The plant samples were fixed first in fixation buffer I (5% glutaraldehyde and 0.1 M phosphate, pH 7.4) for 4 h at room temperature and then in fixation buffer II (2% osmium tetroxide and 0.1 M phosphate, pH 7.4) at 4 °C overnight. After one wash in phosphate buffer and two washes in double-distilled water, the samples were stained for 1 h in 1% (m/v) uranyl acetate. After another wash in double-distilled water, the samples were dehydrated through a gradient of alcohol (from 30%, 50%, 70%, to 85%) and then embedded in Spurr’s resin (Sigma-Aldrich). Ultrathin sections of 70 nm were prepared using the UC7 μLtramicrotome (Leica Microsystem) and mounted on the AG100M molybdenum grids with a single slot (Agar Scientific) to minimize the background for copper. The sections were stained with uranyl acetate and lead citrate, observed, and photographed using a Tecnai G2 20 Twin electron microscope (FEI) at 120 kV. EF-TEM for copper element analysis was carried out on the same specimen using a JEM-2100F field-emission high-resolution transmission electron microscope equipped with the energy filter (JEOL). Three individual leaves were used for each genotype and at least 10 cells were analyzed per sample.
Tecnai g2 20 twin electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Tecnai G2 20 Twin electron microscope is a high-performance transmission electron microscope (TEM) designed for advanced materials and life science research. The instrument provides exceptional imaging and analytical capabilities, enabling users to visualize and characterize samples at the nanoscale level.
Automatically generated - may contain errors
Lab products found in correlation
Em uc7, by Leica (5 mentions)
Eagle 4k ccd digital camera, by Thermo Fisher Scientific (2 mentions)
Spurr s resin, by Merck Group (1 mentions)
Jem 2100f field emission high resolution transmission electron microscope, by JEOL (1 mentions)
Phosphate buffered saline (pbs), by Wuhan Servicebio Technology (1 mentions)
Rnalater solution, by Thermo Fisher Scientific (1 mentions)
Spi pon 812 resin, by SPI Supplies (1 mentions)
Embed 812, by Electron Microscopy Sciences (1 mentions)
15 protocols using tecnai g2 20 twin electron microscope
1
Ultrastructural Analysis of Plant Leaves
2
Ultrastructural Analysis of Soleus Muscle
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Mice were euthanized and perfused with sodium phosphate buffer (PB, 100 mM, pH7.4) and pre-fixed solution (2.5% (vol/vol) glutaraldehyde, 1% paraformaldehyde in PB). Soleus muscle was dissected, cut into small pieces and fixed in the same pre-fixed solution overnight at 4 °C. After rinsing with PB, tissues were immersed in 0.2 M imidazole in PB for 15 min, and then post-fixed with 1% osmium tetraoxide in PB. After rinsing with high-purity water, the samples were stained with 1% aqueous lead at 4 °C overnight. Gradient dehydration was accomplished by incrementing the concentration of acetone and embedded in epoxy resin (60 °C for 24 h). Samples were sectioned using Leica EM UC7 and placed on copper grids. Images were taken on a FEI Tecnai G2 20 Twin electron microscope equipped with an Eagle 4k CCD digital camera (FEI; USA) in a double-blind manner.
3
Ultrastructural analysis of demyelination
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To further evaluate demyelination injury, the mice were anaesthetised at the end of the study, and 1 mm3 of corpus callosum tissue was dissected on ice immediately after death. These samples were immersed in 2.5% glutaraldehyde TEM fixation solution (Servicebio) and 1% osmic acid in 0.1 M PBS (pH 7.4). After fixation and dehydration with ethanol, the tissue was embedded using acetone and embedding medium (Servicebio). Ultrathin sections were prepared using an ultramicrotome (Leica UC7). The ultrastructure of myelinated axons was imaged using the FEI Tecnai G2 20 TWIN electron microscope.
4
Electron Microscopy Sample Preparation
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Cells were collected and washed in 0.1 M sodium cacodylate, then fixed in a solution of 2% glutaraldehyde and 4% paraformaldehyde for 1 h at 4°C. Samples were treated postfixation with 1% OsO4 and 1% K3Fe(CN)6 for 40 min. After three washes with sodium cacodylate, samples were dehydrated with ethanol, treated with propylene oxide, and embedded in Epon. Ultrathin sections were prepared and examined at 200 kV under a Tecnai G220 TWIN electron microscope (FEI, America).
5
Negative Staining of SARS-CoV-2 Spike Proteins
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For the negative-staining study, S2P, S6P, and BD-368-2 Fab were diluted to 0.02 mg/ml using 25 mM HEPES, pH 7.2, 150 mM NaCl. BD-368-2 Fab was then mixed with S2P or S6P in a 1:1 volume ratio and incubated on ice for 3 min or at room temperature for 30 min. The mixture was then applied onto a glow-discharged carbon-coated copper grid (Zhong Jing Ke Yi, Beijing). After 1 min, the excess liquid was removed using a filter paper. The grids were then stained using 1% uranyl acetate for 30 seconds and air-dried. A Tecnai G2 20 Twin electron microscope (FEI) operated at 120 kV was used to examine the grids. Images were recorded using a CCD camera (Eagle, FEI).
