Scanning Transmission Electron Microscopy (STEM) was performed on an aberration corrected FEI Titan G2 60–300, operated at 300 kV. The microscope was equipped with a SuperX EDX spectrometer. The sample preparation of a MSP20 electrode (cycled at 2 mV s−1/−0.2–0.6 V) and removed at 0.6 V was performed with ultramicrotomy after embedding in epoxy (Epofix, Struers, Germany). Ultrathin sections (<70 nm) were produced on the Leica Ultramicrotome UC6 (Leica Microsystems, Vienna, Austria) equipped with a 45° diamond knife (Diatome, Biel, Switzerland). After cutting, the thin sections were transferred to Quantifoil R3/3 TEM grids. Data acquisition and analysis were performed using the Gatan Digital Micrograph (GMS) Suite (v3.42, https://www.gatan.com/products/tem-analysis/gatan-microscopy-suite-software , accessed on: 3 April 2022). For determination of the iodine atomic positions the STEM HAADF, images were background corrected by subtracting a 128 times binned and subsequently rescaled copy of the corresponding.
45 diamond knife
Manufactured by Electron Microscopy Sciences
Sourced in Austria, Switzerland, Germany, United States
The 45° diamond knife is a specialized lab equipment used in electron microscopy and ultramicrotomy. It features a 45-degree angled diamond edge designed for cutting ultra-thin sections of samples for analysis under an electron microscope.
Automatically generated - may contain errors
Lab products found in correlation
Em uc6 ultramicrotome, by Electron Microscopy Sciences (3 mentions)
Tecnai t12 tem, by Thermo Fisher Scientific (2 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (2 mentions)
Ultracut uct ultramicrotome, by Leica (2 mentions)
Ultracut uct, by Electron Microscopy Sciences (2 mentions)
Ultracut uct type 706200, by Leica (2 mentions)
Cm100, by Thermo Fisher Scientific (2 mentions)
Titan g2 60 300, by Thermo Fisher Scientific (1 mentions)
Uc6 ultramicrotome, by Leica (1 mentions)
Autosamdri 815, by Tousimis (1 mentions)
Jem 1400, by JEOL (1 mentions)
Jem 1400 tem, by JEOL (1 mentions)
Veleta ccd camera, by Olympus (1 mentions)
Orius 830, by Ametek (1 mentions)
Edge 3.0 digital microscope, by Dino-Lite (1 mentions)
Fn s2n, by Nikon (1 mentions)
Embed 812 kit, by Electron Microscopy Sciences (1 mentions)
Temcam f416 ccd camera, by TVIPS (1 mentions)
Tecnai g2 spirit transmission electron microscope, by Thermo Fisher Scientific (1 mentions)
Supra 40vp, by Zeiss (1 mentions)
29 protocols using 45 diamond knife
1
Iodine Atomic Positions in MSP20 Electrodes
2
Cell Ultrastructural Analysis by SEM and TEM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For scanning electron microscopy (SEM) analysis, cell suspensions were adhered to 0.2 μm pore polycarbonate track-etched membranes and fixed in 2.5% glutaraldehyde (Electron Microscopy Sciences (EMS), Hatfield, PA) for 2 h. Cells were washed three times in 0.1 M sodium cacodylate buffer (EMS) and gradually dehydrated in an ethanol series (25, 33, 50, 75, 90%), followed by three washes in 100% ethanol (15 min each). After dehydration, the cells were processed in the automatic critical point dryer (CPD) Autosamdri-815 (Tousimis, Rockville, MD), with CO2 as a transitional fluid. The CPD-processed membranes were mounted on aluminum SEM stubs and sputter-coated with carbon. Cells were imaged with a Helios 600 Nanolab dual-beam microscope (FEI, Hillsboro, OR) at 2 kV.
For transmission electron microscopy (TEM) analysis, cell suspensions were pelleted by centrifugation (15,000 × g, 4°C, 10 min), fixed, and dehydrated as described for SEM. After the last ethanol wash, samples were gradually infiltrated in Spurr's low-viscosity embedding media (EMS). After polymerization at 60°C for 24 h, the hardened resin blocks were sectioned on a Leica EM UC6 Ultramicrotome using a 45° diamond knife (Diatome, Hatfield, PA). Seventy-nanometer sections were post-stained with 2% uranyl acetate and Reinold's lead citrate (seven and 3 min, respectively) and imaged in a Tecnai T-12 TEM (FEI) at 120 kV.
