Sample preconditioning before scanning electron microscopy (SEM)
An.Huang
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Abstract
Bacterial samples need to be preconditioned before visualized by scanning electron microscope (SEM). This protocol describes preconditioning methods of bacteria before SEM.
Steps
Bacterial samples preconditioning
Obtain a small quantity of bacteria to be tested from a culture.
Centrifuge the samples at5000rpm,4°C . Resuspend the pellet using 1X phosphate buffed saline (PBS). Repeat this step for three times. Keep the supernatant after the third centrifuge and preserve the samples at 4°C .
SEM sample preconditioning
Wash silica wafers with ddH2O and acetone for three times.
Add samples onto the silica wafer using a pipette tip.
Incubate the silica wafers in fixation solution (2.5% glutaraldehyde in 1X PBS) at4°C overnight.
Incubate the samples in ethanol solution with an increasing concentration (40%, 70%, 96%, 100%) for 1h 0m 0s each concentration.
Dry the samples at room temperature for 1h 0m 0s.
Use a precision etching coating system, sputtering the samples with Au-Pd alloy.
The samples are ready to be visualized using SEM
