For TEM analysis, samples were fixed in Karnovsky’s Fixative consisting of 8% PFA, 25% glutaraldehyde (Agar Scientific, Stansted, UK), and 0.2 M cacodylate buffer (Agar Scientific) for 1 hour before washing in 0.1 M cacodylate buffer. Samples were further fixed in 1% osmium tetroxide (Agar Scientific) for 1 hour, washed in 0.1 M cacodylate buffer before being dehydrated through an ethanol series before embedding in Epon resin (Agar Scientific) containing dodecenylsuccinic anhydride (DDSA), methyl nadic anhydride (MNA) and benzyldimethylamine (BDMA). Ultra-thin (70 nm) sections were obtained using a diamond knife (Agar Scientific) and loaded onto formvar-coated slot grids (Agar Scientific). Grids were then stained with uranyl acetate (Agar Scientific) and lead citrate (Sigma Aldrich) prior to imaging using a Hitachi H7600 electron microscope.
H 7600 electron microscope
Manufactured by Hitachi
Sourced in Japan
The H-7600 is an electron microscope manufactured by Hitachi. It is a tool used for high-resolution imaging and analysis of various samples at the nanoscale level. The core function of the H-7600 is to produce detailed images and data by focusing a beam of electrons onto a sample and detecting the resulting signals.
Automatically generated - may contain errors
Lab products found in correlation
Lkb 3 ultratome, by Leica (5 mentions)
Lead citrate, by Ted Pella (3 mentions)
Glutaraldehyde, by Ted Pella (3 mentions)
Graded ethanol and propylene oxide, by Thermo Fisher Scientific (3 mentions)
Epoxy resin, by Polysciences (2 mentions)
Uranyl acetate, by Ted Pella (2 mentions)
Jem 2010, by JEOL (2 mentions)
Amt em image acquisition software, by Advanced Microscopy Techniques (2 mentions)
Buffered formalin, by Merck Group (2 mentions)
Glutaraldehyde, by Agar Scientific (1 mentions)
Diamond knife, by Agar Scientific (1 mentions)
Lead citrate, by Merck Group (1 mentions)
Uranyl acetate, by Agar Scientific (1 mentions)
Osmium tetroxide, by Agar Scientific (1 mentions)
0.1 m cacodylate buffer, by Agar Scientific (1 mentions)
Epon resin, by Agar Scientific (1 mentions)
Formvar coated slot grids, by Agar Scientific (1 mentions)
Hq silver, by Nanoprobes (1 mentions)
Ultracut uct, by Leica (1 mentions)
Araldite, by Polysciences (1 mentions)
33 protocols using h 7600 electron microscope
1
Transmission Electron Microscopy Sample Preparation
2
Pre-embedding Immuno-EM of GLAST Mice
3
Antibiotic-Induced Morphological Changes in Bacteria
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Morphological changes of E. coli and S. aureus after antibiotic rifampicin and 5-iodoindole treatment were examined as previously described with some modification (Kim et al. 2011 (link)). As mentioned above, bacteria were exposed to rifampicin (100 μg mL−1) and incubated for 30 min at 37 °C, washed, and exposed to 5-iodoindole (2 mM) for 1 h at 37 °C and 250 rpm. Cells were then fixed initially with an aldehyde mixture of 50% glutaraldehyde and 35% formaldehyde, incubated at 4 °C overnight, collected by centrifugation, then fixed with 1% osmium tetroxide overnight at 4 °C, and washed three times. Cell pellets were mixed with 3% agarose and sliced to desired sizes, and slices were dehydrated and embedded in an Epon resin mixture (Hatfield, USA). Slices containing embedded cells were then sectioned using a MT-X ultramicrotome (Tucson, USA), loaded onto TEM copper grids, stained with uranyl acetate, and treated with lead citrate. Scanning electron microscopy was performed using a H-7600 electron microscope (Hitachi, Tokyo) at 80 keV.
4
Ultrastructural Analysis of Murine Aorta
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Two week-old mice were perfusion fixed through the heart, then thoracic aorta segments were isolated and immersion fixed with 1.5% glutaraldehyde and 2.5% paraformaldehyde in 0.1M HCl-Na cacodylate buffer for 2 hours, then post-fixed for 90 minutes at 40°C with 1% osmium tetraoxide in the same buffer, and then incubated with 0.5% tannic acid for 30 minutes at room temperature. After dehydration in graded ethanol, and propylene oxide treatment, specimens were embedded in epoxy resin. Sections were cut with a diamond knife, stained with lead citrate and uranyl acetate and examined with a Hitachi H-7600 electron microscope.
