Automated Comet assay

The comet assay is a sensitive, well established technique for quantifying DNA damage in eukaryotic cells. Compatible with the detection of a wide range of DNA damaging agents, its principle consists in migration of fragmented DNA in an electrophoresis gel (damaged DNA forming the tail of the comet), while intact DNA moves at a slower rate (head of the comet). The percentage of fragmented DNA in the comet tail is a direct measure of DNA damage.

Micronucleus and H2AX assay

In response to clastogen DNA double strand breaks, H2AX is phosphorylated within minutes at Ser 139, and is then referred as γH2AX. Contrary to γH2AX, micronuclei are a late marker of damages either to DNA and be related to aneugens nor clastogens effect. By detecting both H2AX and micronuclei on our high content screening plateform, our assay provide a efficient tool to assess the genotoxicity properties of your compounds.

Carcinogenesis potential of new entity are really important to detect at the early stage of discovery. Cell imaging in HCA/HCS can allow to analysis micronuclei (MN) in a automation manner as we can see in this article on lymphocytes, which are suspension cells, analysed by laser scanning cytometry and high content analysis.

This fluorescent approach allowed clear identification of binucleated cells and detection of MN. Highcontent analysis was developed to further automatically score MN within mono-, tri- and tetra-nucleated cells and to determine the nuclear division index and nuclear circularity values. Importantly, it allows for co-detection of other biomarkers of interest within a single lymphocyte, and further development of this capability is anticipated.

Curated from www.biotechniques.com

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