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PURPOSE: A major obstacle to the clinical implementation of quantitative MR is the lengthy acquisition time required to derive multi-contrast parametric maps. We sought to reduce the acquisition time for QSM and macromolecular tissue volume by acquiring both contrasts simultaneously by leveraging their redundancies. The joint virtual coil concept with GRAPPA (JVC-GRAPPA) was applied to reduce acquisition time further. METHODS: Three adult volunteers were imaged on a 3 Tesla scanner using a multi-echo 3D GRE sequence acquired at 3 head orientations. Macromolecular tissue volume, QSM, R2∗ , T1 , and proton density maps were reconstructed. The same sequence (GRAPPA R = 4) was performed in subject 1 with a single head orientation for comparison. Fully sampled data was acquired in subject 2, from which retrospective undersampling was performed (R = 6 GRAPPA and R = 9 JVC-GRAPPA). Prospective undersampling was performed in subject 3 (R = 6 GRAPPA and R = 9 JVC-GRAPPA) using gradient blips to shift k-space sampling in later echoes. RESULTS: Subject 1's multi-orientation and single-orientation macromolecular tissue volume maps were not significantly different based on RMSE. For subject 2, the retrospectively undersampled JVC-GRAPPA and GRAPPA generated similar results as fully sampled data. This approach was validated with the prospectively undersampled images in subject 3. Using QSM, R2∗ , and macromolecular tissue volume, the contributions of myelin and iron content to susceptibility were estimated. CONCLUSION: We have developed a novel strategy to simultaneously acquire data for the reconstruction of 5 intrinsically coregistered 1-mm isotropic resolution multi-parametric maps, with a scan time of 6 min using JVC-GRAPPA.
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Processamento de Imagem Assistida por Computador , Imageamento por Ressonância Magnética , Encéfalo/diagnóstico por imagem , Humanos , Estudos Prospectivos , Estudos RetrospectivosRESUMO
Due to the technical challenges of large-scale microscopy and analysis, to date only limited knowledge has been made available about axon morphometry (diameter, shape, myelin thickness, volume fraction), thereby limiting our understanding of neuronal microstructure and slowing down research on neurodegenerative pathologies. This study addresses this knowledge gap by establishing a state-of-the-art acquisition and analysis framework for mapping axon morphometry, and providing the first comprehensive mapping of axon morphometry in the human spinal cord. We dissected, fixed and stained a human spinal cord with osmium tetroxide, and used a scanning electron microscope to image the entirety of 23 axial slices, covering C1 to L5 spinal levels. An automatic method based on deep learning was then used to segment each axon and myelin sheath to produce maps of axon morphometry. These maps were then registered to a standard spinal cord magnetic resonance imaging (MRI) template. Between 500,000 (lumbar) and 1 million (cervical) myelinated axons were segmented at each level of this human spinal cord. Morphometric features show a large disparity between tracts, but high right-left symmetry. Our results suggest a modality-based organization of the dorsal column in the human, as it has been observed in the rat. The generated axon morphometry template is publicly available at https://osf.io/8k7jr/ and could be used as a reference for quantitative MRI studies. The proposed framework for axon morphometry mapping could be extended to other parts of the central or peripheral nervous system that exhibit coherently-oriented axons.
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Atlas como Assunto , Axônios/ultraestrutura , Imageamento Tridimensional/métodos , Medula Espinal/ultraestrutura , Idoso , Feminino , Humanos , Processamento de Imagem Assistida por Computador/métodos , Microscopia Eletrônica de Varredura , Bainha de Mielina/ultraestruturaRESUMO
Atlases of the central nervous system are essential for understanding the pathophysiology of neurological diseases, which remains one of the greatest challenges in neuroscience research today. These atlases provide insight into the underlying white matter microstructure and have been created from a variety of animal models, including rats. Although existing atlases of the rat spinal cord provide some details of axon microstructure, there is currently no histological dataset that quantifies axon morphometry exhaustively in the entire spinal cord. In this study, we created the first comprehensive rat spinal cord atlas of the white matter microstructure with quantifiable axon and myelin morphometrics. Using full-slice scanning electron microscopy images and state-of-the-art segmentation algorithms, we generated an atlas of microstructural metrics such as axon diameter, axonal density and g-ratio. After registering the Watson spinal cord white matter atlas to our template, we computed statistics across metrics, spinal levels and tracts. We notably found that g-ratio is relatively constant, whereas axon diameter showed the greatest variation. The atlas, data and full analysis code are freely available at: https://github.com/neuropoly/atlas-rat.
