Gold Nanoparticles for X-ray Microtomography of Neurons.
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.
['Depannemaecker|Damien|D|', 'Santos|Luiz E Canton|LEC|', 'de Almeida|Antonio-Carlos Guimarães|AG|', 'Ferreira|Gustavo B S|GBS|', 'Baraldi|Giovanni L|GL|', 'Miqueles|Eduardo X|EX|', 'de Carvalho|Murilo|M|', 'Costa|Gabriel Schubert Ruiz|GSR|', 'Marques|Marcia J Guimarães|MJG|', 'Scorza|Carla A|CA|', 'Rinkel|Jean|J|0000-0001-9980-0304']
D000818:Animals D001921:Brain D006046:Gold D021621:Imaging, Three-Dimensional D053768:Metal Nanoparticles D051379:Mice D059906:Neuroimaging D009474:Neurons D055114:X-Ray Microtomography
gold nanoparticles immunohistochemistry Brain imaging dual-energy X-ray microtomography synchrotron