Fixation of the specimens was done by phosphate-buffered glutaraldehyde (pH 7.4), and post fixation by 1% osmium tetroxide at 4 °C; then, dehydration and embedding in epoxy resin occurred. Cutting by (Leica ultra-cut UCT), staining was performed by uranyl acetate and lead citrate [28 ]. Examination and photography using (JEOL JEM 1010 transmission electron microscope; Jeol Ltd., Tokyo, Japan) in the Regional Center of Mycology and Biotechnology (RCMB), Al-Azhar University, Egypt.
Jem 1010 transmission electron microscope
Manufactured by JEOL
Sourced in Japan, United States, United Kingdom, Germany, Spain
The JEM 1010 is a transmission electron microscope (TEM) manufactured by JEOL. It is designed to provide high-quality imaging and analysis of sample materials at the nanoscale level. The JEM 1010 utilizes an electron beam to illuminate and interact with the sample, allowing for the observation and study of microscopic structures and features.
Automatically generated - may contain errors
Lab products found in correlation
Amt imaging system, by Advanced Microscopy Techniques (72 mentions)
Ultracut microtome, by Leica (62 mentions)
Em stainer, by Leica (47 mentions)
Ultracut uct ultramicrotome, by Leica (8 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (5 mentions)
Ultracut uct, by Leica (5 mentions)
Ultramicrotome, by Leica (5 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (5 mentions)
Megaview 3 high resolution digital camera and imaging software, by JEOL (4 mentions)
Embed 812, by Electron Microscopy Sciences (3 mentions)
Uranyl acetate, by Merck Group (3 mentions)
Durcupan epoxy resin, by Merck Group (3 mentions)
Embed 812 resin, by Electron Microscopy Sciences (3 mentions)
Megaview 3, by Olympus (3 mentions)
Em uc7, by Leica (3 mentions)
Em grade 50 solution, by Electron Microscopy Sciences (3 mentions)
Picric acid, by Merck Group (3 mentions)
0.1 m cacodylate buffer, by Merck Group (3 mentions)
Em uc7 ultramicrotome, by Leica (3 mentions)
Mega view 3 camera, by Olympus (2 mentions)
291 protocols using jem 1010 transmission electron microscope
1
Ultrastructural Analysis of Biological Specimens
2
Ultrastructural Analysis of Cell Lines
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HCT-116 and RKO (control and treatment) cell lines were centrifuged and the culture medium above the pellets was replaced with 2.7% glutaraldehyde (Electron Microscopy Sciences, Hatfield, USA) in 0.1 M phosphate buffer, pH 7.4. Prefixation was performed at 4 °C for 2 h. The pellets were washed four times with 0.1 M phosphate buffer, and then postfixed for 24 h at 4 °C with 1.5% OsO4 (Sigma-Aldrich) in 0.15 M phosphate buffer, pH 7.4. The cells were embedded in EMBed-812 (Electron Microscopy Sciences, Hatfield, USA), after a previous dehydration in an acetone series. Polymerization of the resin was performed at 60 °C for 72 h. Ultrathin sections were cut with a DiATOME diamond knife (DiATOME, USA) on a Bromma 8800 ULTRATOME III ultramicrotome (LKB, Sweden). They were collected on 300 mesh copper grids (Agar Scientific Ltd., Stansted, UK) and double contrasted with saturated alcoholic uranyl acetate (Merck, Darmstadt, Germany) for 12 min, and 2.8% lead citrate (Fluka AG, Buchs, Switzerland). The sections were examined on a JEOL JEM 1010 transmission electron microscope (JEOL Ltd., Japan) at 80 kV, and images were captured using a Mega VIEW III camera (Olympus, Soft Imaging System, Germany).
3
Endothelial Cell Adhesion to Decellularized Lamina
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For electron microscopy observation, eight endothelial cell seeded laminas were embedded in a fixative consisting of 2% paraformaldehyde and 2% glutaraldehyde (Wako Pure Chemicals) in 0.1 M cacodylate buffer at pH 7.4 for 1 h at 4°C. Next, they were washed, treated with 1% osmium tetroxide in 0.1 M cacodylate buffer and embedded in epoxy resin for standard transmission electron microscopy. Ultrathin sections were stained with uranyl-lead and examined in a JEOL JEM 1010 transmission electron microscope (JEOL USA, Inc., MA, USA) to assess adhesion of endothelial cells to decellularized lamina and possible morphological or differentiation changes in the endothelial cells.
