Specimens for LM and TEM analysis were obtained from the central part of healthy green leaf blades (second or third leaf) and were fixed in 3.5% glutaraldehyde solution in phosphate buffer with the pH of 7.0 for 12 h at room temperature, followed by secondary fixation in 2.5% osmium tetroxide solution. The specimens were rinsed and dehydrated in a graded series of alcohols and acetone, and were embedded in Poly Bed 812 epoxide resin (Polyscience). Microtome sections were prepared in the Leica Ultracut R microtome using Diatome diamond knives. Semi-thin sections, 1.5 μm thick, were placed on slides and stained with toluidine blue and azure B. The specimens were mounted with a drop of glycerin. They were analyzed under the Nikon Eclipse 80i light microscope with compatible hardware and software (NIS ELEMENTS) for digital image recording. Ultra-thin sections, 60-90 nm thick, were mounted on nickel grids with 300 mesh squares. Immediately before examination, saturated aqueous uranyl acetate solution and lead citrate were added to impart contrast to the specimens. The specimens were examined and electronograms were obtained simultaneously under two transmission electron microscopes-JEOL JEM 100S and JEOL 1400. JEOL JEM 100S supports analog image recording, whereas JEOL 1400 is equipped with hardware and iTEM software for recording data files.
Jem 100s
Manufactured by JEOL
Sourced in Japan
The JEM-100S is a transmission electron microscope (TEM) manufactured by JEOL. It is designed to provide high-resolution imaging of samples at the nanoscale level. The JEM-100S is capable of magnifying specimens up to 600,000 times, allowing for detailed observation and analysis of the internal structure and composition of materials.
Automatically generated - may contain errors
Lab products found in correlation
Zetaplus, by Brookhaven Instruments (3 mentions)
Araldite, by Electron Microscopy Sciences (2 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (2 mentions)
Uv 3600, by Shimadzu (1 mentions)
Icp ms, by Agilent Technologies (1 mentions)
So 163, by Kodak (1 mentions)
Nicolet is10 ft ir spectrometer, by Thermo Fisher Scientific (1 mentions)
Brookhaven bi 900at, by Brookhaven Instruments (1 mentions)
Xl30 microscope, by Philips (1 mentions)
Jem 1400plus, by JEOL (1 mentions)
Ultracut s ultramicrotome, by Leica (1 mentions)
D 19 developer, by Kodak (1 mentions)
Zetasizer nano, by Malvern Panalytical (1 mentions)
Optima camera, by Biospace (1 mentions)
Affinity 1, by Shimadzu (1 mentions)
Tga dsc1, by Mettler Toledo (1 mentions)
Nicolet nexus 470 esp, by Thermo Fisher Scientific (1 mentions)
Tecnai tf20 tem, by Thermo Fisher Scientific (1 mentions)
Bx53 polarizing microscope, by Olympus (1 mentions)
Dmx 400 mhz spectrometer, by Bruker (1 mentions)
32 protocols using jem 100s
1
Leaf Ultrastructure Microscopy Protocol
2
Imaging and Characterizing Protein Fibrils
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1 μL of fibril solution was mounted on a carbon-coated palladium grid (400 mesh). The sample was dried at room temperature for 5–10 min, then negatively stained with 2% aqueous uranyl acetate solution (Reachim) and dried with filter paper. The sample grids were analyzed by a TEM (JEOL-JEM-100S, Japan) at an instrumental magnification of 25,000. The dimensions of the fibrils were obtained directly from the micrographs.
3
Morphological Examination of Nanoparticles by TEM
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Morphological examination of the NPs was conducted using a JEM-100S transmission electron microscope (TEM, JEOL Ltd., Akishima-shi, Japan). One drop of NP suspension was placed on a copper grid covered with nitrocellulose membrane and air-dried before observation.
4
Synthesis of LMWC-Coated Silver Nanoparticles
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Four grams of LMWC was mixed with 0.1 g AgNO3 in 50 mL deionized water at 50°C under stirring at 200 rpm overnight in a hood. On the next day, acetone was added slowly with stirring until the solution became cloudy, followed by centrifugation at 2,000 rpm for 30 minutes. The supernatant was transferred to a new tube, dried by vacuum, and dissolved in 2–3 mL deionized water. The concentration of the resulting low molecular weight chitosan-coated silver nanoparticles (LMWC-AgNPs) was determined by UV-Vis (UV3600; Shimadzu Corporation, Kyoto, Japan) and ICP-MS (Agilent Technologies, Richardson, TX, USA). The particle size of LMWC-AgNPs was determined by transmission electron microscopy (TEM) (JEM-100S; JEOL, Tokyo, Japan).
