Cells were fixed for 1h at room temperature in 2.5% glutaraldehyde in culture medium, and postfixed in 2% OsO4 in the same buffer. After serial dehydration in increasing ethanol concentrations, samples were embedded in agar 100 (Agar Scientific Ltd., United Kingdom) and left to polymerize for 2 days at 60°C. Ultrathin sections (50 to 70 nm thick) were collected in Formvar-carbon-coated copper grids by using a Leica EM UC6 ultramicrotome and stained with uranyl acetate and lead citrate. Observations were made on a Tecnai10 electron microscope (FEI), and images were captured with an Olympus VELETA camera and processed with AnalySIS software.
Tecnai 10 electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in Netherlands, Germany
The Tecnai 10 is a transmission electron microscope (TEM) manufactured by Thermo Fisher Scientific. It is designed to produce high-resolution images of small-scale structures and samples. The Tecnai 10 utilizes an electron beam to illuminate and magnify specimens, allowing users to observe details at the nanoscale level.
Automatically generated - may contain errors
Lab products found in correlation
Em uc6 ultramicrotome, by Leica (7 mentions)
Veleta camera, by Olympus (6 mentions)
Agar 100, by Agar Scientific (6 mentions)
Agar 100 resin, by Agar Scientific (4 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (2 mentions)
Uranyl acetate, by Electron Microscopy Sciences (2 mentions)
Osmium tetroxide, by Electron Microscopy Sciences (2 mentions)
Ferrocyanide, by Electron Microscopy Sciences (2 mentions)
Item software, by Olympus (2 mentions)
Em uc7 ultramicrotome, by Leica (2 mentions)
Cd11c magnetic beads, by Miltenyi Biotec (1 mentions)
Ms column, by Miltenyi Biotec (1 mentions)
Minimacs, by Miltenyi Biotec (1 mentions)
Veleta ccd camera, by Olympus (1 mentions)
Item sis software, by Olympus (1 mentions)
Potassium ferrocyanide, by Merck Group (1 mentions)
Uc6 ultramicrotome, by Leica (1 mentions)
Megaview 2 camera, by Adobe (1 mentions)
7600f microscope, by JEOL (1 mentions)
17 protocols using tecnai 10 electron microscope
1
Electron Microscopy Sample Preparation
2
Transmission Electron Microscopy of T Cells
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A total of 27 (WT) and 29 (AMPK-KO) T cells were analyzed by transmission electron microscopy. Naive T cells were maintained 4 days in IL-7-supplemented medium and then fixed overnight in 2.5% glutaraldehyde (Electron Microscopy Sciences, UK) at 4 °C and post-fixed with 1% osmium tetroxide (Electron Microscopy Sciences, UK) and 1.5% ferrocyanide (Electron Microscopy Sciences, UK) in 0.15 M cacodylate buffer. The cells were further stained with 1% uranyl acetate (Electron Microscopy Sciences, UK), serially dehydrated and embedded in epoxy resin (Agar 100 resin; Agar Scientific, UK). Ultrathin 70-nm sections were produced with a Leica EM UC6 ultramicrotome and mounted on copper–Formvar–carbon grids (Electron Microscopy Sciences). Observations were made on a Tecnai 10 electron microscope (FEI, Eindhoven, Netherlands), and images were captured with a Veleta camera and processed with SIS iTEM sofware (Olympus, Germany).
3
Electron Microscopy Tissue Preparation
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Cells were fixed for 1 h at room temperature in 2.5% glutaraldehyde in culture medium, and postfixed in 2% OsO4 in the same buffer. After serial dehydration in increasing ethanol concentrations, samples were embedded in agar 100 (Agar Scientific Ltd., UK) and left to polymerize for 2 days at 60 °C. Ultrathin sections (50–70-nm thick) were collected in Formvar-carbon-coated copper grids by using a Leica EM UC6 ultramicrotome and stained with uranyl acetate and lead citrate. Observations were made on a Tecnai10 electron microscope (FEI), and images were captured with an Olympus VELETA camera and processed with AnalySIS and Adobe Photoshop softwares.
4
Transmission Electron Microscopy Protocol
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TEM analysis was performed as described (Fontaine et al., 2017 (link)). Briefly, cells were fixed for 1h at room temperature in 2.5% glutaraldehyde in culture medium, and postfixed in 2% OsO4 in the same buffer. After serial dehydration in increasing ethanol concentrations, samples were embedded in agar 100 (Agar Scientific Ltd., United Kingdom) and left to polymerize for 2 days at 60°C. Ultrathin sections (50 to 70 nm thick) were collected in Formvar-carbon-coated copper grids by using a Leica EM UC6 ultramicrotome and stained with uranyl acetate and lead citrate. Observations were made on a Tecnai10 electron microscope (FEI), and images were captured with an Olympus VELETA camera and processed with iTEM software (Olympus GMBH, Germany).
5
Electron Microscopy Sample Preparation
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Cells were fixed for 1h at room temperature in 2.5% glutaraldehyde in culture medium, and postfixed in 2% OsO4 in the same buffer. After serial dehydration in increasing ethanol concentrations, samples were embedded in agar 100 (Agar Scientific Ltd., United Kingdom) and left to polymerize for 2 days at 60°C. Ultrathin sections (50 to 70 nm thick) were collected in Formvar-carbon-coated copper grids by using a Leica EM UC6 ultramicrotome and stained with uranyl acetate and lead citrate. Observations were made on a Tecnai10 electron microscope (FEI), and images were captured with an Olympus VELETA camera and processed with AnalySIS software.
6
Mitochondrial Ultrastructural Analysis
7
Ultrastructural Analysis of Zebrafish Skeletal Muscle
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Skeletal muscle of 5 dpf nup88+/+ and nup88-/- zebrafish larvae were analyzed by transmission electron microscopy on longitudinal and transversal ultrathin sections.
