The sperm were collected from the epididymis of 8–12-week-old male mice and fixed in 50 mM sodium cacodylate, pH 7.2, containing 2.5% glutaraldehyde and 2% sucrose, for 2 h at 4 °C, as described previously [26 (link)]. The fixed sperm were then rinsed three times and postfixed in a fixative containing 2% osmium tetroxide for 2 h at 4 °C. The specimens were subsequently dehydrated in ethanol, immersed twice in propylene oxide for 15 min, and then embedded in Epon. Ultrathin sections were prepared by using an ultramicrotome (Reichert Ultracut; Leica AG, Vienna, Austria), and stained with uranyl acetate and lead citrate. The sections were observed using a transmission electron microscope (H-7000; Hitachi High-Tech, Tokyo, Japan).
H 7000
Manufactured by Hitachi
Sourced in Japan, United States, United Kingdom
The H-7000 is a high-performance transmission electron microscope (TEM) manufactured by Hitachi. It is designed for various applications in materials science, nanotechnology, and life sciences. The H-7000 provides high-resolution imaging and analytical capabilities, enabling users to study the structure and composition of materials at the atomic scale.
Automatically generated - may contain errors
Lab products found in correlation
Epon resin 828, by Polysciences (4 mentions)
Lambda 40, by PerkinElmer (3 mentions)
90 plus particle sizer analyzer, by Brookhaven Instruments (3 mentions)
Reichert jung ultracut e, by Leica camera (2 mentions)
0.1 m sodium cacodylate buffer, by Merck Group (2 mentions)
Ultramicrotome, by Reichert Technologies (2 mentions)
Epoxy resin, by TAAB (2 mentions)
Ultracut uct, by Leica camera (2 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (2 mentions)
Osmium tetroxide, by Merck Group (2 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions)
Reichert ultracut s, by Leica Microsystems (2 mentions)
Xr 41, by Advanced Microscopy Techniques (2 mentions)
Quetol 812, by Nisshin EM (2 mentions)
Spurr resin, by Agar Scientific (2 mentions)
Reichert ultracut e ultramicrotome, by Leica Microsystems (2 mentions)
Sis megaview 3 plate camera, by EMSIS (2 mentions)
Eclipse 80i, by Nikon (1 mentions)
Veleta 2 k 2 k ccd side mount camera, by EMSIS (1 mentions)
Zetasizer 3000hs, by Malvern Panalytical (1 mentions)
120 protocols using h 7000
1
Ultrastructure Analysis of Mouse Sperm
2
Microscopic Analysis of Nerve Fiber Regeneration
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After electrophysiological analysis, the rats were euthanized under deep isoflurane anesthesia, and the left sciatic nerve was dissected. Semithin and ultrathin transverse sections were prepared in the middle of the graft according to a previous study29 (link). Images of semithin sections stained with toluidine blue were obtained using a light microscope (ECLIPSE 80i; Nikon, Tokyo, Japan). The number of myelinated nerve fibers was counted using the ImageJ software (National Institutes of Health, Bethesda, MD, USA), and the density was calculated. Ten areas of each ultrathin section stained with uranyl acetate and lead citrate were randomly obtained at 2000-fold magnification using a transmission electron microscope (TEM; H-7000, Hitachi High-Technologies, Tokyo, Japan). The shortest diameter of the myelinated nerve fibers (α) and axons (β) was measured using the ImageJ software, and the myelin sheath thickness of each fiber was calculated using the formula (α − β)/2.
3
Negative Contrast Electron Microscopy of Virions
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10 μL-drops of virion extracts were placed on the 400 mesh copper grids (Formvar/Carbon Square Mesh, UB, Electron Microscopy Sciences, Hatfield, PA, USA) for negative contrast electron microscopy. After 5 min incubation, the virion solution on each grid was removed using filter paper. The grids were stained using 1% uranyl acetate dissolved in distilled water for 30 s followed by drying in the air. Images were taken using Hitachi H-7000 transmission electron microscope (TEM, Hitachi High-Technologies, Schaumburg, IL, USA) equipped with a Veleta (2k × 2k) CCD side mount camera (EMSIS, Munster, Germany) at 100 kV. The length of virions was measured using Image J software (http://rsbweb.nih.gov/ij ) directly from the images taken. For each mutant virus, over 100 flexuous virions were measured and up to 10 longest virions were selected to calculate the mean value.
4
Characterization of Curcumin-Loaded Nanostructured Lipid Carriers
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A Zetasizer 3000HS (Malvern Instruments, Malvern, UK) was employed to characterize Cur-TCA NLCs and Cur NLCs for particle size, polydispersity index (PDI), and ζ-potential. Morphology of the NLCs was observed under transmission electron microscopy (TEM; H-7000; Hitachi, Tokyo, Japan). A previously reported method was used to investigate the EE and DL of NLCs.22
Differential scanning calorimetry (DSC) was done with the aid of a TA 60WS with DSC-60 (Shimadzu) to analyze thermal properties of S100-TCA, Cur, blank TCA50 NLCs, and Cur-TCA50 NLCs. Each sample was sealed in an aluminum crucible and subjected to 50°C–300°C temperatures at 10 K/min in a nitrogen atmosphere. Ultraviolet-visible spectroscopy was utilized to probe the Cur orientation within the hydrophobic core of Cur NLCs or Cur-TCA50 NLCs.23 ,24 Spectra of aqueous suspension of NLCs and methanolic solution of Cur were taken from 200 to 800 nm at 1.5 µg/mL Cur concentration.
