High-throughput sample processing and sample management; the functional evolution of classical cytogenetic assay towards automation.

Ramakumar, Adarsh; Subramanian, Uma; Prasanna, Pataje G.S.

doi:10.1016/j.mrgentox.2015.07.011

Cited by 12 publications

(7 citation statements)

References 23 publications

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“…DCScore would not be as effective as ADCI in a radiation event involving a large number of potentially exposed individuals. Large aperture microscope systems can collect images of multiple metaphase cells 18 , however, they do not analyze them. "CABAS" 19 and "Dose Estimate" 20 software can generate calibration curves and estimate dose, but do not score DCs.…”

Section: Discussionmentioning

confidence: 99%

Expedited Radiation Biodosimetry by Automated Dicentric Chromosome Identification (ADCI) and Dose Estimation

Shirley¹,

Li²,

Knoll

et al. 2017

JoVE

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Biological radiation dose can be estimated from dicentric chromosome frequencies in metaphase cells. Performing these cytogenetic dicentric chromosome assays is traditionally a manual, labor-intensive process not well suited to handle the volume of samples which may require examination in the wake of a mass casualty event. Automated Dicentric Chromosome Identifier and Dose Estimator (ADCI) software automates this process by examining sets of metaphase images using machine learning-based image processing techniques. The software selects appropriate images for analysis by removing unsuitable images, classifies each object as either a centromere-containing chromosome or non-chromosome, further distinguishes chromosomes as monocentric chromosomes (MCs) or dicentric chromosomes (DCs), determines DC frequency within a sample, and estimates biological radiation dose by comparing sample DC frequency with calibration curves computed using calibration samples. This protocol describes the usage of ADCI software. Typically, both calibration (known dose) and test (unknown dose) sets of metaphase images are imported to perform accurate dose estimation. Optimal images for analysis can be found automatically using preset image filters or can also be filtered through manual inspection. The software processes images within each sample and DC frequencies are computed at different levels of stringency for calling DCs, using a machine learning approach. Linear-quadratic calibration curves are generated based on DC frequencies in calibration samples exposed to known physical doses. Doses of test samples exposed to uncertain radiation levels are estimated from their DC frequencies using these calibration curves. Reports can be generated upon request and provide summary of results of one or more samples, of one or more calibration curves, or of dose estimation.

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Section: Discussionmentioning

confidence: 99%

Expedited Radiation Biodosimetry by Automated Dicentric Chromosome Identification (ADCI) and Dose Estimation

Shirley¹,

Li²,

Knoll

et al. 2017

JoVE

View full text Add to dashboard Cite

show abstract

“…Certain studies have reported that DCA is affected not only by radiation exposure but also due to variability by different chemical compounds that induce cancer development (Livingston et al, 2016). At present, one of the limited uses to assess radiation dose to individuals after a mass casualty is throughput; although there have been several reports on automation assays, considerable individual variation by personnel training has been reported (Beaton-Green et al, 2016; Ramakumar et al, 2015). According to the World Health Organization (WHO) BioDoseNet, which is an international network of biodosimetry laboratories, there are two methods for DCA scoring: conventional analysis (C-DCA) and QuickScan analysis (QS-DCA) (Lin et al, 2021).…”

Section: Discussionmentioning

confidence: 99%

“…However, the EPR linear dose-response curves in the signal dose-response were in the range of 5-80 kGy (Miyake et al, 2000). The excessive range with the median lethal dose of radiation (LD50/60), is estimated to be 4.14 Gy, which hinders the use of this technology (Ramakumar et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Intercomparison of DCA and EPR scoring for validation with individual physical dosimetry

Chang,

Cheng,

Lin

et al. 2024

Preprint

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Radiation is widely used in industrial, medical, agricultural, and military applications. The improper use and disposal of manmade radiation sources have resulted in various events that have caused different levels of human radiation exposure. These experiences indicate that estimating only the amount of radioactive contamination and dose after an incident was not sufficient to realize in detail and determine the exact exposure dose owing to the incident. Therefore, a retrospective assessment of radiation exposure and dose reconstruction is a necessary and crucial method for risk analysis. In this studies, we used one healthy volunteer peripheral blood and fingernail samples for the project were irradiated with 0–5 and 0–20 Gy Co-60 irradiation for establish a dose-dependent curve with DCA and EPR dose-response curve assays, respectively. Further, another 20 volunteers participated in this clinical trial; their DCA and EPR scores were calculated, and followed by an evaluation of their equivalent radiation doses and corresponded with their monthly physical dosimetry. Through this study, we established the dose-response curve of DCA and EPR after radiation exposure, and calculated the correlation between physical dosimetry and biological doses by 20 volunteers participating in clinical trials. This evidence will promote estimating the personal absorbed doses and as a reference for medical treatment for the safety of radiation workers and normal populations.

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“…The simplicity of the scoring procedure suggests that the assay is suitable for automation. It has been suggested that the automation of image examination and analysis of chromosomal aberration would aid in speeding up the estimation of dose and better support medical management of exposed individuals ( 4 , 5 , 18 ) . This option has not yet been tested by us but is envisaged as a future project.…”

Section: Discussionmentioning

confidence: 99%

Fluorescence in situ hybridisation for interphase chromosomal aberration-based biological dosimetry

Meher,

Lundholm,

Wojcik

2023

Radiation Protection Dosimetry

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Metaphase spreads stained with Giemsa or painted with chromosome-specific probes by fluorescence in situ hybridisation (FISH) have been in use since long for retrospective dose assessment (biological dosimetry). However, in cases of accidental exposure to ionising radiation, the culturing of lymphocytes to obtain metaphase chromosomes and analysis of chromosomal aberrations is time-consuming and problematic after high radiation doses. Similarly, analysing chromosomal damage in G0/G1 cells or nondividing cells by premature chromosome condensation is laborious. Following large-scale radiological emergencies, the time required for analysis is more important than precision of dose estimate. Painting of whole chromosomes using chromosome-specific probes in interphase nuclei by the FISH technique will eliminate the time required for cell culture and allow a fast dose estimate, provided that a meaningful dose-response can be obtained by scoring the number of chromosomal domains visible in interphase nuclei. In order to test the applicability of interphase FISH for quick biological dosimetry, whole blood from a healthy donor was irradiated with 8 Gy of gamma radiation. Irradiated whole blood was kept for 2 h at 37°C to allow DNA repair and thereafter processed for FISH with probes specific for Chromosomes-1 and 2. Damaged chromosomal fragments, distinguished by extra color domains, were observed in interphase nuclei of lymphocytes irradiated with 8 Gy. These fragments were efficiently detected and quantified by the FISH technique utilising both confocal and single plane fluorescence microscopy. Furthermore, a clear dose-response curve for interphase fragments was achieved following exposure to 0, 1, 2, 4 and 8 Gy of gamma radiation. These results demonstrate interphase FISH as a promising test for biodosimetry and for studying cytogenetic effects of radiation in nondividing cells.

show abstract

High-throughput sample processing and sample management; the functional evolution of classical cytogenetic assay towards automation.

Cited by 12 publications

References 23 publications

Expedited Radiation Biodosimetry by Automated Dicentric Chromosome Identification (ADCI) and Dose Estimation

Expedited Radiation Biodosimetry by Automated Dicentric Chromosome Identification (ADCI) and Dose Estimation

Intercomparison of DCA and EPR scoring for validation with individual physical dosimetry

Fluorescence in situ hybridisation for interphase chromosomal aberration-based biological dosimetry

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