Visualizing the 3D architecture of multiple erythrocytes infected with Plasmodium at nanoscale by focused ion beam-scanning electron microscopy

• Published in: PloS one Vol. 7; no. 3; p. e33445
• Main Authors: Medeiros, Lia Carolina Soares, De Souza, Wanderley, Jiao, Chengge, Barrabin, Hector, Miranda, Kildare
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 14-03-2012; Public Library of Science (PLoS)
• Subjects: Animals; Architecture; Biology; Blood; Cell cycle; Cytoplasm; Developmental stages; Electron microscopy; Electrons; Erythrocytes; Erythrocytes - parasitology; Erythrocytes - ultrastructure; Glycosylated hemoglobin; Hemoglobin; Hemoglobins - metabolism; Image Processing, Computer-Assisted; Imaging, Three-Dimensional - methods; Infections; Intracellular Space - parasitology; Ion beams; Life cycle engineering; Life cycles; Malaria; Malaria - parasitology; Male; Medical imaging; Medicine; Mice; Microscopy, Electron, Scanning - methods; Models, Biological; Models, Molecular; Morphology; Nanoparticles - ultrastructure; Parasites; Physics; Plasmodium; Plasmodium - ultrastructure; Plasmodium chabaudi; Plasmodium falciparum; Plasmodium simium; Proteins; Red blood cells; Saimiri sciureus; Scanning electron microscopy; Statistical analysis; Surface Properties; Three dimensional imaging; Tomography; Trends; Tubules; Visualization (Computer); Brazil; Rio de Janeiro Brazil
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0033445

Different methods for three-dimensional visualization of biological structures have been developed and extensively applied by different research groups. In the field of electron microscopy, a new technique that has emerged is the use of a focused ion beam and scanning electron microscopy for 3D reconstruction at nanoscale resolution. The higher extent of volume that can be reconstructed with this instrument represent one of the main benefits of this technique, which can provide statistically relevant 3D morphometrical data. As the life cycle of Plasmodium species is a process that involves several structurally complex developmental stages that are responsible for a series of modifications in the erythrocyte surface and cytoplasm, a high number of features within the parasites and the host cells has to be sampled for the correct interpretation of their 3D organization. Here, we used FIB-SEM to visualize the 3D architecture of multiple erythrocytes infected with Plasmodium chabaudi and analyzed their morphometrical parameters in a 3D space. We analyzed and quantified alterations on the host cells, such as the variety of shapes and sizes of their membrane profiles and parasite internal structures such as a polymorphic organization of hemoglobin-filled tubules. The results show the complex 3D organization of Plasmodium and infected erythrocyte, and demonstrate the contribution of FIB-SEM for the obtainment of statistical data for an accurate interpretation of complex biological structures.