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| Microscopy and Imaging Methods: |
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Using a 25X/0.8 LD LCI Plan Apochromat objective with Glycerin as immersion medium, a Z-stack comprised of 17 optical sections was collected at an interval of 1.0µm with a 1024x1024 frame size and a 350 nm XY pixel size. The excitation was provided by 488 nm, 561 nm and 633 nm lasers. The raw data was processed using a blind deconvolution algorithm with 10 constrained iterations followed by psuedocoloring and 3D volume rendering. Deconvolution and 3D rendering was performed using 3rd party software. The image was collected using a LSM 710. | | A maximum intensity projection of a deconvolved confocal z-stack of a dissected Drosophila embryo expressing Green Fluorescent Protein (GFP) in all neurons (blue) counter stained for microtubules (green) and actin (red). |