Labeled FSDCs were fixed in 1% OsO4 buffered with PBS for 15 minutes. The cells were washed with distilled water three times. The samples were dehydrated using a gradient series of ethanol and embedded in an Epon-Araldite resin. The samples were sectioned using a diamond knife on a Reichert-Jung Ultracut-E ultramicrotome. 100 nm sections were mounted onto copper grids and coated with carbon. The sections were imaged on a Hitachi 7100 transmission electron microscope.
7100 transmission electron microscope
Manufactured by Hitachi
Sourced in Japan, France, United Kingdom
The Hitachi 7100 transmission electron microscope is a high-performance laboratory instrument designed for the analysis of microscopic samples. It utilizes a focused electron beam to produce detailed images of the internal structure and composition of various materials at the nanoscale level. The Hitachi 7100 provides users with the ability to study the morphology, crystal structure, and elemental distribution of their samples with high resolution and magnification capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Uct ultramicrotome, by Leica (3 mentions)
Ultracut e ultramicrotome, by Reichert Technologies (2 mentions)
Em amw, by Leica (2 mentions)
Veleta 2k x 2k megapixel side mounted tem ccd camera, by Olympus (2 mentions)
Veleta 2 k 2 k megapixel side mounted tem ccd camera, by Olympus (2 mentions)
Ultracut e ultramicrotome, by Leica (1 mentions)
Amt advantage 10 ccd camera system, by Advanced Microscopy Techniques (1 mentions)
Rabbit anti goat 10 nm gold conjugated secondary antibody, by Electron Microscopy Sciences (1 mentions)
Em uc6 ultramicrotome, by Leica (1 mentions)
Reichert ultracut e ultramicrotome, by Leica (1 mentions)
Osmium tetroxide, by TAAB (1 mentions)
Paraformaldehyde (pfa), by TAAB (1 mentions)
0.22 μm pore filter, by Merck Group (1 mentions)
Exosome quick extraction solution, by System Biosciences (1 mentions)
Ultrascan 1000 ccd camera, by Ametek (1 mentions)
Formvar carbon coated 400 mesh copper grids, by Agar Scientific (1 mentions)
23 protocols using 7100 transmission electron microscope
1
Transmission Electron Microscopy of Cells
2
Transmission Electron Microscopy of Exosomes
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Exosomes were analyzed by transmission electron microscopy using negative staining. 2 μl of exosomes was placed on Formvar/carbon coated copper mesh grids, washed three times with PBS, and fixed with 2.0% phosphotungstic acid in aqueous suspension. Samples were examined with a Hitachi 7100 transmission electron microscope.
3
Electron Microscopy of Sympathetic Ganglia
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For electron microscopy, SCGs were dissected with their exiting nerve and immersed in a solution of 2.5% glutaraldehyde in PHEM buffer (1X, pH 7.4) overnight at 4°C. They were then washed in PHEM buffer and post-fixed in a 0.5% osmic acid + 0.8% Potassium ferrocyanide solution for 2 hr at room temperature in the dark. After two washes with PHEM buffer, samples were dehydrated with solutions of increasing ethanol concentration (30–100%). Samples were embedded in EmBed 812 using an Automated Microwave Tissue Processor for Electronic Microscopy, Leica EM AMW. Thin sections (70 nm; Leica-Reichert Ultracut E) were collected at different levels of each block. These sections were counterstained with uranyl acetate and lead citrate and observed using a Hitachi 7100 transmission electron microscope in the Centre de Ressources en Imagerie Cellulaire de Montpellier (France).
4
Transmission Electron Microscopy Sample Preparation
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Cells were fixed in 2% paraformaldehyde (PFA) buffered with PBS at 4 °C overnight, followed by washing with PBS twice. Cells were then fixed in 1% OsO4 buffered with PBS for 1 hr and then washed three times with distilled water. The samples were dehydrated using a gradient series of ethanol and embedded in an Epon-Araldite resin. 100-nm sections were cut using a DDK diamond knife on a Reichert-Jung Ultracut-E ultramicrotome (Leica, Wetzlar, Germany). The sections were not stained with lead citrate or uranyl acetate. The sections were then mounted onto copper grids and imaged on a Hitachi 7100 transmission electron microscope (Pleasanton, CA) operated at 75 kv. Digital images were obtained using an AMT Advantage 10 CCD Camera System (Advanced Microscopy Techniques Corporation, Danvers, MA) and NIH Image software (Bethesda, MD).
