Fei nova nanosem 450

All tissue samples were sputter-coated with 3 $\mathrm{n}$ $\mathrm{m}$ of platinum in high-resolution Turbo-Pumped Sputter Coater Q150T (Quorum Technologies Ltd, Ringmer, UK) before their examination under FEI Nova NanoSEM 450 field emission gun scanning electron microscope (Thermo Fisher Scientific, Brno, Czech Republic). The Navigation Montage option of the SEM software, v. 6.3.4.3233 (Helios NanoLab, Thermo Fisher Scientific, Brno, Czech Republic) was used to map whole tissue sections at low resolution. Final SEM analyses were performed at acceleration voltage ranging from 2 kV to 5 kV and spot size 3 using Everhart–Thornley Detector (ETD), Circular Backscatter Detector (CBS), and Through the Lens Detector (TLD). If specimen charging was experienced, a beam deceleration mode combined with the magnetic immersion final lens [31 (link),32 (link),33 (link)] was used.

SEM images were taken on a scanning electron microscope (FEI Nova NanoSEM 450, Thermo Fisher Scientific, Waltham, MA, USA), at the working distance of 5 mm, operation voltage of 5 kV and spot size of 3.0 nm. XRD patterns were collected on an X-ray diffractometer (BRUKER D8-ADVANCE, Bruker Co., Karlsruhe, Germany) using Cu Kα (λ = 1.5418 Å) radiation, with operation voltage 40 kV and current 40 mA, respectively. TEM images and SAED were observed on a transmission electron microscope (JEOL JEM-2100, JEOL, LTD, Akishima, Japan) at an acceleration voltage of 200 kV. UV–vis absorption spectra were obtained by using an UV-Vis spectrophotometer (PE Lambda 950, PerkinElmer Inc., Waltham, MA, USA) with a wavelength range of 300–800 nm. FTIR spectra were recorded on a Fourier transform infrared spectrometer (NICOLET AVATAR 360, Nicolet Instrument Corp., Richardson, TX, USA) with a wavenumber range of 4000–500 cm⁻¹.

Cells were treated with olaparib-Ga for 48 h, and then fixed, washed, and dehydrated with a graded series of ethanol and finally tert-butanol. After drying, cells were coated with platinum and assessed by SEM (FEI Nova NanoSEM 450; Thermo Fisher Scientific Inc.). The Ga³⁺ elements of the samples were examined using energy-dispersive x-ray spectroscopy (Octane EDS-70; EDAX; Ametek Inc.).

The morphology of the cross-section of surface treated prismatic stone samples (10 × 15 × 10 mm³) was observed by FEG-SEM (FEI Nova NanoSEM 450, Thermo Fisher Scientific, Waltham, MA, USA). Stones treated with PHB- and PHBVV-based formulations applied by poultice on one surface were observed. The cone for back-scattered electrons was set to the widest opening in order to obtain morphological images and, at the same time, the definition of the contrast in function of the chemical composition given by the back-scattered electrons (BSE) mode. The main purpose of the observation of the cross sections was to investigate not only the morphology of the coatings, but also their possible penetration in the porosity of the samples.

The changes in molecular weight size of both HCT and HCT-f samples were determined using gel permeation chromatography (GPC). The samples were coated with gold on the sample surface and observed using the FEI Nova Nano SEM 450 instrument (Thermo Fisher Scientific, Waltham, MA, USA).

Bacteria were cultivated in small-scale under conditions described in Table 1. A volume corresponding approximately to 2 × 10⁹ CFU was pelleted (6000 × g, 15 min, 4°C) and washed twice in 100 mM sodium cacodylate/5 g/L NaCl, pH 7.2. Bacteria were then fixed by 3% glutaraldehyde in the same buffer at RT for 1 h and then at 4°C overnight with slow rotation. Sterility of the fixed solution was checked by aliquot plating. The washed bacterial cells were then allowed to sediment overnight onto poly-L-lysine treated circular coverslips at 4°C. The coverslips with attached bacteria were post-fixed with 1% OsO₄ for 1 h at room temperature and three times washed with ddH₂O. Washed coverslips with the bacteria were dehydrated through an alcohol series (25, 50, 75, 90, 96, 100, and 100%) followed by absolute acetone and critical point dried from liquid CO₂ in a K850 Critical Point Dryer (Quorum Technologies Ltd.). The dried samples were sputter coated with 3 nm of platinum in a Q150T Turbo-Pumped Sputter Coater (Quorum Technologies Ltd.). The final samples were examined in a FEI Nova NanoSEM 450 scanning electron microscope (Thermo Fisher Scientific) at 5 kV using CBS and TLD detectors.

Scanning electron microscope (SEM) was used to investigate the morphology and microstructure of nanoparticles and compare the appearance of ARM-HSA NPs with free artemether. The lyophilized nanoparticles were coated with gold sputter, and analyzed under SEM (FEI-Nova NanoSEM 450, Thermo Fisher, Waltham, MA, USA). For the TEM method, the sample was diluted with distilled water and dried at room temperature by placing it on a carbon-coated copper grid. The dried sample was observed under the TEM (PHILIPS CM300, PHILIPS, Cambridge, MA, USA, and 200 kV). At the end, the diameter, morphology and microstructure of nanoparticles were evaluated by using Image analyzer (Digital Micrograph software 1.81.78, Gatan, Pleasanton, CA, USA).