Optimized, automated assays for HCS

Understanding the mechanisms of cell death can provide key insights for toxicological profiling and drug discovery research. Apoptosis is a form of programmed cell death. A distinctive feature of the early stages of apoptosis is the activation of caspase enzymes, which participate in the cleavage of protein substrates and in the subsequent disassembly of the cell. We offer a caspase assay for HCS that allows the simple detection of active caspases in living cells in real time or in fixed cells.

Typical apoptosis—caspase assay

Live or fixed-cell assay protocol.

Imaging mode

  • Widefield or confocal
  • 10x or 20x magnification

Automatically measured properties

  • Fluorescence intensity, morphology, and count values for each object
  • Fluorescence intensity, morphology, and count values of spots in different cell regions
  • Average fluorescence intensity difference and ratios between different regions for each cell
  • Intensity and count ratio between channels within different regions for each cell

Caspase assay images

Hap1 cells were labeled with CellEvent Caspase-3/7 Green reagent and Hoechst 33342 following treatment with staurosporine. The cells were imaged using a Thermo Scientific CellInsight CX5 high-content platform (row A) and Thermo Scientific HCS Studio Software , which were used to automatically identify (row B) and quantify the intensity of CellEvent green fluorescence from each cell.

Sample experimental data

Dose-dependent increase in the induction of apoptosis, as reported by activation of Caspase 3/7. The sensitivity of two cell types: wild type Hap1 (red) and Hap1 lacking ATG5 (blue), were compared.