Automatic sputter coater

Materials for scanning electron microscopy (SEM) were prepared for observing the intact and cleaned cells in two ways. The intact morphology from the isolates was observed by filtering live cells on a 3-μm polycarbonate membrane and washed with distilled water without any oxidation. The cleaned external and internal structures after the detachment of the organic matter were observed by adding some live cells to an equal volume of H₂O₂ and exposing them to ultraviolet light (312 nm) for 3 h on the UV-transilluminator (WUV-L50; witeg, Wertheim, Germany) and then washing them with distilled water. The cleaned cells were filtered and dried on a 3-μm polycarbonate membrane. The dried polycarbonate membranes were attached to aluminum stubs by using carbon conductive adhesive tapes and coated with 10 nm gold in an automatic sputter coater (Agar Scientific Ltd., Essex, UK). The prepared specimens were examined using FE-SEM (JSM7600F; Jeol, Tokyo, Japan) operating at 10 kV and 8 mm working distance. Valve dimensions from the SEM photographs were measured using ImageJ 1.51 software [16 (link)]. The terminology used for the structures discussed herein follows that of Ross et al. [17 ], Round et al. [18 ] and Theriot & Serieyssol [19 ].

Morphological investigation on P and PP NPs with or without DPH were performed using a Field Emission Scanning Electron Microscope (SEM) equipped with an Energy Dispersive Spectrometer (FESEM/EDS; Zeiss Merlin VP Compact coupled with Oxford Instruments Microanalysis Unit; Carl-Zeiss Strasse, Oberkochen, Germany). Samples were glued by a carbon tape film on a sample holder and then gold-metalized, using an automatic sputter coater (Agar Scientific ltd-Parsonage Lane, Stansed-Essex, UK), with an automatically controlled complete sequence of flush, leak, coat, and vent.
The INCA (Oberkochen, Germany) X-stream pulse processor and the INCA Energy software 5.05 were used to obtain datasets. Operative conditions were reported on each acquisition and were chosen to be consistent with the performed measurement.

The morphology of
the fabricated materials was investigated using a large chamber scanning
electron microscope (Model S-3700 N, Hitachi, Tokyo, Japan). An accelerating
voltage of 15 kV was used. Prior to SEM, the samples were mounted
onto aluminum stubs using carbon tabs and Au coating (Automatic sputter
coater, Agar Scientific, Stansted, UK) at a current of 40 mA for 20
s.

To assess the presence of biofilm‐like structures, nine representative samples were also investigated by scanning electron microscopy. Three parts of removed plates or screws of each patient were prepared for the scanning electron microscopy. The remaining parts of the implants were used for microbiological investigations. For scanning electron microscopy, the implants were immersed in 2 ml of glutaraldehyde 2.5% for fixation. After fixating for 24 hr at 4°C, the implants were dehydrated with an ascending alcohol series (50%–70%–80%–99.9% ethanol). Each step lasted 5 min. After the last step, the implants were placed in an incubator for drying. The dried samples were glued on aluminum pins with Leit‐C (Plano GmbH, Wetzlar, Germany). The pins were sputtered with Au using an automatic sputter coater (Agar Scientific Ltd, Stansted, Great Britain) for 1 min and analyzed by scanning electron microscopy (SEM, JSM‐6010LV, JEOL GmbH, Freising, Germany).

A CCR array was purchased from the JunAN (SL150-18, China) and used as a mold during embossing processing. Silicone elastomer base and curing agent (SYLGARD 184, 1.1KG) chemicals were purchased from Farnell, UK and used as a soft polymer embossing medium. Automatic sputter coater was purchased from the Agar Scientific, UK to make thin Au coating over flexible CCR array. COMSOL Multiphysics 5.2, MATLAB (Math Works, R2013) was used for the numerical simulations and data processing.

The surface morphology of pure curcumin powder, the blank, and curcumin-loaded (7% w/w) scaffolds was studied with a MERLIN™ Scanning Electron Microscope (Jena, Thuringia, Germany). The samples were coated with platinum by sputter-coating before analysis with an automatic sputter coater (Agar Scientific Ltd., Stansted, Essex, UK). An accelerating voltage of 2 kV was used to obtain SEM images.

Unmodified and pGL3-PEI-coated HSA nanoparticles, as well as pGL3-PEI complexes were diluted with purified water (1∶10). An aliquot was placed on an aluminium sample plate and air-dried at room temperature over-night. In order to obtain electrical conductibility for electron microscopy analysis, the sample plates were sputtered with gold for 45 s under argon gas atmosphere (Automatic sputter coater, Agar Scientific, Essex, United Kingdom). Samples were analysed with a field emission electron microscope with upper detector (EM Hitachi S-45000, Hitachi High-Technologies Europe GmbH, Krefeld, Germany) at 15–25 kV.