Multiflow DNA damage assay

A multiflow DNA damage assay indicates where a substance has the potential to be DNA reactive within in vitro mammalian cells. 

DNA damage can occur as single and double strand DNA breaks, alkali labile sites and oxidation. There are three endpoints observed in this assay; the first is p53, a nuclear translocation event. The second is γH2AX, a marker for DNA double strand breaks. The third endpoint is specific phosphorylation of histone H3 in DNA packaging into chromatin, inducing mitotic delay/cytotoxic conditions. 

Additionally, multiflow assays include fluorescent polystyrene “counting beads”, which are used to calculate cell/nuclei densities and hence determine cytotoxicity levels via relative nuclei count (RNC), relative increased nuclei count (RINC) and relative population doubling (RPD). Multiplex machine leaning models can use these endpoints to predict the genotoxic mode of action and classification a compound.