RESUMO
A deeper knowledge of the architecture of the peripheral nerve with three-dimensional (3D) imaging of the nerve tissue at the sub-cellular scale may contribute to unravel the pathophysiology of neuropathy. Here we demonstrate the feasibility of X-ray phase contrast holographic nanotomography to enable 3D imaging of nerves at high resolution, while covering a relatively large tissue volume. We show various subcomponents of human peripheral nerves in biopsies from patients with type 1 and 2 diabetes and in a healthy subject. Together with well-organized, parallel myelinated nerve fibres we show regenerative clusters with twisted nerve fibres, a sprouted axon from a node of Ranvier and other specific details. A novel 3D construction (with movie created) of a node of Ranvier with end segment of a degenerated axon and sprout of a regenerated one is captured. Many of these architectural elements are not described in the literature. Thus, X-ray phase contrast holographic nanotomography enables identifying specific morphological structures in 3D in peripheral nerve biopsies from a healthy subject and from patients with type 1 and 2 diabetes.
Assuntos
Neuropatias Diabéticas/diagnóstico por imagem , Neuropatias Diabéticas/patologia , Holografia , Nervos Periféricos/diagnóstico por imagem , Nervos Periféricos/patologia , Idoso , Estudos de Casos e Controles , Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 2/complicações , Feminino , Holografia/métodos , Humanos , Processamento de Imagem Assistida por Computador , Masculino , Microscopia , Pessoa de Meia-Idade , Nanotecnologia , Microtomografia por Raio-X/métodosRESUMO
Here we report a method for increased resolution of single exposure three modality x-ray images using super-resolution. The three x-ray image modalities are absorption-, differential phase-contrast-, and dark-field-images. To create super-resolution, a non-mechanically movable micro-focus x-ray source is used. A series of almost identical x-ray projection images is obtained while the point source is translated in a two-dimensional grid pattern. The three image modalities are extracted from fourier space using spatial harmonic analysis, also known as the single-shot method. Using super-resolution on the low-resolution series of the three modalities separately results in high-resolution images for the modalities. This approach allows to compensate for the inherent loss in resolution caused by the single-shot method without increasing the need for stability or algorithms accounting for possible motion.