Talos tem

To prepare samples for TEM, 4 μl of either undiluted (1 mg/ml) or diluted (0.1 mg/ml) minicollagen-1 was applied to a 400 mesh copper grid supporting a thin carbon film. The sample was allowed to adhere for 1 min before staining with Uranyl-less stain for 3 min, washing, then blotting to dryness with Whatman #1 filter paper. Samples were imaged on a Ceta camera on a Thermo Fisher Scientific Talos TEM operating at 200 kV and using a 40 μm objective aperture.

The grazing incidence small-angle x-ray scattering (GISAXS) measurements were performed at the synchrotron Elettra, Trieste, Italy at the SAXS beamline using a photon energy of 8 keV and a Pilatus3 1M detector. The films deposited on non-porous Si substrates were used only for the GISAXS measurements. We expect the same structure of the films grown on membranes and SiN_x membrane because they grow with the same mechanism independently on the substrate. The GISAXS maps were numerically analyzed using the procedure described in [43 (link),44 (link),45 (link)].
The transmission electron microscopy (TEM) of the cross-section of the film was performed using a JEOL ARM 200 CF scanning transmission electron microscope (STEM), operated at 200 kV and equipped with a field-emission gun and a high-angle annular dark-field detector (HAADF) for Z-contrast imaging.The TEM of the opening in the SiN_x membrane overgrown by the film was performed using Talos TEM (ThermoFisher Scientific, Hillsboro, OR, USA) at 200 kV.
The scanning electron microscopy (SEM) was performed using the thermal field emission scanning electron microscope (FE SEM, model JSM-7000 F) manufactured by JEOL Ltd (Tokyo, Japan). FE SEM was linked to the EDS/INCA 350 (energy-dispersive X-ray analyzer) manufactured by Oxford Instruments Ltd. (Abingdon, UK).

TEM images were first acquired with a JEOL JEM-1400 Plus working
at 120 kV. For imaging, the as-prepared samples were dispersed in
ethanol and supported on a TEM grid. These images verify the size,
morphology, and dispersion of the Bi/C NPs. High-resolution (HR)-TEM
images were taken from the samples before and after the electrocatalytic
experiments with a Titan 80-300 at 300 kV (Thermo Fisher Scientific).
Energy-dispersive X-ray analysis (EDX) elemental maps were acquired
using a 200 kV Talos TEM (Thermo Fisher Scientific).

CPMV was propagated in black-eyed pea no. 5 plants and purified as previously reported [60 ]. The concentration of the purified CPMV fraction was determined by UV–Vis spectroscopy ((ε of CPMV at 260 nm = 8.1 mL / (mg x cm)), and particle integrity was confirmed by NuPAGE, transmission electron microscopy (TEM), dynamic light scattering (DLS), and size exclusion chromatography (SEC). For NuPAGE analysis, 10 µg heat-denatured samples (mixed with 4 × lithium dodecylsulfate buffer, Thermo Fisher Scientific) were analyzed using a 4–12% NuPAGE gel in 1 × MOPS buffer (Thermo Fisher Scientific). Gels were stained with Coomassie brilliant blue and imaged using a ProteinSimple FluorChem R imager. For TEM, CPMV (2 µg) was applied to a glow-discharged carbon-coated 300-mesh Cu grid and stained with 4 µL 1% (w/v) uranyl acetate (Electron Microscopy Sciences). The samples were imaged using a Talos TEM (Thermo Fisher Scientific) at a nominal magnification of 120,000 ×  DLS measurements were recorded using a Zetasizer Nano ZSP/Zen5600 instrument (Malvern Panalytical) and 100 µg CPMV (1 mg/mL). For SEC, we used a Superose 6 increase 10/300 GL column mounted on an ÄKTA purifier system (GE Healthcare). 100 µg CPMV (1 mg/mL) was analyzed at a flow rate of 0.5 mL/min, and the absorbance was monitored at 260 nm (RNA) and 280 nm (protein).

The as-prepared ZIF-8 NPs were characterized for their physicochemical properties before carrying out interaction studies. The size and morphology of the particles were determined using transmission electron microscopy (TEM), scanning electron microscopy (SEM), and dynamic light scattering (DLS) measurements. A dilute sample of ZIF-8 NPs dispersed in methanol was drop casted on carbon-coated copper grids, followed by air-drying and analysis using TALOS TEM (Thermo Scientific) operating at 200 kV. Nanoparticles dispersed in methanol were drop casted on a glass slide and air-dried, and then SEM analysis was carried out using JEOL JSM-6610 operating at 20 kV. Nanoparticles were well dispersed in water and the Malvern Zetasizer instrument was used for DLS and zeta potential analysis. Elemental composition was verified by energy dispersive spectroscopy (EDS) using JEOL JSM-6610 instrument. Fourier-transform infrared (FT-IR) spectroscopy was used to analyze surface functionalities using Shimadzu IRAffinity-1S spectrophotometer. Shimadzu DTG-60 simultaneous DTA-TG apparatus was used for thermogravimetric analysis (TGA) of nanoparticles in nitrogen atmosphere. The crystallinity of particles was determined using the Rigaku Miniflex benchtop powder X-ray diffraction (XRD) instrument.

Bacteria were grown as overnight cultures. Cells were transferred to a 5 ml tube and centrifuged for 4 min at 800 g. The supernatant was removed and the cell pellets were resuspended twice in 3 ml of Phosphate-buffered saline (PBS, Sigma-Aldrich). A sample with OD₆₀₀ of 1.8 was then taken for vitrification. 5 µl of NiNTA-gold 10 nm as fiducial markers were mixed with the samples before vitrification. 4 µl of the samples were introduced to Lacey grids pre-treated with plasma under vacuum to make their surface hydrophilic. A Vitrobot FEI Eindhoven with 3 s of blot time standard and 1.0 blot force was used to vitrify the samples in liquid ethane. The imaging was performed using a 200 kV FEI Talos TEM equipped with a 16 M pixel CMOS camera.

PXRD data were recorded on a Rigaku Miniflex with Cu source. SEM data for Fig. 1 were recorded on a JEOL JSM 6610LV, whereas data for reduced electrodes were taken with a S-Hitachi-4700-II SEM. Raman data were recorded on a Thermofisher DXR3xi Raman microscope with 532 nm laser excitation. TEM data were recorded on an FEI Talos TEM with an FEI Tecnai F20ST S/TEM. For characterization of reduced electrodes, samples were washed with 0.1 M acetic acid and water following battery disassembly. The freestanding gauze – and if applicable a steel foil support – was inserted directly into SEM and XRD for respective measurements, whereas the sample was scratched from the mesh onto a lacey carbon grid for TEM analysis.