Application of dicentric chromosome assay for evaluation of radioprotective effect

• Published in: Biology methods and protocols Vol. 10; no. 1; p. bpaf058
• Main Authors: Milanová, Marcela, Chmil, Vojtěch, Tichý, Aleš, Lecová, Lenka
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 2025
• Subjects: Amifostine; Chromosome aberrations; Chromosomes; Dosimetry; Ionizing radiation; Lymphocytes; Metaphase; Nuclear accidents & safety; Piperazine; Protocols; Radioprotective agents; Software; chromosome aberrations; amifostine; radioprotective agent; cytogenetics; ionizing radiation
• ISSN: 2396-8923; 2396-8923
• DOI: 10.1093/biomethods/bpaf058

The dicentric chromosome assay is a well-established biodosimetric method used to assess absorbed ionizing radiation doses by detecting dicentric chromosomal aberrations. Here, we present a detailed, reproducible protocol for applying the dicentric chromosome assay for in vitro evaluation of radioprotective agents, including novel piperazine derivatives compared with amifostine and its active metabolite WR-1065. The protocol covers all key steps—blood sample preparation, in vitro irradiation, lymphocyte culture, metaphase preparation, and scoring of dicentric chromosomes. It highlights critical stages that affect data quality and reproducibility. Integrating manual scoring with automated analysis using the Metafer system ensures accurate and efficient assessment. Thus, this protocol bridges the fields of biological dosimetry and preclinical screening of radioprotective agents, providing a reliable framework for emergency radiation dose estimation and the development of new radiation medical countermeasures.