The morphology of all five selected Salmonella phages was analyzed using Transmission electron microscopy (TEM.) To prepare the phages for TEM, 1 ml of phage stocks ≥ 9 log10 PFU/ml was centrifuged at 16 000 × g for 1 h at 4°C (Microfuge R centrifuge, Beckman Coulter Inc.) and washed twice with HEPES buffer (0.1 M, pH 7.4). Copper–rhodium (400-mesh) grids were covered with a thin layer of amorphous carbon made hydrophilic by 45 s vacuum glow-discharge. Phage solution samples (4–6 µl) were placed on individual grids and left for 2 min to adsorb onto the carbon. Excess sample was gently removed by touching filter paper to the edge of the droplet after adsorption was stablished. Excess small molecule contaminants were washed from the grid with three rinses of water. Sample on the grids were stained with 2% (w/v) uranyl acetate. A Tecnai G2 transmission electron microscope (FEI Company, Hillsboro, OR, USA) at Electron Microscopy Unit, University of Guelph (Guelph, ON, Canada) was used for visualization, operating at 200 kV under variable magnification (x 50 000). Images were acquired using the Gatan Ultascan 4 K CCD (Pleasanton, CA, USA) and Gatan Digital Micrograph software imaging system.
Digital micrograph software imaging system
Manufactured by Ametek
Sourced in United States
Digital Micrograph is a software imaging system designed for analysis and processing of digital microscopy data. The software provides a comprehensive suite of tools for image acquisition, enhancement, and quantitative analysis, catering to the needs of scientific researchers and imaging professionals.
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Lab products found in correlation
2 protocols using digital micrograph software imaging system
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Salmonella Phage Morphology Analysis
2
Phage Morphology Visualization by TEM
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Transmission electron microscopy was performed on samples to visualize the effect of heat on phage morphology. Phage suspensions were concentrated by density gradient centrifugation (Cardarelli et al., 2010 (link)). The concentrated phages were diluted 1:20 with double–distilled water and directly negative stained for TEM. Copper–rhodium (400-mesh) grids were covered with a thin layer of amorphous carbon made hydrophilic by 45 s vacuum glow-discharge. Phage solution samples (4–6 μL) were placed on individual grids and left for 2 min to adsorb onto the carbon. Excess sample was gently removed by touching filter paper to the edge of the droplet after adsorption was stablished. Excess small molecule contaminants were washed from the grid with three rinses of water. Sample on the grids were stained with 2% (w/v) uranyl acetate (Cardarelli et al., 2010 (link)). A Tecnai G2 transmission electron microscope (FEI Company, Hillsboro, OR, United States) at Electron Microscopy Unit, University of Guelph (Guelph, ON, Canada) was used for visualization, operating at 200 kV under variable magnification. Images were acquired using the Gatan Ultascan 4K CCD (Pleasanton, CA, United States) and Gatan Digital Micrograph software imaging system.
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