Adult mice received an intraperitoneal overdose of pentobarbital and were then transcardially perfused with 0.9% saline followed by a 1% PFA, 1% glutaraldehyde fixative in 0.1M phosphate buffer (PB). Spinal cords were then removed and post-fixed in the same solution for 24 hours and transferred to 0.1M PB before sectioning. 60μm sections were prepared using a vibrating-blate microtome, post-fixed with osmium tetroxide OsO4, dehydrated and flat-embedded in epoxy resin. At this stage, some images were acquired using light microscopy. Subsequently, ultrathin sections were cut using a Diatome diamond knife and Leica Ultracut S ultramicrotome. Images were acquired at 120kV on a Philips CM100 transmission electron microscope with a Gatan OneView CMOS camera. The appearance of identifiable glomerular structures was interpreted qualitatively.
Cm100 transmission electron microscope
Manufactured by Ametek
Sourced in Netherlands
The CM100 is a transmission electron microscope designed for high-resolution imaging and analysis of materials at the nanoscale. It features a tungsten filament electron gun, a LaB6 electron gun, and advanced imaging and analytical capabilities. The CM100 enables users to visualize and study the internal structure and composition of a wide range of samples, including biological specimens, semiconductors, and advanced materials.
Automatically generated - may contain errors
Lab products found in correlation
Ultracut s ultramicrotome, by Leica (1 mentions)
Oneview cmos camera, by Ametek (1 mentions)
Em uc7 ultramicrotome, by Leica (1 mentions)
Orius sc2001, by Ametek (1 mentions)
Poly bed 812, by Polysciences (1 mentions)
1k ccd camera, by Ametek (1 mentions)
Cu pd tem grids, by Agar Scientific (1 mentions)
Dissection microscope, by Olympus (1 mentions)
Ultracut s, by Leica (1 mentions)
Em uc6 ultramicrotome, by Leica (1 mentions)
Orius ccd camera, by Ametek (1 mentions)
Jsm 7401f microscope, by JEOL (1 mentions)
Digital camera, by Olympus (1 mentions)
Szx7 microscope, by Olympus (1 mentions)
Formvar and carbon coated grid, by Agar Scientific (1 mentions)
Orius sc200 ccd camera, by Ametek (1 mentions)
6 protocols using cm100 transmission electron microscope
1
Ultrastructural Analysis of Mouse Spinal Cord
2
Ultrastructural Analysis of ZIKV Infection
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Vero cells were infected with single-round infectious E103A HA-NS2A ZIKV (derived from trans-complementation as shown in Fig. 2E ) at an MOI of 1. At 24 h p.i., cells were fixed with fixative (50 mM cacodylate buffer [pH 7.3], 2.5% formaldehyde, 0.1% glutaraldehyde, 0.01% picric acid, 0.03% CaCl2) for 2 h at room temperature and then washed twice with 100 mM cacodylate buffer. The monolayers were scraped off and centrifuged at 12,000 rpm for 5 min to pellet cells. The pellets were postfixed with 1% OsO4 in 100 mM cacodylate buffer, washed with electron microscopy (EM)-grade water, and en bloc stained with 2% aqueous uranyl acetate in water for 20 min at 60°C. The pellets were dehydrated in ethanol series (from 50% to 100%) and processed through propylene oxide before being embedded in Poly/Bed 812 (Polysciences). Ultrathin sections (70 nm) were cut on a Leica EM UC7 ultramicrotome (Leica Microsystems), stained with 0.4% lead citrate in water for 3 min, and examined with a Philips CM100 transmission electron microscope at 60 kV. Digital images were acquired with a bottom-mounted charge-coupled device (CCD) camera, Orius SC2001 (Gatan, Pleasanton, CA).
3
Characterizing Polymer Nanoparticles by TEM
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Cu/Pd TEM grids (Agar
Scientific, UK) were coated in-house with a thin carbon film. A single
7 μL droplet of a 0.1% w/w aqueous dispersion of a glutaraldehyde-crosslinked
PHBA latex (or glutaraldehyde-crosslinked PHBA–PNAEP diblock
copolymer nanoparticles) was pipetted onto the carbon-coated grid
and carefully blotted with a filter paper after 1 min. Then, a single
7 μL droplet of a 0.75% w/w aqueous solution of uranyl formate
was pipetted onto the grid for 1 min to stain the deposited particles.
