Monitoring Cell Health in Real-time / Time-lapse Cell-based Assays
Poster Feb 21, 2017
The purpose here is to validate a novel far-red DNA binding viability probe, DRAQ7™, in real-time cell based assays for 2-D and 3-D models. This requires DRAQ7™ to neither synergise nor interfere with toxicants / anti-cancer compounds. It must be capable of addition at any stage of an assay to permit real-time monitoring of membrane integrity loss, retaining its functionality throughout.
As background, DRAQ7™’s DNA-specificity allows monitoring cell-by-cell while spectral properties permit incorporation in multi-colour experiments e.g. with vital probes, mitochondrial health probes or Annexin V. DRAQ7™ has previously been shown to be truly cell impermeant yet retains DNA binding and far-red fluorescence of the DRAQ chromophore which makes it compatible with HCS imagers, fluorescence microscopes and cytometers. The excitation/emission properties are particularly suited to penetrative imaging of multi-cellular structures and thick ex-plant tissue sections, whilst limiting risks of short wavelength DNA damage.
Multiplexing cell-based assays is possible using 3D culture models that are larger and more complex than monolayers
Real-time detection methods to measure live or dead cells provide much flexibility for multiplexing
All multiplexed assay combinations should be verified using appropriate controls for each 3D cell culture model.