Visualizing Oocyte Lipids and Membranes

Young, H₂O₂-treated, and aged oocytes were treated with two fluorescent dyes to visualize natural fatty acids and plasma membrane [40 (link)]. To stain oocyte plasma membranes, CellMask Deep Red Plasma Membrane Stain (C10046, Invitrogen, Grand Island, NY, USA) was used at 2.5 μg/mL in media for 30 min. BODIPY fatty acid 500/510 (D-3823, Invitrogen) was used for staining natural fatty acids in mouse oocytes. These oocytes were washed three times with media and transferred to a glass-bottom confocal dish (SPL Life Sciences, Pocheon, Korea). Live-cell images were captured with an Olympus FV1000 spectral confocal microscope (Olympus, Tokyo, Japan) equipped with a warm plate or with an LSM 710 confocal microscope (Carl Ziess, Oberkochen, Germany).