Flow cytometric detection of micronuclei by combined staining of DNA and membranes

Cytometry
J M Wessels, M Nüsse

Abstract

A new staining method is presented for flow cytometric measurement of micronuclei (MN) in cell cultures and human lymphocytes using membrane-specific fluorescent dyes in addition to DNA staining. Several combinations of fluorescent membrane and DNA dyes were studied for a better discrimination of MN from debris in a suspension of nuclei and micronuclei. For staining of membranes, the lipophilic dyes 2-hydroxyethyl-7,12,17-tris(methoxyethyl)porphycene (HEPn) and 1,6-diphenyl-1,3,5-hexatriene (DPH) were used in combination with ethidium bromide (EB), proflavine (PF), and Hoechst 33258 (HO). Due to their spectral properties, HO or EB combined with HEPn were not as suitable for the discrimination of MN from debris as was HEPn in combination with PF. With HEPn in combination with PF, however, additional noise was found at low fluorescence intensities, probably due to free fluorescent dye molecules in the solution. The optimal simultaneous staining of membranes and DNA was obtained using a combination of DPH and EB. The induction of MN in Chinese hamster and mouse NIH-3T3 cells by UV-B illumination was studied with this new staining technique. UV-B illumination (280-360 nm) induced MN in both cell lines. Chinese hamster cells were fo...Continue Reading

References

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Citations

Nov 1, 1996·Mutation Research·W U MüllerC Streffer
Dec 18, 2010·Mutagenesis·Svetlana AvlasevichStephen Dertinger
Aug 22, 2000·The Journal of Investigative Dermatology·G EmriA A Schothorst
Jan 10, 2008·Mutation Research·Steven M BryceStephen D Dertinger
Mar 21, 2012·Environmental and Molecular Mutagenesis·Magdalena Lukamowicz-RajskaAzeddine Elhajouji
Oct 22, 2010·Environmental and Molecular Mutagenesis·Magdalena LukamowiczAzeddine Elhajouji
Dec 2, 2009·Environmental and Molecular Mutagenesis·Steven M BryceStephen D Dertinger
Mar 8, 2000·Pharmacological Research : the Official Journal of the Italian Pharmacological Society·C FimognariP Hrelia
Aug 18, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Matthew A Rodrigues

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