Sp5 sp8 confocal microscopes

Confocal images were acquired with Leica SP5/SP8 confocal microscopes using a 63× objective. Quantitative analysis for neuron images was performed using ImageJ. Three independent experiments were performed for all neuron data. Primary dendrites were counted at a ring within 10 μm distance from the soma. For counting Shank3 clusters, dendritic fragments beginning at a minimum of 20 µm radial distance from the soma were analysed. For counting postsynaptic Shank3 clusters, only Shank3 clusters distinctly colocalising with PSD-95 were quantified. The counting was performed using the Multi-Point tool of ImageJ. The evaluation of the fluorescence resonance energy transfer was performed using Bitplane Imaris software.

Images of imaginal tissues were acquired using 20x (or 40x oil) objectives on the Leica SP5/SP8 confocal microscopes (maintained with help from Single Cell Facility, D-BSSE, ETHZ), and processed using Fiji/ImageJ or Photoshop. Fiji was also used to determine the number of total clones and those with reporter activity upon blocking different signalling pathways.

Images of eye-antennal imaginal tissues were acquired using 20x or 40x objectives on the Leica SP5/SP8 confocal microscopes and processed using ImageJ or Imaris. Images of adult eyes or transplantation hosts were acquired with Nikon SMZ1270.