Raman spectroscopic histology using machine learning for nonalcoholic fatty liver disease

Helal, Khalifa Mohammad; Taylor, J. Nicholas; Cahyadi, Harsono; Okajima, Akira; Tabata, Kenji; Itoh, Yoshito; Tanaka, Hideo; Fujita, Katsumasa; Harada, Yoshinori; Komatsuzaki, Tamiki

doi:10.1002/1873-3468.13520

Cited by 18 publications

(17 citation statements)

References 31 publications

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“…However, evaluation of myocardial viability for determining the surgical procedures has hitherto been performed only by limited modalities, i.e., myocardial scintigraphy and gadolinium-enhanced magnetic resonance imaging, both of which are utilized only preoperatively and not available for precise intraoperative evaluation [ 11 , 38 ]. Regarding this issue, Raman spectroscopic analysis may be a promising candidate, which provides clues to identify the molecular structure of substances without using any specific chemical labeling, e.g., by fluorescent dyes [ 14 , 15 , 17 , 21 , 29 , 33 ]: specific Raman shifts in Raman spectrum acquired under green light excitation can be used to unveil changes of molecules such as reduced cytochromes c and b, oxygenated myoglobin and lipids [ 1 , 6 , 20 , 31 ]. We have previously demonstrated that myocardial ischemia and infarct could be detected quantitatively by Raman scattering light, which represents the energy states of molecular vibrations [ 26 , 27 , 37 ].…”

Section: Introductionmentioning

confidence: 99%

Raman Spectroscopic Assessment of Myocardial Viability in Langendorff-Perfused Ischemic Rat Hearts

Ikemoto

Hashimoto

Harada

et al. 2021

Acta Histochem. Cytochem

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Spontaneous Raman spectroscopy, which senses changes in cellular contents of reduced cytochrome c, could be a powerful tool for label-free evaluation of ischemic hearts. However, undetermined is whether it is applicable to evaluation of myocardial viability in ischemic hearts. To address this issue, we investigated sequential changes in Raman spectra of the subepicardial myocardium in the Langendorff-perfused rat heart before and during ligation of the left coronary artery and its subsequent release and re-ligation. Under 532-nm wavelength excitation, the Raman peak intensity of reduced cytochrome c at 747 cm −1 increased quickly after the coronary ligation, and reached a quasi-steady state within 30 min. Subsequent reperfusion of the heart after a short-term (30-min) ligation that simulates reversible conditions resulted in quick recovery of the peak intensity to the baseline. Further re-ligation resulted in resurgence of the peak intensity to nearly the identical value to the first ischemia value. In contrast, reperfusion after prolonged (120-min) ligation that assumes irreversible states resulted in incomplete recovery of the peak intensity, and re-ligation resulted in inadequate resurgence. Electron microscopic observations confirmed the spectral findings. Together, the Raman spectroscopic measurement for cytochrome c could be applicable to evaluation of viability of the ischemic myocardium without labeling.

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Section: Introductionmentioning

confidence: 99%

Raman Spectroscopic Assessment of Myocardial Viability in Langendorff-Perfused Ischemic Rat Hearts

Ikemoto

Hashimoto

Harada

et al. 2021

Acta Histochem. Cytochem

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“…Raman microscopy is, therefore, a powerful tool for the investigation of lipids in cells and tissues 10 – 15 . Although some researchers applied the Raman microscopy to NAFLD 16 – 18 , the efficacy of Raman microscopy for the investigation of NAFLD is still under investigation.…”

Section: Introductionmentioning

confidence: 99%

Molecular imaging analysis of microvesicular and macrovesicular lipid droplets in non-alcoholic fatty liver disease by Raman microscopy

Minamikawa

Ichimura‐Shimizu

Takanari

et al. 2020

Sci Rep

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Predominant evidence of non-alcoholic fatty liver disease (NAFLD) is the accumulation of excess lipids in the liver. A small group with NAFLD may have a more serious condition named non-alcoholic steatohepatitis (NASH). However, there is a lack of investigation of the accumulated lipids with spatial and molecular information. Raman microscopy has the potential to characterise molecular species and structures of lipids based on molecular vibration and can achieve high spatial resolution at the organelle level. In this study, we aim to demonstrate the feasibility of Raman microscopy for the investigation of NAFLD based on the molecular features of accumulated lipids. By applying the Raman microscopy to the liver of the NASH model mice, we succeeded in visualising the distribution of lipid droplets (LDs) in hepatocytes. The detailed analysis of Raman spectra revealed the difference of molecular structural features of the LDs, such as the degree of saturation of lipids in the LDs. We also found that the inhomogeneous distribution of cholesterol in the LDs depending on the histology of lipid accumulation. We visualised and characterised the lipids of NASH model mice by Raman microscopy at organelle level. Our findings demonstrated that the Raman imaging analysis was feasible to characterise the NAFLD in terms of the molecular species and structures of lipids.

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“…Minamikawa et al demonstrated the difference in chemistry between large and small LDs in a nonalcoholic steatohepatitis (NASH) model mice [ 20 ]. In our previous study, we combined Raman imaging with machine learning to evaluate NAFLD liver tissue samples in rats and grouped tissues with similar composition, providing a descriptor that enabled inference of tissue states, thus contributing valuable information to histological inspection of NAFLD [ 11 ]. However, it is unclear whether spontaneous Raman microscopy is applicable to the evaluation of the nascent state of NAFLD; the morphological features of NAFLD are not clear in this state and the intensity of spontaneous Raman scattering light is generally weak [ 9 ].…”

Section: Introductionmentioning

confidence: 99%

Label-free Assessment of the Nascent State of Rat Non-alcoholic Fatty Liver Disease Using Spontaneous Raman Microscopy

Takemura

Mochizuki

Harada

et al. 2022

Acta Histochem. Cytochem.

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Spontaneous Raman microscopy, which can detect molecular vibrations in cells and tissues, could be a useful tool for the label-free assessment of non-alcoholic fatty liver disease (NAFLD). However, it is unclear whether it can be used to evaluate the nascent state of NAFLD. To address this, we analyzed the Raman spectra of rat liver tissues in the nascent state of NAFLD upon excitation at 532 nm. Raman and histochemical analyses were performed of liver tissues from rats fed a high-fat, high-cholesterol diet (HFHCD). Raman microscopic imaging analysis of formalin-fixed thin tissue slices showed hepatic steatosis, as revealed by the Raman band at 2,854 cm −1 , whereas lipid droplets were not detectable by hematoxylin-eosin staining of images until 3 days after feeding a HFHCD. Raman signals of retinol at 1,588 cm −1 emitted from hepatic stellate cells were distributed alongside hepatic cords; the retinol content rapidly decreased after feeding a HFHCD, whereas hepatic lipid content increased inversely. Raman microscopic analysis of the surface of fresh ex vivo livers enabled early detection of lipid accumulation after a 1-day feeding a HFHCD. In conclusion, spontaneous Raman microscopy can be applied to the label-free evaluation of the nascent state of NAFLD liver tissues.

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Raman spectroscopic histology using machine learning for nonalcoholic fatty liver disease

Cited by 18 publications

References 31 publications

Raman Spectroscopic Assessment of Myocardial Viability in Langendorff-Perfused Ischemic Rat Hearts

Raman Spectroscopic Assessment of Myocardial Viability in Langendorff-Perfused Ischemic Rat Hearts

Molecular imaging analysis of microvesicular and macrovesicular lipid droplets in non-alcoholic fatty liver disease by Raman microscopy

Label-free Assessment of the Nascent State of Rat Non-alcoholic Fatty Liver Disease Using Spontaneous Raman Microscopy

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