360-ASOD (360° Angular Scanning monitoring Organelle Distribution)

Use the macro "polarisation_polar_dispersion.ijm" to :

• Binarize your labelling (nucleus and endosomes) and find the centroids :

• Calculate the distance between centroids and normalize this distance by (feret/2): the value will be between 0 and 1.
• Transform your image in polar image with the plugin polar_transformer.class inside the macro :

• Calculate the profile in intensity (number of white pixel in the radial image in Y) versus angle

Use the jupyter notebook "Polarisation_Profile_Lowess_STD.ipynb" to:

• Use Lowess function to remove high peak (smoothing)
• Normalize your profile in X (shift the max intensity to 0 (angle) and from -180 to 180) and Intensity by the sum to obtain probability
• Make the mean per condition and display the mean curves overlay per condition
• Calculate the standard deviation for each condition to quantify the dispersion in angle among the mean. As non polarized curved do not go to 0 (offset), the std of these curves should be higher

optional :

• Bin your profile data by 30 degrees to smooth the curves (the peak maximum is originally too high due to high local maximum value)
• Fit the curves with a Gaussian to determine the Y0 per condition (angle = 180 degrees) to obtain a quantification of polarisation from these curves (If the offset is low, the polarisation is high).