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Article Dans Une Revue ACS Chemical Neuroscience Année : 2019

Gold Nanoparticles for X-ray Microtomography of Neurons

E Canton Santos
  • Fonction : Auteur
Gustavo B S Ferreira
  • Fonction : Auteur
Giovanni L Baraldi
  • Fonction : Auteur
Eduardo X Miqueles
  • Fonction : Auteur
∥ Murilo de Carvalho
  • Fonction : Auteur
Gabriel Schubert Ruiz Costa
  • Fonction : Auteur
Marcia J Guimaraẽs Marques
  • Fonction : Auteur
Carla A Scorza
  • Fonction : Auteur
Jean Rinkel
  • Fonction : Auteur

Résumé

Commonly used methods to visualize the biological structure of brain tissues at subcellular resolution are confocal microscopy and two-photon microscopy. Both require slicing the sample into sections of a few tens of micrometers. The recent developments in X-ray microtomography enable three-dimensional imaging at sub-micrometer and isotropic resolution with larger biological samples. In this work, we developed and compared original microtomography methods and staining protocols to improve the contrast for in vitro mouse neuron imaging. Using Golgi's method to stain neurons randomly, we imaged the whole set of mouse brain structures. For specific and nonrandom neuron labeling, we conjugated 20 nm gold nanoparticles to antibodies used in the immunohistochemistry (IHC) method, using anti-NeuN to label specifically neuronal nuclei. We applied an original subtraction dual-energy method for microtomography in the vicinity of the Au L-III absorption edge and compared image reconstructions to confocal microscopy images acquired on the same samples. The results show the possibility to characterize the 3D entire brain structure of mice. They demonstrated a high contrast and neuron detection improvement by applying the dual-energy method coupled to IHC staining.
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Dates et versions

hal-02507506 , version 1 (13-03-2020)

Identifiants

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Damien Depannemaecker, E Canton Santos, Antonio-Carlos Guimaraẽs de Almeida, Gustavo B S Ferreira, Giovanni L Baraldi, et al.. Gold Nanoparticles for X-ray Microtomography of Neurons. ACS Chemical Neuroscience, 2019, ⟨10.1021/acschemneuro.9b00290⟩. ⟨hal-02507506⟩
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