Murine samples were fixed and embedded as above. Tomography experiments based on high-angle annular dark-field (HAADF) imaging in the scanning transmission electron microscopy (STEM) mode were performed at 200 kV on a FEI Tecnai F20 FEG-TEM tilting the sample from −42° to +70° at 2° intervals about a single axis using a Fischione 2020 ultra-high-tilt tomography holder. Using the Inspect 3D software, tilt series were aligned by cross-correlation and reconstructed using the SIRT algorithm (with 20 iterations).
Tecnai f20 feg tem
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Tecnai F20 FEG-TEM is a field emission gun transmission electron microscope (FEG-TEM) manufactured by Thermo Fisher Scientific. It is designed to provide high-resolution imaging and analytical capabilities for a variety of applications in materials science, nanotechnology, and life sciences research.
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Lab products found in correlation
Acs reagent, by Merck Group (1 mentions)
Suprapur, by Merck Group (1 mentions)
Centrifugation, by Merck Group (1 mentions)
Milli q, by Merck Group (1 mentions)
Ht7700 microscope, by Hitachi (1 mentions)
Delsa nano c particle analyzer, by Beckman Coulter (1 mentions)
80 mm2 silicon drift detector, by Oxford Instruments (1 mentions)
Inca 350 edx system, by Oxford Instruments (1 mentions)
Nova nanosem 230, by Thermo Fisher Scientific (1 mentions)
Nova 200 nanolab, by Thermo Fisher Scientific (1 mentions)
Quanta 3d feg sem, by Thermo Fisher Scientific (1 mentions)
Xl30s, by Philips (1 mentions)
Inspect3d, by Thermo Fisher Scientific (1 mentions)
9 protocols using tecnai f20 feg tem
1
Murine Sample Tomography by STEM
2
Dual-Axis Tilt Series TEM Imaging
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A Tecnai F20 FEG TEM (FEI, Eindhoven, The Netherlands) was used operated at 200 kV with digital images recorded via a Gatan 2Kx 2 K CCD camera during tomographic data acquisition. Dual-axis tilt series were collected by tilting the specimen in the microscope from − 60° to + 60° with 1° increments. Once the first tilt series were recorded around axis A, the specimen was manually rotated 90° for the second tilt series around axis B. Images of the raw stacks were collected using a binned pixel size of 0.888 nm.
3
Manganese Oxide Synthesis and Characterization
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Manganese oxide was prepared as described before (McKenzie, 1971 (link); Zhang et al., 2015 (link)). Briefly, 0.2 mol of KMnO4 (ACS reagent, Sigma-Aldrich, St. Louis, MO, United States) was weighted in 350 ml of ultrapure water and boiled. While boiling, 50 ml of concentrated HCl (Suprapur, Sigma-Aldrich, St. Louis, MO, United States) was added drop-wisely. After all the HCl was added, the solution was left to boil for an additional 30 min. After the solution cooled, the brown precipitate was harvested by centrifugation (Thermo Fischer Scientific, Waltham, MA, United States) (3500 rpm for 10 min) and the pellet washed with ultrapure water (18 MΩ, Milli-Q, Millipore, Burlington, MA, United States) until the pH of the solution was neutral. An additional step of washing was applied to remove any K+, Cl- as well as aqueous Mn ions, by dialysing the oxide against ultrapure water using a 14 kDa molecular weight cut-off dialysis membrane (Spectrum Labs, Phoenix, AZ, United States). After the washing steps, manganese oxide was dried over night at 60°C. To assess particle size and morphology, transmission electron microscopy (TEM) (Tecnai F20 FEG-TEM, FEI Company, Hillsboro, OR, United States) analysis was performed at 200 kV. The particles were first sonicated in ethanol then casted onto a mesh TEM grid (C-Formvar 200, Ted Pella, United States) for imaging.
4
Murine Sample Tomography by STEM
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Murine samples were fixed and embedded as above. Tomography experiments based on high-angle annular dark-field (HAADF) imaging in the scanning transmission electron microscopy (STEM) mode were performed at 200 kV on a FEI Tecnai F20 FEG-TEM tilting the sample from −42° to +70° at 2° intervals about a single axis using a Fischione 2020 ultra-high-tilt tomography holder. Using the Inspect 3D software, tilt series were aligned by cross-correlation and reconstructed using the SIRT algorithm (with 20 iterations).
