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RABiT-II-DCA: A Fully-automated Dicentric Chromosome Assay in Multiwell Plates.

Authors
  • Royba, Ekaterina1, 2
  • Repin, Mikhail1
  • Pampou, Sergey2
  • Karan, Charles2
  • Brenner, David J1
  • Garty, Guy1
  • 1 Center for Radiological Research.
  • 2 Columbia Genome Center High-Throughput Screening Facility, Columbia University Medical Center, New York, New York 10032.
Type
Published Article
Journal
Radiation Research
Publisher
BioOne (Radiation Research Society)
Publication Date
Sep 01, 2019
Volume
192
Issue
3
Pages
311–323
Identifiers
DOI: 10.1667/RR15266.1
PMID: 31295087
Source
Medline
Language
English
License
Unknown

Abstract

We developed a fully-automated dicentric chromosome assay (DCA) in multiwell plates. All operations, from sample loading to chromosome scoring, are performed, without human intervention, by the second-generation Rapid Automated Biodosimetry Tool II (RABiT-II) robotic system, a plate imager and custom software, FluorQuantDic. The system requires small volumes of blood (30 µl per individual) to determine radiation dose received as a result of a radiation accident or terrorist attack. To visualize dicentrics in multiwell plates, we implemented a non-classical protocol for centromere FISH staining at 37°C. The RABiT-II performs rapid analysis of chromosomes after extracting them from metaphase cells. With the use of multiwell plates, many samples can be screened at the same time. Thus, the RABiT-II DCA provides an advantage during triage when risk-based stratification and medical management are required for a large population exposed to unknown levels of ionizing radiation.

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