To determine ultrastructural alterations of M. canis prior to and following FSH1 gene- knockdown, SEM and TEM were used to observe the cultured fungal colonies. Samples were prepared for transmission electron microscopy as described by Basma et al (32 (link)). Electron micrographs were generated using a Hitachi HT7700 electron microscope (Hitachi, Ltd.) at 100 kV. Samples were prepared for scanning electron microscopy as described by Wang et al (33 (link)). Dried colonies were coated with platinum-vanadium and observed under a Supra55 scanning electron microscope (Carl Zeiss AG) at 15 kV. All assays were performed in triplicate.
Ht7700 electron microscope
Manufactured by Hitachi
Sourced in Japan, France, Germany
The HT7700 is an electron microscope manufactured by Hitachi. It is designed for high-resolution imaging and analysis of a wide range of materials and samples. The HT7700 utilizes an electron beam to generate magnified images, allowing for detailed observation and investigation of the microscopic structure of various specimens.
Automatically generated - may contain errors
Lab products found in correlation
2kx2k veleta ccd camera, by Olympus (6 mentions)
Glutaraldehyde, by Merck Group (4 mentions)
Em uc6 microtome, by Leica (3 mentions)
Em uc6, by Leica (3 mentions)
Paraformaldehyde (pfa), by Nacalai Tesque (2 mentions)
Uc7 ultramicrotome, by Leica (2 mentions)
X maxn65t ccd camera, by Oxford Instruments (2 mentions)
Zen2 program, by Zeiss (2 mentions)
Ultramicrotome, by Leica (2 mentions)
Veleta camera, by Olympus (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Escalab 250xi, by Thermo Fisher Scientific (2 mentions)
Lx 112, by Ladd Research Industries (2 mentions)
Supra 55 scanning electron microscope, by Zeiss (1 mentions)
Glutaraldehyde, by Fujifilm (1 mentions)
Can get signal immunostain solution b, by Toyobo (1 mentions)
Tecnai 10 transmission electron microscope, by Philips (1 mentions)
Megaview 3 digital camera, by EMSIS (1 mentions)
S 3400n scanning electron microscope, by Hitachi (1 mentions)
Digital sight ds u3, by Nikon (1 mentions)
128 protocols using ht7700 electron microscope
1
Ultrastructural Analysis of M. canis Fungus
2
Ultrastructural Imaging of HepG2 Cells
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HepG2 cells were fixed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer at 4°C for 2 h, followed by 1% osmium tetroxide in 0.1 M sodium cacodylate buffer for 2 h at room temperature. Then, they were stained using 1% uranyl acetate for 1 h at room temperature, dehydrated in an increasing ethanol series and embedded in epoxy resin. The cells were subsequently imaged using a Hitachi-HT7700 electron microscope (Hitachi, Ltd.).
3
Evaluating Hepatic Steatosis and Mitochondrial Changes
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The hepatic tissues were stained with H&E and examined using light microscopy. Liver steatosis stages were evaluated semiquantitatively according to the NASH-Clinical Research Network criteria [15 (link)]. The amount of steatosis (percentage of hepatocytes containing fat droplets) was scored as 0 (<5%), 1 (5%–33%), 2 (>33%–66%), and 3 (>66%). Liver sections were fixed with 2.5% glutaraldehyde and 4% paraformaldehyde, then stained with lead-uranyl acetate, and changes in mitochondrial morphology were observed with a Hitachi HT7700 electron microscope (Hitachi, Tokyo, Japan). Lipid extraction was performed using the Folch et al.’s method [16 (link)]. Hepatic TG and cholesterol contents were measured using commercially available kits (Wako Pure Chemical Industries, Osaka, Japan). Malonyl-CoA concentration was determined using the kit (Cusabio, Wuhan, China).
4
Transmission Electron Microscopy of Liposomes
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TEM analysis was performed on SIM-LCL and SIM-DOX-LCL prepared at different PL and drug ratios. For this analysis, negative staining method with ammonium molybdate was used, but, prior to this procedure, liposomes were firstly treated with osmium tetroxide 4% in order to obtain a better contrasting image [45 (link),46 (link)]. The technique implied the dilution of liposomal suspension (200µL) in PBS (1 to 7 v/v) and the addition of 1.25 µL osmium tetroxide 4%. The obtained suspension was kept for 1 h at 4 °C. After this period, the liposomal dilution was centrifugated for 5 min at 12,000× g, and the sediment was resuspended in 200 µL ultrapure water. From the final suspension, a volume of 5 µL was deposited on carbon-coated copper grids for 2 to 5 min, after which the excess of the sample was removed using filter paper. The sample was then treated with 5% ammonium molybdate and 1% trehalose and left to dry at room temperature in a desiccator. After 24 h, the samples were analyzed using a Hitachi HT7700 electron microscope (Hitachi Ltd., Tokyo, Japan) equipped with a camera.
