The interaction between the bacterial consortium and the FOC in in vitro biocontrol dishes was investigated by scanning electron microscopy. The part of the mycelium that develops towards the bacterial streak was sampled with the head of 1000 µL sterile pipette tips with the aid of a Greenough stereo microscope, Leica S8 APO with 8:1 apochromatic zoom. The samples were fixed overnight with a 2.5% glutaraldehyde solution in 0.05 M phosphate buffer (pH 7.3), washed with distilled water and dehydrated with a few drops of hexamethyldisilazane (HMDS—Sigma-Aldrich, St. Louis, MO, USA). The dried samples were fixed with carbon tape (Agar Scientific, Stansted, UK) on stubs and coated with chromium for SEM observations (Gemini SEM 500 SEM—Zeiss, Oberkochen, Germany). Acquisitions were performed with an acceleration voltage of 5 kV and type II secondary electrons (SE2 signal).
Gemini sem 500 sem
Manufactured by Zeiss
Sourced in Germany
The Gemini SEM 500 is a scanning electron microscope (SEM) produced by Zeiss. It provides high-resolution imaging capabilities for a wide range of materials and applications. The Gemini SEM 500 utilizes a field emission electron source and advanced optical components to deliver detailed, high-quality images at magnifications up to 1,000,000x.
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Lab products found in correlation
3 protocols using gemini sem 500 sem
1
Scanning Electron Microscopy of Bacterial-Fungal Interactions
2
Actinomycetes Inhibit Fungal Pathogens
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The effects of actinomycetes strains on F. oxysporum f. sp. radicis-lycopersici (FORL) and R. solani (RHS) were investigated by scanning electron microscopy (SEM), using a Gemini SEM 500 SEM (Zeiss, Oberkochen, Germany). Small pieces of agar from the actinomycete/fungus interaction zone and the fungus growth control were taken and mounted on adhesive tape. Freshly collected samples were observed using a specific stage for Peltier technology, with a working temperature of −1 °C. Micrographs acquisition was performed with a working distance of 8.1 mm, an acceleration voltage of 10 kV, and using a backscattered electrons detector (BSD4 signal).
3
Automated STEM Imaging of Nanoparticle Internalization
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Previously observed TEM grids were automatically imaged in the STEM imaging mode of the GeminiSEM 500 SEM (20 kV, 20 µm aperture and high current mode) driven by Atlas 5 software (Carl Zeiss Microscopy). An automated mosaic process of acquisition (stitching of 25 adjacent ROI of 24.6 x 24.6 mm FOV) generated a large high-resolution map (FOV 100 x 100 mm, 1,7
Gpix, 3 nm / pixel). All images were acquired by mixing electrons collected simultaneously in Bright Field and in High Angular Annular Dark Field modes.
Two of these maps were examined for CS and for CF-H4-CS, representing more than 40 cells per condition. The nanoparticles were divided in 5 classes, depending on their location: endosome, multi-vesicular body (MVB), autophagosome, rupture of endosomal membrane, and cytosol. Two manual counting were realized using FIJI ImageJ software 34 (link) and Cell Counter plugin (Total of events: 820 for CS and 834 for CF-H4-CS). Data were analyzed using the χ 2 test of independence in Excel, according to the formula in the equation below (A ij : observed values and E ij theoretical values):
Gpix, 3 nm / pixel). All images were acquired by mixing electrons collected simultaneously in Bright Field and in High Angular Annular Dark Field modes.
Two of these maps were examined for CS and for CF-H4-CS, representing more than 40 cells per condition. The nanoparticles were divided in 5 classes, depending on their location: endosome, multi-vesicular body (MVB), autophagosome, rupture of endosomal membrane, and cytosol. Two manual counting were realized using FIJI ImageJ software 34 (link) and Cell Counter plugin (Total of events: 820 for CS and 834 for CF-H4-CS). Data were analyzed using the χ 2 test of independence in Excel, according to the formula in the equation below (A ij : observed values and E ij theoretical values):
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