ABSTRACT
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.
Subject(s)
Brain/ultrastructure , Imaging, Three-Dimensional/methods , Metal Nanoparticles , Neuroimaging/methods , Neurons/ultrastructure , X-Ray Microtomography/methods , Animals , Gold , MiceABSTRACT
We describe a new approach for the inversion of the generalized attenuated radon transform in X-ray fluorescence computed tomography (XFCT). The approach consists of using the radon inverse as an approximation for the actual one, followed by an iterative refinement. Also, we analyze the problem of retrieving the attenuation map directly from the emission data, giving rise to a novel alternating method for the solution. We applied our approach to real and simulated XFCT data and compared its performance to previous inversion algorithms for the problem, showing its main advantages: better images than those obtained by other analytic methods and much faster than iterative methods in the discrete setting.
Subject(s)
Algorithms , Image Processing, Computer-Assisted/methods , Spectrometry, X-Ray Emission/methods , Tomography, X-Ray/methods , Models, Theoretical , Phantoms, ImagingABSTRACT
X-ray fluorescence computed tomography (XFCT) is a relatively new synchrotron-based imaging modality aiming at reconstructing the distribution of nonradiative elements within a sample irradiated with high-intensity monochromatic x-rays. In a recent paper La Rivière (2004 Phys. Med. Biol. 49 2391-405) presented an approximated inversion method based on reducing the problem to the inversion of the exponential radon transform. In this paper we compare La Rivière's results with recently derived 'exact' analytic formulae for the generalized attenuated radon transform. We present numerical experiments with real and simulated data.