5
Ultrastructural Analysis of Synaptic Profiles
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Ultrastructure was analyzed with two electron microscopes (EM). One EM is a JEOL JEM 100S transmission electron microscope with which areas of interest were photographed at 15,000–40,000 magnification with high-resolution film (Kodak SO-163; Kodak, Rochester, NY, USA). The negatives were scanned at 1200–2000 pixels/inch (ScanMaker 800, Microtek, Santa Fe Springs, CA, USA) to produce digital images. A second EM is a Phillips CM10 equipped with a digital camera for direct capture of digital images. Synapses were identified by the presence of vesicles in the presynaptic profile, a clear synaptic cleft and the presence of a postsynaptic density. Tracer-labeled profiles were readily identified by the presence of DAB precipitate. GABA-positive (GABA+) profiles were easily distinguished from GABA-negative profiles by a distinct difference in the density of overlying gold particles (Nakamoto et al., 2013a (link)). GABA immunostaining was also readily distinguished from the DAB label (Nakamoto et al., 2013b (link)).
6
Characterization of Nanoparticle Properties
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Particle size and zeta potential of these two NPs were measured using a Brookhaven BI-900AT instrument/zeta potential analyzer (Brookhaven Instruments Corporation, Holtsville, NY, USA). Fourier transform infrared (FTIR) spectra were recorded using a Nicolet iS10 FTIR spectrometer (Thermo Scientific, Swedesboro, NJ, USA). Morphology studies of the NPs were carried out using transmission electron microscopy (TEM) (JEM-100s; JEOL, Tokyo, Japan).
7
TEM Imaging of Sal-Doc Nanoparticles
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A drop of Sal-Doc SE-NP and Sal-Doc NR-NP suspension was dripped on a nitrocellulose-covered copper grid respectively and air-dried before observation under JEM-100S transmission electron microscope (TEM) (JEOL Ltd., Akishima-shi, Japan).
8
Microscopic Evaluation of Nanocomposites
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The morphological evaluation of NCs has been investigated by a scanning electron microscope (SEM) (Philips XL 30 microscope, Hillsboro, USA). Raw ETO MC powder and processed NC powder were placed on a double-sided tape, then coated with a 30 nm layer of gold under vacuum (10–6 Pa) for 2 min, then observed using SEM at an accelerating voltage of 15 kV under vacuum. Also, transmission electron microscopy (TEM) was performed to understand the surface morphology and structure of ETO NCs from the dispersion in the aqueous polymeric solution. A drop of the ETO NCs solution was put on a copper grid with Formvar films Cu 200 Mesh. Then, negative staining was performed by adding a drop of uranyl acetate solution (1% w/v). The excess fluid was removed with filter paper. The grids were examined under a transmission electron microscope (JEM-100S, JEOL, Tokyo, Japan) at accelerating voltage of 80 kV.
9
Ultrastructural Analysis of Secretory Granules
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Ultra-thin slices were used for electron microscopy. They were contrasted with uranyl acetate and lead citrate. The study was carried out on the electron microscope JEM-100S (JEOL, Japan). For ultrastructural analysis, whole samples were photographed under a magnification of 14.000. Verification of the secretory granules on the ultrastructural level was made using the uranaffin reaction. For this purpose samples were fixed with 3% glutarate aldehyde on the 0.1M cacodylic buffer (pH = 7.2) for 90 minutes. Subsequently, 1mm cut blocks were rinsed with 0.9% sodium chloride solution 3 times for 15 minutes each. The rinsed blocks were placed into 4% uranyl acetate water solution (pH = 3.9) at 4°C for 18 hours. After rinsing, the blocks were dehydrated in the alcohols and placed in EPON resin.
10
Transmission Electron Microscopy Specimen Preparation
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Pellets were fixed with 2.5% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in 0.1 cacodylate buffer, pH 7.3, for 1 h at room temperature, rinsed twice for 10 min in 0.1 cacodylate buffer and postfixed with 1% osmium tetroxide in the same buffer for 1 h at 4 °C (Hayat, Basic techniques for transmission electron microscopy, Academic Press, Inc., Orlando, FL, USA, 1986). Pellets were dehydrated in ascending alcohols, treated with propylene oxide and embedded in Araldite (Electron Microscopy Sciences, Hatfield, PA, USA). Ultrathin sections of the samples were cut on a Top Ultra 150 ultramicrotome (Pabish, Germany) and collected on 300-mesh copper grids. The grids were stained with uranyl acetate and lead citrate and examined at 80 kV using a Jeol JEM 100S transmission electron microscopy.
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