Samples of Decaisnea insignis fruits at different developmental stages were prepared by pre-fixing them with 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.0) for 48 h, and post-fixing in 1% osmium tetroxide overnight at 4 °C. After dehydration using a graded ethanolic series, the samples were treated with tertiary butyl alcohol two times, and then placed under −20 ℃ overnight and vacuumed for 24 h in succession. The dried samples were then mounted on stubs, coated with gold for 1 min using a Hitachi E-102 Ion sputter (Hitachi High-Technologies Corporation, Tokyo, Japan), and observed using a Hitachi TM-1000 tabletop scanning electron microscope (Hitachi, Tokyo, Japan).
E102 ion sputter
Manufactured by Hitachi
Sourced in Japan
The E102 Ion Sputter is a laboratory equipment used for sputter coating. It is designed to deposit thin, uniform metal or carbon coatings on samples for scanning electron microscopy (SEM) and similar applications.
Automatically generated - may contain errors
Lab products found in correlation
Tm 1000 tabletop scanning electron microscope, by Hitachi (2 mentions)
S 4500, by Hitachi (1 mentions)
Su8230, by Hitachi (1 mentions)
Jsm 7900f, by JEOL (1 mentions)
S 2500, by Hitachi (1 mentions)
S 3400 sem, by Hitachi (1 mentions)
Smile view software, by JEOL (1 mentions)
Crossbeam 550, by Zeiss (1 mentions)
Em cpd030, by Leica camera (1 mentions)
Jem 1400 electron microscope, by JEOL (1 mentions)
7 protocols using e102 ion sputter
1
Scanning Electron Microscopy of Decaisnea insignis
2
Scanning Electron Microscopy of Decaisnea insignis
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Samples of Decaisnea insignis fruits at different developmental stages were prepared by pre-fixing them with 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.0) for 48 h, and post-fixing in 1% osmium tetroxide overnight at 4 °C. After dehydration using a graded ethanolic series, the samples were treated with tertiary butyl alcohol two times, and then placed under −20 ℃ overnight and vacuumed for 24 h in succession. The dried samples were then mounted on stubs, coated with gold for 1 min using a Hitachi E-102 Ion sputter (Hitachi High-Technologies Corporation, Tokyo, Japan), and observed using a Hitachi TM-1000 tabletop scanning electron microscope (Hitachi, Tokyo, Japan).
3
Microstructural Analysis of Experimental Specimens
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The top surfaces of the specimens from each experimental group were investigated for microstructural analysis using scanning electron microscopy (SEM: S-4500 Hitachi, Tokyo, Japan). The specimens were coated with a thin layer of Pt (E102 Ion Sputter, Hitachi, Tokyo, Japan). The surfaces were investigated at an accelerating voltage of 5 kV, emission current of 8 μA and working distance of 10 mm. In addition, one of the fractured specimens from each experimental group was cross-sectioned, polished and argon-ion milled (Cross Section Polisher, SM-09010; JEOL, Tokyo, Japan). A thin layer of Pt was coated on the samples prior to examination in backscattered electron imaging mode using a field-emission-gun SEM (FE-SEM; Hitachi SU8230, Hitachi) operated at 15 kV. The elemental distributions for the samples were determined using energy dispersive X-ray spectroscopy (EDS).
4
Scanning Electron Microscopy of COV434 Cells
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The morphological characteristics of the COV434 cells were determined by scanning electron microscopy (SEM; Technex, Tokyo, Japan), where the cells were incubated according to the methods described above. Next, the cells were fixed with 2% glutaraldehyde in 0.2 M cacodylate buffer (pH 7.4) for 2 hr at 4°C. The cells were washed with PBS and dehydrated using a graded series of ethanol (50, 70, 90, 95 and 100%), 50% tert-butyl alcohol (in ethanol) and 100% tert-butyl alcohol. Then, the dehydrated cells were lyophilized. Samples were coated with palladium and platinum using an E102 Ion sputter (Hitachi, Tokyo, Japan), followed by observation using SEM.
