Nanoparticle size was determined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) imaging. SEM images of Nano-ZnO and M-Nano-ZnO powder samples were collected using a JSM-7100F Field Emission Scanning Electron Microscope (Jeol, Tokyo, Japan) operating at an accelerating voltage of 5 kV, with low probe current (1 or 2) and a working distance of 4 or 6 mm as required. TEM images were collected using a JEM 2100 Plus with LaB6 (Jeol, Tokyo, Japan). Samples were then prepared for TEM analysis as follows: ground nanoparticles (1 mg) were added to deionised water (10 mL). The sample was sonicated for 30 min with occasional shaking. Parafilm was placed on filter paper in a petri-dish. A copper TEM grid was placed on top of the parafilm and a 4 µL drop of the relevant sample was placed onto the TEM grid. The lid of the petri-dish was replaced, and the sample was left for 24 h to allow the deionised water to evaporate prior to analysis.
Jsm 7100f field emission scanning electron microscope
Manufactured by JEOL
Sourced in Japan
The JSM-7100F is a field emission scanning electron microscope (FE-SEM) manufactured by JEOL. It is designed to provide high-resolution imaging and analysis of a wide range of materials and samples. The FE-SEM uses a field emission gun to produce a high-brightness electron beam, which allows for high-resolution imaging at low accelerating voltages. The instrument is equipped with various detectors, such as secondary electron and backscattered electron detectors, to capture different types of information about the sample surface.
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6 protocols using jsm 7100f field emission scanning electron microscope
1
Nanoparticle Characterization using SEM and TEM
2
Scanning Electron Microscopy of Fungal Co-Cultures
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The co-cultures were prepared as above and an additional time point of 8 h was included to assess the interactions during germination of fungal conidia. At the end of each incubation period, the co-cultures were washed with PBS and fixed in 3% (v/v) glutaraldehyde (ProSciTech, Australia) in PBS. The samples were dehydrated using a series of ethanol concentrations (30–100%) and dried to the critical point using a K850 critical point dryer (Emitech). Dried specimens were mounted on the specimen stubs and coated with gold particles using K550 gold splutter coater unit (Emitech) and visualized using JSM-7100F Field Emission Scanning Electron Microscope (Jeol) at 10 kV working voltage. At least 10 microscopy fields were studied per each sample, representative images are shown.
3
Characterizing CaCO3 Crystal Structures
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CaCO3 crystal samples were prepared via depositing
the dried powder onto carbon tape upon the aluminum sample stub to
image their internal structure, and the crystals were cracked via mechanical force. Capsule samples were prepared via depositing 30 μL of coated-crystal suspension
onto a circular glass slide, followed by the addition of 30 μL
of 50 mM EDTA pH 7.0 for capsule formation (the same conditions for
typical capsule preparation throughout the study). The resulting capsules
were then washed with water repeatedly and frozen for freeze-drying.
The slides with resulting freeze-dried capsules were sputtered with
10 nm of gold and imaged using the JSM-7100F field-emission scanning
electron microscope (JEOL, USA). Both crystals and capsules were imaged
using a probe current of 1 μA and an accelerating voltage of
2 kV.
the dried powder onto carbon tape upon the aluminum sample stub to
image their internal structure, and the crystals were cracked via mechanical force. Capsule samples were prepared via depositing 30 μL of coated-crystal suspension
onto a circular glass slide, followed by the addition of 30 μL
of 50 mM EDTA pH 7.0 for capsule formation (the same conditions for
typical capsule preparation throughout the study). The resulting capsules
were then washed with water repeatedly and frozen for freeze-drying.
The slides with resulting freeze-dried capsules were sputtered with
10 nm of gold and imaged using the JSM-7100F field-emission scanning
electron microscope (JEOL, USA). Both crystals and capsules were imaged
using a probe current of 1 μA and an accelerating voltage of
2 kV.
4
MDCK Cell Monolayer Preparation for SEM
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MDCK monolayers grown on cover slides covered with a thin layer of collagen were fixed for 1 h with 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.2. Samples were washed in the same buffer and dehydrated in crescent concentrations of ethanol to reach 100%. Monolayers were incubated for 20 min with hexamethyldisilazane (HMDS; 692 Polysciences, Warrington, PA). After a second 10-min incubation in HMDS, the excess of reactive was eliminated and left to air-dry for 30 min. Samples were mounted in specimen metal mounts and gold coated in an ion sputtering device (JFC-1100; JEOL, Peabody, MA). Samples were examined in a JEOL JSM-7100-F field emission scanning electron microscope.
5
Cell Morphology Examination via SEM
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To examine cell morphology isolates were grown in 10 ml of nutrient broth (same as for NA excluding agar) overnight at 40 °C for Bowen and Sydney basin isolates and 30 °C for Surat basin isolates with shaking (140 rpm) or until visible turbidity was observed. 200 µl of broth culture was then spotted onto poly-L-lysine (mol. wt. 150,000–300,000) coated glass coverslips for 5 min. Broth culture was then removed and the coverslips incubated in 3% glutaraldehyde in 0.01 M phosphate buffer (pH 7.4) for 24 hours at room temperature before being washed three times, each for 10 min with 0.01 M phosphate buffer. The coverslips were then incubated for 10 min in each of 20%, 50%, 70%, 80% and 90% ethanol solutions, twice with 100% ethanol before final dehydration with a Leica EM CPD300 critical point dryer. Coverslips were mounted on metal stubs, coated with 20 nm of gold with an Emitech K550 gold sputter coater unit and imaged using a JEOL JSM-7100F field emission scanning electron microscope operating at 5 kV accelerating voltage.
6
Biopolymer Characterization Using SEM
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A pinch of Grewia spp. biopolymer was suspended in 100 µL ethanol and agitated for proper dispersion of the biopolymer. This was closely followed by the addition of 900 µL of MilliQ water followed by vortexing and sonication for 30 s. The sample was then placed on aluminium stubs and allowed to dry in desiccator overnight. SEM Images were collected with a JEOL, JSM-7100F Field Emission Scanning Electron Microscope (Model SM 71031SE2A) Japan.
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