups.
4
Quantitative Analysis of Hippocampal CA1 Neuron Morphology
5
Dendritic Morphometry of CA1 Pyramidal Neurons
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To be included in the analysis, a loaded neuron had to satisfy the following criteria: (1) reside within the pyramidal layer of the CA1 as defined by cytoarchitectural characteristics; (2) demonstrate complete filling of dendritic tree, as evidenced by well-defined endings; and (3) demonstrate intact tertiary branches, with the exception of branches that extended beyond 50 μm in radial distance from the cell soma9 (link),32 (link),33 (link). Neurons meeting these criteria were reconstructed in three-dimensions (3D) with a 40 × /1.4 N.A., Plan-Apochromat oil immersion objective on a Zeiss Axiophot 2 microscope equipped with a motorized stage, video camera system, and Neurolucida morphometry software (MBF Bioscience, Williston, VT). Using Neurolucida Explorer software (MBF Bioscience) total dendritic length, number of intersections, and the length of dendritic material per radial distance from the soma, in 30 μm increments were analyzed in order to assess neuronal morphological diversity and potential differences among animals34 (link). Personnel undertaking the reconstructions were blinded to the group genotypes.
6
3D Reconstruction of Pyramidal Neurons
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Pyramidal neurons were included for reconstruction if they were within layer III of the PFC, were completely filled, and showed intact tertiary branches within at least 50 µm of the soma. Such neurons were reconstructed in 3 dimensions under a 40×/1.3 N.A. Plan-Neofluar oil immersion objective on a Zeiss Axiophot microscope equipped with a motorized stage (Ludl Electronic Products, Hawthorne, NY), a Microfire video camera (Optronics, Tulsa, OK), and Neurolucida morphometry software (v. 11.11.3, MBF Bioscience, Williston, VT). Using Neurolucida Explorer software (MBF Bioscience), total dendritic length and number of intersections per 30-µm incremental radial distance from the soma were analyzed by the Sholl method [58 (link)].
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