The enamel blocks were ultrasonically washed in distilled water three times, dehydrated in a series of ethanol solutions, dried in a desiccator and finally sputter-coated with carbon. The surface morphology of the enamel blocks was then examined under a scanning electron microscopy (SEM) (Hitachi S-4800 FEG Scanning Electron Microscope, Hitachi Ltd., Tokyo, Japan) at 5 kV in high-vacuum mode. An elemental analysis was then carried out to study the fluoride (F) ions on the enamel lesion surface by an energy-dispersive X-ray spectroscopy (EDX) under SEM. The elemental analysis was performed by measuring three areas (5 × 5 μm2) on the surface of each enamel block, and the mean weight percentages of F were calculated.
S 4800 feg scanning electron microscope
Manufactured by Hitachi
Sourced in Japan
The S-4800 FEG Scanning Electron Microscope is a high-resolution imaging instrument produced by Hitachi. It utilizes a field emission gun source to generate a finely focused electron beam for scanning samples. The microscope is designed to capture detailed surface topography and compositional information of specimens at the nanometer scale.
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14 protocols using s 4800 feg scanning electron microscope
1
Enamel Surface Morphology and Fluoride Analysis
2
Scanning Electron Microscopy of Biofilm
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Scanning electron microscopy (SEM) was used to monitor the topographical features of the biofilm. In preparation for SEM, the dentine blocks with biofilm were rinsed in 4% formaldehyde followed by 1% PBS; they were then placed in 1% osmium tetroxide solution for 60 min. The blocks were washed in distilled water and dehydrated in a series of ethanol solutions at increasing concentrations (70% for 10 min, 95% for 10 min, and 100% for 20 min). The blocks were dried in a desiccator and sputter-coated with gold. The surface topographies of biofilms were studied under SEM (Hitachi S-4800 FEG Scanning Electron Microscope, Hitachi Ltd., Tokyo, Japan) at 12 kV in high-vacuum mode [25 (link)]. Two dentine blocks per group were used in this qualitative experiment.
3
Ultrastructural Analysis of Samples
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Two blocks from each group were fixed in 2.5% glutaraldehyde and were dehydrated using ethanol solutions (70% for 10 min, 85% for 10 min, 95% for 10 min, and 100% for 20 min). They were critical-point dried and sputter-coated with gold to study the surface morphology under SEM (Hitachi S-4800 FEG Scanning Electron Microscope; Hitachi Ltd., Tokyo, Japan).
4
Dentine Surface Morphology and Topography
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Four dentine blocks (two for the examination of the surface morphology and two for the examination of the cross-section topography) from each group were fixed in a 2.5% glutaraldehyde solution at 4 °C for 8 h. They were then rinsed with deionized water ultrasonically and dehydrated with ethanol. The blocks were placed in a desiccator for critical-point drying and were sputter coated with carbon prior to observation with scanning electron microscopy (SEM; Hitachi S-4800 FEG Scanning Electron Microscope, Hitachi Ltd., Tokyo, Japan) at 5 kV in high-vacuum mode.17 (link)
5
Scanning Electron Microscopy of Dehydrated Specimens
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Four specimens from each group were fixed in 2.5% glutaraldehyde overnight. They were ultrasonically cleaned three times with distilled water and then dehydrated using ethanol solutions with different concentrations (75%, 80%, 95%, and 100%). The dehydrated specimens were further dried in a critical-point dryer (Leica EM CPD300, Wetzlar, Germany) and sputter-coated with carbon. Afterwards, the morphology of the specimen surface was observed under a scanning electron microscope (SEM, Hitachi S-4800FEG scanning electron microscope, Hitachi Ltd., Tokyo, Japan).
6
SEM Analysis of S. mutans Biofilm
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SEM (Hitachi S-4800 FEG Scanning Electron Microscope; Hitachi, Tokyo, Japan) was employed to evaluate the morphology of the S. mutans biofilm (n = 2 for each group). Each dentin block was fixed in a 2.5% glutaraldehyde solution for 4 h at 4 °C. Following dehydration, all the blocks were critical point dried in a desiccator and coated using a sputter coater [14 (link)].
7
Scanning Electron Microscopy of Scaffolds
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After mechanical characterization, the bisected scaffold samples were fixed with glutaraldehyde (G6257, Sigma) at 2.5% (v/v) in PBS at 4°C overnight. After washing with PBS, the samples were dehydrated using an increasing concentration gradient of ethanol and then subjected to critical point drying. Dehydrated samples were fractured, mounted and sputtered coated with gold, and then observed with a Hitachi S4800 FEG scanning electron microscope.
8
Scanning Electron Microscopy Analysis of Dentin Surface
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SEM (Hitachi S-4800 FEG Scanning Electron Microscope; Hitachi, Tokyo, Japan) was employed to evaluate the dentin surface morphology (n = 2 for each group) and cross-sectional topography (n = 2 for each group). Four blocks from each group were ultrasonically cleaned in distilled water to remove the biofilm prior to its fixation in 2.5% glutaraldehyde at 4 °C for 4 h. Of the four blocks, two were fractured in half in liquid nitrogen for an observation of their cross-sectional topography. A series of ethanol solutions was used to dehydrate the blocks. Subsequently, the blocks were dried in a desiccator and coated with carbon using a sputter coater [14 (link)].
9
SEM Imaging of NET Formation
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To observe the formation of NET, control and infected Nϕs were xed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate-HCL buffer pH 7.4 at 1, 6 and 21hpi. Fixed cells on coverslip were washed in cacodylate buffer with 0.1M sucrose to remove excess xative and post-x in osmium tetroxide for 1h at room temperature. The cells were then dehydrated using increased concentration of ethanol each for 15 mins from 30%, 50%, 70%, 90%, to 100%. The cells were dried in a critical point dryer using liquid carbon dioxide as transitional uid and the coverslips were then mount on the specimen holders. A thin layer (100-200Å) of metallic lm was coated on the specimen surface using a vacuum evaporator. The samples were kept in a desiccator until viewing. Hitachi S-4800 FEG scanning electron microscope (Electron Microscope Unit, HKU) was used to examine these samples, with the EMAX energy software.
10
Mineral Analysis of Tooth-Restoration Interfaces
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Each sample was sectioned along the long axis of the tooth, midway across the restoration using a water-cooled copper disc [27] . One half of the sample was used for elemental analysis with scanning electron microscopy with energy-dispersive X-ray spectroscopy (SEM/EDX: Hitachi S-4800 FEG Scanning Electron Microscope, Hitachi Ltd., Tokyo, Japan), and the other half was used for Fourier transform infrared (FTIR) spectroscopy (UMA 500, Bio-Rad Laboratories, Hercules, CA, USA). The objective of SEM/EDX analysis was to evaluate changes in mineral content along the material-root junction. The cross-section surfaces of the restored teeth were treated with 1% acetic acid for 5 seconds and washed ultrasonically using deionized water to remove the smear layer [28] . The prepared teeth were examined under a scanning electron microscope under operating conditions of 5 kV. Lines with 500 µm length [28] and 50 µm away from the root surfaces with analysis commencing at the restoration-root junction. The lines were analyzed by means of line-scan in terms of phosphorus, calcium, fluoride and silver ion levels.
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