Asked 22nd Jan, 2018

Does anyone have any advice on fixing leaf tissue for SEM to observe structural changes?

Looking through the literature, there are many slightly different techniques for fixing biological specimens using different pre-post fixatives, mainly paraformaldehyde, gluteraldehyde, cacodylate and osmium tetroxide followed by critical point drying but all of the papers use slightly different concentrations with little justification as to why. If anyone has access to a solid protocol for this it would be greatly appreciated.

All Answers (2)

22nd Jan, 2018
Vladimir Dusevich
University of Missouri - Kansas City
Any specimen preparation may induce artifacts. So, if you need to observe a leaf surface I would strongly recommend ESEM which does not demand any specimen preparation at all. If your task is to observe wax on a leaf and you do not have a ESEM at hand, simple air drying may be the best way to go (to avoid disturbing wax with organic solvents). Otherwise you can use various protocols (nobody can claim to have the very best recipe). Bear in mind that Os treatment is optional for SEM and in most (all?) cases can be omitted. CPD drying can be substituted with HMDS treatment.
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