# Application of dicentric chromosome assay for evaluation of radioprotective effect

## TL;DR

This paper describes a detailed protocol for using the dicentric chromosome assay to evaluate the effectiveness of radioprotective agents in a lab setting.

## Contribution

The study introduces a reproducible protocol integrating manual and automated methods for assessing radioprotective agents using dicentric chromosome analysis.

## Key findings

- The protocol includes steps for blood sample preparation, irradiation, and dicentric chromosome scoring.
- Manual and automated scoring methods are combined for accurate and efficient assessment.
- The protocol supports preclinical screening of novel piperazine derivatives as potential radioprotectants.

## Abstract

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.