5
Comprehensive Characterization of PVP Emulsions
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TEM was performed on a Hitachi HT-7700 microscope equipped with a LaB6 filament, operated at 100 kV. High-resolution TEM (HRTEM) was performed on an FEI Tecnai F20 FEG-TEM, operating at 200 kV. The PVP emulsion was visualized by TEM after staining with Lugol’s iodine (5% I2 + 10% KI). Typically, 160 μL of DEA and 40 μL of Lugol’s iodine were added to 16 mL of 10 × diluted PVP/seeds in sequence with stirring. After stirring for 1 h, the resulting emulsion was dropped onto a copper grid and dried for TEM imaging. SEM and EDS were performed on a field-emission JEOL JSM-7800F microscope operating at 15 kV. DLS was performed on a Beckman Coulter Delsa Nano C particle analyzer. XRD patterns were recorded on a Rigaku SmartLab Powder X-ray diffractometer equipped with a Cu Kα X-ray source and a D/teX Ultra detector, scanning at a rate of 5° min–1 over a range of 10–90° (2θ). UV-vis spectroscopy was recorded on an Ocean Optics HR2000 + ES UV-vis-NIR spectrophotometer with a DH-2000-Bal light source. ICP-MS was performed on an Agilent 7500ce.
6
Nanomaterial Characterization using TEM
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TEM was conducted at 200 kV using a Tecnai F20 FEG-TEM fitted with a high angle annular dark field (HAADF) detector (FEI, Eindhoven, The Netherlands) and an INCA 350 EDX system with a 80 mm2 silicon drift detector (Oxford Instruments, Abingdon UK). Images were recorded using an Orius SC600A CCD camera (Gatan Inc., Abingdon, UK).
7
Microstructural Characterization of Ti-12Mo Alloy
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The binary Ti-12 wt.%Mo (hereafter referred to as Ti-12Mo) alloy was fabricated by vacuum arc melting using pure Ti and Mo. Following this, they were solution-treated (ST) at 900 °C for 30 min and subsequently water-quenched. Microstructural characterization was performed using scanning electron microscopy (SEM), FEI NovaNano SEM230, and transmission electron microscopy (TEM), FEI Tecnai F20-FEG TEM operating at 200 kV. As the EBSD analysis was carried out after deformation to failure, the analyzed regions were intentionally selected away from the fracture surface, more towards the grip section, in order to get a better indexing in EBSD, and a schematic showing the location from which the analysis was done is shown in the Supplementary Figure. Site-specific TEM foils were prepared via a FEI Nova NanoLab 200™ focused ion beam (FIB). The sample was made in such a way that half the foil was from the β-matrix and the other half from the β-twin. The location of this has also been shown in the aforementioned schematic. Dog-bone shaped tensile specimens of gage length ~5 mm, width ~1 mm and thickness ~0.8 mm were used for the mechanical testing, which was done under uniaxial tension at a strain rate of 10−3 s−1 at 25 °C. Details of the setup for the mechanical testing are presented elsewhere.
8
Characterization of Alloy Microstructure Dynamics
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SEM characterization was performed using a JEOL 7001F and a Philips XL30S devices. BSE images based on the diffraction contrast were taken to identify the evolution of grains and twins for the different processing schedules. EBSD measurements were performed using AztecHKL and TSL/OIM software. The data obtained were analyzed using HKL/Channel 5 and TSL/OIM. Texture analysis was conducted using JTEX60 . SEM samples were prepared by grinding with SiC paper up to 2400-grit, followed by polishing with a colloidal silica suspension for 2 h. EBSD samples were subsequently polished by argon ion beam using a GATAN precision etching-coating system to remove the strained layer. TEM characterization was carried out using an FEI Tecnai F20 FEG TEM operating at an acceleration voltage of 200 kV. TEM foils were prepared by focused ion beam (FIB) lift-out performed on an FEI Quanta 3D FEG SEM. The area of the SEM and EBSD observation was 3 mm away from the center of the specimen. TEM foils were lifted out at this location, as well.
9
Electron Tomography of Gold Nanoparticles
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Electron tomography tilt series data was acquired using an FEI Tecnai F20 FEG-TEM operated at 200 kV. Gold nanoparticles were prepared by a modified seed-mediated growth process as reported by Bandhopadhyay et al. (2017) . The sample was drop cast onto 200 mesh copper TEM grids coated with a continuous carbon film. A tilt series of HAADF STEM images was acquired from -68° to +64° in 2° increments. The tilt series image stack was first aligned using cross-correlation methods followed by alignment of the tilt axis, performed using Inspect3D (FEI).
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