Fig. 2

Overview of the screening strategy for the study of essential genes. a Homozygous phb-2(tm2998);Phsp6::GFP mutants are sorted at L2 stage from a mixed population of balanced heterozygous phb-2(tm2998)/mIn1 animals into multiwell plates using the COPAS Biosort “worm sorting”. b Next, bacteria containing the OrthoList RNAi sublibrary are added to the wells and worms are incubated at 20 °C. c When worms reach the desired stage, they are imaged in brightfield and fluorescent channels using an automated microscope, IN Cell Analyzer (GE Healthcare). d Employing a user-defined image segmentation protocol, hits can be defined based on different measurements like reporter expression or size of the worms in comparison with the control. e Finally, hits can be analysed by building genetic networks based on predicted and described protein interactions in different organisms