5
Zirconia Filler Characterization and Fracture Analysis
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The particle size and distribution of the zirconia filler and the fracture surface of the specimens after the three-point bending tests were analyzed by scanning electron microscopy (SEM; JSM-7900F, JEOL, Tokyo, Japan). The specimens were attached to an aluminum stub and their surfaces were coated with a thin layer of platinum (E102 Ion Sputter, Hitachi, Tokyo, Japan). The samples were then analyzed at an acceleration voltage of 5 k, a discharge current of 8 μA, and a working distance of 6–10 mm. Microscopic images were acquired at a magnification of 40,000× (10 wt.%) for particle size, 3000× (0 (control), 3, 5, and 10 wt.%) for filler distribution, and 150× (5 and 10 wt.%) for the fracture surface of the specimens after the three-point bending tests. The average particle size was determined based on the SEM images using Fiji-ImageJ software (National Institutes of Health, Bethesda, MD, USA) with the linear intercept method. Particle size distribution was measured for a minimum of 600 particles.
6
Leaf Epidermal Characteristics Analysis
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Mature leaves were selected and peeled by a razor blade. The epidermis was stained with Safranin-O for 10‒15 min, and then dehydrated in an ethyl alcohol series, finally mounted with DePeX for a permanent slide. The stomatal index was calculated according to the method described by Salisbury (1927 (link)).
Leaf surfaces were also studied intensively by the scanning electron microscope (SEM). Small pieces of leaf (5 × 5 mm2) were dehydrated in ethanol series and sonicated to remove unwanted parts from the surfaces. After that, dried samples were coated with platinum-palladium in a sputter coater (Hitachi E-102 Ion Sputter). The cuticular patterns were observed and imaged under a Hitachi S-2500 scanning electron microscope.
Leaf surfaces were also studied intensively by the scanning electron microscope (SEM). Small pieces of leaf (5 × 5 mm2) were dehydrated in ethanol series and sonicated to remove unwanted parts from the surfaces. After that, dried samples were coated with platinum-palladium in a sputter coater (Hitachi E-102 Ion Sputter). The cuticular patterns were observed and imaged under a Hitachi S-2500 scanning electron microscope.
7
Ultrastructural Analysis of Fungal Hyphae
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For transmission electron microscopy analysis, hyphae were cultured on PDA medium for 18 hr and prefixed with 2% (w/v) glutaraldehyde containing 0.1 M phosphate buffer (pH 7) at 4 °C for 10 hr, followed by staining with 1% osmium tetroxide in phosphate buffer at room temperature for an additional hour. After dehydration, tissues were embedded in epoxy resin. Ultrathin sections were stained with uranyl acetate and lead citrate and visualised under a JEM 1400 electron microscope (JEOL, Japan). The cell wall thickness was measured using SMile View software (JEOL, Japan).
For SEM analysis, hyphae were cultured on PDA medium at 37 °C for 48 hr, and agar blocks containing fungal cells were fixed as described for the transmission electron microscopy analysis. The blocks were then dehydrated by passage through a graded series of ethanol solutions, replaced with isoamyl acetate, and dried using the critical‐point method (EM CPD030; Leica, Germany). After sputter coating with platinum‐palladium (using an E102 Ion sputter; Hitachi, Japan), samples were visualised using an S‐3400 SEM (Hitachi, Japan). The proportion of spike area to the total surface area of each conidium was measured using Image J. For field emission‐scanning electron microscopy analysis, dehydrated samples were visualised on poly‐L‐lysine coated silicon wafers under CrossBeam 550 (Zeiss).
For SEM analysis, hyphae were cultured on PDA medium at 37 °C for 48 hr, and agar blocks containing fungal cells were fixed as described for the transmission electron microscopy analysis. The blocks were then dehydrated by passage through a graded series of ethanol solutions, replaced with isoamyl acetate, and dried using the critical‐point method (EM CPD030; Leica, Germany). After sputter coating with platinum‐palladium (using an E102 Ion sputter; Hitachi, Japan), samples were visualised using an S‐3400 SEM (Hitachi, Japan). The proportion of spike area to the total surface area of each conidium was measured using Image J. For field emission‐scanning electron microscopy analysis, dehydrated samples were visualised on poly‐L‐lysine coated silicon wafers under CrossBeam 550 (Zeiss).
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