5
Exosome Characterization by TEM
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Exosomes were suspended in 1 × PBS and fixed with 2% paraformaldehyde. The fixed sample was absorbed onto formvar-coated copper grids for 20 min in a dry environment and fixed in 1% glutaraldehyde for 5 min. After being rinsed in distilled water, samples were stained with uranyl oxalate for 5 min followed by methyl cellulose uranyl acetate for 10 min on ice. Excess liquid was wicked off the grid, and grids were stored at room temperature until imaging. Imaging was performed using a Hitachi H7600 electron microscope.
6
Electron Microscopy of Corneal Ultrastructure
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The cornea was prepared via prefixation using 2% glutaraldehyde at 4°C, washing overnight at 4°C, postfixation using 2% osmium tetraoxide for 2 h at 4°C, dehydration, substitution using propylene oxide for 30 min and a mixture of propylene oxide and epoxy resin for 2 h, embedding, and sectioning. Ultrathin sections were mounted on 200-mesh copper grids and stained with 2% uranyl acetate for 15 min and then with lead solution for 5 min. Sections were observed with a Hitachi H-7600 electron microscope at 100 kV.
7
Imaging Rigid Protein Filaments
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STEM100Cu Elastic carbon substrate grids (Ohkenshoji Co., Ltd.) were glow discharged (5 mA, 45 s) and used within one hour. 5 µl of the incubated reaction mixture was applied to the glow discharged grids and allowed to absorb for 1 min. The reaction mixture was then blotted away using filter paper and 5 µl of 1% uranyl acetate applied to the grid. Staining was allowed to proceed for 1 min before being blotted by filter paper. Prepared grids were then dried overnight in a dry box before imaging. Grids were observed with a Hitachi H-7600 electron microscope operated at 100 kV and at a nominal magnification of ×40,000. Films were scanned in 7 µm steps with a Zeiss Z/I Imaging PhotoScan 2000 scanner. Fifty filament images were extracted, unbent and averaged. A Fourier pattern was calculated. Many obvious layer lines were observed, indicating the filament structure is rigid and uniform.
8
Ultrastructural Analysis of Bursal Tissue
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Tissue samples were fixed in 4% phosphate buffered glutaraldehyde at 4 °C overnight. PBS removed the excess fixative and the samples postfixed in 1% osmium-tetroxide for 2 h. After dehydration in graded ethanol, the bursal tissue was embedded in a mixture of araldite and epoxy-resin (Polysciences, Warrington, PA, USA). Ultrathin sections were contrasted by lead citrate and uranyl-acetate, studied using a Hitachi H-7600 electron microscope.
9
TEM Imaging of Nanoparticle-Treated Biofilms
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Synthesized nanoparticle-treated and untreated biofilms of MSSA 6538 were examined by TEM. Initially, overnight grown cells in 96-well plates were incubated with nanodispersions for 24 h without shaking at 37°C. For prefixation, cells were then treated with aldehyde mixture (glutaraldehyde 2.5% and formaldehyde 2%) and kept overnight at 4°C. After washing, cells were harvested by sonication to disrupt biofilms, immediately postfixed using 2% osmium tetroxide overnight at 4°C, and washed with 0.2 M phosphate buffer. Cell blocks were made using 2% agarose in sterile tubes, which were then sliced to the desired size. Specimens were dehydrated in an ethanol series and embedded in an Epon resin mixture (Electron Microscopy Sciences, Hatfield, PA, USA), which was treated at different temperatures to ensure complete polymerization. Thin sections were obtained using a MT-X ultramicrotome (Boeckeler Instruments, Tucson, AZ, USA) and loaded onto TEM copper grids. Finally, sections were stained with 1% uranyl acetate followed by lead citrate to remove any traces of moisture. Microscopy was performed using an H-7600 electron microscope (Hitachi Ltd., Tokyo, Japan) at 120-keV.
10
Electron Microscopy of Arterial Samples
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After fixation of arterial samples in 2.5% glutaraldehyde (TED PELLA, CA, USA) in PBS (pH 7.2), specimens were post-fixed in 1% osmium tetroxide (Heraeus, Hanau, Germany), dehydrated in graded ethanol and propylene oxide (Acros Organics, USA), and then embedded in Epoxy resin (mix with Nadic Methyl Anhydride (NMA) and Dodecenyl Succinic Anhydride (DDSA) and DMP-30, all reagents from Polysciences (PA, USA). Serial ultrathin sections were cut using an LKB-III ultratome (LEICA, Wetzlar, Germany). Ultrathin sections were stained with uranyl acetate (TED PELLA, CA, USA) and lead citrate (TED PELLA, CA, USA) and were examined with the aid of a Hitachi H7600 electron microscope (Hitachi, Japan) at an accelerating voltage of 100 kV.
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