RP‐CARS reveals molecular spatial order anomalies in myelin of an animal model of Krabbe disease

Vito, Giuseppe de; Cappello, Valentina; Tonazzini, Ilaria; Cecchini, Marco; Piazza, Vincenzo

doi:10.1002/jbio.201500305

Cited by 17 publications

(20 citation statements)

References 59 publications

(101 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Applications in biomedical optics are also promising, following the growing interest in monitoring lipid organization in myelin, a possible marker for early detection of neurodegenerative diseases [20][21][22]. Adapting polarized structural imaging for in vivo imaging is accessible, as shown already by in vivo CARS imaging [36]; however, it would necessitate the implementation of epi-detection modalities.…”

Section: Discussionmentioning

confidence: 99%

“…Myelin structural imaging has been demonstrated in fixed tissues in mouse brain [18,19] and sciatic nerve [20], as well as in the spinal cord of mice [17,21] and anesthetized zebrafish [22]. Being able to monitor lipid order without the need of fluorescent labels ultimately brings new knowledge on lipid packing properties, which is critical for the structural integrity of myelin in the central and peripheral nervous systems, with consequences in neuropathologies [20][21][22]. More generally, lipid packing plays a role in the maintenance of skin lipid barriers, in cell signaling, and in cell mechanics.…”

Section: Introductionmentioning

confidence: 99%

“…Yet currently developed techniques are not able to address fast time scale dynamics, the main reason being the requirement of point scanning modalities. Typical molecular order imaging is performed at the rate of minutes for sequential polarization tuning on images of 150 × 150 pixels [17,19,21,22], to tens of seconds in schemes using synchronization between line scanning and polarization rotation [20]. This limits the current applicability to minute-scale processes, with the obvious inconvenience of sensitivity to sample motion and lack of dynamics information.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

High-speed polarization-resolved coherent Raman scattering imaging

Hofer¹,

Balla²,

Brasselet³

2017

Optica

View full text Add to dashboard Cite

Polarization-resolved coherent Raman scattering (polar-CRS) provides rich information on molecular orientational organization, with the strong advantages of being a label-free and chemically specific imaging method. Its implementation, however, strongly reduces the imaging acquisition rate, due to limits imposed by polarization tuning. Here we demonstrate fast-polar-CRS imaging based on combined electro-optic polarization and acousto-optic amplitude modulations, applicable to both stimulated Raman scattering and coherent anti-Stokes Raman scattering imaging. The proposed scheme adds polarization information without compromising the capacities of regular CRS intensity imaging; increases the speed of orientational imaging by two orders of magnitude as compared with previous approaches; and does not require post-processing analyses. We show that this method permits sub-second time-scale imaging of lipid order packing and local lipid membrane deformations in artificial lipid multilayers, but also in red blood cell ghosts, demonstrating its high sensitivity down to a single lipid bilayer membrane.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

High-speed polarization-resolved coherent Raman scattering imaging

Hofer¹,

Balla²,

Brasselet³

2017

Optica

View full text Add to dashboard Cite

show abstract

“…Genomic DNA was extracted from the clipped tails of mice by Proteinase K lysis buffer 20 as previously described 21 . The genetic status of each mouse was determined from the genome analysis of the twitcher mutation, as reported in Sakai and coworkers 22 .…”

Section: Methodsmentioning

confidence: 99%

Size and specimen-dependent strategy for x-ray micro-ct and tem correlative analysis of nervous system samples

Parlanti

Cappello

Brun

et al. 2017

Sci Rep

View full text Add to dashboard Cite

Correlative approaches are a powerful tool in the investigation of biological samples, but require specific preparation procedures to maintain the strength of the employed methods. Here we report the optimization of the embedding protocol of nervous system samples for a correlative synchrotron X-ray computed microtomography (micro-CT) and transmission electron microscopy (TEM) approach. We demonstrate that it is possible to locate, with the micrometric resolution of micro-CT, specific volumes of interest for a further ultrastructural characterization to be performed with TEM. This approach can be applied to samples of different size and morphology up to several cm. Our optimized method represents an invaluable tool for investigating those pathologies in which microscopic alterations are localized in few confined regions, rather than diffused in entire tissues, organs or systems. We present a proof of concept of our method in a mouse model of Globoid Cells Leukodistrophy.

show abstract

“…CARS microscopy is frequently employed in myelin imaging, thanks to the strong CARS signal obtained by targeting its extremely abundant CH 2 bonds. Consequently, myelin has been studied using CARS imaging under normal physiologic [24,25] and pathologic [26][27][28][29] conditions.…”

Section: Introductionmentioning

confidence: 99%

Effect of scattering on coherent anti-Stokes Raman scattering (CARS) signals

Ranasinghesagara¹,

Vito²,

Piazza³

et al. 2017

Opt. Express

Self Cite

View full text Add to dashboard Cite

Abstract:We develop a computational framework to examine the factors responsible for scattering-induced distortions of coherent anti-Stokes Raman scattering (CARS) signals in turbid samples. We apply the Huygens-Fresnel Wave-based Electric Field Superposition (HF-WEFS) method combined with the radiating dipole approximation to compute the effects of scatteringinduced distortions of focal excitation fields on the far-field CARS signal. We analyze the effect of spherical scatterers, placed in the vicinity of the focal volume, on the CARS signal emitted by different objects (2µm diameter solid sphere, 2µm diameter myelin cylinder and 2µm diameter myelin tube). We find that distortions in the CARS signals arise not only from attenuation of the focal field but also from scattering-induced changes in the spatial phase that modifies the angular distribution of the CARS emission. Our simulations further show that CARS signal attenuation can be minimized by using a high numerical aperture condenser. Moreover, unlike the CARS intensity image, CARS images formed by taking the ratio of CARS signals obtained using xand y-polarized input fields is relatively insensitive to the effects of spherical scatterers. Our computational framework provide a mechanistic approach to characterizing scattering-induced distortions in coherent imaging of turbid media and may inspire bottom-up approaches for adaptive optical methods for image correction.

show abstract

RP‐CARS reveals molecular spatial order anomalies in myelin of an animal model of Krabbe disease

Cited by 17 publications

References 59 publications

High-speed polarization-resolved coherent Raman scattering imaging

High-speed polarization-resolved coherent Raman scattering imaging

Size and specimen-dependent strategy for x-ray micro-ct and tem correlative analysis of nervous system samples

Effect of scattering on coherent anti-Stokes Raman scattering (CARS) signals

Contact Info

Product

Resources

About