Computational protocol: Brain Tumor promotes axon growth across the midline through interactions with the microtubule stabilizing protein Apc2

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Protocol publication

[…] Phenotypes were analyzed and images were acquired using a spinning disk confocal system (Perkin Elmer) built on a Nikon Ti-U inverted microscope using a Nikon OFN25 60x 40x or 10x objective with a Hamamatsu C10600-10B CCD camera and Yokogawa CSU-10 scanner head with Volocity imaging software. Images were processed using ImageJ and Adobe Illustrator software. For fluorescence quantification of GFP antibody staining in embryos, ten embryos per genotype (+/+; UAS-Apc2GFP or brat(-); UAS-Apc2GFP, +/+; src64bGFP or brat(-); scr64bGFP) were imaged using identical settings. Max projections were generated using ImageJ. After subtracting the staining background, the average pixel intensity was measured on twelve to sixteen clusters of EW neurons or across five regions within longitudinal axons for each embryo. The values from the five to ten embryos for each phenotype were averaged. […]

Pipeline specifications

Software tools ImageJ, Adobe Illustrator
Applications Miscellaneous, Microscopic phenotype analysis
Organisms Drosophila melanogaster
Diseases Brain Neoplasms, Adenomatous Polyposis Coli