To obtain large numbers of organisms, trophozoites of Acanthamoeba were grown in my laboratory in 500 ml antibiotic-free peptone-yeast extract glucose broth for 42 hours in 5 litre flasks on a shaker set at 100 rpm inside an incubator. Further culture-related details, including a comment on how cysts were obtained, are given in the following publication (I think it can be downloaded from my RG profile): Niszl, I.A. & Markus, M.B. 1998. Anti-Acanthamoeba activity of contact lens solutions. British Journal of Ophthalmology 82: 1033-1038. As regards electron microscopy, we used a particular technique for embedding. If you are unable to access this article, I could provide it: Niszl, I.A. & Markus, M.B. 1989. Processing of free-living amoebae for transmission electron microscopy. Stain Technology 64: 259-260.

Thank you 

I would be more than gratefully if you can supply me with the article 

Certainly. The article is in my office. I'll find it when next I'm there. Am away from home at present.