Monitoring aging‐associated structural alterations in <scp><i>Caenorhabditis elegans</i></scp> striated muscles via <scp>polarization‐dependent second‐harmonic generation</scp> measurements

Tsafas, Vassilis; Giouroukou, Konstantina; Kounakis, Konstantinos; Mari, Meropi; Fotakis, C.; Tavernarakis, Nektarios; Filippidis, George

doi:10.1002/jbio.202100173

Cited by 3 publications

(1 citation statement)

References 31 publications

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“…S3 ). We may alternatively detect a violation of cylindrical symmetry by fitting P-SHG data with a theoretical equation based on trigonal symmetry 37 or by looking for discrepancies between φ 2 and φ 4 (see Supplementary Material) 38 , 39 . Application of an R 2 threshold is anyway easy to implement and efficient to eliminate unreliable P-SHG data 40 .…”

Section: Discussionmentioning

confidence: 99%

Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy

Raoux¹,

Chessel²,

Mahou³

et al. 2023

Light Sci Appl

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A key property of the human cornea is to maintain its curvature and consequently its refraction capability despite daily changes in intraocular pressure. This is closely related to the multiscale structure of the corneal stroma, which consists of 1–3 µm-thick stacked lamellae made of thin collagen fibrils. Nevertheless, the distribution, size, and orientation of these lamellae along the depth of the cornea are poorly characterized up to now. In this study, we use second harmonic generation (SHG) microscopy to visualize the collagen distribution over the full depth of 10 intact and unstained human corneas (500–600 µm thick). We take advantage of the small coherence length in epi-detection to axially resolve the lamellae while maintaining the corneal physiological curvature. Moreover, as raw epi-detected SHG images are spatially homogenous because of the sub-wavelength size of stromal collagen fibrils, we use a polarimetric approach to measure the collagen orientation in every voxel. After a careful validation of this approach, we show that the collagen lamellae (i) are mostly oriented along the inferior–superior axis in the anterior stroma and along the nasal-temporal axis in the posterior stroma, with a gradual shift in between and (ii) exhibit more disorder in the anterior stroma. These results represent the first quantitative characterization of the lamellar structure of the human cornea continuously along its entire thickness with micrometric resolution. It also shows the unique potential of P-SHG microscopy for imaging of collagen distribution in thick dense tissues.

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Section: Discussionmentioning

confidence: 99%

Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy

Raoux¹,

Chessel²,

Mahou³

et al. 2023

Light Sci Appl

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Extraction of collagen morphological features from second‐harmonic generation microscopy images viaGLCMandCTanalyses: A cross‐laboratory study

Cicchi,

Baria,

Mari

et al. 2024

Journal of Biophotonics

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Second‐harmonic generation (SHG) microscopy provides a high‐resolution label‐free approach for noninvasively detecting collagen organization and its pathological alterations. Up to date, several imaging analysis algorithms for extracting collagen morphological features from SHG images—such as fiber size and length, order and anisotropy—have been developed. However, the dependence of extracted features on experimental setting represents a significant obstacle for translating the methodology in the clinical practice. We tackled this problem by acquiring SHG images of the same kind of collagenous sample in various laboratories using different experimental setups and imaging conditions. The acquired images were analyzed by commonly used algorithms, such as gray‐level co‐occurrence matrix or curvelet transform; the extracted morphological features were compared, finding that they strongly depend on some experimental parameters, whereas they are almost independent from others. We conclude with useful suggestions for comparing results obtained in different labs using different experimental setups and conditions.

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Comparison between Cylindrical, Trigonal, and General Symmetry Models for the Analysis of Polarization-Dependent Second Harmonic Generation Measurements Acquired from Collagen-Rich Equine Pericardium Samples

et al. 2022

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Polarization-dependent second harmonic generation (PSHG) microscopy is used as an innovative, high-resolution, non-destructive, and label-free diagnostic imaging tool to elucidate biological issues with high significance. In the present study, information on the structure and directionality of collagen fibers in equine pericardium tissue was collected using PSHG imaging measurements. In an effort to acquire precise results, three different mathematical models (cylindrical, trigonal, and general) were applied to the analysis of the recorded PSHG datasets. A factor called the “ratio parameter” was calculated to provide quantitative information. The implementation of the trigonal symmetry model to the recorded data led to the extraction of improved results compared with the application of the widely used cylindrical symmetry model. The best outcome was achieved through the application of the general model that does not include any kind of symmetry for the data processing. Our findings suggest that the trigonal symmetry model is preferable for the analysis of the PSHG datasets acquired from the collagenous tissues compared with the cylindrical model approach although an increased computational time is required.

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Monitoring aging‐associated structural alterations in Caenorhabditis elegans striated muscles via polarization‐dependent second‐harmonic generation measurements

Cited by 3 publications

References 31 publications

Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy

Unveiling the lamellar structure of the human cornea over its full thickness using polarization-resolved SHG microscopy

Extraction of collagen morphological features from second‐harmonic generation microscopy images viaGLCMandCTanalyses: A cross‐laboratory study

Comparison between Cylindrical, Trigonal, and General Symmetry Models for the Analysis of Polarization-Dependent Second Harmonic Generation Measurements Acquired from Collagen-Rich Equine Pericardium Samples

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