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

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

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

5 Scopus citations

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).

Original languageEnglish (US)
Title of host publicationNanomaterial Interfaces in Biology
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages179-198
Number of pages20
ISBN (Print)9781627034616
DOIs
StatePublished - Jan 1 2013

Publication series

NameMethods in Molecular Biology
Volume1025
ISSN (Print)1064-3745

Keywords

  • Electron tomography (ET)
  • Energy dispersive X-ray spectroscopy (EDXS) chemical analysis
  • HeLa
  • Immunocytochemistry
  • Iron oxide nanoparticles (IONPs) loaded nanogel
  • Scanning transmission electron microscopy (STEM)
  • Stimuli-responsive polymeric nanoparticles
  • Tokuyasu cryosectioning technique

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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