For transmission electron microscopy (TEM) analysis, cell suspensions were pelleted by centrifugation (15,000 × g, 4°C, 10 min), fixed, and dehydrated as described for SEM. After the last ethanol wash, samples were gradually infiltrated in Spurr's low-viscosity embedding media (EMS). After polymerization at 60°C for 24 h, the hardened resin blocks were sectioned on a Leica EM UC6 Ultramicrotome using a 45° diamond knife (Diatome, Hatfield, PA). Seventy-nanometer sections were post-stained with 2% uranyl acetate and Reinold's lead citrate (seven and 3 min, respectively) and imaged in a Tecnai T-12 TEM (FEI) at 120 kV.
3
Transmission Electron Microscopy Sample Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell monolayers were fixed for 2 h at 4°C with 0.1 M PBS containing 2.5% glutaraldehyde. Fixed samples were rinsed with PBS and further fixed with 1% OsO4 for 0.5 h at 4°C. The samples were rinsed with distilled water and dehydrated by sequential incubation with an acetone series (30%, 50%, 70%, 80%, 90%, 95%, 100%, and 100%, 5 min each). Samples were infiltrated with Araldite 502/Embed 812 by gradually increasing the concentration of acetone (25% and 50%, 20 min; 75%, 30 min; 100%, 20 h) and then polymerized at 60°C for 72 h. Embedded samples were sectioned using an UC6 ultramicrotome (Leica Biosystems) equipped with a 45° diamond knife (Diatome) to obtain 70-nm ultrathin sections. The grids were stained at RT with 2% aqueous uranyl acetate (10 min) and Reynolds lead citrate (5 min) before imaging. Imaging was performed at 80 kV on a JEM-1400 (JEOL) transmission electron microscope.
4
Ultrastructural Analysis of Mouse Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were fixed via cardiac perfusion at 37 °C with 2% paraformaldehyde (freshly made), 2.5% glutaraldehyde in pH 7.4, 0.15 M sodium cacodylate buffer with 0.03% calcium chloride. Fat pads were removed and immersion fixed overnight at 4 °C. Tissues were washed five times with ice-cold buffer consisting of 0.15 M sodium cacodylate, 0.03% calcium chloride, and postfixed with ice-cold 2% OsO4, 0.8% KFe(CN)3, 0.03% CaCl2 in 0.15 M sodium cacodylate for 1 h on ice, washed three times with ice-cold distilled water and stained overnight with 2% uranyl acetate at 4 °C. Tissues were dehydrated in an ethanol series, infiltrated and embedded in Durcupan ACM resin (Fluka). Ultrathin 80-nm-thick sections were made using a Leica EM UC6 ultramicrotome and Diatome 45° diamond knife. Sections were imaged by an FEI Spirit transmission electron microscope operated at 80 kV with a Teitz TemCam F224 2k by 2k CCD camera.
5
Transmission Electron Microscopy Virion Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For TEM, 70 nm thick sections were cut from the Epon block with a 45° diamond knife (Diatome, Bern, Switzerland) mounted on a Ultracut UCT ultramicrotome (Leica, Wetzlar, Germany) and then collected on freshly glow-discharged 300 mesh copper grids coated with a carbon-reinforced formvar film. Samples were imaged with a JEM-1400 TEM (Jeol, Tokyo, Japan) operating at 120 kV acceleration voltage and images were acquired with a Veleta CCD camera (Olympus, Tokyo, Japan). The diameters of virions were measured in the TEM images using ImageJ (1.53q, [43 (link)]).
6
Cryogenic Bacterial Fixation and TEM Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Concentrated bacteria grown in liquid broth were directly filled into 3 nm carriers (Leica, Germany) and vitrified with Leica EM HMP100. Freeze substitution was carried out as previously described (Bleck et al., 2010 (link)). The infiltration at -90°C lasted for 48 h before the temperature was raised with 5°C per h until reaching -30°C. Samples were kept at -30°C for 3 h before the staining solution was washed out 3 times and replaced with acetone. The bacteria were infiltrated with increasing epoxy Embed812-DER 736 resin (EMS, Hatfield, PA) concentration (1:2. 1:1, 2:1 and 1:0) for 2 h each before final exchange to 100% epoxy resin overnight was done and polymerized at 60°C for 48 h. Epoxy resin embedded bacteria were sectioned with a Leica FC6 ultramicrotome and 45° diamond knife (Diatome, Nidau, Switzerland) into 80 nm thin sections, which were placed on formvar coated copper grids and counterstained with lead citrate. The samples were examined with a Jeol 1230 TEM operating at 80 kV at 40,000x magnification. Micrographs were recorded with Gatan Orius 830 with 2048 x 2048 pixels charge-coupled device (CCD) camera using the Digital Micrograph software.
