High Content Ratio Imaging for Apoptosis Detection Figure 4. The HCS platform is adaptable for high content apoptosis analysis. OVCAR8 cells treated with a random panel of drugs in duplicates were imaged at the time points: 0 hr, 6 hr, 18 hr, 36 hr, 48 hr post drug treatment. Simple polygon segmentation was applied (A) and segmentation parameters were optimized to pick out intact cells as well as apoptotic bodies. Montages of 161 and 363 are shown to highlight the possibility of choosing different montage types based on the type of experiment or to increase the number of cells for analysis. Bar chart showing time kinetics of FRET ratio loss upon drug treatment at the indicated time points (B). The same area and same cells are represented over a period of time indicating potential for readout of kinetics. doi:10.1371/journal.pone.0020114.g004 ison with the end point, fixed cell, cell proliferation assays used in conventional anticancer drug screening, this platform gives a more physiologically relevant mechanistic picture of the drug activity in the initial screening stage itself. The possibility of on- the-fly acquisition-cum-segmentation-cum-analysis also highlights its superiority over other drug screening platforms. Also, the methodology is simple, in comparison to the enormity of information that is obtained. Further, as tumor etiology varies PLoS ONE | www.plosone.org 9 May 2011 | Volume 6 | Issue 5 | e20114