Use of A Fluorescent Stain for Visualization of Nuclear Material in Living Oocytes and Early Embryos

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Abstract
Hoechst dyes 33342 and 33258 were used to visualize pronuclei and nuclei of early preimplantation embryos. Murine one-cell zygotes exposed to dye stained rapidly over a range of concentrations (0, 0.02, 0.04, 0.1 or 0.2 μg/50 μl of media). Development to morula and blastocyst in vitro was reduced (39/70, 56%; p < 0.05) compared to controls (44/57, 77%) but not completely blocked. Porcine and bovine zygotes and embryos could also be stained but required incubation times up to 4 hr. Porcine embryos exposed to Hoechst 33342 had limited (p < 0.01) in vitro development (29/74, 39%) compared to unstained controls (49164, 76%). Hoechst dyes stain embryos from different species but suitably adjusted incubation times are required. Limited preimplantation development in vitro may be expected following staining and exposure to ultraviolet light.