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Axônios/ultraestrutura , Neurônios/ultraestrutura , Medula Espinal/ultraestrutura , Algoritmos , Animais , Atlas como Assunto , Feminino , Processamento de Imagem Assistida por Computador , Masculino , Ratos Sprague-Dawley , Substância Branca/ultraestruturaRESUMO
BACKGROUND AND PURPOSE: The stiffness of large arteries and increased pulsatility can have an impact on the brain white matter (WM) microstructure, however those mechanisms are still poorly understood. The aim of this study was to investigate the association between central artery stiffness, axonal and myelin integrity in 54 cognitively unimpaired elderly subjects (65-75 years old). METHODS: The neuronal fiber integrity of brain WM was assessed using diffusion tensor metrics and magnetization transfer imaging as measures of axonal organization (Fractional anisotropy, Radial diffusivity) and state of myelination (Myelin volume fraction). Central artery stiffness was measured by carotid-femoral pulse wave velocity (cfPWV). Statistical analyses included 4 regions (the corpus callosum, the internal capsule, the corona radiata and the superior longitudinal fasciculus) which have been previously denoted as vulnerable to increased central artery stiffness. RESULTS: cfPWV was significantly associated with fractional anisotropy and radial diffusivity (pâ¯<â¯0.05, corrected for multiple comparisons) but not with myelin volume fraction. Findings from this study also show that improved executive function performance correlates with Fractional anisotropy positively (pâ¯<â¯0.05 corrected) as well as with myelin volume fraction and radial diffusivity negatively (pâ¯<â¯0.05 corrected). CONCLUSIONS: These findings suggest that arterial stiffness is associated with axon degeneration rather than demyelination. Controlling arterial stiffness may play a role in maintaining the health of WM axons in the aging brain.
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Envelhecimento , Artérias/diagnóstico por imagem , Axônios , Função Executiva/fisiologia , Imageamento por Ressonância Magnética/métodos , Bainha de Mielina , Análise de Onda de Pulso/métodos , Rigidez Vascular , Substância Branca/diagnóstico por imagem , Idoso , Envelhecimento/patologia , Envelhecimento/fisiologia , Axônios/patologia , Imagem de Tensor de Difusão/métodos , Feminino , Humanos , Masculino , Bainha de Mielina/patologia , Rigidez Vascular/fisiologia , Substância Branca/patologiaRESUMO
The fiber g-ratio is the ratio of the inner to the outer diameter of the myelin sheath of a myelinated axon. It has a limited dynamic range in healthy white matter, as it is optimized for speed of signal conduction, cellular energetics, and spatial constraints. In vivo imaging of the g-ratio in health and disease would greatly increase our knowledge of the nervous system and our ability to diagnose, monitor, and treat disease. MRI based g-ratio imaging was first conceived in 2011, and expanded to be feasible in full brain white matter with preliminary results in 2013. This manuscript reviews the growing g-ratio imaging literature and speculates on future applications. It details the methodology for imaging the g-ratio with MRI, and describes the known pitfalls and challenges in doing so.
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Imagem de Difusão por Ressonância Magnética/métodos , Fibras Nervosas Mielinizadas , Neuroimagem/métodos , Substância Branca/anatomia & histologia , Substância Branca/diagnóstico por imagem , Imagem de Difusão por Ressonância Magnética/normas , Humanos , Fibras Nervosas Mielinizadas/ultraestrutura , Neuroimagem/normasRESUMO
PURPOSE: Recent MRI techniques have been introduced that can extract microstructural information in the white matter, such as the density or macromolecular content. Translating quantitative MRI to the clinic raises many challenges in terms of acquisition strategy, modeling of the MRI signal, artifact corrections, and metric extraction (template registration and partial volume effects). In this work, we investigated the scan-rescan repeatability of several quantitative MRI techniques in the human spinal cord. METHODS: AxCaliber metrics, macromolecular tissue volume, and the fiber g-ratio were estimated in the spinal cord of eight healthy subjects, scanned and rescanned the same day in two different sessions. RESULTS: Scan-rescan repeatability deviation was 3% for all metrics, in average in the white matter of all subjects. Intraclass correlation coefficient was up to 0.9. A three-way analysis of variance showed significant effects of white matter pathway, laterality, and subject. CONCLUSION: The present study suggests that quantitative MRI gives stable measurements of white matter microstructure in the spinal cord of healthy subjects. Our findings remain to be evaluated in diseased populations. Magn Reson Med 79:2759-2765, 2018. © 2017 International Society for Magnetic Resonance in Medicine.
