Workflows for In-depth High Content Analysis

High content screening (HCS) is a rapid and flexible tool for studying biological processes in great detail at a high throughput. As a consequence, HCS campaigns generate large amounts of data that need to be analyzed. The initial analysis of acquired images can include object segmentation, calculation of object properties and object classification into different populations. Screening campaigns typically contain between 10 and a few hundred microplates with individual treatment conditions distributed among the different plates. Therefore, the identification of parameters with suitable Z’ scores and the analysis of plate-to-plate variability are mandatory steps in the analysis of screening data. These analysis steps are performed during secondary analysis. Once a compound treatment generates a cellular reaction, the images from the selected hit wells can be analyzed in more detail. This could include the calculation of 3D representations from acquired image stacks, the generation of movies from time lapse experiments or the preparation of images for publication, e.g. after image deconvolution.