Local assessment of myelin health in a multiple sclerosis mouse model using a 2D Fourier transform approach

Bégin, Steve; Bélanger, Erik; Laffray, Sophie; Aubé, Benoît; Chamma, Émilie; Bélisle, Jonathan M.; Lacroix, Steve; Côté, Daniel

doi:10.1364/boe.4.002003

Cited by 24 publications

(23 citation statements)

References 31 publications

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“…Such debris, reported in the literature 7,8 , are found in two categories: "non filled debris", which show no lipid signal at their center, and filled debris that are pure lipids volumetric structures. We observed an increased amount of damaged or vesiculated myelin with the progression of the disease as previously reported 13,32 .…”

Section: B Relation Between Lipid Packing Orientational Order and Mysupporting

confidence: 89%

“…In CARS, a pump field (frequency ω p ) and a Stokes field (ω s ) interact with the sample to generate an anti-Stokes field at frequency 2ω p − ω s , which is enhanced when ω p − ω s is tuned to the resonant frequency of a Raman band [23][24][25] . By addressing lipid molecules by their CH vibration bonds, which are highly abundant in the myelin sheath, CARS microscopy has been used to image myelin in the mouse brain 26,27 , nerves of the peripheral nervous system 28,29 , as well as spinal cord tissues 13,14,27,[30][31][32][33][34] including during the progression of EAE 14,27,32 . Using CARS images, the commonly used parameter in the assessment of the myelin health by its morphology is the g-ratio, defined as the ratio between the inner and outer diameter of myelin fibers 14,[28][29][30]33,35 .…”

Section: Introductionmentioning

confidence: 99%

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Lipid order degradation in autoimmune demyelination probed by polarization resolved coherent Raman microscopy

Gasecka

Jaouën

Bioud

et al. 2017

Preprint

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Myelin around axons is currently widely studied by structural analyses and large scale imaging techniques, with the goal to decipher its critical role in neuronal protection. While there is strong evidence that in myelin, lipid composition and lipid membrane morphology are affected during the progression of neurodegenerative diseases, there is no quantitative method yet to report its ultrastructure in tissues at both molecular and macroscopic levels, in conditions potentially compatible with in vivo observations. In this work, we study and quantify molecular order of lipids in myelin at sub-diffraction scales, using label-free polarization resolved Coherent Anti Stokes Raman (PR-CARS), which exploits CARS sensitivity to coupling between light polarization and oriented molecular vibrational bonds. Importantly, the method does not use any a priori parameters in the sample such as lipid type, orientational organization and composition. We show that lipid molecular order of myelin in the mouse spinal cord is significantly reduced throughout the progression of experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, even in myelin regions that appear morphologically unaffected. This technique permits to unravel molecular-scale perturbations of lipid layers at early stage of the demyelination progression, while the membrane architecture at the mesoscopic scale (here about 100 nm) seems much less affected. Such information cannot be brought by pure morphological observation and opens new prospectives towards molecular-scale understanding of neurodegenerative diseases.peer-reviewed)

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Section: B Relation Between Lipid Packing Orientational Order and Mysupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Lipid order degradation in autoimmune demyelination probed by polarization resolved coherent Raman microscopy

Gasecka

Jaouën

Bioud

et al. 2017

Preprint

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“…For each position, the sample is imaged at various depths ( [46] to generate a 3D mosaic. However, because of the high scattering coefficient of myelin, there is usually only one optimal plane extracted from the 3D mosaic to form the 2D mosaic [44] suitable for the segmentation analysis. A complete mosaic of 5000 × 7000 pixels (typical) is typically acquired in 3.5 hours.…”

Section: Large-scale Mosaic Acquisitions Of Spinal Cord Cross Sectionsmentioning

confidence: 99%

“…the contrast can be tuned to the myelin lipid content [33][34][35][36]. Since then, it has been used to visualize demyelination [37][38][39][40][41], and we have proposed several techniques to characterize myelin morphology [42][43][44].…”

