Which steps should I follow to dehydration and drying before SEM evaluation. The device i’ve use for this study is a new-generation product of Hitachi btw.
Specimen preparation depends not only on material, but on your task too. What is a purpose of your SEM study?
The purpose of my study is to observe the effectiveness of different irrigation solutions in removing smear layer.
The easiest way is to observe smeared/etched surface from the top, to see if smear plugs in tubules are dissolved. You do not need any fixation, just dehydration in graded solutions of alcohol (for example, 33%, 66%, 85%, 95%, 2*100%). About 1 hour in each of solutions and overnight in the last 100% solution. Then air dry for about 24 hours.
There is another approach - to observe collagen network in an etched layer, but for this you need additional steps - fixation and specimen fracture after dehydration.
I've tried 10 minutes for each of solutions before, and it didn't work. But 1 hour seems to me very long time period. Are you sure the smear layer will not damaged by this?
I did not understand what "does not work" means. Actually you can skip all the steps but the last one (100% alcohold for 1-13 hours) and air drying. You may have more cracks, but otherwise you should get the same results.
Are you looking for tubule's openings?
i mean we couldn't get any SEM images, SEM analyses system didn't work because of samples are not dry enough. i don't know much about the technical details about the device...
(yeah we are looking for dentin tubules)
To get rid of moisture in SEM use smaller pieces - hot whole teeth, but cut off a place of interest and process it.
@Vladimir Dusevich thanks for your advice. I'll try what you suggested.
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