Tcnai g2

Somatic embryo explants generated by DEC, DSEI, ECS and ECB were collected on the 5th day. The samples were pre-fixed in 3% glutaraldehyde in phosphate buffer [pH 7.4] for at least 3 hours at 4°C. After rinsing 3 times with phosphate buffer, samples were fixed with 1% osmium tetroxide for 1–2 hours, dehydrated in an increasing concentration series of acetone, and embedded in epoxy resin. Ultrathin sections were obtained with an ultramicrotome (Leica Ultracut UCT) and contrasted with uranyl acetate and lead citrate. Image collection was performed with a transmission electron microscope (FEI/Philips TCNAI G2) at 50 kV.

The morphology and size of BMNPs were checked by transmission electron microscopy (TEM, FEI/Philips TCNAI G2) at an accelerating voltage of 200 kV and direct magnification of×250,000. The composition and phase of BMNPs were recognized by powder X-ray diffraction (XRD) on an D/max-rB X-ray diffractometer (Rigaku, Japan) using Cu Kα radiation (λ=1.5418 Å). Fourier transform infrared (FT-IR) spectra of the nanoparticles were recorded in the range of 350-7800 cm⁻¹ using FT-IR spectroscopy (Nicolette-6700). Room temperature-magnetic experiments were performed on a vibrating sample magnetometer (Model 3900, Princeton Measurements Corporation, sensitivity is 5.0×10−10 Am2). The hysteresis loop was measured between +500 and −500 mT with an average time of 400 ms. Saturation magnetization (Ms) and saturation remanence (Mrs) were determined after correction for paramagnetic phases.

Mitochondrial morphology in L02 cells and liver samples were observed by transmission electron microscope (FEI/Philips TCNAI G2, Eindhoven, The Netherlands).