4
Immunogold Labeling of Mitochondrial Proteins
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Isolated mitochondria from the cultured cells were fixed in 2% paraformaldehyde (#15700, EMS) and 0.25% glutaraldehyde (#16000, EMS) for 1 h at 4 °C, and centrifuged at 12,000 g for 10 min at 4 °C. The pellet was resuspended in 10 μl of 2% liquid agarose in PBS. After solidification, the specimens were fixed again in 2% paraformaldehyde and 0.25% glutaraldehyde for 30 min at 4 °C. The fixed specimens in agarose were dehydrated through an increased ethanol series, and infiltrated in a series of increasing LR White resin (#14381, EMS) and decreasing ethanol at 4 °C. The samples were placed in gelatin capsule and polymerized at 60 °C for 24 h. Sections were cut on a ultramicrotome (Leica, Buffalo Grove, IL, USA) and placed on 300 mesh nickel grids (#EMS300-Ni, EMS). The sections were labelled with a combination of anti-SLC4A11 and anti-TOM20 for overnight at 4 °C and incubated with two different secondary antibodies conjugated to different sizes of colloidal gold particles (10 nm; goat-anti-rabbit against anti-SLC4A11 (#25109, EMS), and 25 nm goat-anti-mouse against anti-TOM20 (#25136, EMS)). Thereafter, the sections were washed on large droplets of 0.1% Tween 20 in PBS. The rinsed sections were stained with 2% uranyl acetate for 5 min in the dark, rinsed in H2O, and then observed under a JEOL JEM 1010 transmission electron microscope (JEOL, Peabody, MA, USA).
5
Transmission Electron Microscopy of Exosomes
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Exosomes to be examined by transmission electron microscopy (TEM) were isolated and loaded on to a carbon‐coated electron microscopy grid. The samples were fixed with 2% glutaraldehyde and 2% paraformaldehyde in 0.1 mol/L sodium cacodylate buffer at pH 7.3 for 3 hours at room temperature. After air drying, samples were mounted on specimen stubs and visualized using a JEOL JEM‐1010 transmission electron microscope (JEOL Ltd., Tokyo, Japan).
6
TEM Analysis of S. aureus Inhibition
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S. aureus ATCC 6538 cells were treated with an NS oil sample at Minimum Inhibitory Concentrations (MIC), this bacterial pathogen was identified due to its apparent and clear diameter of the zone of inhibition detected when treated with an NS oil sample among other pathogens tested, selected for a TEM. For the TEM preparation, from 24-h-old cultures grown on MHB medium, bacteria were extracted by centrifugation (at 4000 rpm for 10 min); the samples were then cleaned with distilled water, fixed in 3% glutaraldehyde, rinsed in phosphate buffer, and post-fixed in potassium permanganate solution for five minutes. at ambient temperature. The samples were dehydrated for 15 min in each ethanol dilution, ranging from 10 to 90%, and then for 30 min in absolute ethanol. Over a graded sequence, samples were infiltrated with epoxy resin and acetone until they were finally laid in plain resin. Using copper grids, extremely thin pieces were gathered. After that, sections were thrice stained with lead citrate and uranyl acetate. Stained sections were examined via JEOL-JEM 1010 transmission electron microscope at 70 kV at Alexandria University [39 –41 (link)].
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S. aureus ATCC 6538 cells were treated with an NS oil sample at Minimum Inhibitory Concentrations (MIC), this bacterial pathogen was identified due to its apparent and clear diameter of the zone of inhibition detected when treated with an NS oil sample among other pathogens tested, selected for a TEM. For the TEM preparation, from 24-h-old cultures grown on MHB medium, bacteria were extracted by centrifugation (at 4000 rpm for 10 min); the samples were then cleaned with distilled water, fixed in 3% glutaraldehyde, rinsed in phosphate buffer, and post-fixed in potassium permanganate solution for five minutes. at ambient temperature. The samples were dehydrated for 15 min in each ethanol dilution, ranging from 10 to 90%, and then for 30 min in absolute ethanol. Over a graded sequence, samples were infiltrated with epoxy resin and acetone until they were finally laid in plain resin. Using copper grids, extremely thin pieces were gathered. After that, sections were thrice stained with lead citrate and uranyl acetate. Stained sections were examined via JEOL-JEM 1010 transmission electron microscope at 70 kV at Alexandria University [39 –41 (link)].
