At the end of differentiation (on day 30), neurite analysis was performed on iPSC‐derived neurons differentiated on top of PA6, either treated or not with L‐Dopa and Carbidopa for 10 or 24 days (early L‐Dopa), fixed and stained for TH. We randomly selected a minimum of 10 DAn per iPSC line (in the only condition that were isolated from surrounding DAn, so that neurites could be unambiguously attributed to a single DAn), using a Carl Zeiss LSM880 confocal microscope and analyzed with the FIJI® is Just ImageJ™ plugin NeuronJ to determine the number and length of neurites per cell.
For fiber density quantification, we generated a mask using ImageJ to delimit the area of the image occupied by TH+ fibers which did not include nuclei (TH− stained area). The area was corrected by the number of TH+ neurons present in each image (Ishikawa et al, 2016 (link); Prots et al, 2018 (link)).
For fiber density quantification, we generated a mask using ImageJ to delimit the area of the image occupied by TH+ fibers which did not include nuclei (TH− stained area). The area was corrected by the number of TH+ neurons present in each image (Ishikawa et al, 2016 (link); Prots et al, 2018 (link)).
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