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Imagem de Difusão por Ressonância Magnética/métodos , Interpretação de Imagem Assistida por Computador/métodos , Medula Espinal/química , Medula Espinal/diagnóstico por imagem , Adolescente , Adulto , Algoritmos , Feminino , Humanos , Masculino , Bainha de Mielina/química , Reprodutibilidade dos Testes , Processamento de Sinais Assistido por Computador , Adulto JovemRESUMO
Diffusion magnetic resonance imaging (MRI) methods for axon diameter mapping benefit from higher maximum gradient strengths than are currently available on commercial human scanners. Using a dedicated high-gradient 3T human MRI scanner with a maximum gradient strength of 300 mT/m, we systematically studied the effect of gradient strength on in vivo axon diameter and density estimates in the human corpus callosum. Pulsed gradient spin echo experiments were performed in a single scan session lasting approximately 2h on each of three human subjects. The data were then divided into subsets with maximum gradient strengths of 77, 145, 212, and 293 mT/m and diffusion times encompassing short (16 and 25 ms) and long (60 and 94 ms) diffusion time regimes. A three-compartment model of intra-axonal diffusion, extra-axonal diffusion, and free diffusion in cerebrospinal fluid was fitted to the data using a Markov chain Monte Carlo approach. For the acquisition parameters, model, and fitting routine used in our study, it was found that higher maximum gradient strengths decreased the mean axon diameter estimates by two to three fold and decreased the uncertainty in axon diameter estimates by more than half across the corpus callosum. The exclusive use of longer diffusion times resulted in axon diameter estimates that were up to two times larger than those obtained with shorter diffusion times. Axon diameter and density maps appeared less noisy and showed improved contrast between different regions of the corpus callosum with higher maximum gradient strength. Known differences in axon diameter and density between the genu, body, and splenium of the corpus callosum were preserved and became more reproducible at higher maximum gradient strengths. Our results suggest that an optimal q-space sampling scheme for estimating in vivo axon diameters should incorporate the highest possible gradient strength. The improvement in axon diameter and density estimates that we demonstrate from increasing maximum gradient strength will inform protocol development and encourage the adoption of higher maximum gradient strengths for use in commercial human scanners.
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Axônios , Corpo Caloso/anatomia & histologia , Imagem de Difusão por Ressonância Magnética/métodos , Humanos , Processamento de Imagem Assistida por Computador/métodos , Cadeias de Markov , Modelos Estatísticos , Método de Monte CarloRESUMO
The ability to characterize white matter microstructure non-invasively has important applications for the diagnosis and follow-up of several neurological diseases. There exists a family of diffusion MRI techniques, such as AxCaliber, that provide indices of axon microstructure, such as axon diameter and density. However, to obtain accurate measurements of axons with small diameters (<5µm), these techniques require strong gradients, i.e. an order of magnitude higher than the 40-80mT/m currently available in clinical systems. In this study we acquired AxCaliber diffusion data at a variety of different q-values and diffusion times in the spinal cord of five healthy subjects using a 300mT/m whole body gradient system. Acquisition and processing were optimized using state-of-the-art methods (e.g., 64-channel coil, template-based analysis). Results consistently show an average axon diameter of 4.5+/-1.1µm in the spinal cord white matter. Diameters ranged from 3.0µm (gracilis) to 5.9µm (spinocerebellar tracts). Values were similar across laterality (left-right), but statistically different across spinal cord pathways (p<10(-5)). The observed trends are similar to those observed in animal histology. This study shows, for the first time, in vivo mapping of axon diameter in the spinal cord at 300mT/m, thus creating opportunities for applications in spinal cord diseases.
