Embryos (from morula to hatching stage) were loaded with 0.5 μM CoroNa Green AM (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions and a previous report [12 (link)] with slight modifications, at 37°C for 30 min. Subsequently, intracellular and/or blastocoel fluorescence intensity was measured using an LSM780 confocal laser-scanning microscope (Carl Zeiss AG, Jena, Germany). The excitation and emission wavelengths were 488 and 510–520 nm, respectively (Fig 1A ). The same experiments were repeated at least three times.
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