6
Transmission Electron Microscopy of Carbon-Coated Polymeric Micelles
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For the TEM visualization, 15 μL of CCPM dispersion (1250×
diluted in milliQ water) was dropped onto a layer of activated carbon
film supported on a 100-mesh hexagonal copper grid and incubated for
15 min. CCPMs/polymers not bound to the activated carbon film were
washed away with several drops of PBS pH 7.4. Subsequently, the sample
was fixated using 1% glutaraldehyde in PBS for 10 min and the grid
was extensively washed with several drops of milliQ water. Next, 150
μL of a negative staining solution (a mixture of 2% uranyl oxalate
and 0.15% methylcellulose in an ammonium acetate buffer pH 7.0) was
applied for a 15 min incubation, after which the excess was blotted
away using a filter paper and the sample was air-dried for at least
1 h at RT. Images were obtained on an FEI Tecnai G2 20 TWIN electron
microscope, which was operated at an acceleration voltage of 120 kV
and a spot size of 3. Images were recorded using RADIUS software.
diluted in milliQ water) was dropped onto a layer of activated carbon
film supported on a 100-mesh hexagonal copper grid and incubated for
15 min. CCPMs/polymers not bound to the activated carbon film were
washed away with several drops of PBS pH 7.4. Subsequently, the sample
was fixated using 1% glutaraldehyde in PBS for 10 min and the grid
was extensively washed with several drops of milliQ water. Next, 150
μL of a negative staining solution (a mixture of 2% uranyl oxalate
and 0.15% methylcellulose in an ammonium acetate buffer pH 7.0) was
applied for a 15 min incubation, after which the excess was blotted
away using a filter paper and the sample was air-dried for at least
1 h at RT. Images were obtained on an FEI Tecnai G2 20 TWIN electron
microscope, which was operated at an acceleration voltage of 120 kV
and a spot size of 3. Images were recorded using RADIUS software.
7
Ultrastructural Analysis of Rectal Tissue
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The rectal tissue obtained by resection was soaked in RNAlater solution Ambion company overnight, the solution was discarded, and the tissue was frozen at −80°C. The rectal tissue was cut into 1-mm-thick sections and fixed in 2.5% glutaraldehyde and 1% osmium tetroxide in a biosafety cabinet with level 2 protection, and subsequently dehydrated using different ascending concentrations of alcohol (30% to 100%), and immersed and embedded in epoxy resin. Ultrathin sections (80–100 nm) were prepared on formvar-coated copper grids (200 mesh) Yasheng Electronic Technology Co., Ltd. The virions were observed with a Tecnai G2 20 Twin electron microscope (FEI Company) under 200 kV.
8
Reovirus Infection of Cell Lines
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DLD-1, PC-3, NCI-H460, and CIK cells were incubated with 1 pfu/cell of reovirus at 4°C for 2 hours. Then, the cells were washed with PBS twice and incubated at 37°C for 12 hours. Following incubation, the cells were harvested and fixed in 2.5% glutaraldehyde in 0.1M sodium phosphate buffer (pH 7.4) at room temperature for 2 hours. The cells were then washed once in sodium phosphate buffer and dehydrated using a graded series of ethanol. After dehydration, the samples were subjected to two changes of propylene oxide and embedded in epoxy resin. After ultra-microtome sectioning, sections were analyzed on a FEI Tecnai G2 20 TWIN electron microscope operating at 200 kV.
9
Electron Microscopy of EBOV Infected Cells
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Ultrathin-section electron microscopy was performed as described previously (6 (link)). Briefly, Huh7 cells were cultured in 0.4-μm-pore-size Transwells (Corning), in which 2 × 105 cells were seeded on polycarbonate membrane for each chamber and infected with EBOV at an MOI of 0.1. At 72 h after infection, cells were fixed with 4% of paraformaldehyde for at least 30 min and stored with the fix buffer at 4°C overnight. Cells and polycarbonate membranes were removed from Transwell chambers as a complete monolayer. The cell monolayer was postfixed with 1% osmium tetroxide in 0.1 M PBS for 30 min and dehydrated with a series of ethanol gradients followed by propylene oxide before being embedded in resin. Thin sections were stained with 2% uranyl acetate and Reynolds lead citrate for 15 min and processed for TEM imaging under an FEI Tecnai G2 20 Twin electron microscope.
10
Ultrastructural Analysis of Cells
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Cells were washed twice with 0.1 mol/L sodium cacodylate buffer (pH = 7.4) followed by fixation with 2.5% glutaraldehyde in the same buffer for 2 h, and then post-fixed with 1% OsO4 for 1 h at room temperature. After rinsing several times in cacodylate buffer and distilled water, the cells were incubated in 0.1% tannic acid (in cacodylate buffer) for 30 min, and stained in 1% uranyl acetate for 1 h. They were washed again in distilled water and dehydrated in a graded ethanol series and embedded in SPI-Pon 812 resin (SPI Supplies, PA, USA). Ultrathin (70 nm) sections were cut using an ultramicrotome (UC7, Leica Microsystem), and collected on copper grids with a single slot, stained with uranyl acetate and lead citrate. Then the sections were observed under a Tecnai G2 20 TWIN electron microscope at 120 kV and photographed with an Eagle (4k×4k) digital camera (FEI, Oregon, USA).
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