Differential scanning calorimetry (DSC) was done with the aid of a TA 60WS with DSC-60 (Shimadzu) to analyze thermal properties of S100-TCA, Cur, blank TCA50 NLCs, and Cur-TCA50 NLCs. Each sample was sealed in an aluminum crucible and subjected to 50°C–300°C temperatures at 10 K/min in a nitrogen atmosphere. Ultraviolet-visible spectroscopy was utilized to probe the Cur orientation within the hydrophobic core of Cur NLCs or Cur-TCA50 NLCs.23 ,24 Spectra of aqueous suspension of NLCs and methanolic solution of Cur were taken from 200 to 800 nm at 1.5 µg/mL Cur concentration.
5
TEM Imaging of Nanoparticle Morphology
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In conjunction with the phosphotungstic acid-based negative dyeing method, transmission electron microscopy (TEM) was used to observe the morphology of the prepared NPs. In brief, a droplet of NP suspension was placed on a copper grid, the excess liquid was drained onto filter paper, and the grid was dried at room temperature. The copper grid was stained for 2 minutes in a 2% phosphotungstic acid solution, and then the morphology of the NPs was observed using TEM (H-7000, Hitachi Company, Tokyo, Japan).
6
Micelle Length Distribution Characterization
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Bright-field transmission electron microscopy (TEM) images were taken at the nanoimaging facility of the chemistry department of the University of Toronto using a Hitachi H-7000 instrument (Hitachi High-Tech Corporation, Tokyo, Japan). Samples were prepared by placing one drop of solution on a Formvar carbon-coated grid, touching the edge of the droplet with a filter paper to remove excess liquid and allowing the grid to dry.
For each sample, micelle length distributions were determined by tracing more than 200 micelles using the software ImageJ (NIH, Laboratory for Optical and Computational Instrumentation, LOCI, University of Wisconsin, Madison, WI, US). Error bars were calculated using the standard error of the mean, s.e.m., obtained with a 99% confidence interval.
For each sample, micelle length distributions were determined by tracing more than 200 micelles using the software ImageJ (NIH, Laboratory for Optical and Computational Instrumentation, LOCI, University of Wisconsin, Madison, WI, US). Error bars were calculated using the standard error of the mean, s.e.m., obtained with a 99% confidence interval.
7
Ultrastructural Analysis of Salivary Glands
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At the end of the experiment, a transcardial perfusion using 2.5% glutaraldehyde and 4% paraformaldehyde mixed fixative in 0.1 M mol phosphate buffer (PB, pH 7.4) was performed on anesthetized rats. Next, SMG and SSG tissues were rapidly isolated and fixed in 2.5% glutaraldehyde at 4°C for 24 h, followed by 1% osmium tetroxide at 4°C for 3 h. After gradient alcohol dehydration, specimens were infiltrated and embedded using epoxy resin. A transmission electron microscope (TEM) (HITACHI H-7000, Tokyo, Japan) was then used to observe the ultrastructure in SMG and SSN tissues, including acinar cells, ductal cells, and secretory granules of the SMG and axon of the SSN. Moreover, at least 100 axon diameters of the SSN were determined in each group using ImageJ software in accordance with a previous study (Ito et al., 2016 (link)). Subsequently, the G-ratio was calculated using the following formula: G-ratio = inner axonal diameter (d)/(outer axonal diameter) (D), as shown in Figure 5C .
8
Negative Staining and TEM Imaging of Vesicles
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A drop of vesicle suspension was dropped and adsorbed onto a copper grid covered in carbon film. Subsequently, a 1% (w/v) solution of phosphotungstic acid (PTA) was used to negatively stain the copper grids. The dyed grids containing the samples (CUN, GIQ9, AER-200 and CUN-S-SNEDDS) were then dried and examined by TEM (Hitachi H-7000). The instrument had a digital camera and operated at a voltage of 200 kV (Mega View II, Olympus, Tokyo, Japan) [46 (link)].
9
TEM Preparation of Cellular Samples
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Cells were pelleted at 250 g for 5 min, washed twice with PBS, and then twice with 0.1 M sodium cacodylate buffer, pH 7.2 (Sigma-Aldrich), before primary fixation in 2% glutaraldehyde and 2% paraformaldehyde (Electron Microscopy Sciences) in cacodylate buffer overnight at 4°C. Cells were then secondarily fixed in 2% osmium tetroxide (Sigma-Aldrich) in cacodylate buffer for 2 h at room temperature. The cell pellet was then dehydrated through an ethanol gradient (30, 60, 90, and 100%) followed by propylene oxide. Subsequently, cells were embedded in Low Viscosity Resin (TAAB). Sections were cut on a microtome (Reichert-Jung Ultracut E; Leica), stained with uranyl acetate and lead citrate, and then viewed on a transmission electron microscope (H-7000; Hitachi). Images were taken with a camera (ORCA-HRL; Hamamatsu Photonics) and processed using AMT version 6 (AMT Imaging).
10
Characterization of Drug-Loaded HSA Nanoparticles
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The mean particle size, size distribution, zeta potential, and polydispersity index (PDI) of drug-loaded HSA NPs were measured with a Zetasizer nano ZS (Malvern, Worcestershire, UK) by scattering angle of 90° at 25°C. The drug-loaded HSA NPs were diluted with double-distilled water before the measurement, and all measurements were performed at least in triplicate. The shape and size were also observed by transmission electron microscopy (TEM), using Hitachi H-7000 (Hitachi, Tokyo, Japan). The purified NPs were diluted with water to allow clearer pictures to be taken. Samples were prepared by placing a drop on carbon-coated copper grids and sponging off the excess with filter paper. Then, the samples were stained with uranyl acetate (2% aqueous solution) for 3 minutes and dried at room temperature.28 (link)
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