5
Immunogold Labeling of Galectin-7
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Cells were fixed in a 0.1% (v/v) gluteraldehyde and 4% (v/v) paraformaldehyde solution and embedded in the low viscosity embedding Spurr media. Ultrathin sections were cut, placed on nickel grids and incubated in sodium metaperiodate. Samples were blocked in 1% (v/v) BSA for 5 min, incubated 60 min in a goat anti-human gal-7 polyclonal antibody (1:150) and 60 min in a rabbit anti-goat 10 nm gold-conjugated secondary antibody (1:20, Electron Microscopy Sciences, Hatfield, PA). Each section were counterstained with uranyle acetate and lead citrate and visualized using a Hitachi 7100 transmission electron microscope.
6
Exosome Morphology Visualization by TEM
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Exosomes isolated from MHCC97-H and MHCC97-L cells were identified for morphology by transmission electron microscopy (TEM) as previously described (19 (link)). In brief, exosomes were transferred to a copper grid coated with 0.125% Formvar in chloroform immediately after isolation. Then the grids were stained with 1% (v/v) uranyl acetate in double-distilled water right before examination. A Hitachi 7100 transmission electron microscope was applied for imaging.
7
Ultrastructural Analysis of Drosophila Larval Brains
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Third instar larvae were dissected in PBS 1x, EDTA 1 mM. Larval brains were then fixed overnight in 5% glutaraldehyde in 0.1 M phosphate buffer (NaH2PO4·2H2O and Na2HPO4·2H2O). They were then rinsed in phosphate buffer and post-fixed in a 1% osmic plus 0.8% potassium ferrocyanide for 2 h in the dark at room temperature. After two rinses in a phosphate buffer, the brains were dehydrated in a graded series of ethanol solutions (30–100%). The brains were embedded in EMBed 812 DER 736. Thin sections (85 nm; Leica-Reichert Ultracut E) were collected at different levels of each block. These sections were counterstained with uranyl acetate and lead citrate and observed using a Hitachi 7100 transmission electron microscope in the RIO imaging facility (C. Cazevieille, Montpellier, France).
8
Transmission Electron Microscope Preparation
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Cells were immersed in a solution of 2.5% glutaraldehyde in Sorensen's buffer (0.1M, pH 7.4) overnightat4 °C. After a rinse in Sorensen's buffer, cells were postfixed in a 0.5% osmic acid for 2 h in the dark at room temperature. After two rinses in Sorensen's buffer, the cells were dehydrated in a graded series of ethanol solutions (30-100%). The cells were embedded in EMbed 812 using a Leica EM AMW automated microwave tissue processor for electron microscopy. Thin sections (70 nm, obtained with a Leica-Reichert Ultracut E microtome) were collected at different levels of each block. These sections were counterstained with uranyl acetate and observed with a Hitachi 7100 transmission electron microscope at the Centre de Ressources en Imagerie Cellulaire de Montpellier, Montpellier, France.
9
TEM Analysis of Spinal Cord Injury
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The transmission electron microscopy images were obtained from healthy (control) and hemisected (3 days after SCI) spinal cord tissue ipsilateral to the injury at segment L5, lamina VIII, where most of the locus coeruleus axons terminate (Clark and Proudfit, 1991) . The surgical procedure and tissue preparation were performed as described above. The spinal cord slices were fixed in 4% paraformaldehyde, postfixed in 1% OsO 4 (Taab, Aldermaston, Berkshire, UK) for 20 min, dehydrated in a graded ethanol series and embedded in Taab 812 (Taab). During dehydration, the sections were treated with 1% uranyl acetate in 50% ethanol for 20 min. Ultrathin sections were cut with a Leica EM UC6 ultramicrotome (Leica Microsystems, Vienna, Austria) and analysed using a Hitachi 7100 transmission electron microscope (Hitachi, Tokyo, Japan) equipped with a Veleta side-mounted TEM CCD camera (Olympus, Tokyo, Japan). Contrast and brightness of electron micrographs were edited using Adobe Photoshop CS3 (Adobe Systems, San Jose, CA, USA).
10
Ultrastructural Analysis of Renal Cortex
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Renal cortexes were fixed with 2% glutaraldehyde in 0.1 mol/L phosphate buffer at 4°C for 120 min. They were then sectioned into ultrathin slices and were double stained with 4% uranyl acetate and lead citrate. The ultrathin slices were observed by a Hitachi 7100 transmission electron microscope (Hitachi High Technologies, Tokyo, Japan).
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