After disassembly, the microstructure of the fracture surfaces was observed through SEM. Surfaces were coated with gold using a sputter coater (EMS550X, Electron Microscopy Sciences, Hatfield, PA, USA) under a vacuum of 10−1 mbar, at 25 mA for 2 min. A high-performance JSM-6610LV SEM (JEOL Ltd., Tokyo, Japan) was employed to capture images, at an acceleration voltage of 15 kV.
Ems 550x
Manufactured by Electron Microscopy Sciences
Sourced in United States
The EMS 550X is a high-resolution scanning electron microscope (SEM) designed for advanced materials analysis and characterization. It features a stable and reliable electron optical column, delivering high-quality imaging capabilities across a wide range of magnification levels. The core function of the EMS 550X is to provide detailed, high-resolution visualization and analysis of micro- and nano-scale structures and features in various sample types.
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4 protocols using ems 550x
1
Fracture Surface Microstructure Analysis
2
Scanning Electron Microscopy of Starved Cells
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Morphological changes between 6-months starved and non-starved cells were observed under a scanning electronic microscope (SEM) as previously described with minor modification (Arias et al., 2012 (link)). Briefly, 10 μL of bacteria was fixed in 2.5% glutaraldehyde (v/v) on 8 × 8 mm glass slides at 4°C overnight. Then, samples were dehydrated in a graded ethanol series (50, 70, 90, and 100%), coated with gold palladium alloy in an EMS 550X (Electron Microscopy Science), and examined under Zeiss EVO 50 electronic microscope (Zeiss, Germany).
3
Scanning Electron Microscopy of Bacteria
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Exponential phase bacteria (1×109 CFU/mL) were exposed to AgCNTs at approximately 4× MIC for 4 hours at 37°C, before being centrifuged at 2,000× g (Sorvall ST 40R; Thermo Fisher Scientific) for 10 minutes. The pellets were then washed in 0.1% phosphate-buffered saline (PBS) and fixed overnight in a mix of 2.5% glutaraldehye and 1% formaldehyde in 0.1% PBS. Samples were treated with 1% osmium tetroxide in 0.1% PBS, before stepwise dehydration in increasing concentrations of ethanol in water. The dehydrated samples (5 µL each) were then placed on SEM stubs (Electron Microscopy Sciences, Hatfield, PA, USA), and air-dried and sputter-coated with gold in a sputter-coat device (EMS 550X; Electron Microscopy Sciences) prior to performing SEM (EVO 50 variable pressure scanning; Carl Zeiss Meditec AG, Jena, Germany) analysis.
4
Scanning Electron Microscopy Preparation Protocol
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Clusters were fixed in 2% PFA and 2% glutaraldehyde in 0.1 M PBS on a 13 mm diameter 2 μm pore polycarbonate filter. They were then rinsed with 0.1 M PBS and distilled water. Samples were sequentially dried with hexamethyldisilazane (HMDS, Electron Microscopy Sciences), 1:1 100% ethanol and HDMS, and 100% HDMS. After HDMS evaporation, samples were coated with platinum in an EMS 550X sputter coater (Electron Microscopy Sciences) and imaged with a scanning electron microscope camera (JSM-6610, JEOL Ltd.) under high vacuum mode.
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