Figure 4
Comparison of HCS platform to standard cell biology assays, MTS and flow cytometry.
(a) Illustrates schematic of experimental design. Cells were admixed at a starting ratio of 1:1 H3255 to H3255R-RFP cells and treated with erlotinib. Following a 72-hour incubation, cell populations were analyzed via MTS, flow cytometry, or our HCS approach. The table summarizes important features of each experimental design including processing time and data outputs. (b) Viability curve displaying the data generated from each assay. The flow cytometry curve remains constant across drug concentrations (at approximately 100% viability) due to loss of dead cells during sample processing and the acquisition of relative counts (not absolute counts) of live cells. (c) Top – Population percentages of H3255 live, H3255 dead, H3255R live and H3255R dead as determined by flow cytometry and HCS analyses. Bottom – Absolute cell counts generated by HCS over time and in response to erlotinib treatment. (d) Box plot showing the distribution of H3255 and H3255R nuclear area in response to increasing concentrations of erlotinib.
