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Immunocytochemistry, Electron Tomography, and Energy Dispersive X-ray Spectroscopy (EDXS) on Cryosections of Human Cancer Cells Doped with Stimuli Responsive Polymeric Nanogels Loaded with Iron Oxide Nanoparticles

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Nanomaterial Interfaces in Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1025))

Abstract

The cryosectioning technique is an alternative method for preparing biological material for Transmission Electron Microscopy (TEM). We have applied this technique to study the mechanism of cell internalization of stimuli-responsive polymeric nanogels exploited as cargo nanovectors. With respect to conventional TEM processing, cryosectioning technique better preserves the morphology of solvent-sensitive nanogels and enhances the visibility of membrane-bounded organelles inside the cell cytoplasm. In this chapter we describe the protocols we have established to perform Electron Microscopy (EM)-immunocytochemistry, Electron Tomography (ET), and Energy Dispersive X-ray Spectroscopy (EDXS) chemical analysis in Scanning TEM (STEM) on cryosections of HeLa cells treated with pH-responsive nanogels hosting short interference RNA (siRNAs) and iron oxide nanoparticles (IONPs).

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Author information

Authors and Affiliations

  1. Istituto Italiano di Tecnologia, Genoa, Italy

    Roberto Marotta, A. Falqui, A. Curcio, A. Quarta & Teresa Pellegrino

  2. National Nanotechnology Laboratory of CNR-NANO, Lecce, Italy

    A. Quarta & Teresa Pellegrino

Authors
  1. Roberto Marotta
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  2. A. Falqui
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  3. A. Curcio
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  4. A. Quarta
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  5. Teresa Pellegrino
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Editor information

Editors and Affiliations

  1. Dipto di Ingegneria Meccanica, Università degli Studi di Brescia, Brescia, Italy

    Paolo Bergese

  2. Dept of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Kimberly Hamad-Schifferli

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Marotta, R., Falqui, A., Curcio, A., Quarta, A., Pellegrino, T. (2013). Immunocytochemistry, Electron Tomography, and Energy Dispersive X-ray Spectroscopy (EDXS) on Cryosections of Human Cancer Cells Doped with Stimuli Responsive Polymeric Nanogels Loaded with Iron Oxide Nanoparticles. In: Bergese, P., Hamad-Schifferli, K. (eds) Nanomaterial Interfaces in Biology. Methods in Molecular Biology, vol 1025. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-462-3_14

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  • DOI: https://doi.org/10.1007/978-1-62703-462-3_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-461-6

  • Online ISBN: 978-1-62703-462-3

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