Figure 1

Workflow of the computational analysis of images from a high-content, genome-wide RNAi screen in Drosophila cell culture. Raw images of S2R+ cells stained for F-actin, microtubules and DNA were analyzed computationally to calculate the total monolayer area and mean cell area in each image (see Materials and methods for details). Image analysis scores were then normalized across screen plates to create z-scores (see Materials and methods for details). Hits were selected using a z-score threshold of -2 for mean cell area, and a monolayer area z-score of -2 to exclude images with low cell number where small cell size may reflect viability defects (green squares). This, approach yielded 73 putative hits (red triangles), which were examined for known physical and genetic interactions in FLIGHT [20].