![]() Some of plugins are neuron-based, allowing for the semi-automatic tracing of neurite extension (e.g. The use of image analysis software should overcome these problems and several plugins have been developed to help in this task. Moreover, visual selection of neurons by the investigator may lead to selection bias. The morphometric description of cell cultures is tedious when done by hand, confining analysis to a number of cells that may not be representative of the whole neuronal culture. Eventually, one neurite acquires enhanced growth capabilities and becomes an axon (Stage 3). These protrusions undergo successive elongation and retraction phases with no net outgrowth. Few hours later the neurites sprouting begins (Stage 2). Hippocampal neurons begin to form lamellipodia right after adhesion to the substrate (Stage 1). Stages of early development of hippocampal neurons in culture. (2017-CE11-0026 MAMAs).Ĭompeting interests: The authors have declared that no competing interests exist.įig 1. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the manuscript and its Supporting Information files.įunding: This work was supported by Institut National de la Santé et de la recherche Médicale, Commissariat à l'Energie Atiomique et aux Energies Alternatives, Université Grenoble Alpes and by awards from the French Agence Nationale de la Recherche to A.A. Received: Accepted: JPublished: July 16, 2020Ĭopyright: © 2020 Boulan et al. Gilestro, Imperial College London, UNITED KINGDOM (2020) AutoNeuriteJ: An ImageJ plugin for measurement and classification of neuritic extensions. ![]() Overall the use of AutoNeuriteJ will provide rapid, unbiased and accurate measurement of neuron morphologies.Ĭitation: Boulan B, Beghin A, Ravanello C, Deloulme J-C, Gory-Fauré S, Andrieux A, et al. Moreover, by analyzing mouse neurons deficient for the microtubule associated protein 6 (MAP6) and wild type neurons we illustrate that AutoNeuriteJ is capable to detect subtle phenotypic difference in axonal length. In these experiments measurement of more than 5000 mouse neurons per conditions was obtained within a few hours. We showed that AutoNeuriteJ is able to detect variations of neuritic growth induced by several compounds known to affect the neuronal growth. To automate these measurements, we wrote AutoNeuriteJ, an imageJ/Fiji plugin that measures and classifies neurites from a very large number of neurons. This task usually requires labor-intensive measurements and the classification of numerous neurites from large numbers of neurons in culture. 13, 2018, 10:04 a.m.Morphometry characterization is an important procedure in describing neuronal cultures and identifying phenotypic differences. If you decide to do this, remember to discard the first row of the results. I often duplicate the first slice for just this purpose. As an alternative to 9, if you can discard the information on your first slice, then when selecting plants, press "return" instead of "space".this will draw the ROI.(If you choose to save as tiff, the whole stack will be saved as a multi-image tiff). You can then save this top slice as a jpg file. YOU DO NOT WANT TO DO THIS BEFORE CLICKING MULTI, BECAUSE THE LABELS WILL BE PART OF THE IMAGE AND GET COUNTED. To create a key: click label all ROIs.Select all ROIs from the multi measure window.Start the multi measure plugin Plugins>multi measure.Experiment with the correct setting for the radius (by double clicking in the icon in the tool bar) to make an appropriately sized selection. Click on the circle tool on the tool bar (probably the rightmost tool the icon is a circle drawn in blue, (not an oval in black).Go to Analyze>Set Measurements and select Integrated Density as your measurement.You will need to do this for each session. IF you click the global box, these settings will be applied to any additional images that you open in this session. Set Distance in pixels and Known Distance to 1. Image>Look Up Tables>Fire (or whatever you like). Also check the the other functions available in Image > Stacks. You can use the slider bar on the bottom to move through the stack. If you want it loaded by default see instructions for start-up macros in the macros pulldown menu, or on the imageJ website). (This will need to be done every time you start the program. Install the circle tool: Plugins>Macros>Install>Tools>CircleTool.txt.You may want to increase the memory allocated to ImageJ: Edit>Options>Memory.
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