Rapid Micronucleus MoA (Mode of action) is a ready to use kit for the fast identification of chromosomal breakage (clastogenic effect) or damage to the spindle apparatur (aneugenic effect) in dividing mammalien cells (primary cells like human lymphocytes or TK6 cell line). The test kit was developed to detect the genotoxic potential of chemical substances.
The ready to use test kit Rapid Micronucleus MoA detects aberrations like micronuclei (MN), nucleoplasmic bridges (NPBs), nuclear buds (NBUDs), aneugens and clastogens according to OECD TG473 or OECD TG487. Nucleoplasmic bridges and nuclear buds have been shown to be a associated with chromosomal instability and chromosomal aberrations.
Cells are prepared as per standard protocols according to OECD TG 487 or TG 473 and the slides are sequentially treated with a series of solutions provided with the kit “Rapid Micronucleus MoA”, including a human-specific, highly sensitive centromere probe and a counterstaining solution containing DAPI (4’,6-diamidino-2-phenylindole). Slides are sequentially washed with PBS and an ethanol series after treatment. Centromere probes are then added to the area of interest and the slide is heated at 80°C for 3 minutes and transferred to a humidified chamber for 20 minutes. Washes and a 5-minutes counterstaining step follow prior to the addition of the mounting solution.
Using a 40× magnification objective it will be possible to easily identify micronuclei and assess whether a centromere is present. Automated reading and archiving can be obtained with the slide scanning systems, e.g. Metafer. Slides are usually ready for evaluation within 1 hour. The application of FISH probes allows directly to distinguish micronuclei originating either from chromosome loss (aneugenic compounds) or breakage (clastogenic compounds).
FISH probe technology as used in the Rapid Micronucleus MoA Kit can also be used for cytogenetic analysis in research or in physical dosimetry, i.e. people exposed in medical or natural radiation.
Characteristics for the Rapid Micronucleus MoA Test Kit
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