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Imaging of Extracellular Vesicles Derived from Plasmodium falciparum–Infected Red Blood Cells Using Atomic Force Microscopy

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Malaria Immunology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2470))

Abstract

Malaria is one the most devastating infectious diseases in the world: of the five malaria-associated parasites, Plasmodium falciparum and P. vivax are the most pathogenic and widespread, respectively. P. falciparum invades human red blood cells (RBCs), releasing extracellular vesicles (Pf-EV) carrying DNA, RNA and protein cargo components involved in host-pathogen communications in the course of the disease. Different strategies have been used to analyze Pf-EV biophysically and chemically. Atomic force microscopy (AFM) stands out as a powerful tool for rendering high quality images of extracellular vesicles. In this technique, a sharp tip attached to a cantilever reconstructs the topographic surface of the extracellular vesicles and probes their nano-mechanical properties based on force–distance curves. Here, we describe a method to separate Pf-EV using differential ultracentrifugation, followed by nanoparticle tracking analysis (NTA) to quantify and estimate the size distribution. Finally, the AFM imaging procedure on Pf-EV adsorbed on a Mg2+-modified mica surface is detailed.

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Acknowledgments

We greatly appreciate Dr. Sidney R. Cohen for his critical review of the manuscript.

Author information

Authors and Affiliations

  1. Department of Chemical Research Support, Weizmann Institute of Science, Rehovot, Israel

    Irit Rosenhek-Goldian

  2. Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel

    Paula Abou Karam, Neta Regev-Rudzki & Alicia Rojas

  3. Laboratory of Helminthology, Centro de Investigaciones en Enfermedades Tropicales, University of Costa Rica, San José, Costa Rica

    Alicia Rojas

Authors
  1. Irit Rosenhek-Goldian
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  2. Paula Abou Karam
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  3. Neta Regev-Rudzki
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  4. Alicia Rojas
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Corresponding author

Correspondence to Alicia Rojas .

Editor information

Editors and Affiliations

  1. Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark

    Anja Tatiana Ramstedt Jensen

  2. Centre for Medical Parasitology, Department of Immunology and Microbiology, University of Copenhagen, Copenhagen, Denmark

    Lars Hviid

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Rosenhek-Goldian, I., Abou Karam, P., Regev-Rudzki, N., Rojas, A. (2022). Imaging of Extracellular Vesicles Derived from Plasmodium falciparum–Infected Red Blood Cells Using Atomic Force Microscopy. In: Jensen, A.T.R., Hviid, L. (eds) Malaria Immunology. Methods in Molecular Biology, vol 2470. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2189-9_12

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  • DOI: https://doi.org/10.1007/978-1-0716-2189-9_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2188-2

  • Online ISBN: 978-1-0716-2189-9

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