Use of 2-color flow cytometry to assess radiationinduced geotoxic damage on cho-ki cells
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2017
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INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE
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The micronucleus assay is an important technique used to evaluate genotoxic damage of chemical or physical
agents (as ionizing radiations) on cells, based on quantification of cells bearing micronuclei, which are fragments
derived from damage (breakage) of the DNA. Currently, this technique was updated to an automated approach
that relies on plasma membrane dissolution to analyze fluorescent dye-labelled nuclei and micronuclei by flow
cytometry. Cell suspensions were irradiated in PBS by a ⁶⁰Co source in doses between 0 and 16Gy, and incubated
by 48h. Cell membranes were lysed in the presence of SYTOX Green and EMA dyes, so EMA-stained nuclei
could be discriminated as from dead cells, and nuclei and micronuclei could be quantified. Amounts of
micronuclei (percent of events) in the samples, were found to be proportional to radiation doses, and could be
fitted to a linear-quadratic model (R² = 0.993). Only higher doses (8 and 16Gy) and positive control could induce
relevant increases in micronucleus amounts. The incorporation EMA showed an increase in irradiated cells. Midto
high doses (4, 8 and 16Gy) induced reduction of cell proliferation. Experiments showed the suitability of the
technique to replace traditional microscopy analysis in evaluation of the effects of ionizing radiations on cells,
with possibility to use in biological dosimetry.
Como referenciar
CARVALHO, LUMA R. de; BONFIM, LETICIA; VIEIRA, DANIEL P. Use of 2-color flow cytometry to assess radiationinduced geotoxic damage on cho-ki cells. In: INTERNATIONAL NUCLEAR ATLANTIC CONFERENCE, October 22-27, 2017, Belo Horizonte, MG. Proceedings... Rio de Janeiro, RJ: Associação Brasileira de Energia Nuclear, 2017. Disponível em: http://repositorio.ipen.br/handle/123456789/28296. Acesso em: 10 May 2024.
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