Cells were cultured in a six-well plate and then fixed with 2.5% glutaraldehyde in PBS at 4°C for 4 hours. Following this, the cells were washed three times with PBS for 15 minutes each. Post-fixation was then conducted using 1% OsO4 in PBS for 1.5 hours, followed by three more washes with PBS for 15 minutes. Subsequently, a graded series of ethanol solutions (30%, 50%, 70%, and 80%) was used for dehydration for 15 minutes, followed by a graded series of acetone solutions (90% and 95%) for 15 minutes. This was followed by dehydration in absolute acetone for 20 minutes twice. The cells were then placed in a mixture of absolute acetone and the final Spurr resin in a 1:1 ratio for 1 hour at room temperature and transferred to a 1:3 mixture of absolute acetone and the final resin mixture for 2 hours. Finally, the cells were transferred to a mixture of the final Spurr resin and absolute acetone overnight. After the embedding process was completed, ultrathin sections were cut and stained, and images of the samples were obtained using a transmission electron microscope (H-7650, Hitachi, Japan).
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