Recently I encounter the problem about how to make a sample of the protoplast and make sure they are in shape without any breakup? It was really hard to see a complete protoplast with the method I used currently to make the sample.
Dear Tao-Ho,
I guess you'll be / are not alone with this problem. Not being a "plant electron microscopist" (only due to my interest) I would like to give you only some hints first (not knowing the extent of your special knowledge, so apologize if you know the following at all / already).
Tonoplast/vacuole ultrastructural preservation in plant ("equivalent image") usually is said to be (but it might be that in some cases / under certain circumstances you'll have included some = 1-3 cells lacking fixation artifacts).At least you would need a certain time to find such intact plant cells in your preparation.
Chemical fixation without appropriate adjustment of fixative (pH as well as tonicity) will result either in swelling or shrinking.
Despite a for the first time was demonstrated eventually by KAUFMAN BP and DE,DN (1956) Fine structure of Chromosome,J.Biophys.Biochem.2 Suppl, 419 by means of electron microscopy, other papers on Tonoplast in general by conventional EM seem to be very rare. Some more articles exist using more modern (and perhaps mandatory) preparations like: Fixation with the aid of Microwave application, cryofixation and cryosectioning, HPF(High power freezing) & FF+FS (Freeze Fracture, Freeze substitution).
If you are not experienced with this I would like to point you at least to a Google search, using the search phrase:
< Arabidopsis AND Tonoplast AND "electron microsc*" > (use as it stands without the parentheses < > ).
You'll find a lot of electron microscopic work done on Arabidopsis (not having checked whether all of them include also preservation of the , most of them (at least out of the first 20 results) were published in JEM (Journal of Electron Microscopy, Tokyo, which has changed its name several months ago to "MICROSCOPY").
Good luck and very best regards,
Wolfgang
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