5
Transmission Electron Microscopy of Cells
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Cells were fixed in 2% glutaraldehyde (FUJIFILM Wako Pure Chemical Corporation, 072–02262), 2% paraformaldehyde (Nacalai Tesque, Inc., 30525–89-4) buffered with 0.1 M PB, pH 7.2. Cells were then post-fixed with 1% OsO4 and embedded in Epon 812 (Oken Shoji, 02–1002) after dehydration. Ultrathin sections, 60 nm, were cut with an ultramicrotome (UC6, Leica Microsystems, Vienna Austria), stained with uranium acetate and lead citrate and examined with a Hitachi HT7700 electron microscope (Hitachi, Tokyo, Japan).
6
Transmission Electron Microscopy Imaging
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Materials were directly adsorbed onto a carbon film membrane on a 300-mesh copper grid, stained with 1% uranyl acetate, dissolved in distilled water, and dried at room temperature. Grids were examined with Hitachi HT7700 electron microscope operated at 80 (Elexience – France), and images were acquired with a charge-coupled device camera (AMT).
7
Immunoelectron Microscopy of Mitochondria
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Immunoelectron microscopy using ultrathin cryosections was performed as previously described [39 (link)]. Briefly, conditioned media-derived pellets containing DsRed2-mitochondria fixed in 4% paraformaldehyde (Nacalai Tesque, Inc., 30525–89-4) in 0.1 M PB (pH7.2) overnight at 4°C were washed with PBS, embedded in 12% gelatin (Rousselot) in 0.1 M PB, immersed in 2.3 M sucrose in 0.1 M PB overnight and plunged into liquid nitrogen. Sections approximately 60 nm thick were cut with a cryo-ultramicrotome (UC7/FC7, Leica Microsystems, Vienna, Austria) and reacted overnight at 4°C with mouse anti-TOMM20 antibody (1:50) (Santa Cruz Biotechnology, sc-17764) and 1 µg/ml rabbit anti-mRFP antibody (kindly gifted by Dr. Hiroyuki Hioki, Juntendo University, Japan) [40 (link)], followed by donkey anti-mouse IgG conjugated with 12 nm colloidal gold particles (1:40; Jackson ImmunoResearch Laboratories, Inc., 115–205-146) and donkey anti-rabbit IgG conjugated with 6 nm colloidal gold particles (1:40; Jackson ImmunoResearch Laboratories, Inc., 711–195-152). The grids with sections were embedded in a thin layer of 2% methylcellulose (Sigma-Aldrich, M-6385) with 0.4% uranyl acetate (pH 4.0), air-dried, and observed with a Hitachi HT7700 electron microscope (Hitachi, Tokyo, Japan).
8
Mitochondrial Morphology of Articular Chondrocytes
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At culture day 6, primary articular chondrocytes were fixed in 2.5% glutaraldehyde in PBS for 60 min at room temperature. The mitochondrial morphology of primary articular chondrocytes was evaluated using a Hitachi HT7700 electron microscope (HITACHI high-tech, Tokyo, Japan).
9
Visualizing Yeast Pfk2p-GFP Localization
10
Ultrastructural Analysis of Semen Samples
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Semen samples of normal controls and patients (F1: II-2 and F2: II-2) were treated as previously described.14 (link) Samples were fixed with glutaraldehyde (Sigma-Aldrich, St. Louis, Missouri, USA) and osmium tetroxide, followed by OsO4 and sucrose, and dehydrated using graded ethanol. Subsequently, the samples were embedded in Epon 812, dodecenylsuccinic anhydride, methylnadic anhydride and dimethylaminomethyl phenol. Ultrathin (70–90 nm) sections were contrasted with uranyl acetate and lead citrate. An HT7700 Hitachi electron microscope and a MegaView III digital camera (Munster, Germany) were used for image capturing.
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