1230 tem

Young adult wild-type N2 and ptrn-1(tm5597) animals were prepared as described (Cueva et al., 2012 (link)). Briefly, animals were frozen in an EMPACT2 high-pressure freezer system, and a Leica AFS freeze substitution apparatus (Vienna, Austria) was used to preserve in 2% glutaraldehyde plus 1% osmium tetroxide and embed in Epon/Araldite. A Leica Ultracut S microtome equipped with a diamond knife was used to cut 50-nm serial sections, which were collected on Formvar-coated copper slot grids. The grids were poststained to enhance contrast in 3.5% uranyl acetate (30 s) and Reynold’s lead citrate preparation (3 min). The grids were imaged on a transmission electron microscope (JEOL TEM 1230, Tokyo, Japan), and images were acquired with an 11 megapixel bottom-mounted cooled CCD camera (Orius SC1000, Gatan, Pleasanton, CA).

The morphology and structure of the selected TCR–SPLs (F3 OA 1) were studied using TEM (TEM 1230; JEOL, Tokyo, Japan). A drop of the TCR–SPL suspension was employed to immerse the copper grid for 1 min. Consequently, the particles on the grid were stained with 2% (w/v) phosphotungstic acid solution for 10 s and cleansed two times with distilled water for 1 s. The grids were dehydrated in a dust-free atmosphere for 30 min and then studied by TEM accurately at an acceleration voltage of 100 Kv.

Mid-log-phase cultures were applied to glow-discharged carbon-coated grids, stained with 1.5% uranyl acetate, blotted, and air dried. Images were taken at 80 kV on a JEOL TEM1230 transmission electron microscope equipped with a Gatan 967 slow-scan, cooled CCD camera.

T. cruzi trypomastigotes (5x10⁷) were treated with DMS (1, 2 or 4 µM) and incubated for 24 h at 37 °C. After incubation, parasites were fixed for 1 h at room temperature with 2% formaldehyde and 2.5% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA) in sodium cacodylate buffer (0.1 M, pH 7.2) for 1 h at room temperature. After fixation, parasites were processed for transmission electron microscopy as previously described^{50 (link)}. Images were obtained in a JEOL TEM-1230 transmission electron microscope.

The RAW264.7 cells were collected and washed with PBS 3 times then fixed in 2.5% glutaraldehyde solution and osmium tetroxide, dehydrated through a graded ethanol series to 100%, and polymerized in epoxy resin. Ultrathin sections were collected, stained, and imaged using a transmission electron microscope (JEOL, transmission electron microscopy [TEM]-1230).

The powder X-ray diffraction (XRD) study of the obtained materials was carried out using a PANalytical X'Pert Pro diffractometer. Fourier transform infrared spectroscopy (FTIR) in KBr pellets was recorded using a Nicolet 470 Nexus instrument (Thermo Scientific, USA). Magnetic properties of nanoparticles were measured with an EV9 vibrating sample magnetometer. In order to determine the saturation magnetization (M_s), magnetic hysteresis loop experiments were performed in a magnetic field (H) of 20 kOe. Transmission electron microscopy (TEM) was used to examine the size and morphology of the biosynthesized MNPs. The images were done by TEM 1230 JEOL (Tokyo, Japan) with an acceleration voltage of 80 kV. The JSM-6100 (JEOL, Japan) scanning electron microscope (SEM) equipped with an Oxford Instruments INCA energy-dispersive X-ray spectrometer (EDX) was used to study the chemical composition of solids. Textural parameters of the magnetic materials were determined from the N₂ adsorption/desorption isotherms recorded at 77 K with an ASAP 2020 apparatus. The specific surface area (S_BET) of the samples was determined by the BET method.^{36 (link)} X-ray photoelectron spectroscopy (XPS) spectra were measured with an Axis Ultra DLD electron spectrometer (Kratos Analytical, UK) using a monochromated Al Kα source that was operated at 150 W.