FIVE STEPS TO PHENOTYPIC DATA FROM AUTOMATED QUANTITATIVE IMAGING
In the simple schematic example shown to the right, a two channel assay is performed.
The first channel is occupied by the nuclear marker Hoechst. This channel is used primarily to focus the imaging system in the correct Z plane where the cells are located. The Hoechst channel is also used to count cells and quantify other nuclear properties such as size, shape and texture – these features can be used to detect cell cycle, toxicity and stress.
The second channel is depicted in green and simulates a cytoplasmic protein that forms puncta in the cytoplasm and nuclear compartments.