Label-free imaging and quantitative chemical analysis of Alzheimer’s disease brain samples with multimodal multiphoton nonlinear optical microspectroscopy

Lee, Janghyuk; Kim, Dae Hwan; Song, Woo Keun; Oh, Myoung-Kyu; Ko, Do‐Kyeong

doi:10.1117/1.jbo.20.5.056013

Cited by 30 publications

(27 citation statements)

References 27 publications

(36 reference statements)

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“…[58][59][60] In this paper, we show that these techniques produce images with high contrast for the amyloids. This can be attributed to the previously demonstrated multiphoton absorption (MPA), [61] being as effective for insulin fibers as for typical two-photon dyes.…”

Section: Introductionmentioning

confidence: 73%

“…Enhanced intrinsic fluorescence, strong MPA and intrinsic molecular ordering are a combination of features that likely distinguishes amyloids from most other structures in normal tissue. [58][59][60] A visualization strategy including all these features would be a helpful tool to selectively find and characterize amyloid deposits in, for example, brain tissue from patients with Alzheimer's disease, without the use of any labels or probes. As a result, non-destructive identification of amyloids in such tissues becomes possible without introduction of any contaminating agents.…”

Section: Resultsmentioning

confidence: 99%

“…As a result, non-destructive identification of amyloids in such tissues becomes possible without introduction of any contaminating agents. This subsequently allows chemical analyses of the identified amyloid structures by vibrational sum-frequency scattering spectroscopy, [70] coherent anti-stokes Raman scattering microscopy, [60,71] and time-of-flight secondary ion mass spectrometry, [72] among other techniques. The fundamental optical properties of amyloids discussed and presented in this paper are essential for continuing efforts with the goal of providing innovative strategies for the study of amyloid fiber structures.…”

Section: Resultsmentioning

confidence: 99%

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Label-free imaging of amyloids using their intrinsic linear and nonlinear optical properties

Johansson¹,

Koelsch²

2017

Biomed. Opt. Express

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The optical properties of amyloid fibers are often distinct from those of the source protein in its non-fibrillar form. These differences can be utilized for label-free imaging or characterization of such structures, which is particularly important for understanding amyloid fiber related diseases such as Alzheimer's and Parkinson's disease. We demonstrate that two amyloid forming proteins, insulin and β-lactoglobulin (β-LG), show intrinsic fluorescence with emission spectra that are dependent on the excitation wavelength. Additionally, a new fluorescence peak at about 430 nm emerges for β-LG in its amyloid state. The shift in emission wavelength is related to the red edge excitation shift (REES), whereas the additional fluorescence peak is likely associated with charge delocalization along the fiber backbone. Furthermore, the spherulitic amyloid plaque-like superstructures formed from the respective proteins were imaged label-free with confocal fluorescence, multiphoton excitation fluorescence (MPEF), and second-harmonic generation (SHG) microscopy. The latter two techniques in particular yield images with a high contrast between the amyloid fiber regions and the core of amorphously structured protein. Strong multiphoton absorption (MPA) for the amyloid fibers is a likely contributor to the observed contrast in the MPEF images. The crystalline fibrillar region provides even higher contrast in the SHG images, due to the inherently ordered non-centrosymmetric structure of the fibers together with their non-isotropic arrangement. Finally, we show that MPEF from the insulin spherulites exhibits a spectral dependence on the excitation wavelength. This behavior is consistent with the REES phenomenon, which we hypothesize is the origin of this observation. The presented results suggest that amyloid deposits can be identified and structurally characterized based on their intrinsic optical properties, which is important for probe-less and label-free identification and characterization of amyloid fibers in vitro and in complex biological samples. A. 82(12), 4245-4249 (1985). 8. L. C. Serpell, "Alzheimer's amyloid fibrils: structure and assembly," Biochim. Biophys. Acta 1502(1), 16-30 (2000

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Section: Introductionmentioning

confidence: 73%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Label-free imaging of amyloids using their intrinsic linear and nonlinear optical properties

Johansson¹,

Koelsch²

2017

Biomed. Opt. Express

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“…1B). The interplay between proteins and lipids has important implications in cancer (36,37) and Alzheimer's disease (38,39). For the cell-silent window, many emerging vibrational tags reside in this region (40)(41)(42)(43)(44), allowing imaging of metabolic dynamics of small metabolites and also super multiplexed imaging (45,46).…”

Section: Development Of Raman-tailored Reagents For Clearing-enhancedmentioning

confidence: 99%

Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

Wei

Shi

Shen

et al. 2019

Proc. Natl. Acad. Sci. U.S.A.

