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The micronucleus assay on cryopreserved whole blood

Renu Dayal (UGent) , Elien Beyls (UGent) , Anne Vral (UGent) and Ans Baeyens (UGent)
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Abstract
The in vitro cytokinesis-block micronucleus (CBMN) assay is a widely used technique in radiobiology research, biological dosimetry, genotoxicity studies, and in vitro radiosensitivity testing. This cytogenetic method is based on the detection of micronuclei in binucleated cells resulting from chromosomal fragments lagging during cell division. Fresh whole blood samples are the most preferred sample type for the CBMN assay. However, the disadvantages of working with fresh blood samples include immediate processing after blood collection and the limited number of repeated analyses that can be performed without extra blood sampling. As the need for fresh blood samples can be logistically challenging, CBMN assay on cryopreserved whole blood samples would be of great advantage, especially in large-scale patient studies. This paper describes a protocol to freeze whole blood samples and to perform the CBMN assay on these frozen blood samples. Blood samples from healthy volunteers have been frozen and thawed at different time points and then, subjected to a modified micronucleus assay protocol. The results demonstrate that this optimized procedure allows the performance of the CBMN assay on frozen blood samples. The described cryopreservation protocol may also be very useful for other cytogenetic assays and a variety of functional assays requiring proliferating lymphocytes.
Keywords
General Immunology and Microbiology, General Biochemistry, Genetics and Molecular Biology, General Chemical Engineering, General Neuroscience

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MLA
Dayal, Renu, et al. “The Micronucleus Assay on Cryopreserved Whole Blood.” JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, no. 204, 2024, doi:10.3791/65855.
APA
Dayal, R., Beyls, E., Vral, A., & Baeyens, A. (2024). The micronucleus assay on cryopreserved whole blood. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, (204). https://doi.org/10.3791/65855
Chicago author-date
Dayal, Renu, Elien Beyls, Anne Vral, and Ans Baeyens. 2024. “The Micronucleus Assay on Cryopreserved Whole Blood.” JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, no. 204. https://doi.org/10.3791/65855.
Chicago author-date (all authors)
Dayal, Renu, Elien Beyls, Anne Vral, and Ans Baeyens. 2024. “The Micronucleus Assay on Cryopreserved Whole Blood.” JOVE-JOURNAL OF VISUALIZED EXPERIMENTS (204). doi:10.3791/65855.
Vancouver
1.
Dayal R, Beyls E, Vral A, Baeyens A. The micronucleus assay on cryopreserved whole blood. JOVE-JOURNAL OF VISUALIZED EXPERIMENTS. 2024;(204).
IEEE
[1]
R. Dayal, E. Beyls, A. Vral, and A. Baeyens, “The micronucleus assay on cryopreserved whole blood,” JOVE-JOURNAL OF VISUALIZED EXPERIMENTS, no. 204, 2024.
@article{01HR1TAZCNTKW156G4RATCT39Z,
  abstract     = {{The in vitro cytokinesis-block micronucleus (CBMN) assay is a widely used technique in radiobiology research, biological dosimetry, genotoxicity studies, and in vitro radiosensitivity testing. This cytogenetic method is based on the detection of micronuclei in binucleated cells resulting from chromosomal fragments lagging during cell division. Fresh whole blood samples are the most preferred sample type for the CBMN assay. However, the disadvantages of working with fresh blood samples include immediate processing after blood collection and the limited number of repeated analyses that can be performed without extra blood sampling.

As the need for fresh blood samples can be logistically challenging, CBMN assay on cryopreserved whole blood samples would be of great advantage, especially in large-scale patient studies. This paper describes a protocol to freeze whole blood samples and to perform the CBMN assay on these frozen blood samples. Blood samples from healthy volunteers have been frozen and thawed at different time points and then, subjected to a modified micronucleus assay protocol. The results demonstrate that this optimized procedure allows the performance of the CBMN assay on frozen blood samples. The described cryopreservation protocol may also be very useful for other cytogenetic assays and a variety of functional assays requiring proliferating lymphocytes.}},
  articleno    = {{e65855}},
  author       = {{Dayal, Renu and Beyls, Elien and Vral, Anne and Baeyens, Ans}},
  issn         = {{1940-087X}},
  journal      = {{JOVE-JOURNAL OF VISUALIZED EXPERIMENTS}},
  keywords     = {{General Immunology and Microbiology,General Biochemistry, Genetics and Molecular Biology,General Chemical Engineering,General Neuroscience}},
  language     = {{eng}},
  number       = {{204}},
  pages        = {{10}},
  title        = {{The micronucleus assay on cryopreserved whole blood}},
  url          = {{http://doi.org/10.3791/65855}},
  year         = {{2024}},
}

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