For TEM, a TiO2 NPs suspension (500 µg/mL; 5 µL) was applied onto glow-discharge-activated [22 (link)] carbon-coated 400-mesh copper grids (G400, SPI Supplies, Structure Probe, Inc., West Chester, PA, USA). Nanoparticles were sedimented for 1 min and the remaining solution was then blotted with filter paper and the grids were air-dried. A Philips CM100 electron microscope (Philips EO, Eindhoven, The Netherlands; Thermo Fisher Scientific) equipped with a Veleta slow-scan CCD camera (EMSIS GmbH, Muenster, Germany) was used to examine the grids. TEM images were processed in the proprietary iTEM software (EMSIS GmbH, Muenster, Germany).
Cm100 electron microscope
Manufactured by Philips
Sourced in Germany
The Philips CM100 is a transmission electron microscope designed for high-resolution imaging and analysis of materials at the nanoscale. It features an electron beam that is accelerated to a high voltage, allowing for the observation of fine structural details. The CM100 is capable of magnifying specimens up to 600,000 times, enabling detailed examination of micro- and nanostructures.
Automatically generated - may contain errors
Lab products found in correlation
Formvar ni grids, by Electron Microscopy Sciences (2 mentions)
Ultracut e, by Reichert Technologies (2 mentions)
Embed 812, by Electron Microscopy Sciences (2 mentions)
Epon lx 112, by Ladd Research Industries (2 mentions)
Amt software, by Advanced Microscopy Techniques (2 mentions)
Glutaraldehyde, by Merck Group (2 mentions)
Ultracut s ultramicrotome, by Philips (2 mentions)
Poly bed 812, by Polysciences (2 mentions)
Uranyl acetate, by Ted Pella (2 mentions)
Formvar support film, by SPI Supplies (2 mentions)
Spurr resin, by Merck Group (2 mentions)
Megaview g2 ccd camera, by EMSIS (2 mentions)
Tecnai g2 tem, by Thermo Fisher Scientific (2 mentions)
Ultracut s, by Reichert Technologies (2 mentions)
Veleta slow scan ccd camera, by EMSIS (1 mentions)
Item software, by EMSIS (1 mentions)
Prism 6.0 for windows, by GraphPad (1 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (1 mentions)
Osmium tetroxide, by Electron Microscopy Sciences (1 mentions)
Spurr s epoxy resin, by Electron Microscopy Sciences (1 mentions)
73 protocols using cm100 electron microscope
1
Characterization of TiO2 Nanoparticles by TEM
2
Ultrastructural Analysis of Infarcted Rat Myocardium
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The rats anesthetized and ventilated by precooled PBS (pH7.4) and 2.5 % glutaraldehyde. Subsequently, a tissue block measuring 1 mm on each side was acquired from the adjacent region of the infarct and the corresponding region of rats that underwent a sham operation. The tissue block was placed in a solution of 2.5 % glutaraldehyde at a temperature of 4 °C for a duration of 4 h. Following this, the samples were fixed using a combination of 1 % osmium tetraoxide and 0.1 M phosphate buffer (PB, pH7.4) at room temperature for 2 h. Subsequently, the samples underwent dehydration using a series of graded ethanol solutions and were then embedded in SPI PON 812 epoxy resin at a temperature of 37 °C overnight. Polymerization took place at a temperature of 60° Celsius for a duration of 48 h. The Leica UC7 Ultrathin Cutting Machine was utilized to cut ultra-thin sections (60–80 nm), which were then stained with a mixture of 2 % uranyl acetate and lead citrate for a period of 15 min. Finally, the Phillips CM 100 electron microscope was used for observation.
3
Ultrastructural Analysis of Pancreas
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Hematoxylin and eosin staining of paraffin sections was performed following standard protocol. Electron microscopy was performed as described previously [24 (link)]. Briefly, pancreas was minced with razor blade and fixed for 2 hours with a mixture of 2% glutaraldehyde and 2% formaldehyde in phosphate-buffered saline (PBS), post-fixed for 45 minutes with 1% OsO4, and then dehydrated and embedded in Epon. Ultrathin sections were stained with uranyl acetate and lead citrate; images were recorded digitally using a Philips CM-100 electron microscope.
4
Transmission Electron Microscopy of OMVs
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Five microliters of OMV sample was applied onto glow-discharge activated formvar-/carbon-coated 300 Mesh copper grids (Benada and Pokorný, 1990 (link)). After 60 s of adsorption, the grids were negatively stained with 1% ammonium molybdate, pH 6.5. Alternatively, a mixture of 1% ammonium molybdate and 0.1% trehalose was used (Harris et al., 1995 (link)). The grids were examined in Philips CM100 Electron Microscope (Philips EO, now Thermo Fisher Scientific) at 80 kV. The digital images were recorded using MegaViewII or MegaViewIII slow scan CCD cameras (Sis GmbH; Olympus, now EMSIS GmbH). All digital images were processed in the AnalySis3.2 Pro software suite (Version Build 788, 2003) using standard modules (shading correction and digital contrast enhancement).
