Alcian blue staining to track the intracellular fate of hyaluronic-acid-based nanoparticles at transmission electron microscopy

  • Flavia Carton Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of Verona, Italy. https://orcid.org/0000-0001-6310-9079
  • Mathieu Repellin Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of Verona, Italy. https://orcid.org/0000-0003-0613-3667
  • Giovanna Lollo Laboratory of Automatic Control, Chemical and Pharmaceutical Engineering (LAGEPP), French National Centre for Scientific Research (CNRS), UMR 5007, Claude Bernard University Lyon 1, Villeurbanne, France.
  • Manuela Malatesta | manuela.malatesta@univr.it Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of Verona, Italy. https://orcid.org/0000-0001-8196-9232

Abstract

The main step in the assessment of nanomaterial safety and suitability for biomedical use is the location and the dynamic tracking of nanoparticles (NPs) inside cells or tissues. To precisely investigate the uptake mechanisms and intracellular fate of NPs, transmission electron microscopy is the technique of choice; however, the detection of NPs may sometimes be problematic. In fact, while NPs containing strongly electron dense (e.g., metal) components do not require specific detection methods at the ultrastructural level, organic NPs are hardly detectable in the intracellular environment due to their intrinsic moderate electron density. In this study, the critical-electrolyte-concentration Alcian Blue method set up by Schofield et al. in 1975 was applied to track hyaluronic-acid-based NPs in muscle cells in vitro. This long-established histochemical method proved to be a powerful tool allowing to identify not only whole NPs while entering cells and moving into the cytoplasm, but also their remnants following lysosomal degradation and extrusion.

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Published
2019-12-12
Section
Technical Notes
Keywords:
nanocarriers, hyaluronate, histochemistry, ultrastructure
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How to Cite
Carton, F., Repellin, M., Lollo, G., & Malatesta, M. (2019). Alcian blue staining to track the intracellular fate of hyaluronic-acid-based nanoparticles at transmission electron microscopy. European Journal of Histochemistry, 63(4). https://doi.org/10.4081/ejh.2019.3086