Oct 11, 2022
Sample preconditioning before scanning electron microscopy (SEM)
- 1XJTLU
Protocol Citation: An.Huang 2022. Sample preconditioning before scanning electron microscopy (SEM). protocols.io https://dx.doi.org/10.17504/protocols.io.36wgqj465vk5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 10, 2022
Last Modified: October 11, 2022
Protocol Integer ID: 71066
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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.
Bacterial samples preconditioning
Bacterial samples preconditioning
10m
10m
Obtain a small quantity of bacteria to be tested from a culture.
Centrifuge the samples at5000 rpm, 4°C, 00:10:00 . 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 .
10m
SEM sample preconditioning
SEM sample preconditioning
2h
2h
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 01:00:00 each concentration.
1h
Dry the samples at room temperature for 01:00:00 .
1h
Use a precision etching coating system, sputtering the samples with Au-Pd alloy.
Note
If samples contain gold, sputter the samples with chromium.
The samples are ready to be visualized using SEM