Eftem

All chemicals and reagents purchased from sigma Aldrich and Daejung chemicals South Korea, and TCI chemicals, Japan without further purifications. Absorption spectra and drug release study were recorded in 1 cm quartz cuvettes using Evolution™ 60 UV–visible Spectrophotometer (Thermo Fischer Scientific, USA). Emission spectra were obtained on a Jasco-FP 6500 spectrofluorometer. Particle size was analyzed by Dynamic Light Scattering Spectrophotometer (Otsuka Electronics Co., Ltd, Japan). Zeta potential was measured by electrophoretic light scattering (ELS) spectrometer (Otsuka Electronics Co., Ltd, Japan). The morphology of the Ru-1@TPP-PEG-Biotin SAN was analyzed using Energy-Filtering Transmission Electron Microscope (EF-TEM, Carl Zeiss, LIBRA 120, Germany). Fourier transform infrared (FTIR) spectroscopy experiments performed using a JASCO, FT/IR-4200 instrument. The human breast cancer cell line MCF-7 and hepatoma cell line HepG2 were supplied from the Korean Cell Line Bank (KCLB, Korea). Quantum dot conjugation kit (Invitrogen, USA) was used to conjugate the antibody with a quantum dot.

Light microscopy was performed as described before [72 (link)]. Cell size and number determination covered all parts of the leaf sparing cells surrounding the primary vein and at the edge of the leaves. For transmission electron microscopy, leaf samples were fixed in 2.5% glutaraldehyde, 0.1 M cacodylate buffer (pH 7.4), 5 mM calcium chloride for 4 h at 4°C, and post-fixed with 1% Os0₄ and 0.8% K₃Fe(CN)₆ for 2 h at 4°C. The samples were washed with water and post-stained with 2% aqueous uranyl acetate for 2 h. Subsequently, the tissue was dehydrated in a series of ethanol and propylene oxide and embedded in Spurr’s low viscosity epoxy resin. Ultrathin sections (60–70 nm) were cut with a Leica UC6 ultramicrotome using a diamond knife, stained with uranyl acetate and lead citrate and examined on an energy-filtering transmission electron microscope (EFTEM, Zeiss) at 120 kV.

Oenothera leaf pieces of 2 mm² were fixed with 2.5% [w/v] glutaraldehyde and 2% [w/v] paraformaldehyde in 0.2 M sodium cacodylate buffer (pH 7.4) for at least 4 h at 4°C. Samples were then post-fixed in 1% [w/v] OsO₄ at 4°C overnight, stained in 2% [w/v] aqueous uranyl acetate for 2 h and dehydrated gradually in 30%, 50%, 70%, 80%, 90%, and 100% [v/v] ethanol followed by washing in 100% [v/v] propylene oxide two times. The samples were then embedded in low-melting Spurr epoxy resin (Agar Scientific), degassed and cured at 65°C for 24 h. Thin sections (60–70 nm) were obtained with a Leica Ultracut UC 6 (Leica Microsystems), mounted on 150-mesh nickel grids, counterstained with uranyl acetate followed by lead citrate, and examined with a transmission electron microscope at 120 kV (EFTEM, Zeiss).