usually carbon is used as protective layer for nanoparticles from oxidation. these NPs are observed by HRTEM to find lattice spacing. Is C film transparent for electron beam, if not transparent then how e-beam can peneterate through the protection layer and give correct d-spacing
Check Monte Carlo simulations done by Vladmir Stray (Page 212) in the following link:
http://cdn.intechopen.com/pdfs/14013.pdf
In addition to "transparent" there are such things as "semitransparent" and "almost transparent".
Higher accelerating voltage, more transparent C layer. If you want to see it you may want to use as low voltage as possible. You have to use substrate with holes in it, so it will be possible to see particles on edges, not on substrate itself (it will be definitely thicker than 5 nm and will reduce contrast).
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