110

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Three-dimensional visualization of tissue structures using optical microscopy facilitates the understanding of biological functions. However, optical microscopy is limited in tissue penetration due to severe light scattering. Recently, a series of tissue-clearing techniques have emerged to allow significant depth-extension for fluorescence imaging. Inspired by these advances, we develop a volumetric chemical imaging technique that couples Raman-tailored tissue-clearing with stimulated Raman scattering (SRS) microscopy. Compared with the standard SRS, the clearing-enhanced SRS achieves greater than 10-times depth increase. Based on the extracted spatial distribution of proteins and lipids, our method reveals intricate 3D organizations of tumor spheroids, mouse brain tissues, and tumor xenografts. We further develop volumetric phasor analysis of multispectral SRS images for chemically specific clustering and segmentation in 3D. Moreover, going beyond the conventional label-free paradigm, we demonstrate metabolic volumetric chemical imaging, which allows us to simultaneously map out metabolic activities of protein and lipid synthesis in glioblastoma. Together, these results support volumetric chemical imaging as a valuable tool for elucidating comprehensive 3D structures, compositions, and functions in diverse biological contexts, complementing the prevailing volumetric fluorescence microscopy.

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“…In recent years, CRS has emerged as a powerful approach for label-free, biochemical imaging of neuronal structures and for dynamic monitoring of functional parameters such as membrane potential 4 and neurotransmitter concentrations 5 . In addition, CRS has been used in fundamental research on neurodegenerative diseases, to characterize Alzheimer's pathology 6,7,8 , Parkinson's Disease 9 and multiple sclerosis 10 , or to study peripheral nerve degeneration in ALS 11 . Currently, there is a major push to develop label-free histopathological analyses based on CRS (sometimes integrated in a multi-modal approach with other nonlinear optical imaging techniques such as two-photon fluorescence or harmonic-generation microscopy) 12 , with major efforts in brain cancer 13,14,15 , other cancers 16,17,18 and a range of other diseases 19,20 .…”

Section: Introductionmentioning

confidence: 99%

Label-free characterization of Amyloid-β-plaques and associated lipids in brain tissues using stimulated Raman scattering microscopy

Schweikhard¹,

Baral

Krishnamachari³

et al. 2019

Preprint

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The brains of patients with neurodegenerative diseases such as Alzheimer's Disease (AD) often exhibit pathological alterations that involve abnormal aggregations of proteins and lipids. Here, we demonstrate that high-resolution, label-free, chemically-specific imaging using Stimulated Raman Scattering Microscopy (SRS) provides novel insights into the biophysical properties and biochemical composition of such pathological structures. In brain slices of a mouse model of AD, SRS reveals large numbers of Amyloid-β plaques that commonly form a characteristic, three-dimensional core-shell structure, with a fibrillar proteinaceous core surrounded by a halo-like shell of lipid-rich deposits. SRS spectroscopic imaging allows for a clean, label-free visualization of the misfolded (β-sheet) Amyloid-β content in the plaque core. Surrounding lipid-rich deposits are found to contain comparatively high concentrations of membrane lipids (sphingomyelin, phosphatidylcholine), but lower levels of cholesterol than healthy white matter structures. Overall, the SRS spectra of plaque-associated lipids closely resemble those of nearby neurites, with the notable difference of a higher degree of lipid unsaturation compared to healthy brain structures. We hypothesize that plaque-associated lipid deposits may result from neuritic dystrophy associated with AD, and that the observed increased levels of unsaturation could help identify the kinds of pathological alterations taking place. Taken together, our results highlight the potential of Stimulated Raman Scattering microscopy to contribute to a deeper understanding of neurodegenerative diseases.

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Label-free imaging and quantitative chemical analysis of Alzheimer’s disease brain samples with multimodal multiphoton nonlinear optical microspectroscopy

Cited by 30 publications

References 27 publications

Label-free imaging of amyloids using their intrinsic linear and nonlinear optical properties

Label-free imaging of amyloids using their intrinsic linear and nonlinear optical properties

Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

Label-free characterization of Amyloid-β-plaques and associated lipids in brain tissues using stimulated Raman scattering microscopy

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