5
Ultrastructural cell imaging protocol
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Cells were fixed in serum-free medium with 2.5% glutaraldehyde and 3% formaldehyde for 2 h at room temperature, washed with PBS and incubated with 2% osmium tetroxide and 1% K-hexacyanoferrate in H2O for 1h at 4°C. After washing with H2O, uranyl-acetate (2% in H2O) was added and incubated at 4°C overnight. Cells were scraped after washing with H2O, pelleted, dehydrated by sequential incubation in 20%, 50%, 70%, 90%, and three times 100% acetone/H2O for 30 min each, infiltrated with EMbed-812 (Electron Microscopy Science) according to the manufacturer's protocol, and allowed to polymerize for 24 h at 60°C. The embedded cell pellets were cut into 60-70 nm thin sections using an ultramicrotome (UltracutE, Reichert-Jung), collected on carbon-coated Formvar-Ni grids (Electron Microscopy Science), and stained for 10 min in 4% uranyl acetate and 2 min with lead citrate. Images were acquired on a CM100 electron microscope (Philips).
6
Ultrastructural Analysis of PyMT-Derived Tumors
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Tumors of different sizes from PyMT-VE-CadhSnail1CT and PyMT-VE-CadhSnail1KO mice were dissected and fixed with p-formaldehyde (2%) plus glutaraldehyde (2%) in 0.2 M cacodylate buffer. Samples were stored in cacodylate buffer until processing. After post-fixation with 2% osmium tetroxide, samples were dehydrated and embedded in Epon LX112 (Ladd Research Industries). Semi-thin sections (approximately 1 μm-thick) were cut and stained with toluidine blue. Ultrathin sections (60-80 nm) were obtained in an ultramicrotome, placed on parlodion/carbon-coated nickel grids and stained with lead citrate and uranyl acetate. The grids were examined using a Phillips CM100 electron microscope.
7
Melanosome Volume Estimation in RPE Cells
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Ocular tissue was micro-dissected and prepared as described above for electron microscopy. From each animal, two tissue blocks were chosen by random sampling for further analyses. Using a Philips CM 100 electron microscope, 5–8 pictures of RPE cells from the same section were obtained at a magnification of ×5,700. Melanosome volume was estimated by point-sampling of linear intercepts using the mathematical principle of volume-weighted mean volume as previously described (Sorensen 1991 (link)). In short, assuming isotropy of the melanosomes their mean volume was estimated using the formula:
Σl03: Sum of cubed intercept lengths; P(mel): Total number of melanosomes intercepted; Ln: Length of ruler.
All graphs were constructed using software (GraphPad Prism 6.0 for Windows, San Diego, CA, USA).
Σl03: Sum of cubed intercept lengths; P(mel): Total number of melanosomes intercepted; Ln: Length of ruler.
All graphs were constructed using software (GraphPad Prism 6.0 for Windows, San Diego, CA, USA).
8
Immunogold Labeling of Transfected Neuro-2a Cells
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Transfected Neuro-2a cells were grown in 10-cm dishes, transiently transfected with the NP∆ constructs and grown until 80% confluence. Cells were fixed in 3% formaldehyde and 0.2% glutaraldehyde for 2 h at room temperature, then scraped, pelleted, resuspended, and washed three times in PBS, incubated with 50 mM NH4Cl in PBS for 30 min, washed three times in PBS, resuspended in 2% warm agarose, and left to solidify on ice. Agarose pieces were dehydrated, infiltrated with LR gold resin (London Resin), and allowed to polymerize for 1 d at −10°C. Sections of 60–70 nm were collected on carbon-coated Formvar Ni-grids, incubated with rabbit anti-His6 antibodies in PBS, 2% BSA, 0.1% Tween-20 for 2 h, washed with PBS, and incubated with 10-nm colloidal gold-conjugated goat anti-rabbit immunoglobulin antibodies in PBS, 2% BSA, and 0.1% Tween-20 for 90 min. Grids were washed five times for 5 min in PBS and then five times in H2O, before staining for 10 min in 2% uranyl acetate. Sections were imaged with a Phillips CM100 electron microscope.
9
Exosome Visualization and Characterization
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Exosomes were fixed in 2% paraformaldehyde (w/v) in 200 mM phosphate buffer (pH 7.4), overlaid on a Formvar carbon-coated grid (FCF400-NI-50; Electron Microscopy Sciences, Hatfield, PA, USA) and left to dry. After 3 washes in phosphate buffer, the exosomes were further fixed in 1% glutaraldehyde for 5 min, washed in distilled water, and stained with aqueous uranyl oxalate (pH 7) for 5 min. The exosomes were then stained with saturated aqueous uranyl acetate, and samples were embedded in 0.4% uranyl acetate and 1.8% methylcellulose on ice for 10 min. Excess liquid was then absorbed with a Whatman filter (Sigma-Aldrich). The grids were dried at room temperature for 5 min before analyses. Exosome samples were visualized with the CM100 electron microscope (Philips, Eindhoven, The Netherlands).
In parallel, an aliquot of exosome samples was run on a Nanosight NS500 system (Nanosight Ltd., Amesbury, UK), and size distribution was analyzed using the NTA 1.3 software.
In parallel, an aliquot of exosome samples was run on a Nanosight NS500 system (Nanosight Ltd., Amesbury, UK), and size distribution was analyzed using the NTA 1.3 software.
10
Kidney Tissue Preparation for Electron Microscopy
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Kidney tissue specimens were fixed, embedded and stained as described previously (Liu et al., 2014 (link)). Ultrathin sections were cut using a Philips CM100 electron microscope.
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