Cells were processed for electron microscopy and stained for lipids by the imidazole-buffered osmium tetroxide procedure (Angermuller and Fahimi, 1982 (link)). Briefly, Caco-2/TC7 cells were fixed for 60 min with 2.5% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) and then incubated for 30 min in 0.2 M imidazole buffer (pH 7.4) to which 4% aqueous osmium tetroxide was added immediately before use. All material was dehydrated with ethanol and embedded in Epon 812. Ultrathin sections were counterstained with 3% lead citrate for 60 min and examined with a Jeol 100CX-II electron microscope.
100 cxii electron microscope
Manufactured by JEOL
Sourced in United Kingdom, Japan, United States
The JEOL 100-CXII is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of materials at the nanoscale. It features a LaB6 electron source and a sophisticated optical system to produce high-quality, high-magnification images. The 100-CXII is equipped with advanced analytical capabilities, including energy-dispersive X-ray spectroscopy (EDS) and electron energy-loss spectroscopy (EELS), enabling comprehensive characterization of specimen composition and structure.
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Lab products found in correlation
Axio observer z1, by Zeiss (3 mentions)
Epoxy resin, by TAAB (3 mentions)
Item software, by Olympus (2 mentions)
Megaview 3 digital camera, by Olympus (2 mentions)
V750 pro scanner, by Epson (1 mentions)
Em bed, by Electron Microscopy Sciences (1 mentions)
Fcf 200 cu, by Electron Microscopy Sciences (1 mentions)
Formvar coated 200 mesh copper grid, by Electron Microscopy Sciences (1 mentions)
18 protocols using 100 cxii electron microscope
1
Electron Microscopy Lipid Staining
2
Transmission Electron Microscopy of Pichia pastoris
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EM of P. pastoris cells was performed as previously described (Levi et al., 2010 (link)). Briefly, a 50-ml culture of yeast cells was grown in rich glucose medium to an OD600 of ∼0.5. The culture was concentrated to a volume of <5 ml with a bottle-top vacuum filter and was fixed by adding 40 ml of ice-cold 50 mM KPi, pH 6.8, 1 mM MgCl2, and 2% glutaraldehyde and leaving on ice for 1 h. The cells were washed repeatedly and then resuspended in 0.75 ml of 4% KMnO4 and mixed for 1 h at room temperature. The cells were washed, resuspended in 0.75 ml of 2% uranyl acetate, and mixed for 1 h at room temperature. Finally, the cells were washed and dehydrated and embedded in Spurr’s resin. The resin was polymerized for 2 d at 68°C. Thin sections were stained with uranyl acetate and lead citrate and were viewed with a JEOL 100CX II electron microscope.
3
Ultrastructural Analysis of HepaRG Cells
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HepaRG cells were fixed by the addition of 2.5% glutaraldehyde for 30 min. After fixation, the specimens were rinsed with 0.2 M Na cacodylate buffer and post-fixed with 2% osmium tetroxide for 30 min. After further rinsing, the samples were dehydrated, infiltrated with a mixture of acetone-eponate (50/50) and embedded in DMP30-Eponate. Ultrathin sections were examined with a JEOL 100CX II electron microscope42 (link).
4
Measuring Cuticle Thickness in Frieseomelitta varia Guards
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To test whether the darker Frieseomelitta varia guards had a thicker cuticle, we caught one guard and one forager from nine different colonies (nine guards, nine foragers) and sectioned the cuticle in three different parts of the clypeal area of the head to measure the thickness of the cuticle using transmission electron microscopy (Jeol-Jem-100cx-II Electron Microscope). Thinly cut head slices were fixed in glutaraldehyde 5% and washed three times in sodium cacodylate 0.1 M. Subsequently, the thickness of the cuticle was measured using ImageJ 1.46.
5
Ultrastructural Imaging of Brain Capillaries
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Tissue samples from each brain region were cut into 1 mm × 1 mm blocks and post fixed in an aqueous solution of 1% osmium tetroxide and 1% lanthanum nitrate for 1 h. The samples were contrast stained en-block with 1% Uranyl acetate for 1 h followed by sequential dehydration in ethanol and propylene oxide. Samples were then embedded in Epon 812 acrylic resin and ultrathin sections of tissue were placed on copper grids for imaging with a JEOL 100CXII electron microscope. Capillaries within the tissue were imaged at a 10,000× magnification. Only capillaries which entirely fit within the image were selected for analysis. Five images per region were captured by a microscopist blinded to the nature of the samples. Film containing the images was developed and digitized using an EPSON V750 pro scanner.
