Gold Nanoparticles for X-ray Microtomography of Neurons.

Autor:	Depannemaecker D; UNIFESP , Universidade Federal de São Paulo/Escola Paulista de Medicina (UNIFESP/EPM) , São Paulo , São Paulo , Brazil., Santos LEC; UNIFESP , Universidade Federal de São Paulo/Escola Paulista de Medicina (UNIFESP/EPM) , São Paulo , São Paulo , Brazil., de Almeida AG; UFSJ , Universidade Federal de São João , São João del-Rei , Minas Gerais , Brazil., Ferreira GBS; IFGW-UNICAMP , Institute of Physics at the University of Campinas , Campinas , São Paulo , Brazil., Baraldi GL; IFGW-UNICAMP , Institute of Physics at the University of Campinas , Campinas , São Paulo , Brazil., Miqueles EX; LNLS-CNPEM , Brazilian Synchrotron Light Laboratory , Campinas , São Paulo , Brazil., de Carvalho M; LNLS-CNPEM , Brazilian Synchrotron Light Laboratory , Campinas , São Paulo , Brazil.; LNBio-CNPEM , Brazilian Biosciences National Laboratory , Campinas , São Paulo , Brazil., Costa GSR; LNLS-CNPEM , Brazilian Synchrotron Light Laboratory , Campinas , São Paulo , Brazil., Marques MJG; UNIFESP , Universidade Federal de São Paulo/Escola Paulista de Medicina (UNIFESP/EPM) , São Paulo , São Paulo , Brazil., Scorza CA; UNIFESP , Universidade Federal de São Paulo/Escola Paulista de Medicina (UNIFESP/EPM) , São Paulo , São Paulo , Brazil., Rinkel J; IFGW-UNICAMP , Institute of Physics at the University of Campinas , Campinas , São Paulo , Brazil.
Jazyk:	angličtina
Zdroj:	ACS chemical neuroscience [ACS Chem Neurosci] 2019 Aug 21; Vol. 10 (8), pp. 3404-3408. Date of Electronic Publication: 2019 Jul 16.
DOI:	10.1021/acschemneuro.9b00290
Abstrakt:	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.
Databáze:	MEDLINE
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