Section: Introductionmentioning

confidence: 99%

Automated method for the segmentation and morphometry of nerve fibers in large-scale CARS images of spinal cord tissue

Bégin

Dupont-Therrien

Bélanger

et al. 2014

Biomed. Opt. Express

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A fully automated method for large-scale segmentation of nerve fibers from coherent anti-Stokes Raman scattering (CARS) microscopy images is presented. The method is specifically designed for CARS images of transverse cross sections of nervous tissue but is also suitable for use with standard light microscopy images. After a detailed description of the two-part segmentation algorithm, its accuracy is quantified by comparing the resulting binary images to manually segmented images. We then demonstrate the ability of our method to retrieve morphological data from CARS images of nerve tissue. Finally, we present the segmentation of a large mosaic of CARS images covering more than half the area of a mouse spinal cord cross section and show evidence of clusters of neurons with similar g-ratios throughout the spinal cord.

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“…In CARS, a pump field (frequency u p ) and a Stokes field (u s ) interact with the sample to generate an anti-Stokes field at frequency 2u p -u s , which is enhanced when u p -u s is tuned to the resonant frequency of a Raman band (23)(24)(25). By addressing lipid molecules by their CH vibration bonds, which are highly abundant in the myelin sheath, CARS microscopy has been used to image myelin in the mouse brain (26,27), nerves of the peripheral nervous system (28,29), and spinal cord tissues (13,14,27,(30)(31)(32)(33)(34) including during the progression of EAE (14,27,32). Using CARS images, the commonly used parameter in the assessment of the myelin health by its morphology is the g ratio, defined as the ratio between the inner and outer diameter of myelin fibers (14,(28)(29)(30)33,35).…”

Section: Introductionmentioning

confidence: 99%

Lipid Order Degradation in Autoimmune Demyelination Probed by Polarized Coherent Raman Microscopy

Gasecka

Jaouën

Bioud

et al. 2017

Biophysical Journal

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Myelin around axons is currently widely studied by structural analyses and large-scale imaging techniques, with the goal to decipher its critical role in neuronal protection. Although there is strong evidence that in myelin, lipid composition, and lipid membrane morphology are affected during the progression of neurodegenerative diseases, there is no quantitative method yet to report its ultrastructure in tissues at both molecular and macroscopic levels, in conditions potentially compatible with in vivo observations. In this work, we study and quantify the molecular order of lipids in myelin at subdiffraction scales, using label-free polarization-resolved coherent anti-Stokes Raman, which exploits coherent anti-Stokes Raman sensitivity to coupling between light polarization and oriented molecular vibrational bonds. Importantly, the method does not use any a priori parameters in the sample such as lipid type, orientational organization, and composition. We show that lipid molecular order of myelin in the mouse spinal cord is significantly reduced throughout the progression of experimental autoimmune encephalomyelitis, a model for multiple sclerosis, even in myelin regions that appear morphologically unaffected. This technique permits us to unravel molecular-scale perturbations of lipid layers at an early stage of the demyelination progression, whereas the membrane architecture at the mesoscopic scale (here ∼100 nm) seems much less affected. Such information cannot be brought by pure morphological observation and, to our knowledge, brings a new perspective to molecular-scale understanding of neurodegenerative diseases.

show abstract

Local assessment of myelin health in a multiple sclerosis mouse model using a 2D Fourier transform approach

Cited by 24 publications

References 31 publications

Lipid order degradation in autoimmune demyelination probed by polarization resolved coherent Raman microscopy

Lipid order degradation in autoimmune demyelination probed by polarization resolved coherent Raman microscopy

Automated method for the segmentation and morphometry of nerve fibers in large-scale CARS images of spinal cord tissue

Lipid Order Degradation in Autoimmune Demyelination Probed by Polarized Coherent Raman Microscopy

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