The use of acridine orange and ethidium bromide to determine the viability of pre-implantation mouse embryos cultured in vitro |
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| Authors: | S M Swanson A Ijaz M L Fahning |
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| Affiliation: | 1. Michigan Technological University, Department of Chemistry, Houghton, MI 49931, USA;2. North Carolina A&T State University, Physics Department and Computational Science and Engineering Department, Greensboro, NC 27411, USA;1. Instituto de Biologia do Exército, Rio de Janeiro, Brazil;2. Laboratório de Diagnósticos por DNA, DECOL, IBRAG, Universidade do Estado do Rio de Janeiro, Brazil;1. Department of Microbiology and Parasitology, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil;2. Department of Biochemistry and Molecular Biology, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil;3. Laboratory of Veterinary Pathology, Instituto Federal Catarinense (IFC), Concórdia, SC, Brazil;4. Department of Clinical and Toxicological Analysis, Universidade Federal de Santa Maria, Santa Maria, RS, Brazil;5. Laboratory of Nanosciences, Centro Universitário Franciscano, Santa Maria, RS, Brazil;6. Department of Animal Science, Universidade do Estado de Santa Catarina (UDESC), Chapecó, SC, Brazil;7. Laboraroty of General Biochemistry and Microorganisms, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Mato Grosso do Sul (UFMS), Campo Grande, MS, Brazil;1. Division of Immunology, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, India;2. Department of Biotechnology, National Institutes of Pharmaceutical Education and Research, Hajipur 844102, India;3. Dept. of Clinical Medicine, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, India;4. Dept. of Molecular Biology, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, India;5. Department of Bioinformatics, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, India;6. Department of Pathology, Rajendra Memorial Research Institute of Medical Sciences, Patna 800007, India |
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| Abstract: | Pre-implantation mouse embryos at the morula or early blastocyst stage were stained with acridine orange (AO) or ethidium bromide (EB) in order to assess their viability. There was a significant difference (p < 0·05) between the mortality rate of embryos stained with the two concentrations of AO tested (0·125 mg/ml and 0·250 mg/ml) and the unstained control. There was also a significant difference (p < 0·05) between embryos exposed to the dye for 2 min and the unstained control (40·7% vs 16%). Embryos stained with AO showed a light-to bright-green fluorescence with pronounced red-tinged areas within the cells. It was concluded that AO would not be suitable for assessing the viability of mouse embryos because it produced a similar coloured fluorescence in both dead and live cells and was toxic. Neither of the two concentrations of EB (20 μg/ml or 40 μg/ml) nor the two exposure times (3 min or 5 min) tested significantly increased the mortality rate of mouse embryos over that for the unstained control. EB produced green fluorescence in live cells and red fluorescence in dead cells of the stained embryos. EB does not appear to be toxic to pre-implantation mouse embryos, thus it may be a potentially useful tool for assessing their viability. |
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