Semi Manual segmentation of myelin profiles and g-ratio quantification is done in three steps
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Automaticaly segment Inner profiles: Open the image , run "CountMayelinatedAxons.ijm" macro It will ask you to draw a line along the scalebar, and select the relavant part of the image for analysis and
will find candidates of inner part of the mayelinated axons -
Manualy draw the Outer profiles, and correct Inner profiles if needed: Open the image , drag and drop the FILINMAE_InnerRoiSet.zip file into Fiji Draw the outer parts of the malinated axon and add them to the RoiManager (see details in ManualEditingInstructions_ForGRatio.txt) There may be non-relevant detections, but don't bother to delete them, as the next step will include only matched pairs of inner&outer ROIs Save the ROI file when done to Results\FileName_InnerOuterRoiSet.zip
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Match Inner and Outer Profiles and calculate g-ratio Open the image, run the "GratioOfManualOuter_MayelinatedAxons.ijm" macro it will open the Roi file from step 2, find pairs of inner/outer ROIs and for each pair calculate the g-ratio
Written by: Ofra Golani at MICC Cell Observatory, Weizmann Institute of Science
In collaboration with Anya Vainshtein, Christina Katanov and Elior Peles, Weizmann Institute of Science
Software package: Fiji (ImageJ)
Workflow language: ImageJ macro
Used in :
N-Wasp regulates oligodendrocyte myelination
Christina Katanov, Nurit Novak, Anya Vainshtein, Jeffery L Dupree, and Elior Peles