A Scios FIB.SEM (ThermoFisher) was used. It included an Everhart–Thornley Detector (ETD), an in-lens detector of backscattered electrons (BSE), and an in-the-column detector of secondary electrons (SE). The ETD was used for imaging of sample surfaces. The BSE detector was used for imaging cellular ultrastructure in 3D. The in-the-column SE detector was used to enhance fiducial marker contrast. The FIB was equipped with a Ga-ion source; for the 3D acquisition, a current of 0.4 nA and an acceleration voltage of 30 kV were used. The 3D acquisition in the FIB.SEM was controlled by the Slice&View software version 3 (Thermo Fischer).
Fib sem
Manufactured by Thermo Fisher Scientific
The FIB-SEM is a focused ion beam scanning electron microscope (FIB-SEM) that combines a focused ion beam and a scanning electron microscope in a single instrument. The FIB-SEM is used for high-resolution imaging, precise materials modification, and site-specific sample preparation.
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Lab products found in correlation
Scios fib sem, by Thermo Fisher Scientific (1 mentions)
Scios 2, by Thermo Fisher Scientific (1 mentions)
Aquilos dual beam instrument, by Thermo Fisher Scientific (1 mentions)
Cryo fib autogrids, by Thermo Fisher Scientific (1 mentions)
2010f tem, by JEOL (1 mentions)
Scios, by Thermo Fisher Scientific (1 mentions)
Leap 5000 xr, by Cameca (1 mentions)
9 protocols using fib sem
1
Cellular Ultrastructure Imaging via FIB-SEM
2
Ion Irradiation of Copper TEM Grids
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Conventional copper TEM grids (Omniprobe Cu TEM grids) were irradiated using different ion species (Ga and Ne), different incident ion energies (30, 25, and 2 keV), different doses (3371 and 2247 ions/nm2), as well as different incidence angles (0° incidence and glancing angle irradiation). The irradiation experiments were performed using the Zeiss Orion Nanofab Helium Ion Microscope (Ne) as well as the Thermo Fisher, Scios (Ga) FIB/SEM.
3
Morphology Study of LNMO Electrodes
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After electrochemical cycling, the cells were disassembled in an argon-filled glovebox. The post-cycled LNMO electrodes and lithium metal disks were washed several times in DMC to remove residual electrolyte salts. The washed LNMO electrodes were then dried in the glovebox at room temperature. The surface morphologies of the LNMO electrodes caused by cycling were imaged using a scanning electron microscope (FIB-SEM, Thermo Fisher, Scios2). The uniformity and thickness of the surface layer on the LNMO electrodes that were cycled in the electrolyte without and with additives were examined using transmission electron microscopy (TEM, FEI Tecnai Spirit) at 120 kV.
4
Cryo-FIB Milling of Yeast Cells
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4 μl of the cultured yeast cells were added to a glow-discharged (30 s at −30 mA) copper R2/2 holey carbon grid (Quantifoil Micro Tools GmbH). Then the grid was rapidly plunge-frozen in liquid ethane using a homemade plunge freezer device and stored in liquid nitrogen until used. Cryo-FIB milling was performed as previously described (Hariri et al., 2019 (link)). Briefly, grids were mounted in notched cryo-FIB Autogrids (Thermo Fisher Scientific), then loaded into a shuttle under cryogenic conditions, and transferred into an Aquilos dual-beam instrument equipped with a cryo-stage (FIB/SEM; Thermo Fisher Scientific). The sample surface was sputter-coated with platinum for 20 s at 30 mA current and then coated with a layer of organometallic platinum using the gas injection system for 6 s at a distance of 1 mm before milling. The stage was then tilted to 10–18° (so that the bulk-mill-holes lined up in front and behind the cell) and the cell was milled with 30 kV gallium ion beams of 100 pA current for rough milling and 10 pA for polishing until the final lamella was 100–200 nm thick.
5
Scanning Electron Microscopy of Transwell
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Empty Transwell™ inserts were exposed for 30 min as described for Transwell™ inserts containing the tetraculture system. Afterwards samples were metallized with a 20 nm gold film under vacuum. Scanning electron microscopy was done with a FIB-SEM (FEI, Eindhoven, The Netherlands) at 25 kV and 25 mA.
6
Nanostructure Imaging and Analysis
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SEM images and SEM-EDS spectroscopic data were obtained for nanostructures without removing them from the growth substrate, using a FEI FIB/SEM working at 5-20 kV. TEM samples were prepared on copper grids by either dry or wet transfer. For dry transfer, copper grids were gently dragged over the growth substrates to collect the grown material. For wet transfer, the growth substrates were sonicated briefly in iso-propanol solution and the solution was drop-cast onto copper grids. Imaging and selected area diffraction were carried out on a JEOL 2010F TEM operated at 200 keV. Diffraction patterns of ZnO for different zone axes were simulated using Crystal Maker. We note that because of their large sizes, Ge catalysts were easily broken off during the transfer processes. As a result, many TEM images in this study show only ZnO stems.