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Imaging Bacterial Microcompartments In Vivo and In Vitro
7
Cellular Uptake of Varying Size AuNPs
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A total of 5 × 105 cells were plated in 6 well cell culture plates and cultured for 24 h. Cells were then exposed for 3 h to AuNPs. Four nm size AuNPs were added at the concentrations of 3.12 × 1013 p/ml, while 14 nm size AuNPs were used at 7.2 × 1011 p/ml. Following exposure to AuNPs, cells were washed twice in 1x phosphate-buffered saline (PBS), and prepared for TEM as previously described.50 (link) In brief, cells were fixed in in 2.5% glutaraldehyde in 0.1 M Phosphate Buffer, pH 7.4 (Electron Microscopy Sciences, Hatfield, PA) for 1 h at room temperature. Cells were then scraped of the plates, centrifuged at a low speed and suspended in 2.5% glutaraldehyde. Samples were further processed at the Michigan State University (MSU) Center for Advanced Microscopy by post-fixation in 1% osmium tetroxide, rinsing in distilled water, and dehydration through a graded series of acetone. At the end, samples were embedded in epoxy resin and cut into 70 nm sections that were then analyzed and photographed by JEOL 100CXII electron microscope.
8
Electron Microscopy Specimen Preparation
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Briefly, the cells were harvested by trypsinization and fixed with 3% glutaraldehyde at 4°C for 3–4 hours followed by washing with 0.1 M sodium cacodylate buffer. The cell pellets were then fixed in Osmium tetroxide for 1 hour at 4°C in dark, subjected to dehydration by passing through different grades of alcohol and then mounted with Araldite resin. The ultrathin sections (∼60–70 nm) were mounted on formvar coated copper grids. These sections were stained with uranyl acetate solution and counterstained with lead citrate. Electron micrographs were captured on a Jeol 100-CXII electron microscope (Jeol, UK) using Olympus camera and iTEM software.
9
Ultrastructural Examination of Rabbit Eyelid Development
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Small pieces 2.0–3.0 mm long from the specimens were placed on 2.5% cold glutaraldehyde in phosphate buffer (pH 7.2) for 24 h. The pieces were washed twice in 0.1-M phosphate buffer and then post-fixed in 1% osmium tetraoxide, in the same buffer. The post-fixed pieces were dehydrated in graded alcohols and embedded in Araldite resin. Semithin sections (1 μm) in thickness will be stained with 1% toluidine blue.
Specimens from the upper eyelids of three rabbits at one, 2 weeks, and 1 month postnatal were used for semithin sections, which were fixed in 2.5% cold glutaraldehyde in phosphate buffer (pH 7.2) for 24 h. The pieces were washed twice in 0.1-M phosphate buffer and then post-fixed in 1% osmium tetraoxide, in the same buffer. The post-fixed pieces were dehydrated in graded alcohols and embedded in Araldite resin. Semithin sections were cut at (1 μm) in thickness and stained with 1% toluidine blue [16 (link)]. Ultra-thin sections obtained from a Reichert ultra-microtome were stained with uranyl acetate and lead citrate [17 (link)]. Transmission electron microscopy (TEM) images were captured by the JEOL-100CXII electron microscope (JEOL, Tokyo, Japan) at the Electron Microscopy Unit of Assiut University.
Specimens from the upper eyelids of three rabbits at one, 2 weeks, and 1 month postnatal were used for semithin sections, which were fixed in 2.5% cold glutaraldehyde in phosphate buffer (pH 7.2) for 24 h. The pieces were washed twice in 0.1-M phosphate buffer and then post-fixed in 1% osmium tetraoxide, in the same buffer. The post-fixed pieces were dehydrated in graded alcohols and embedded in Araldite resin. Semithin sections were cut at (1 μm) in thickness and stained with 1% toluidine blue [16 (link)]. Ultra-thin sections obtained from a Reichert ultra-microtome were stained with uranyl acetate and lead citrate [17 (link)]. Transmission electron microscopy (TEM) images were captured by the JEOL-100CXII electron microscope (JEOL, Tokyo, Japan) at the Electron Microscopy Unit of Assiut University.
10
Tissue Fixation for Ultrastructural Analysis
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Tissue fragments of spleen, heart, brain, liver and head kidney were fixed in Karnovsky’s solution (2% formaldehyde plus 2.5% glutaraldehyde in phosphate buffer 0.1 M, pH 7.0) for 24 h at 4 ºC, bathed three times in phosphate buffer 0.1 M, pH 7.0, post-fixed in 1% osmium tetroxide in sodium cacodylate buffer (0.1 M, pH 7.2–7.4) for 1 h, and then processed for TEM [20 ]. The sections were examined using a JEOL-100CXII electron microscope (Jeol, Peabody, MA, USA).
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