Em400t

Subcellular quantification was performed using a transmission electron microscope (Philips EM 400 T, Kassel, Germany) by meander-shaped counting of 100 gold particles (12 nm) within the epithelium. Specificity of ERp57-antibody was assessed by evaluating the labelling of goblet cell granules as negative control. For quantifying the intracellular distribution of ERp57 within the epithelium, enterocytes and RACE were screened in a meander-shaped manner and 100 gold particles (12 nm) were counted. To investigate the concentration of ERp57 in the ER, 10 standardized images (magnification × 17,700) of enterocytes and RACE of each patient were taken. To evaluate the ER volume and the concentration of ERp57 within this compartment, a standardized grid (square side length: 4000 µm) and the following formulas were used to calculate organelle circumference (OC) and labelling density (LD) as adapted from Griffiths [24 ].

G = number of gold particles, GF = Grid factor (4,000), Ism = Intersections ER membrane, ISlu = Intersections ER lumen, M = Magnification (× 24,000).

Freshly isolated, 4- and 14-day cultured c-kit⁺ cells were recovered from the scaffolds as already described, pelleted and fixed in 3 % glutaraldehyde, and processed for transmission electron microscopy (TEM) as described previously [13 (link)]. Cells were visualized using a transmission electron microscope at 60 kV (EM 400 T, Philips, Eindhoven, The Netherlands).

Samples were stained with potassium permanganate (KMnO₄) and examined using transmission electron microscopy (TEM). The pulp from 2 mm inside of the peel to 0.5 cm outside of the stone was cut with a sharp blade to generate blocks of 0.5 cm × 0.2 cm × 0.2 cm. The blocks were immediately placed into fixation solution containing 3.5% glutaraldehyde and 0.2 M phosphate (pH 7.2), fixed under low vacuum for 1 h, followed by atmospheric pressure for 2.5 h. The samples were then washed with phosphate buffer and fixed in 1% osmium tetroxide (in 0.2 M phosphate buffer, pH 7.2) at 4°C overnight. The samples were subsequently dehydrated in a series of Spurr resin-water mixtures (1:2, 1:1 and 2:1), supplied by Marsys-Tech Co. Ltd., (Beijing, China, catalog number S2690), before being treated twice with pure Spurr resin. They were then embedded and aggregated, and cut into ultrathin sections with an LKB-2188 microtome. The ultrathin sections were collected with a 100 - mesh copper net. Some of the sections were double stained with uranyl acetate and lead citrate. The remaining sections were stained with 1% KMnO₄ solution (in 0.1% sodium citrate) for 3 min (Yin et al., 2003 (link); Day et al., 2005 (link)). The stained sections were examined and photographed under a TEM (Philips EM-400T). The experiment was repeated six times, with one sample observed each time.

Brain tissues were collected from patients who underwent neurosurgery and immediately fixed in a solution containing 4% paraformaldehyde and 0.5% glutaraldehyde/0.2 M cacodylate. The samples were then post-fixed with 1% osmium tetroxide and embedded in EmBed812. Ultrathin sections were stained with uranyl acetate and lead citrate and examined using a Philips EM-400T transmission electron microscope at 80 kV with the TIA software.

To synthesize 5% of TiO₂ NP, the technique previously described by Venkatasubbu et al. (2012) was utilized. The characterization of the TiO₂ NP and the uniform distribution of the nanoparticles were analysed using samples for transmission electron microscopy (TEM, JEOL, Tokyo, Japan). The TEM analysis was undertaken by placing TiO₂ nanoparticles onto a carbon-coated copper grid that had been left at room temperature overnight. The images were captured using a Philips EM400T operating at 200 kV, with a magnification of roughly 50 nanometres (nm) and a point-to-point resolution of 2 A° [9 (link)].