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

test

Use the macro "polarisation_polar_dispersion.ijm" to :

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

binarize

  • 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 :

Polar Transform

  • 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 Superposition
  • 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).