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. 2020 Aug 4;8:764. doi: 10.3389/fcell.2020.00764

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Copyright © 2020 Javvaji, Dhali, Francis, Kolte, Mech, Roy, Mishra and Bhatta.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Nitroblue tetrazolium (NBT) staining for assessing intracellular ROS in embryos. Ovine embryos (8-cell, morula and degenerating) were produced in vitro and stained with NBT. The dark purple/blue spots of formazan precipitates represent ROS. The level of intracellular ROS (% NBT stained area, mean ± SE) was determined in five individual embryos at each developmental stage. (A) Unstained degenerating embryo; (B) NBT-stained 8-cell embryo; (C) NBT-stained morulae; (D) NBT-stained degenerating embryo; (E) level of intracellular ROS in embryos at different developmental stages. Values without a common superscript (a,b) above error bar differ significantly (P < 0.05). BF, bright-field image; PH, phase-contrast image. Scale bar = 50 μm.