I wanted to compare the effect of partial dehydration of decellularised scaffolds on structure by imaging the surface with SEM and then measure fiber diameter using Image J,, But my problem is that SEM process involve dehydrating the samples before coating them .
I have tried an Environmental SEM to avoid the dehydration step but unfortunately the resolution was really bad .
Can Any one suggest a technique to compare my hydrated scaffold with the partially dehydrated ones?
Best option is cryo SEM. In this no need to dehydrate your samples.
Hello,
I am curious about the ESEM. High resolution of some hydrated samples can be difficult especially with a tungsten gun ESEM. Did you use field emission ESEM? What is the expected diameter of your fibres wet and dry?
The expected fiber diamter is > 10 microns. I have used
Philips XL30 ESEM at variable water vapour pressures and voltages (v. poor results)
Hitachi S-3400N variable pressure SEM at various pressures and voltages (better but still poor results, plus it dried out the wet sample)
I believe what you said is true regarding the ESEM.
Dear MAmatha,
I am trying to locate a cryo SEM in my institute ,, But I wanted to clarify the cryo SEM,, What is the maximum magnification it can go ? have you tried it on natural tissue?
We have sold our XL30 ESEM back to FEI and are waiting for a field emission ESEM. The latter can be used at much lower voltages and would be better for your sample, I expect. Modern cryo systems on a field emission SEM should give excellent results at high magnifications. I found a paper on Google from 2000 stating interpretable biological structures can be seen at 1-2nm.
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