Individual variations in the micronucleus assay for biological dosimetry after high dose exposure

Kacprzak, Justyna; Kuszewski, Tomasz; Lankoff, Anna; Müller, Wolfgang‐Ulrich; Wójcik, Andrzej; Lisowska, Halina

doi:10.1016/j.mrgentox.2013.04.017

Cited by 16 publications

(32 citation statements)

References 17 publications

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“…6D) increased and was sufficiently separated at 6, 8 and 10 Gy. This increase, which is also in agreement with the results available in the literature (36, 42, 48), could indicate the delay in cell cycle progression in the presence of DNA damage, which influences the production of BN cells. The monotonically increasing dose dependence of the ratio of mononucleated to binucleated lymphocytes confirms that this endpoint could be taken into account as an additional parameter for dose-assessment purposes at higher doses.…”

Section: Discussionsupporting

confidence: 92%

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Fast Image Analysis for the Micronucleus Assay in a Fully Automated High-Throughput Biodosimetry System

et al. 2014

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Section: Discussionsupporting

confidence: 92%

“…This variability could reasonably be anticipated. It is well know that there exists a considerable variation in the baseline micronuclei frequency and acute radiosensitivity (46–48). In part this is due to variations in demographics, such as age, gender (46), smoking status, occupational exposure (49), as well as health status (50).…”

Section: Discussionmentioning

confidence: 99%

Fast Image Analysis for the Micronucleus Assay in a Fully Automated High-Throughput Biodosimetry System

et al. 2014

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“…The rate of MN per BNC as a function of dose demonstrates some variability between donors. This variability in the CBMN assay has been well documented in the literature and Kacprzak et al showed that for five donors, the rate of MN per 1,000 BNC was relatively consistent at 0 Gy but had an approximate range of 0.2‐0.6 at 5 Gy. While the variability between donors across the dose range has been reduced in this work to 15% compared to 26% in our previous analysis template , the need for generating a calibration curve that is based on the rate of MN per BNC from multiple donors is demonstrated.…”

Section: Discussionmentioning

confidence: 53%

Optimized automated data analysis for the cytokinesis‐block micronucleus assay using imaging flow cytometry for high throughput radiation biodosimetry

Rodrigues

Probst²,

Beaton-Green

et al. 2016

Cytometry Pt A

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The cytokinesis-block micronucleus (CBMN) assay is a well-established technique that can be employed in triage radiation biodosimetry to estimate whole body doses of radiation to potentially exposed individuals through quantitation of the frequency of micronuclei (MN) in binucleated lymphocyte cells (BNCs). The assay has been partially automated using traditional microscope-based methods and most recently has been modified for application on the ImageStream X (IS X ) imaging flow cytometer. This modification has allowed for a similar number of BNCs to be automatically scored as compared to traditional microscopy in a much shorter time period. However, the MN frequency measured was much lower than both manual and automated slide-based methods of performing the assay. This work describes the optimized analysis template which implements newly developed functions in the IDEAS V R data analysis software for the IS X that enhances specificity for BNCs and increases the frequency of scored MN. A new dose response calibration curve is presented in which the average rate of MN per BNC is of similar magnitude to those presented in the literature using automated CBMN slide scoring methods. In addition, dose estimates were generated for nine irradiated, blinded samples and were found to be within 60.5 Gy of the delivered dose. Results demonstrate that the improved identification accuracy for MN and BNCs in the IS X -based version of the CBMN assay will translate to increased accuracy when estimating unknown radiation doses received by exposed individuals following large-scale radiological or nuclear emergencies.

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“…In addition to the 72 h culture time requirement, typical standard CBMN protocols involve culturing 500 mL to 2 mL of peripheral blood in a large volume of culture medium (5-20 mL) (McNamee et al 2009;IAEA 2011;Kacprzak et al 2013;Lyulko et al 2014). A reduction in the initial blood culture volume may eliminate the need for peripheral blood sampling by venipuncture, which is invasive and may not be possible depending on the nature of the emergency and the state of the patient.…”

mentioning

confidence: 99%

Validation of the Cytokinesis-block Micronucleus Assay Using Imaging Flow Cytometry for High Throughput Radiation Biodosimetry

Rodrigues¹,

Beaton-Green²,

Wilkins³

2016

Health Physics

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The cytokinesis-block micronucleus assay can be employed in triage radiation biodosimetry to determine the dose of radiation to an exposed individual by quantifying the frequency of micronuclei in binucleated lymphocyte cells. Partially automated analysis of the assay has been applied to traditional microscope-based methods, and most recently, the assay has been adapted to an automated imaging flow cytometry method. This method is able to automatically score a larger number of binucleated cells than are typically scored by microscopy. Whole blood samples were irradiated, divided into 2 mL and 200 μL aliquots, cultured for 48 h and 72 h, and processed to generate calibration curves from 0-4 Gy. To validate the method for use in radiation biodosimetry, nine separate whole blood samples were then irradiated to known doses, blinded, and processed. Results indicate that dose estimations can be determined to within ±0.5 Gy of the delivered dose after only 48 h of culture time with an initial blood volume of 200 μL. By performing the cytokinesis-block micronucleus assay using imaging flow cytometry, a significant reduction in the culture time and volume requirements is possible, which greatly increases the applicability of the assay in high throughput triage radiation biodosimetry.

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Individual variations in the micronucleus assay for biological dosimetry after high dose exposure

Cited by 16 publications

References 17 publications

Fast Image Analysis for the Micronucleus Assay in a Fully Automated High-Throughput Biodosimetry System

Fast Image Analysis for the Micronucleus Assay in a Fully Automated High-Throughput Biodosimetry System

Optimized automated data analysis for the cytokinesis‐block micronucleus assay using imaging flow cytometry for high throughput radiation biodosimetry

Validation of the Cytokinesis-block Micronucleus Assay Using Imaging Flow Cytometry for High Throughput Radiation Biodosimetry

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