7
Transmission Electron Microscopy of Magnetite Nanoparticle-Treated Cells
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Immediately after the incubation with the Fe3O4 MNPs having a concentration of 0.2 mg/mL (after 4 h and 24 h respectively) the cells were processed for TEM. They were prefixed directly in the culture flask with 2.7% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in 0.1 M phosphate buffer (pH 7.4) at 4 °C for 1.5 h. After a centrifugation at 1500 RPM for 10 min the cells were washed four times with 0.1 M phosphate buffer (pH 7.4), and then post-fixed for 1.5 h with 1.5% osmium tetroxide (Sigma-Aldrich) at 4 °C. They were next dehydrated in acetone series (30% to 100%), infiltrated and embedded in Epon 812 resin (Fluka, Buchs, Switzerland). The blocks polymerized for 72 h at 60 °C were trimmed and cut with glass knives on a Bromma 8800 ULTRATOME III (LKB, Stockholm, Sweden). The ultrathin sections (60–80 nm) were collected on 3 mm copper grids (with formvar film), and contrasted for 7 min with uranyl acetate (Merck, Billerica, MA, USA). The samples were examined on the JEOL-JEM 1010 transmission electron microscope (Jeol Ltd., Tokyo, Japan), equipped with a Mega VIEW III camera (Olympus, Soft Imaging System, Münster, Germany) and operating at 80 kV.
8
Ultrastructural Analysis of Cochlea
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Samples were prepared for TEM using procedures modified from previous publications (Lang et al., 2015) (link).
Briefly, deeply anesthetized mice were cardiac perfused with a mixture of 10 mL saline and 0.1% sodium nitrite solution, followed by 15 mL of a fixative solution containing 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4). The same fixative solution was used to perfuse the excised cochleae through the round window and for further immersion overnight at 4C. Cochleae were then decalcified using 0.12M EDTA solution at room temperature for 2-3 days with a magnetic stirrer. Then, cochleae were fixed using a solution containing 1% osmium tetroxide and 1.5% ferrocyanide for 2 hours in the dark. They were then dehydrated and embedded in Epon LX 112 resin. Semi-thin sections for pre-TEM observation of AN orientation were cut at 1-m thickness and stained with toluidine blue. Once a coronal plane for a given cochlear turn was seen, ultra-thin sections at 70-nm thickness were cut and stained with uranyl acetate and lead citrate. These ultra-thin sections were examined using a JEOL JEM-1010 transmission electron microscope (JEOL USA, Inc., Peabody, MA).
Briefly, deeply anesthetized mice were cardiac perfused with a mixture of 10 mL saline and 0.1% sodium nitrite solution, followed by 15 mL of a fixative solution containing 4% paraformaldehyde and 2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4). The same fixative solution was used to perfuse the excised cochleae through the round window and for further immersion overnight at 4C. Cochleae were then decalcified using 0.12M EDTA solution at room temperature for 2-3 days with a magnetic stirrer. Then, cochleae were fixed using a solution containing 1% osmium tetroxide and 1.5% ferrocyanide for 2 hours in the dark. They were then dehydrated and embedded in Epon LX 112 resin. Semi-thin sections for pre-TEM observation of AN orientation were cut at 1-m thickness and stained with toluidine blue. Once a coronal plane for a given cochlear turn was seen, ultra-thin sections at 70-nm thickness were cut and stained with uranyl acetate and lead citrate. These ultra-thin sections were examined using a JEOL JEM-1010 transmission electron microscope (JEOL USA, Inc., Peabody, MA).
9
Ultrastructural Analysis of Cultured Epidermal Tissue
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Cultured epidermal tissue on adult ostracod carapaces at three culture time points (0, 1 and 2 wk, n = 3 at each time point) were removed from culture and incubated at 4°C overnight in Karnovsky fixative (2% paraformaldehyde, 2.5% glutaraldehyde, in 0.1 M cacodylate buffer). Fixed carapace-epidermal valves were transferred to 1.5% potassium ferricyanide/1% osmium tetroxide in 0.1 M cacodylate buffer for 1 h and washed in distilled water, before 1-h immersions in 1% aqueous osmium tetroxide and 1% aqueous uranyl acetate, with thorough washing in distilled water after each. After an additional 1-h incubation in Walton’s lead aspartate (Walton 1979) at 60°C with further washes, samples were dehydrated through serial ethanols to 100%, immersed in propylene oxide, before Durcapan resin infiltration and embedding over 2 d. The resin was hardened at 60°C for 48 h and ultrathin sections (∼100 nm thick) were cut and viewed on G300 copper grids in a JEM 1010 transmission electron microscope (Jeol, Welwyn Garden City, UK).
10
Transmission Electron Microscopy of Biological Samples
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2.5 µl of dialysed sample was applied to freshly glow-discharged, carbon-coated 400 mesh copper grids (Veco, Nisshin, Tokyo, Japan), blotted and stained with 2% uranyl acetate solution. Grids were screened using a JEM1010 transmission electron microscope (JEOL, Tokyo, Japan) at 100 kV and images were acquired using a 2k × 2k slow-scan CCD camera (Gatan, Pleasanton, CA). Images were recorded at a magnification of 40,000×, which corresponds to a pixel size of 6 Å using a 2 s exposure.
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