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Imagem de Difusão por Ressonância Magnética/métodos , Processamento de Imagem Assistida por Computador/métodos , Medula Espinal/citologia , Substância Branca/citologia , Adulto , Feminino , Humanos , MasculinoRESUMO
The myelin g-ratio, defined as the ratio between the inner and the outer diameter of the myelin sheath, is a fundamental property of white matter that can be computed from a simple formula relating the myelin volume fraction to the fiber volume fraction or the axon volume fraction. In this paper, a unique combination of magnetization transfer, diffusion imaging and histology is presented, providing a novel method for in vivo magnetic resonance imaging of the axon volume fraction and the myelin g-ratio. Our method was demonstrated in the corpus callosum of one cynomolgus macaque, and applied to obtain full-brain g-ratio maps in one healthy human subject and one multiple sclerosis patient. In the macaque, the g-ratio was relatively constant across the corpus callosum, as measured by both MRI and electron microscopy. In the human subjects, the g-ratio in multiple sclerosis lesions was higher than in normal appearing white matter, which was in turn higher than in healthy white matter. Measuring the g-ratio brings us one step closer to fully characterizing white matter non-invasively, making it possible to perform in vivo histology of the human brain during development, aging, disease and treatment.
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Axônios/ultraestrutura , Encéfalo/ultraestrutura , Imagem de Difusão por Ressonância Magnética/métodos , Bainha de Mielina/ultraestrutura , Adulto , Animais , Corpo Caloso/ultraestrutura , Humanos , Macaca fascicularis , Fenômenos Magnéticos , Masculino , Camundongos Mutantes Neurológicos , Esclerose Múltipla/patologiaRESUMO
PURPOSE: With current gold standard treatment, which associates maximum safe surgery and chemo-radiation, the large majority of glioblastoma patients relapse within a year in the peritumoral non contrast-enhanced region (NCE). A subpopulation of glioblastoma stem-like cells (GSC) are known to be particularly radio-resistant and aggressive, and are thus suspected to be the cause of these relapses. Previous studies have shown that their distribution is heterogeneous in the NCE compartment, but no study exists on the sensitivity of medical imaging for localizing these cells. In this work, we propose to study the magnetic resonance (MR) signature of these infiltrative cells. METHODS: In the context of a clinical trial on 16 glioblastoma patients, relative Cerebral Blood Volume (rCBV) and Apparent Diffusion Coefficient (ADC) were measured in a preoperative diffusion and perfusion MRI examination. During surgery, two biopsies were extracted using image-guidance in the hyperintensities-FLAIR region. GSC subpopulation was quantified within the biopsies and then cultivated in selective conditions to determine their density and aggressiveness. RESULTS: Low ADC was found to be a good predictor of the time to GSC neurospheres formation in vitro. In addition, GSCs were found in higher concentrations in areas with high rCBV. CONCLUSIONS: This study confirms that GSCs have a critical role for glioblastoma aggressiveness and supports the idea that peritumoral sites with low ADC or high rCBV should be preferably removed when possible during surgery and targeted by radiotherapy.
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We developed a Monte Carlo simulator for diffusion-weighted imaging sequences which displays the motion of water molecules and computes the dynamic phase dispersion due to the applied motion probing gradients. This simulator can be used to validate the analytical equations of diffusion models and understand their limitations due to their approximations. Here, we introduce the software and some specific use cases. The software can be downloaded from the following website: https://www.nirs.qst.go.jp/amr_diag.
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Membrana Celular/metabolismo , Simulação por Computador , Imagem de Difusão por Ressonância Magnética/métodos , Movimento (Física) , Água/metabolismo , Difusão , Humanos , Método de Monte CarloRESUMO
Ischemic stroke mostly affects the primary motor cortex and descending motor fibres, with consequent motor impairment. Pre-clinical models of stroke with reproducible and long-lasting sensorimotor deficits in higher-order animals are lacking. We describe a new method to induce focal brain damage targeting the motor cortex to study damage to the descending motor tracts in the non-human primate. Stereotaxic injection of malonate into the primary motor cortex produced a focal lesion in middle-aged marmosets (Callithrix jacchus). Assessment of sensorimotor function using a neurological scale and testing of forelimb dexterity and strength lasted a minimum of 12 weeks. Lesion evolution was followed by magnetic resonance imaging (MRI) at 24 h, 1 week, 4 and 12 weeks post-injury and before sacrifice for immunohistochemistry. Our model produced consistent lesions of the motor cortex, subcortical white matter and caudate nucleus. All animals displayed partial spontaneous recovery with long lasting motor deficits of force (54% loss) and dexterity (≈ 70% loss). Clearly visible T2 hypointensity in the white matter was observed with MRI and corresponded to areas of chronic gliosis in the internal capsule and lenticular fasciculus. We describe a straightforward procedure to reproducibly injure the motor cortex in the marmoset monkey, causing long-lasting motor deficits. The MRI signature reflects Wallerian degeneration and remote injury of corticospinal and corticopontine tracts, as well as subcortical motor loops. Our model may be suitable for the testing of therapies for post-stroke recovery, particularly in the chronic phase.