Excess stain was removed using a vacuum hose. A Philips CM100 transmission
electron microscope equipped with a Gatan 1k CCD camera was used to
image the stained samples at an accelerating voltage of 100 kV and
a beam current of 3 mA.
Scientific, UK) were coated in-house with a thin carbon film. A single
7 μL droplet of a 0.1% w/w aqueous dispersion of a glutaraldehyde-crosslinked
PHBA latex (or glutaraldehyde-crosslinked PHBA–PNAEP diblock
copolymer nanoparticles) was pipetted onto the carbon-coated grid
and carefully blotted with a filter paper after 1 min. Then, a single
7 μL droplet of a 0.75% w/w aqueous solution of uranyl formate
was pipetted onto the grid for 1 min to stain the deposited particles.
Excess stain was removed using a vacuum hose. A Philips CM100 transmission
electron microscope equipped with a Gatan 1k CCD camera was used to
image the stained samples at an accelerating voltage of 100 kV and
a beam current of 3 mA.
4
Visualizing and Characterizing Extracellular Vesicles
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Isolated EVs were visualized and characterized using TEM. The EVs' pellet was resuspended in 100 μL of dPBS, and 10 μL of the sample was further diluted with 90 μL of dPBS. Ten μL of the diluted EVs was placed onto a parafilm strip and then incubated with a graphene oxide on a holey carbon copper mesh grid (Electron Microscopy Sciences, #GOHC300Cu10) for 5 minutes. Excess sample was removed using filter paper, and the grid was subsequently incubated with 1 drop of uranyl acetate (Electron Microscopy Sciences, SKU #22400). Any excess uranyl acetate was removed using filter paper. Images of the EVs were captured using a Philips CM-100 transmission electron microscope at 60 kV, equipped with an Orius SC2001 digital camera (Gatan).
5
Anther Morphology Analysis of Arabidopsis Flowers
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Flower images were taken using an Olympus dissection microscope with an Olympus digital camera. Alexander solution and staining were performed as described (Alexander, 1969 (link)). Photography was performed with an Olympus SZX7 microscope. Transverse sections of anthers of 17–20mm buds treated or not with 100mM uniconazole and/or 100mM NBD for 4 d. Sections were stained with 1% safranine O in 50% ethanol. Mature flowers of Arabidopsis were fixed in 2.5% glutaraldehyde and 4% paraformaldehyde in 0.1M sodium phosphate buffer, pH 7.0 at room temperature for 4h. After three 20min buffer rinses, the samples were post-fixed in 1% OsO4 in the same buffer for 4h at room temperature and then rinsed in three 20min changes of buffer. Samples were dehydrated in an ethanol series and propylene oxide, embedded in Spurr’s resin, and sectioned with a Lecia Reichert Ultracut S or Lecia EM UC6 ultramicrotome. The ultra-thin sections (70–90nm) were stained with uranyl acetate and lead citrate. Sections were observed using a Philips CM 100 transmission electron microscope at 80kV and the images were obtained with a Gatan Orius CCD camera. For SEM, pollen grains were coated with platinum particles (JFC-1600) for 30 s at 20 mA and viewed under a JSM-7401F microscope (JEOL).
6
Fibril Characterization by TEM
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Lyophilized powder was dissolved in 0.1 M phosphate buffer (pH 5) to a final concentration of 1 mg/mL. Prepared sample was filtered through 0.2 mm filter (Iso-Disc TM ) and incubated at 50 °C and 300 rpm for 96 h. Undiluted sample was applied to Formvar and carbon-coated grid (Agar Scientific, Stansted, UK) and left to adsorb for 3 min. The excess of sample was soaked away and stained with 1% (w/v) water solution of uranyl acetate. The excess of stain was removed immediately. The fibrils were observed with a Philips (Amsterdam, the Netherlands) CM 100 transmission electron microscope at 80 kV. Images were recorded by Gatan Orius SC200 CCD camera and Digital Micrograph software 3.1, (Gatan Inc., Washington, DC, USA).
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