Megaview g2 camera

Average hydrodynamic diameter (nm), polydispersity index (PdI) and ζ-potential (mV) of glycerosomes were measured by Dynamic and Electrophoretic Light Scattering, DLS-ELS (Zetasizer Nanoseries ZS90) by Malvern instrument (Worcestershire, UK) at 25 °C, with a scattering angle of 90 °C [26 (link)]. Glycerosomes were diluted using ultrapure water before measurements, in order to achieve a suitable scattering intensity. Successively, glycerosomes were observed by Transmission Electron Microscope, TEM (CM12 TEM, PHILIPS, Eindhoven, The Netherlands) equipped with an OLYMPUS Megaview G2 camera and with an accelerating voltage of 80 kV. A drop of sample, diluted 5-fold in water, was applied and dried by desiccation on a carbon film copper grid and it was counterstained with 1% (w/v) of phosphotungstic acid solution for 3 min. Then, the sample was examined at different amplifications.

Morphologic analysis of K-NLCs was carried out by transmission electron microscopy (TEM) on a TEM CM12 PHILIPS equipped with an OLYMPUS Megaview G2 camera at an accelerating voltage of 80 keV [41 (link)]. For the analyses, freeze-dried K-NLCs were dispersed in ultrapure water at the initial concentration and obtained nanoparticle dispersion was placed on a carbon film-covered copper grid. Subsequently, the excess of the sample was blotted from the grid with filter paper to obtain a thin film stained with a phosphotungstic acid solution (1% w/v) in distilled water. The analysis of the samples was performed 3 min after the staining.

MCF-7, MCF-7/Fulv, and MCF-7/Tam cells were cultured in a six-well plate at a density of 1 × 10⁵ cells/well for 24 h and then a standard procedure was applied (21 (link)). Briefly, cells were washed once with PBS and then fixed with 2.5% glutaraldehyde at 4°C for 2 h. After washing with PBS, cells were dehydrated by using a gradient of ethanol and finally incubated with 100% acetone for 15 min and embedded in SPURR resin. Ultrathin sections were taken and stained with uranyl acetate and lead citrate, and images were captured by using a JEOL 100S transmission electron microscope equipped with an Olympus MegaView G2 camera.

For electron microscopy studies, brains were processed as previously described. In brief, brain samples were fixed by immersion in 4% glutaraldehyde, buffered to pH 7.3 with Millonig's fluid, post‐fixed in 1% osmium tetroxide in the same buffer, and dehydrated in acetone for embedding in Araldite (Sigma). Ultrathin sections were obtained using an OM‐U3 ultramicrotome (Reichert, Vienna, Austria), double‐stained with uranyl acetate and lead citrate, and examined using a JEM 1010 electron microscopy (JEOL, Tokyo, Japan) at the ICTS Spanish National Centre for Electron Microscopy. Images were recorded using a MegaView G2 camera using iTEM Imaging Platform software (Olympus, Münster, Germany).

The morphology of the synthesized AuNPs with sizes of 10 nm and 30 nm was analyzed by transmission electron microscopy (TEM). On a copper grid covered in a Formvar-carbon membrane (300 mesh, Ted Pella Inc., Redding, CA, USA)), 5–10 L of an aqueous dispersion was first applied and air dried at room temperature. Ultrastructural and morphometric analysis of the gold nanoparticles was performed using a TEM model JEM 1010 (Jeol, Tokyo, Japan) equipped with a MEGAVIEW G2 camera and the cooperating iTEM Digital Imaging Solutions program (Olympus Soft Imaging Solutions GmbH, Münster, Germany). The average diameter of the AuNPs reached about 10.5 nm +/− 1.3 nm and 30.9 nm +/− 1.8. The UV–VIS measurements were performed using a Jasco 650v (ABL&E-JASCO, Kraków, Poland).

The nerve samples collected from each group were processed for TEM according to the chemical fixation and embedding protocol (Hayat, 2000) [44 ]. Briefly, they were prefixed for 2 h in glutaraldehyde 2.7% in phosphate buffer 0.1 M, pH 7.4 and postfixed for 1.5 h in OsO₄ 1.5% in phosphate buffer 0.15 M, pH 7.4. Then, the samples were dehydrated in ethanol solution of increasing concentrations (30 minutes each), and infiltrated with EMBED 812. The sections double contrasted for 10 minutes with 13% uranyl acetate and for 5 min with 2.8% lead citrate were examined on a Jeol JEM 1011 transmission electron microscope (Jeol, Tokyo, Japan), equipped with a Mega View G2 camera (Olympus Soft Imaging Solutions, Münster, Germany).
Nerve samples from the CMNPs treatment group were analyzed using a Hitachi HD-2700 scanning transmission electron microscope (Hitachi High-Tech, Krefeld, Germany), which was equipped with an X-Max 1160 energy-dispersive X-ray spectroscopy (EDX) detector (Oxford Instruments, Wiesbaden, Germany) to characterize their compositional structure. The EDX is considered a valuable instrument in every research that necessitates elemental determination, either endogenous or exogenous, in the tissue, cell, or other new types of analyses.