Simplified Specimen Preparation for Electron Microscopic Studies of an Intra-erythrocytic Parasite, Anaplasma marginale

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Abstract
The method included dehemoglobinization of the parasitized bovine erythrocytes by dialysis against a hypotonic solution of a volatile salt (0.25% ammonium acetate). The hemolyzed stromata were then transferred to Formvar-coated grids by gravity sedimentation from pipetted droplets. Metal shadowing with germanium was used to enhance the specimen contrast. The parasites were readily recognizable with the electron microscope. A semiquantitative experiment showed that about 2 out of 3 organisms were retained within their host stroma. The parasites exhibited a complex morphology which usually consisted of a large (0.8-1.3[mu]) electron dense body (frequently subdivided) attached to a sac-like projection. Occasionally, the sac structure appeared to form an anastomosis between 2 dissimilar large bodies. Small rectangular extensions at the terminus of the sac were shown to be associated with subunits of the large electron dense bodies. A possible mode of reproduction of the parasite was suggested by this association.

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