7
Characterization of Lignin Nanoparticle Photonics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A JSM-7401F field emission scanning electron microscope was used to image LNPs and LNPs photonic crystal samples. An accelerating voltage of 1–2 kV and a working distance of 2–15 mm were used during measurement. Some samples were coated for 60–120 s with gold using a JFC-1200 fine coater before the SEM study. The gold particles added by sputtering are about 5–15 nm in size and they were added to increase the contrast in the images. Annular dark field (ADF) scanning transmission electron microscopy (STEM) images were obtained using a ThermoFisher Themis Z double aberration-corrected TEM operated at 300 kV with a convergence angle of 21 mrad and a dwell time of 3 µs. The material was sectioned to 200 nm thickness using ultramicrotomy using a Leica Ultracut UCT with a 45° diamond knife (Diatome) after the crystalline lignin nanoparticles first had been embedded in Agar low viscosity resin to facilitate sectioning.
A Nikon FN-S2N (Japan) microscope was used to image the rectangular platelets of LNPs and record a movie of the assembly and rearrangement of LNPs during evaporation. A Dino-Lite Edge 3.0 digital microscope was used to image the photonic crystals of LNPs.
A Nikon FN-S2N (Japan) microscope was used to image the rectangular platelets of LNPs and record a movie of the assembly and rearrangement of LNPs during evaporation. A Dino-Lite Edge 3.0 digital microscope was used to image the photonic crystals of LNPs.
8
Sample Preparation for TEM Analysis
9
Ultrastructural Analysis of Coral Polyps
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Single polyps were micro-dissected, orientated, and embedded in 1.5% low melting agarose to ease handling of these small tissue samples and to preserve any fragile areas with tissue damage. The agarose was cut into cubes, which were placed in 1% osmium tetroxide in distilled water for 1 h. A series of washes in distilled water (4 × 10 min) and ethanol (3 × 10 min in 50, 70, 90 and 100%) followed, before the samples were embedded in Spurr’s resin. Sections (both 70 nm and 500 nm in thickness) were taken from the mouth region of the polyp using a 45° diamond knife (Diatome, Hatfield, PA, USA). The pharynx of the polyp was chosen as the area of interest, because this is where pathogens were observed to accumulate, both in this study, and in previous studies [37 (link), 39 ]. The resulting thin sections (70 nm) were placed on Formvar/C–coated copper grids, for Scanning Transmission Electron microscopy (STEM), while semi-thin sections (500 nm) were mounted on round glass slides (10 mm) for NanoSIMS imaging.
10
STEM Imaging of Pepto-Bismol Bismuth Subsalicylate
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In order to facilitate STEM imaging of crystalline BSS, a small amount (∼20 μg) of the washed and dried powder from the Pepto-Bismol original liquid formulation (BSS-PB) was embedded in a resin (LR White) in a gelatin capsule (size 00) and then hardened at 60 °C for 24 h. Ultra-thin sectioning, with an estimated section thickness of 40 nm, was later carried out using a Leica Ultracut UCT with a 45° diamond knife from Diatome. The sections were then transferred to carbon-coated copper grids (EMS-CF150-Cu-UL).
STEM images of BSS-PB were obtained using a Thermo Fisher Themis Z double aberration-corrected TEM. The microscope was operated at an accelerating voltage of 300 kV. The images were acquired using a beam current of 10 pA, a convergence angle of 16 mrad and a dwell time of 8 µs. iDPC and ADF images were obtained simultaneously. The ADF detector was set at a collection angle of 25–153 mrad. The iDPC images were formed using a segmented ADF detector. A high-pass filter was applied to the iDPC images to reduce low-frequency contrast. The lattice averaged potential maps were obtained by crystallographic image processing using the software CRISP50 (link).
STEM images of BSS-PB were obtained using a Thermo Fisher Themis Z double aberration-corrected TEM. The microscope was operated at an accelerating voltage of 300 kV. The images were acquired using a beam current of 10 pA, a convergence angle of 16 mrad and a dwell time of 8 µs. iDPC and ADF images were obtained simultaneously. The ADF detector was set at a collection angle of 25–153 mrad. The iDPC images were formed using a segmented ADF detector. A high-pass filter was applied to the iDPC images to reduce low-frequency contrast. The lattice averaged potential maps were obtained by crystallographic image processing using the software CRISP50 (link).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!