Label‐free identification of myopathological features with coherent anti‐Stokes Raman scattering

Niedieker, Daniel; Großerüschkamp, Frederik; Schreiner, Anja; Barkovits, Katalin; Kötting, Carsten; Marcus, Katrin; Gerwert, Klaus; Vorgerd, Matthias

doi:10.1002/mus.26140

Cited by 7 publications

(6 citation statements)

References 16 publications

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“…The laser power at the sample was 28 mW (Pump) and 21 mW (Stokes). Integration times of 34 s per image with a pixel dwell time of 32 µs, 1024 × 1024 pixels and a spatial resolution of 300 nm were used [54]. After the label-free detection of the lipid and protein distribution, immunofluorescent stainings were performed on the same sections (Online Resource Fig.…”

Section: Coherent Anti-stokes Raman Scatteringmentioning

confidence: 99%

The subcellular arrangement of alpha-synuclein proteoforms in the Parkinson’s disease brain as revealed by multicolor STED microscopy

et al. 2021

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Various post-translationally modified (PTM) proteoforms of alpha-synuclein (aSyn)—including C-terminally truncated (CTT) and Serine 129 phosphorylated (Ser129-p) aSyn—accumulate in Lewy bodies (LBs) in different regions of the Parkinson’s disease (PD) brain. Insight into the distribution of these proteoforms within LBs and subcellular compartments may aid in understanding the orchestration of Lewy pathology in PD. We applied epitope-specific antibodies against CTT and Ser129-p aSyn proteoforms and different aSyn domains in immunohistochemical multiple labelings on post-mortem brain tissue from PD patients and non-neurological, aged controls, which were scanned using high-resolution 3D multicolor confocal and stimulated emission depletion (STED) microscopy. Our multiple labeling setup highlighted a consistent onion skin-type 3D architecture in mature nigral LBs in which an intricate and structured-appearing framework of Ser129-p aSyn and cytoskeletal elements encapsulates a core enriched in CTT aSyn species. By label-free CARS microscopy we found that enrichments of proteins and lipids were mainly localized to the central portion of nigral aSyn-immunopositive (aSyn+) inclusions. Outside LBs, we observed that 122CTT aSyn+ punctae localized at mitochondrial membranes in the cytoplasm of neurons in PD and control brains, suggesting a physiological role for 122CTT aSyn outside of LBs. In contrast, very limited to no Ser129-p aSyn immunoreactivity was observed in brains of non-neurological controls, while the alignment of Ser129-p aSyn in a neuronal cytoplasmic network was characteristic for brains with (incidental) LB disease. Interestingly, Ser129-p aSyn+ network profiles were not only observed in neurons containing LBs but also in neurons without LBs particularly in donors at early disease stage, pointing towards a possible subcellular pathological phenotype preceding LB formation. Together, our high-resolution and 3D multicolor microscopy observations in the post-mortem human brain provide insights into potential mechanisms underlying a regulated LB morphogenesis.

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Section: Coherent Anti-stokes Raman Scatteringmentioning

confidence: 99%

The subcellular arrangement of alpha-synuclein proteoforms in the Parkinson’s disease brain as revealed by multicolor STED microscopy

et al. 2021

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“…Recently, Karuna et al reported label-free three-dimensional quantitative imaging of chemical composition of the human osteosarcoma cell division by using the hyperspectral CARS microscopy 186 . Niedieker et al applied CARS imaging to characterize the myopathological features in muscles from patients with different myopathies, and to identify the distinct specific disease features 187 . Hirose et al developed a CARS rigid endoscope using two individual optical fibers to image rat sciatic nerves and also rabbit prostatic fascia without sample slicing, which revealed the potential application of the CARS endoscope to robot-assisted surgery 188 .…”

Section: Coherent Anti-stokes Raman Scattering Microscopic Imagingmentioning

confidence: 99%

Recent advances in nonlinear optics for bio-imaging applications

Zhang

Liu²,

Ren

et al. 2020

Opto-Electronic Advances

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Nonlinear optics, which is a subject for studying the interaction between intense light and materials, has great impact on various research fields. Since many structures in biological tissues exhibit strong nonlinear optical effects, nonlinear optics has been widely applied in biomedical studies. Especially in the aspect of bio-imaging, nonlinear optical techniques can provide rapid, label-free and chemically specific imaging of biological samples, which enable the investigation of biological processes and analysis of samples beyond other microscopy techniques. In this review, we focus on the introduction of nonlinear optical processes and their applications in bio-imaging as well as the recent advances in this filed. Our perspective of this field is also presented.

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“…Surface‐enhanced Raman significantly boosts the signal but would require injection of nanoparticles into the tissue of interest 54 . Coherent anti‐Stokes Raman spectroscopy uses two beams of light to excite the sample and has been shown to effectively map out protein distributions in human muscle biopsy samples from a range of diseases 55 …”

Section: Discussionmentioning

confidence: 99%

Fiber optic Raman spectroscopy for the evaluation of disease state in Duchenne muscular dystrophy: An assessment using the mdx model and human muscle

et al. 2022

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Introduction/Aims: Raman spectroscopy is an emerging technique for the evaluation of muscle disease. In this study we evaluate the ability of in vivo intramuscular Raman spectroscopy to detect the effects of voluntary running in the mdx model of Duchenne muscular dystrophy (DMD). We also compare mdx data with muscle spectra from human DMD patients.Methods: Thirty 90-day-old mdx mice were randomly allocated to an exercised group (48-hour access to a running wheel) and an unexercised group (n = 15 per group). In vivo Raman spectra were collected from both gastrocnemius muscles and histopathological assessment subsequently performed. Raman data were analyzed using principal component analysis-fed linear discriminant analysis (PCA-LDA). Exercised and unexercised mdx muscle spectra were compared with human DMD samples using cosine similarity.Results: Exercised mice ran an average of 6.5 km over 48 hours, which induced a significant increase in muscle necrosis (P = .03). PCA-LDA scores were significantly different between the exercised and unexercised groups (P < .0001) and correlated significantly with distance run (P = .01). Raman spectra from exercised mice more closely resembled human spectra than those from unexercised mice.Discussion: Raman spectroscopy provides a readout of the biochemical alterations in muscle in both the mdx mouse and human DMD muscle.

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Label‐free identification of myopathological features with coherent anti‐Stokes Raman scattering

Abstract: CARS enables the label-free identification of distinct myopathological features, possibly paving the way for subsequent proteomic, metabolic, and genomic analyses. Muscle Nerve 58: 457-460, 2018.

Cited by 7 publications

References 16 publications

The subcellular arrangement of alpha-synuclein proteoforms in the Parkinson’s disease brain as revealed by multicolor STED microscopy

The subcellular arrangement of alpha-synuclein proteoforms in the Parkinson’s disease brain as revealed by multicolor STED microscopy

Recent advances in nonlinear optics for bio-imaging applications

Fiber optic Raman spectroscopy for the evaluation of disease state in Duchenne muscular dystrophy: An assessment using the mdx model and human muscle

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