Surface morphology of the films was analyzed using ultra-high resolution field emission scanning electron microscope (UHR-FESEM, MERLINN/344999-9001-030, Zeiss, Germany). The films were mounted onto stubs (12 mm diameter) using double-sided adhesive carbon tape. The films were then sputter coated with gold for 5 min (QUORUM Sputter Coater Q150R S, Quorum, UK) before placing in the chamber of the microscope and images acquired using SmartTiff software. Images were taken at magnification power of 100×, 500×, 1000×, 2000× and 3000×.
Q150r s sputter coater
Manufactured by Quorum Technologies
Sourced in United Kingdom
The Q150R S is a sputter coater designed for depositing thin films of conductive materials onto samples. It features a compact, benchtop design and supports a variety of substrates. The coater utilizes a high-vacuum system to create an inert environment for the sputtering process.
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9 protocols using q150r s sputter coater
1
Surface Morphology Analysis of Thin Films
2
Surface Morphology Analysis of Films
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Scanning electron microscopy was used to analyze the film’s surface morphology using an ultra-high-resolution field-emission scanning electron microscope (UHR-FESEM, MERLIN/344999-9001-030, Zeiss, Aalen, Germany). Each film was sliced into a 3 × 3 mm piece, which was then attached to a stub via double-sided adhesive carbon tape. The samples were then placed in the microscope chamber after being subjected to a 5-min gold sputter coating procedure (QUORUM Sputter Coater Q150R S, Quorum, Lewes, UK), followed by SEM examination at a 10-KV accelerating voltage. Using the smartTiff tool, the photographs of corresponding parts were taken at magnification powers of 100, 500, 1000, 2000, and 3000× [34 (link)].
3
Characterization of Chitosan-Gelatin Films
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The sectional morphologies of the chitosan–gelatin composite films were imaged using an SEM facility (Phenom, Eindhoven, Netherlands) operated at a voltage of 10 kV. The cryo-fractured sectional surfaces were obtained by liquid nitrogen, and coated with gold for 90 s using a Q-150R-S sputter-coater (Quorum Technologies Ltd, East Sussex, UK).
4
Correlative SBF-SEM Imaging of Cell-Cell Contacts
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Prior to SBF-SEM, the block was sputter-coated with 5–10 nm platinum using a Q150R S sputter coater (Quorum Technologies). SBF-SEM data was collected using a 3View2XP (Gatan Inc.) attached to a Sigma VP SEM (Zeiss). The microscope was operated at 1.8kV with 30-μm aperture, using Focal Charge Compensation mode (Deerinck et al., 2018 (link)). Inverted backscattered electron images were acquired every 50 nm, at a resolution of 8.44 nm/pixel. Acquired images were imported into Fiji (Schindelin et al., 2012 (link)) and aligned using the Register Virtual Stack Slices (Arganda-Carreras et al., 2006 ) and in TrakEM2 using manual landmarks (Cardona et al., 2012 (link)). 63 × Z stack was matched to the aligned SBF-SEM data using BigWarp (Bogovic et al., 2016 ) based on nuclei positions and overlaid to the EM data after applying thin-plate spline transformation. Cells in the region of expanded cell-cell contacts were selected and segmented using the TrakEM2. Cell-cell contacts were segmented if the plasma membranes of two neighboring cells were closely apposed with spacing less than or equal to 25 nm based on the length of homo dimer of PECAM extracellular domains (Jiang et al., 2016 (link)). 3D reconstructions were made in the 3dmod program.
5
Optimized Nano-ESI QToF Protein Analysis
6
Soil Surface Morphology and Elemental Analysis
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The FESEM and EDX analyses were conducted to analyze the morphology on the soil surface and the element composition of the untreated and treated soil specimen using the dispersive X-ray spectrometer The dry specimen was mounted on the platinum sample with adhesive carbon tape before being sputter-coated using the Q150R S Sputter coater (Quorum, Quorum Technologies Ltd, East Sussex, UK). The sputter coater is used to improve the surface electrical conductivity and thus reduces the charging effect. The micrographs of the specimen were obtained and reviewed under SU 8010 FESEM (Hitachi SU8010, Oxford-Horiba Inca XMax50, Hitachi High- Technologies, Tokyo, Japan).
7
Serial Block Face SEM Imaging
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Prior to commencement of a SBF SEM imaging run, the sample was coated with a 2 nm layer of platinum to further enhance conductivity using a Q150R S sputter coater (Quorum Technologies). SBF SEM data were collected using a 3View2XP (Gatan) attached to a Sigma VP SEM (Zeiss). Inverted backscattered electron images were acquired through the entire extent of the ROI. For each 50 nm slice, a low-resolution overview image (pixel size of ∼50 nm using a 1.5 µs dwell time) and several high-resolution images of the different regions of interest (indicated magnification ∼5000×, pixel size of 6-7 nm using a 1.5 µs dwell time) were acquired. The overview image was used to relocate the ROI defined by the confocal images of the sections. The SEM was operated in variable pressure mode at 5 Pa. The 30 µm aperture was used, at an accelerating voltage of 2 kV. Typically, between 300 and 1000 slices were necessary for an entire ROI. As data were collected in variable pressure mode, only minor adjustments in image alignment were needed, particularly where the field of view was altered in order to track the cell of interest.
8
Surface Morphology and Porosity Analysis
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The surface morphologies of RH and SRHCs were observed by using scanning electron microscope (SEM) S-3400N Hitachi. Before the analysis begun, the sample was first placed on a carbon tape and coated with gold using a Q 150R S sputter coater from Quorum. Then, the tape was placed in the SEM sample holder and further analyzed. Images obtained were under vacuum mode of 15 kV acceleration voltage. SEM S-3400N Hitachi was integrated with EDX. Thus, SEM and EDX analysis was done simultaneously. EDX detected the elements present on selected point on the sample surface. The operating condition of EDX was similar to SEM. Two points were selected for analysis, one at the outer surface of RH and SRHCs, and the second point was at the inner surface of the materials.
2.4.5 Specific surface area and pore volume Specific surface area and pore volume of RH and SRHCs were determined by physical adsorption and desorption of nitrogen using ASAP 2020 Micromeritics. Prior to analysis, the sample was degassed at temperature 110˚C for 3h. The resulting isotherms were analyzed using Brunauer-Emmett-Teller (BET) model to calculate the specific surface area. Meanwhile, total pore volume of the sample was evaluated from isotherms using Barrett-Joyner-Halenda (BJH) model.
2.4.5 Specific surface area and pore volume Specific surface area and pore volume of RH and SRHCs were determined by physical adsorption and desorption of nitrogen using ASAP 2020 Micromeritics. Prior to analysis, the sample was degassed at temperature 110˚C for 3h. The resulting isotherms were analyzed using Brunauer-Emmett-Teller (BET) model to calculate the specific surface area. Meanwhile, total pore volume of the sample was evaluated from isotherms using Barrett-Joyner-Halenda (BJH) model.
9
SEM Analysis of Polypyrrole Particles
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The surface morphology of the PPy particles was investigated using a Philips XL30S field emission gun scanning electron microscope (Netherlands) at an accelerating voltage of 5 kV.
The samples for SEM were mounted on aluminium studs using adhesive graphite tape and lightly sputter-coated with platinum using a Quorum Q150RS Sputter Coater.
The samples for SEM were mounted on aluminium studs using adhesive graphite tape and lightly sputter-coated with platinum using a Quorum Q150RS Sputter Coater.
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