Phenotypic screen readout

High-content screen (HCS) is a phenotypic screening that monitors multiple cellular parameters simultaneously. HCS employs fluorescence-based reagents (antibodies, dyes that bind or localize to a given cell component, sensors) to generate a multicolor fluorescence readout that is usually recorded using automated optical image acquisition devices. 

The siRNA screening is performed in duplicate in 96-well plates as shown in Fig. 1. The first replica is used for viability readout and the second for phenotypic readout where we look at spindle structure by using a high content screening platform. The small molecule library screening is also performed in duplicate with the first replica used for viability readout. In contrast to the siRNA... 

Recently, new comprehensive approaches have been developed that use compound libraries for screens in whole living cells using specific phenotype readouts. Some of the applications of this type of approach have been called chemical genetics or chemogenomics [9-27]. In many instances it is probably more accurate to call... 

Figure 17.1 A schematic depicts the flowchart of chemical genetics screening approach. A primary cell-based assay that captures pathways or phenotypic readouts is established and validated to screen a compound library. Owing to the frequent off-target effects of primary screen compounds, it is essential to implement counter screens and secondary screens to filter nonspecific hits to arrive at a group of high-confidence hit compounds. In silica methods for scaffold hopping and compound similarity searching can be utilized to select groups of similar molecules to generate SAR data to better understand the relevant war...

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