SEM-EDS Bacteria image library

Description

In-Vitro Pb Uptake by Bacteria in Liquid Cultures-Scanning Electron Microscopy (SEM-EDX) Analysis. B. paramycoides, B. proteolityticus and B. wiedmannii were able to immobilise Pb on their cell wall.

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Batch studies were conducted using 50 mL Falcon tubes that contained 25 mL GY rich medium supplemented with 100 mg L-1 of Pb (Pb(NO3)2). All tubes were inoculated with 1 mL of a bacterial cell suspension (optical density at 600 nm=1), which was previously grown in GY medium at 28°C for 24 h. Samples were incubated at 28°C and 100 rpm on an orbital shaker. For the visualization of bacterial structure, bacterial pellets were washed 3 times with 0.01 M phosphate-buffered saline buffer (PBS, pH 7.0) to remove unbound metals, sugars and proteins. Pellets were resuspended in 2% glutaraldehyde for 1h at room temperature. Afterwards, bacterial samples were centrifuged for 3 min at 3000 rpm and pellets were washed 3 times with milli-Q water. One uL of sample was placed on a sample holder, in carbon conductive tape. Then, samples were coated 30 seg with a 15 nm gold layer and analysed using a Scanning Electron Microscope (Phenom™ ProX Desktop, Thermofisher) with an integrated energy-dispersive X-ray diffraction (EDS) detector. Images were taken using an accelerating voltage of 15 kV.

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