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An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
Published on: May 13, 2019
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CytoRADx: A High-Throughput, Standardized Biodosimetry Diagnostic System Based on the Cytokinesis-Block Micronucleus
Chris Capaccio1, Jay R Perrier1,2, Lídia Cunha2
1ASELL, LLC, Owings Mills, Maryland.
Radiation Research
|September 13, 2021
Summary
A new automated system, CytoRADx™, offers rapid and accurate radiation biodosimetry for mass casualty events. This high-throughput test quantifies DNA damage in blood cells, providing crucial diagnostic information for medical resource allocation.
Area of Science:
- Biomedical engineering
- Radiation oncology
- Genetics and genomics
Background:
- Accurate radiation biodosimetry is critical for managing mass casualty events following radiological disasters.
- Current diagnostic methods lack FDA clearance and high-throughput capabilities, hindering timely medical response.
- The cytokinesis-block micronucleus (CBMN) assay is a known method for assessing DNA damage but requires significant time and expertise.
Purpose of the Study:
- To develop and validate a high-throughput, automated system for quantitative radiation biodosimetry.
- To address the limitations of the classical CBMN assay in terms of speed, standardization, and labor requirements.
- To provide a reliable diagnostic tool for assessing radiation exposure in diverse populations during emergencies.
Main Methods:
- Adaptation and refinement of the cytokinesis-block micronucleus (CBMN) assay into the automated CytoRADx™ System.
- Standardized reagent kits, optimized protocols, and automated microscopy (RADxScan™ Imager) for high-throughput analysis.
- Quantitative measurement of radiation dose-dependent micronuclei formation in human and non-human primate lymphocytes from whole blood samples.
Main Results:
- The CytoRADx™ System demonstrated accurate and precise dose measurements (0-8 Gy) with 90% of results within 1 Gy of the delivered dose.
- Validation performed on 636 human and 445 non-human primate samples, including in vivo studies.
- System performance showed repeatability across laboratories, operators, and instruments, with statistically similar results across diverse populations and stable dose calculations for up to 28 days.
Conclusions:
- The CytoRADx™ System represents a significant advancement in radiation biodosimetry, enabling rapid and accurate diagnoses in large-scale emergency scenarios.
- The automated, standardized approach overcomes limitations of traditional methods, facilitating efficient allocation of scarce medical resources.
- The enhanced CBMN assay technology offers potential for future diagnostic applications in toxicology and oncology.