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Modelos Animais de Doenças , Força da Mão/fisiologia , AVC Isquêmico/induzido quimicamente , AVC Isquêmico/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Malonatos/toxicidade , Animais , Callithrix , Feminino , Seguimentos , Masculino , Malonatos/administração & dosagem , Reprodutibilidade dos Testes , Técnicas Estereotáxicas/normasRESUMO
OBJECTIVE: Central artery stiffness is a confirmed predictor of cardiovascular health status that has been consistently associated with cognitive dysfunction and dementia. The European Society of Hypertension has established a threshold of arterial stiffness above which a cardiovascular event is likely to occur. However, the threshold at which arterial stiffness alters brain integrity has never been established. METHODS: The aim of this study is to determine the arterial stiffness cut-off value at which there is an impact on the white matter microstructure. This study has been conducted with 53 cognitively elderly without dementia. The integrity of the white matter was assessed using diffusion tensor metrics. Central artery stiffness was evaluated by measuring the carotid-femoral pulse wave velocity (cfPWV). The statistical analyses included 4 regions previously denoted vulnerable to increased central arterial stiffness (the corpus callosum, the internal capsule, the corona radiata and the superior longitudinal fasciculus). RESULTS: The results of this study call into question the threshold value of 10â¯m/s cfPWV established by the European Society of Hypertension to classify patients in neuro-cardiovascular risk groups. Our results suggest that the cfPWV threshold value would be approximately 8.5â¯m/s when the microstructure of the white matter is taken as a basis for comparison. CONCLUSIONS: Adjustment of the cfPWV value may be necessary for a more accurate distinction between lower and higher risk group of patients for white matter microstructural injury related to arterial stiffness. Targeting the highest risk group for prevention methods may, in turn, help preserve brain health and cognitive functions.
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Envelhecimento/patologia , Envelhecimento/fisiologia , Velocidade da Onda de Pulso Carótido-Femoral/normas , Função Executiva/fisiologia , Hipertensão/diagnóstico , Rigidez Vascular/fisiologia , Substância Branca/anatomia & histologia , Idoso , Imagem de Tensor de Difusão , Feminino , Humanos , Masculino , Substância Branca/diagnóstico por imagemRESUMO
Cardiac-related spinal cord motion affects diffusion-weighted (DWI) signal. The goal of this study was to further quantify the specific detrimental effect of cord translational motion on the DWI signal in order to make better informed decisions about the cost-benefit of cardiac gating. We designed an MRI-compatible phantom mimicking the spinal cord translational motion. Cardiac-gated DWI data were acquired by varying the trigger delay and the b-values. Evaluation of the effect of motion on the DWI signal was done by computing the apparent diffusion coefficient (ADC) along (z-direction) and orthogonal (y- and x-directions) to the phantom. The computed ADCs of the phantom moving along Z were similar for the three orthogonal diffusion-encoding directions, with an average value of 1.65·10-9â¯, 1.66·10-9 and 1.65·10-9â¯m2/s along X, Y and Z respectively. DW phase images on the other hand showed the expected linear relationship with phantom velocity. Pure translational motion has minor effect on the diffusion-weighted magnitude signal. The sudden signal drop typically observed in in vivo spinal cord DWI is likely not caused by translational motion of the spinal cord, and possibly originates from non-rigid compression/stretching of the cord and/or from intra-voxel incoherent motion (IVIM).
