Figure 9.

Seed point alignment for filament reconstruction

(A) In order to track filament length, number of filaments and number of bifurcations, Imaris places seed points (this is done automatically, by clicking on the blue forward button, see red box and arrow) along the astrocyte filaments based on IHC signal and surface reconstruction.

(B) Seed points appear mainly at the initial segment of the filament and increase in density in close proximity of a bifurcation. Usually, this feature does not need to be adjusted manually, but should be checked if obtained filament parameters seem inaccurate. As for the starting point placement, the threshold can be adjusted manually and should align nicely with the filament projections as depicted in the image.

(C) Seed points should align with the reconstructed filaments as shown in the overlay image.