7
Nanoscale Element Distribution via APT
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APT analyses were applied to the as-prepared and heat-treated samples to investigate the element distribution in three dimensions up to a sub-nanometer spatial resolution. Needle-shaped specimens required for APT were fabricated by lift-outs and annular milled in a FEI Scios focused ion beam/scanning electron microscope (FIB/SEM). The APT characterizations were performed in a local electrode atom probe (CAMECA LEAP 5000 XR). The specimens were analyzed at 65 K in voltage mode with a pulse repetition rate of 200 kHz, a pulse fraction of 20%, and an evaporation detection rate of 0.3% atom per pulse. The spatial resolution of the APT could reach ~0.2 nm. Imago Visualization and Analysis Software (IVAS) version 3.8 was used for creating the 3D reconstructions and data analyses.
8
Characterization of Anoxic Fe-Oxides
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Anoxically preserved water column and sediment samples were preserved using a 1% osmium tetroxide solution buffered with 0.1 M PIPES at pH 6.8. Filters were rinsed gently with MQ water and then dried using an ethanol dehydration series. The filters were critical-pointdried using a Samdri795 from Toosimis Research Corporation. Finally, the filters were attached to a 12.5 mm stub and coated with 5 nm of iridium to ensure conductivity. The filters were imaged on a Helios FIB-SEM (FEI, Helios NanoLab 650) equipped with field emission gun.
Fe(III) oxyhydroxides were confirmed through energy-dispersive X-ray spectroscopy (EDS) and elemental compositions determined based on X-ray fluorescence at the relevant emission energies for Fe, C and O. Multiple points were measured for each surface found. To verify that the micro-chemical analyses of the SEM-EDS accurately differentiate magnetite from other Feoxide phases, we analyzed two pure Fe-mineral standards; magnetite (Fe 3 O 4 ) and goethite (FeO(OH)). On each standard we collected over 10 distinct EDS spots and compiled their Fe:O stoichiometries (wt%). We then performed bootstrap resampling of the mean Fe:O compositions for these standard minerals. We also obtained EDS spectra on framboids collected in the water column and sediments of LM and LT (n = 17) and performed bootstrapped resampling of their mean Fe:O values.
Fe(III) oxyhydroxides were confirmed through energy-dispersive X-ray spectroscopy (EDS) and elemental compositions determined based on X-ray fluorescence at the relevant emission energies for Fe, C and O. Multiple points were measured for each surface found. To verify that the micro-chemical analyses of the SEM-EDS accurately differentiate magnetite from other Feoxide phases, we analyzed two pure Fe-mineral standards; magnetite (Fe 3 O 4 ) and goethite (FeO(OH)). On each standard we collected over 10 distinct EDS spots and compiled their Fe:O stoichiometries (wt%). We then performed bootstrap resampling of the mean Fe:O compositions for these standard minerals. We also obtained EDS spectra on framboids collected in the water column and sediments of LM and LT (n = 17) and performed bootstrapped resampling of their mean Fe:O values.
9
Characterization of Anoxic Fe-Oxides
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Anoxically preserved water column and sediment samples were preserved using a 1% osmium tetroxide solution buffered with 0.1 M PIPES at pH 6.8. Filters were rinsed gently with MQ water and then dried using an ethanol dehydration series. The filters were critical-pointdried using a Samdri795 from Toosimis Research Corporation. Finally, the filters were attached to a 12.5 mm stub and coated with 5 nm of iridium to ensure conductivity. The filters were imaged on a Helios FIB-SEM (FEI, Helios NanoLab 650) equipped with field emission gun.
Fe(III) oxyhydroxides were confirmed through energy-dispersive X-ray spectroscopy (EDS) and elemental compositions determined based on X-ray fluorescence at the relevant emission energies for Fe, C and O. Multiple points were measured for each surface found. To verify that the micro-chemical analyses of the SEM-EDS accurately differentiate magnetite from other Feoxide phases, we analyzed two pure Fe-mineral standards; magnetite (Fe 3 O 4 ) and goethite (FeO(OH)). On each standard we collected over 10 distinct EDS spots and compiled their Fe:O stoichiometries (wt%). We then performed bootstrap resampling of the mean Fe:O compositions for these standard minerals. We also obtained EDS spectra on framboids collected in the water column and sediments of LM and LT (n = 17) and performed bootstrapped resampling of their mean Fe:O values.
Fe(III) oxyhydroxides were confirmed through energy-dispersive X-ray spectroscopy (EDS) and elemental compositions determined based on X-ray fluorescence at the relevant emission energies for Fe, C and O. Multiple points were measured for each surface found. To verify that the micro-chemical analyses of the SEM-EDS accurately differentiate magnetite from other Feoxide phases, we analyzed two pure Fe-mineral standards; magnetite (Fe 3 O 4 ) and goethite (FeO(OH)). On each standard we collected over 10 distinct EDS spots and compiled their Fe:O stoichiometries (wt%). We then performed bootstrap resampling of the mean Fe:O compositions for these standard minerals. We also obtained EDS spectra on framboids collected in the water column and sediments of LM and LT (n = 17) and performed bootstrapped resampling of their mean Fe:O values.
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