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Diaminobenzidine Photooxidation to Visualize Fluorescent Nanoparticles in Adhering Cultured Cells at Transmission Electron Microscopy

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Histochemistry of Single Molecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2566))

Abstract

Visualizing nanoparticles made of organic material (e.g., polysaccharides, proteins, non-osmiophilic lipids) inside cells and tissues at transmission electron microscopy is a difficult task due to the intrinsic weak electron density of these nanoconstructs, which makes them hardly distinguishable in the biological environment. We describe here a simple protocol to apply photooxidation to fluorescently labeled nanoparticles administered to cultured cells in vitro. The conversion of the fluorescent signal into a granular electron-dense reaction product through light irradiation in the presence of diaminobenzidine makes the nanoparticles clearly visible at the ultrastructural level. Our procedure proved to be reliable with various fluorophores and may be applied to any cell type.

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Author information

Authors and Affiliations

  1. Department of Neurosciences, Biomedicine and Movement Sciences, Anatomy and Histology Section, University of Verona, Verona, Italy

    Manuela Costanzo & Manuela Malatesta

Authors
  1. Manuela Costanzo
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  2. Manuela Malatesta
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Correspondence to Manuela Malatesta .

Editor information

Editors and Affiliations

  1. Department of Biology and Biotechnology, University of Pavia, PAVIA, Italy

    Carlo Pellicciari

  2. Department of Biology and Biotechnology, University of Pavia, PAVIA, Italy

    Marco Biggiogera

  3. Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, VERONA, Italy

    Manuela Malatesta

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Costanzo, M., Malatesta, M. (2023). Diaminobenzidine Photooxidation to Visualize Fluorescent Nanoparticles in Adhering Cultured Cells at Transmission Electron Microscopy. In: Pellicciari, C., Biggiogera, M., Malatesta, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 2566. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2675-7_27

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  • DOI: https://doi.org/10.1007/978-1-0716-2675-7_27

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2674-0

  • Online ISBN: 978-1-0716-2675-7

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