Briefly, larvae were fixed in 2.5% glutaraldehyde and 0.1 M sodium cacodylate buffer, pH 7.4 overnight at 4°C and post-fixed in 1% osmium tetroxide, 1.5% ferrocyanide in 0.15 M cacodylate buffer for 1 h at RT. After serial dehydration in increasing ethanol concentrations, samples were embedded in agar 100 (Agar Scientific Ltd., UK) and left to polymerize for 2 days at 60°C. Ultrathin sections (80 nm thick) were collected using a Leica EM UC6 ultramicrotome and stained with uranyl acetate and lead citrate. Images were recorded on a Tecnai10 electron microscope (FEI) equipped with an Olympus VELETA camera and processed using the AnalySIS software.
Briefly, larvae were fixed in 2.5% glutaraldehyde and 0.1 M sodium cacodylate buffer, pH 7.4 overnight at 4°C and post-fixed in 1% osmium tetroxide, 1.5% ferrocyanide in 0.15 M cacodylate buffer for 1 h at RT. After serial dehydration in increasing ethanol concentrations, samples were embedded in agar 100 (Agar Scientific Ltd., UK) and left to polymerize for 2 days at 60°C. Ultrathin sections (80 nm thick) were collected using a Leica EM UC6 ultramicrotome and stained with uranyl acetate and lead citrate. Images were recorded on a Tecnai10 electron microscope (FEI) equipped with an Olympus VELETA camera and processed using the AnalySIS software.
8
Isolation and Characterization of Pulmonary Alveolar Macrophages
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Pulmonary alveolar macrophages were obtained by homogenization and filtration of lungs, density gradient centrifugation (1.085 g/cm3 nycodenz, 1700xg during 30 min) and CD11c specific Magnetic associated cell sorting (MACS) by using MiniMACS, MS column and CD11c+ magnetic beads (Miltenyl Biotec).
Purified alveolar macrophages were fixed 2 hours in 2% glutaraldehyde in 0.1 M cacodylate buffer at 4°C, washed 3 times (4000xg, 5 min) with 0.2 M cacodylate buffer then post-fixed in 2% osmium tetroxide in 0.1 M cacodylate buffer for 1 hour at RT. After 3 washes, samples were serially dehydrated in ethanol (30% EtOH first 5 min, followed by 10 min–the same for 50%, 70%, 85%, 100% EtOH). Propylene oxide was then added 4x5 min at RT and progressively embedded in epoxy resin (Agar 100 resin; Agar Scientific, United Kingdom) (75,25, 50:50 and then 25:75% if propylene oxide/resin). Ultrathin 50-nm sections were obtained, mounted on copper-Formvar-carbon grids (EMS, United Kingdom), and stained with uranyl acetate and lead citrate by standard procedures. A Tecnai 10 electron microscope (FEI, Eindhoven, The Netherlands) was used for observations, and images were captured with a Veleta charge-coupled-device (CCD) camera and processed using the AnalySIS and Adobe Photoshop software programs.
Purified alveolar macrophages were fixed 2 hours in 2% glutaraldehyde in 0.1 M cacodylate buffer at 4°C, washed 3 times (4000xg, 5 min) with 0.2 M cacodylate buffer then post-fixed in 2% osmium tetroxide in 0.1 M cacodylate buffer for 1 hour at RT. After 3 washes, samples were serially dehydrated in ethanol (30% EtOH first 5 min, followed by 10 min–the same for 50%, 70%, 85%, 100% EtOH). Propylene oxide was then added 4x5 min at RT and progressively embedded in epoxy resin (Agar 100 resin; Agar Scientific, United Kingdom) (75,25, 50:50 and then 25:75% if propylene oxide/resin). Ultrathin 50-nm sections were obtained, mounted on copper-Formvar-carbon grids (EMS, United Kingdom), and stained with uranyl acetate and lead citrate by standard procedures. A Tecnai 10 electron microscope (FEI, Eindhoven, The Netherlands) was used for observations, and images were captured with a Veleta charge-coupled-device (CCD) camera and processed using the AnalySIS and Adobe Photoshop software programs.
9
Visualization of Globular Nanostructures
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Example 12
The nanostructures were diluted 30× with MilliQ water and 3 μl of sample was applied on a carbon 400 mesh copper grid that has been subjected to glow discharging. Samples were negatively stained with UAR-EMS. Subsequently, the grids were washed with ultrapure water and imaged using an FEI Tecnai 10 electron microscope run at 100 keV accelerating voltage. Images were acquired using a 2 k×2 k Veleta CCD camera (Olympus Soft Imaging System). Several globular nanostructures with a diameter larger than 8 nm were observed. The nanostructures contain a central part (dark core) and a peripheral part (white ring). See
10
Liposome Visualization and Characterization by TEM
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Here, 20 µL ALs and HTCC-ALs were deposited on Formvar carbon-coated electron-microscopy grids. Then, 1–2 µL glutaraldehyde 25% v:v was added to fix the liposomes. The preparation was left overnight at 4°C. Grids were transferred onto a drop of distilled water for washing and left for 2 minutes (three times). For contrasting and embedding, grids were placed onto a drop of methylcellulose–uranyl acetate (ratio 9:1 m/m) mixture for 10 minutes in an ice bath. The grids were removed and the excess fluid blotted by gently pushing the loop sideways on filter paper. A thin film was left over the section side of the grids. Observations were performed using a Tecnai 10 electron microscope (FEI, Hillsboro, OR, USA) operating at an accelerating voltage of 100 kV. Images were analyzed and processed using transmission electron microscopy (TEM) analysis (Olympus Soft Imaging Solutions, Münster, Germany).
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