Abstract
Intravital fluorescence microscopy is an invaluable tool to study a dynamic phenomenon through its direct observation in living organisms. This technique can combine qualitative and quantitative analysis and has been capital to address long-standing questions about Plasmodium biology. Beyond a descriptive view of the parasite life cycle, the possibility to image infection in transgenic animals in which a specific cell type, molecule or process is labeled opens new possibilities to study host cell–parasite interactions in cellular and molecular details. An additional layer of refinement can be achieved with the use of fluorescent knockout mutants (parasite, mice, or both) to dissect the molecular basis of the process of interest. Here, we present a basic protocol for imaging the sporozoite behavior in the liver, emphasizing the detection of the sporozoite’s ability to traverse host cells.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Grewe BF et al (2010) High-speed in vivo calcium imaging reveals neuronal network activity with near-millisecond precision. Nat Methods 7:399–405
Keller PJ et al (2008) Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy. Science 322:1065–1069
Petty HR (2007) Fluorescence microscopy: established and emerging methods, experimental strategies, and applications in immunology. Microsc Res Tech 70:687–709
Frevert U et al (2006) Nomadic or sessile: can Kupffer cells function as portals for malaria sporozoites to the liver? Cell Microbiol 8:1537–1546
Amino R et al (2008) Host cell traversal is important for progression of the malaria parasite through the dermis to the liver. Cell Host Microbe 3:88–96
Ishino T et al (2004) Cell-passage activity is required for the malarial parasite to cross the liver sinusoidal cell layer. PLoS Biol 2:E4
Mota MM et al (2001) Migration of Plasmodium sporozoites through cell before infection. Science 291:141–144
Sturm A et al (2009) Alteration of the parasite plasma membrane and the parasitophorous vacuole membrane during exo-erythrocytic development of malaria parasites. Protist 160:51–63
Xu Y et al (2010) Neuropilin-2 mediates VEGF-C-induced lymphatic sprouting together with VEGFR3. J Cell Biol 188:155–130
Thiberge S et al (2007) In vivo imaging of malaria parasites in the murine liver. Nat Protoc 2:1811–1818
Acknowledgments
We thank S. Shorte and the Imagopole for the support with confocal microscopy; C. Bourgouin and the CEPIA for mosquito rearing. This work was supported by funds from Pasteur Institute and the French National Research Agency (JCJC PlasmoPEP). J.T. was supported by a fellowship from the Fundação para Ciência e Tecnologia (SFRH/BPD/48340/2008) and P.F. by a Ph.D. fellowship from the Direction Générale de l’Armement.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Tavares, J., Formaglio, P., Medvinsky, A., Ménard, R., Amino, R. (2012). Imaging Sporozoite Cell Traversal in the Liver of Mice. In: Ménard, R. (eds) Malaria. Methods in Molecular Biology, vol 923. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-026-7_28
Download citation
DOI: https://doi.org/10.1007/978-1-62703-026-7_28
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-025-0
Online ISBN: 978-1-62703-026-7
eBook Packages: Springer Protocols