Multiwell-based G0-PCC assay for radiation biodosimetry

Area of Science:

• Cytogenetics
• Radiation Biology
• Biophysics

Background:

• Cytogenetic biodosimetry, essential for assessing radiation exposure, traditionally requires over 48 hours for cells to reach mitosis.
• This delay hinders the timely administration of critical radiation countermeasures, which are most effective within 24 hours post-exposure.
• There is a significant need for accelerated biodosimetry methods for effective management of radiation emergencies.

Purpose of the Study:

• To develop a faster cytogenetic biodosimetry technique for large-scale radiation emergencies.
• To integrate the G0-Premature Chromosome Condensation (G0-PCC) technique with the Rapid Automated Biodosimetry Tool-II (RABiT-II).
• To enable same-day radiation damage quantification.

Main Methods:

• Incorporation of the G0-Premature Chromosome Condensation (G0-PCC) technique with the Rapid Automated Biodosimetry Tool-II (RABiT-II).
• Optimization of Calyculin A (Cal A) concentration to enhance the yield of highly-condensed G0-PCC cells (hPCC).
• Development of a Convolutional Neural Network (CNN) for automated quantification of radiation-induced chromosome fragmentation in hPCC cells.

Main Results:

• A lower concentration of Calyculin A (Cal A) than recommended increased the yield of hPCC cells.
• Calyculin A alone induced chromosome compaction in G0 cells, suggesting a distinct mechanism from mitotic kinase-driven condensation.
• A correlation between radiation dose and hPCC chromosome fragmentation was identified, enabling automated quantification.

Conclusions:

• The combined G0-PCC and RABiT-II method offers a significantly faster alternative for cytogenetic biodosimetry.
• This approach addresses the critical need for same-day assessment of radiation exposure in emergency situations.
• The automated quantification using CNN enhances the efficiency and reliability of biodosimetry.