Fig. 6

Combined green/red image analysis protocol. a Images acquired in brightfield, green (GFP) and red (Nile Red) channels with the IN Cell Analyzer 2000 and processed using Developer Toolbox software (GE Healthcare). A balanced heterozygous animal expressing pharyngeal GFP has been identified (red arrow) within a population of homozygous phb-2(tm2998) mutants. The heterozygous animal exhibits increased Nile Red staining in comparison to the homozygous phb-2(tm2998) mutants. b Segmentation analysis output of the green/red image analysis for balanced mutants. Worm intensity subtracting the background intensity in the green image (Int Worm-Bkgd FITC), identification of green head (Green Head ID) and worm intensity subtracting the background intensity in the red image (Int Worm-Bkgd Red) are depicted in the analysis output. Target 12, with a green head ID greater than 0, corresponds to the heterozygous worm. Additional measures like area, major axis length, X/Y position and form factor can be added per user requirement