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Imagem de Difusão por Ressonância Magnética/métodos , Coração/fisiologia , Imagens de Fantasmas , Medula Espinal/diagnóstico por imagem , Humanos , Movimento (Física) , Reprodutibilidade dos TestesRESUMO
HIGHLIGHTS AxonPacking: Open-source software for simulating white matter microstructure.Validation on a theoretical disk packing problem.Reproducible and stable for various densities and diameter distributions.Can be used to study interplay between myelin/fiber density and restricted fraction. Quantitative Magnetic Resonance Imaging (MRI) can provide parameters that describe white matter microstructure, such as the fiber volume fraction (FVF), the myelin volume fraction (MVF) or the axon volume fraction (AVF) via the fraction of restricted water (fr). While already being used for clinical application, the complex interplay between these parameters requires thorough validation via simulations. These simulations required a realistic, controlled and adaptable model of the white matter axons with the surrounding myelin sheath. While there already exist useful algorithms to perform this task, none of them combine optimisation of axon packing, presence of myelin sheath and availability as free and open source software. Here, we introduce a novel disk packing algorithm that addresses these issues. The performance of the algorithm is tested in term of reproducibility over 50 runs, resulting density, and stability over iterations. This tool was then used to derive multiple values of FVF and to study the impact of this parameter on fr and MVF in light of the known microstructure based on histology sample. The standard deviation of the axon density over runs was lower than 10-3 and the expected hexagonal packing for monodisperse disks was obtained with a density close to the optimal density (obtained: 0.892, theoretical: 0.907). Using an FVF ranging within [0.58, 0.82] and a mean inter-axon gap ranging within [0.1, 1.1] µm, MVF ranged within [0.32, 0.44] and fr ranged within [0.39, 0.71], which is consistent with the histology. The proposed algorithm is implemented in the open-source software AxonPacking (https://github.com/neuropoly/axonpacking) and can be useful for validating diffusion models as well as for enabling researchers to study the interplay between microstructure parameters when evaluating qMRI methods.
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Characterizing precisely the microstructure of axons, their density, size and myelination is of interest for the neuroscientific community, for example to help maximize the outcome of studies on white matter (WM) pathologies of the spinal cord (SC). The existence of a comprehensive and structured database of axonal measurements in healthy and disease models could help the validation of results obtained by different researchers. The purpose of this article is to provide such a database of healthy SC WM, to discuss the potential sources of variability and to suggest avenues for robust and accurate quantification of axon morphometry based on novel acquisition and processing techniques. The article is organized in three sections. The first section reviews morphometric results across species according to range of densities and counts of myelinated axons, axon diameter and myelin thickness, and characteristics of unmyelinated axons in different regions. The second section discusses the sources of variability across studies, such as age, sex, spinal pathways, spinal levels, statistical power and terminology in regard to tracts and protocols. The third section presents new techniques and perspectives that could benefit histology studies. For example, coherent anti-stokes Raman spectroscopy (CARS) imaging can provide sub-micrometric resolution without the need for fixation and staining, while slide scanners and stitching algorithms can provide full cross-sectional area of SC. In combination with these acquisition techniques, automatic segmentation algorithms for delineating axons and myelin sheath can help provide large-scale statistics on axon morphometry.
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Segmenting axon and myelin from microscopic images is relevant for studying the peripheral and central nervous system and for validating new MRI techniques that aim at quantifying tissue microstructure. While several software packages have been proposed, their interface is sometimes limited and/or they are designed to work with a specific modality (e.g., scanning electron microscopy (SEM) only). Here we introduce AxonSeg, which allows to perform automatic axon and myelin segmentation on histology images, and to extract relevant morphometric information, such as axon diameter distribution, axon density and the myelin g-ratio. AxonSeg includes a simple and intuitive MATLAB-based graphical user interface (GUI) and can easily be adapted to a variety of imaging modalities. The main steps of AxonSeg consist of: (i) image pre-processing; (ii) pre-segmentation of axons over a cropped image and discriminant analysis (DA) to select the best parameters based on axon shape and intensity information; (iii) automatic axon and myelin segmentation over the full image; and (iv) atlas-based statistics to extract morphometric information. Segmentation results from standard optical microscopy (OM), SEM and coherent anti-Stokes Raman scattering (CARS) microscopy are presented, along with validation against manual segmentations. Being fully-automatic after a quick manual intervention on a cropped image, we believe AxonSeg will be useful to researchers interested in large throughput histology. AxonSeg is open source and freely available at: https://github.com/neuropoly/axonseg.
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We provide a detailed morphometric analysis of eight transmission electron micrographs (TEMs) obtained from the corpus callosum of one cynomolgus macaque. The raw TEM images are included in the article, along with the distributions of the axon caliber and the myelin g-ratio in each image. The distributions are analyzed to determine the relationship between axon caliber and g-ratio, and compared against the aggregate metrics (myelin volume fraction, fiber volume fraction, and the aggregate g-ratio), as defined in the accompanying research article entitled 'In vivo histology of the myelin g-ratio with magnetic resonance imaging' (Stikov et al., NeuroImage, 2015).