Search bioRxivSearch

Biology subjects

Repin, M.

Publications and source records attributed to Repin, M..

2 recordsLinked to original sources

The use of a centrifuge-free RABiT-II system for high-throughput micronucleus analysis

The cytokinesis-block micronucleus (CBMN) assay is considered as the most suitable biodosimetry method for automation. Previously, we automated this assay on a commercial robotic biotech high-throughput system (RABiT-II) adopting both a traditional and an accelerated micronucleus protocol, both using centrifugation steps for lymphocyte harvesting and washing, after whole blood culturing. Here we describe further development of our accelerated CBMN assay protocol for using on High Throughput/High Content Screening (HTS/HCS) robotic systems without a centrifuge. This opens the way for implementation of the CBMN assay on a wider range of commercial automated HTS/HCS systems and thus increases the potential capacity of dose estimates following a mass-casualty radiological event.

cell biology

A new approach to automated CBMN scoring following high doses

In recent years we have automated the CBMN assay using microvolumes of blood, processed in multiwell plates. We have seen that at doses above 6 Gy the detected yield of micronuclei actually declines with dose, likely because of mitotic delay, preventing cells from forming micronuclei and also, when using one color imaging, resulting in many false binucleated cells, consisting of two randomly-adjacent nuclei. By using the inverse mitotic index (the ratio of mononuclear to binuclear cells) to adjust the micronucleus yield we were able to obtain a monotonic increasing dose response curve at doses of up to at least 10 Gy from the same samples which generated dose-response curve with a peak near 6 Gy, when scored using the traditional micronucleus yield.

cell biology