SEM was used to observe biofilm formation and effects of antimicrobial drugs on biofilm formation12 (link). In order to observe the morphological changes associated with biofilms following moxifloxacin treatment, biofilms were allowed to grow on sterile flat-bottom 6-well polystyrene tissue culture plates (LabServ, Thermo Fisher Scientific, USA) in the presence or absence of moxifloxacin treatment. Fresh Cation-Adjusted Mueller Hinton Broth (CAMHB) without any antibiotic was used as a control. The samples were fixed with 2.5% glutaraldehyde for at least 4 h at 4 °C. After washing with phosphate buffered saline (PBS), the samples were dehydrated in a series of ethanol solutions of increasing concentration (50 to 100%). The cells were subsequently washed with pure acetone, followed by isoamyl acetate. They were then dried with a critical point dryer (Hitachi, Japan). The dried samples were coated with 15-nm gold using an argon automatic sputter coater (Hitachi, Japan). After processing, the samples were viewed with a Philips XL30CP scanning electron microscope.
Xl30cp scanning electron microscope
Manufactured by Philips
The XL30CP scanning electron microscope is a versatile laboratory instrument designed for high-resolution imaging and analysis of microscopic samples. It utilizes a focused beam of electrons to scan the surface of a specimen, generating detailed information about its topography and composition.
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2 protocols using xl30cp scanning electron microscope
1
Observing Antimicrobial Effects on Biofilms
2
Biofilm Imaging in Porous Media
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Confocal laser scanning microscopy (CLSM) was used to observe the extent of biofilm growth on porous media after termination of experiments. Biofilm samples grown on sand were washed with phosphate buffered saline (PBS) and stained with 4',6-diamidino-2-phenylindole (DAPI) stain at a concentration of 300 µg/ml, made from 10 mg/ml stock solution (Biotium), using PBS as diluent. Samples were fixed onto glass slides using SlowFade Gold Antifade Reagent (Invitrogen), and air-dried. Glass beads were too big to mount on glass slides, therefore 0.5 g of beads were vortexed with PBS and stained with DAPI stain, and a drop was placed on a glass slide and left to air-dry. Once dried, the samples were covered with glass cover slips and fixed with nail varnish. Samples were viewed using a Leica SP5 Confocal Laser Microscope. Stacks of images were processed to 3D images using ImageJ software. Scanning electron microscopy (SEM) images were taken of porous media from the top and bottom sections of each column. Porous media was transferred with a spatula onto 1 cm diameter SEM stubs, and gold sputtered using a BAL-TEC SCD 050 Sputter Coater. Samples were viewed with a Philips XL30CP scanning electron microscope in secondary electron imaging mode.
Surfaces of porous media were visualized using a Nikon SMZ800 stereo microscope with attached Nikon Coolpix 995 digital camera.
Surfaces of porous media were visualized using a Nikon SMZ800 stereo microscope with attached Nikon Coolpix 995 digital camera.
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