The use of flow cytometry enables accurate simultaneous evaluation of multiple analytes, in a statistically relevant sample size in a short time. This application note discusses the use of the Invitrogen Attune NxT Flow Cytometer with Autosampler to evaluate the effectiveness of several cancer drugs in inducing apoptosis in cancer cells, with dose response curves.
Jurkat cells, comprising human T lymphocyte cells used to study acute T cell leukemia, were grown and treated in 96-well plates with various concentrations of different cancer drugs as described below. These cells were evaluated for toxicity using an apoptosis reporter assay and analyzed on the Attune NxT Flow Cytometer. Cell toxicity was measured using the fluorogenic substrate Invitrogen CellEvent Caspase-3/7 Green Detection Reagent, which reacts with the enzymes caspase-3 and caspase-7, which are activated during apoptosis.
The ability to add the CellEvent Caspase-3/7 Green Detection Reagent directly to the live cells in the medium without the need to wash, fix, or permeabilize the cells is ideal for this endpoint assay experiment.
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