<i>In situ</i> assessment of mitochondrial respiratory activity and lipid metabolism of mouse oocytes using resonance Raman spectroscopy

Ishigaki, Mika; Kashiwagi, Shinsuke; Wakabayashi, Satoru; Hoshino, Yumi

doi:10.1039/d1an01106e

Cited by 6 publications

(3 citation statements)

References 35 publications

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“…The PC-3 loadings are positively correlated with bands from the methyl and methylene motions from proteins and peptides at 1448 cm −1 , 1299 cm −1 , and 1378 cm −1 , suggesting high content of methylated amino acids in the cytoplasm of AVAC and IPMC. Moreover, a strong positive correlation is also visible at 1128 cm −1 , which was found to be a spectroscopic marker of reduced cytochrome C in mitochondria [ 49 ]. The negative correlation of PC-3 loadings, which is characteristic of spectra acquired from cytoplasmic areas of cPDAC, is observed for the band from the C-C 6 H 5 stretching in Phe and tryptophan (Trp) at 1208 cm −1 indicating the presence of Phe- and Trp-rich proteins in cPDAC cells cytoplasm.…”

Section: Resultsmentioning

confidence: 99%

Combined analytical approach empowers precise spectroscopic interpretation of subcellular components of pancreatic cancer cells

Szymoński,

Skirlińska-Nosek,

Lipiec

et al. 2023

Anal Bioanal Chem

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The lack of specific and sensitive early diagnostic options for pancreatic cancer (PC) results in patients being largely diagnosed with late-stage disease, thus inoperable and burdened with high mortality. Molecular spectroscopic methodologies, such as Raman or infrared spectroscopies, show promise in becoming a leader in screening for early-stage cancer diseases, including PC. However, should such technology be introduced, the identification of differentiating spectral features between various cancer types is required. This would not be possible without the precise extraction of spectra without the contamination by necrosis, inflammation, desmoplasia, or extracellular fluids such as mucous that surround tumor cells. Moreover, an efficient methodology for their interpretation has not been well defined. In this study, we compared different methods of spectral analysis to find the best for investigating the biomolecular composition of PC cells cytoplasm and nuclei separately. Sixteen PC tissue samples of main PC subtypes (ductal adenocarcinoma, intraductal papillary mucinous carcinoma, and ampulla of Vater carcinoma) were collected with Raman hyperspectral mapping, resulting in 191,355 Raman spectra and analyzed with comparative methodologies, specifically, hierarchical cluster analysis, non-negative matrix factorization, T-distributed stochastic neighbor embedding, principal components analysis (PCA), and convolutional neural networks (CNN). As a result, we propose an innovative approach to spectra classification by CNN, combined with PCA for molecular characterization. The CNN-based spectra classification achieved over 98% successful validation rate. Subsequent analyses of spectral features revealed differences among PC subtypes and between the cytoplasm and nuclei of their cells. Our study establishes an optimal methodology for cancer tissue spectral data classification and interpretation that allows precise and cognitive studies of cancer cells and their subcellular components, without mixing the results with cancer-surrounding tissue. As a proof of concept, we describe findings that add to the spectroscopic understanding of PC. Graphical Abstract

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Section: Resultsmentioning

confidence: 99%

Combined analytical approach empowers precise spectroscopic interpretation of subcellular components of pancreatic cancer cells

Szymoński,

Skirlińska-Nosek,

Lipiec

et al. 2023

Anal Bioanal Chem

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“…Cytochrome c also shows relatively strong and sharp resonant Raman peaks by measuring with a 532 nm excitation. It is useful to explore the cellular distribution of mitochondria on Raman microscopy [84][85][86]. Cytochrome c is a key molecule that maintains respiratory function and cell apoptosis.…”

Section: Molecular Fingerprints Possibly Associated With Diseases On ...mentioning

confidence: 99%

Practices, Potential, and Perspectives for Detecting Predisease Using Raman Spectroscopy

Oshima

Haruki

Koizumi

et al. 2023

IJMS

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Raman spectroscopy shows great potential for practical clinical applications. By analyzing the structure and composition of molecules through real-time, non-destructive measurements of the scattered light from living cells and tissues, it offers valuable insights. The Raman spectral data directly link to the molecular composition of the cells and tissues and provides a “molecular fingerprint” for various disease states. This review focuses on the practical and clinical applications of Raman spectroscopy, especially in the early detection of human diseases. Identifying predisease, which marks the transition from a healthy to a disease state, is crucial for effective interventions to prevent disease onset. Raman spectroscopy can reveal biological processes occurring during the transition states and may eventually detect the molecular dynamics in predisease conditions.

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“…Raman spectroscopy is a powerful tool expected to revolutionize chick sex determination because it can provide information about biological molecules, such as proteins, lipids, saccharides, and DNA/RNA, in a nondestructive and noninvasive manner 3 , 4 . Thus, Raman spectroscopy is suitable for analyzing living organisms, leading to its widespread adoption across various biological and medical applications 5 – 12 . Especially, hemes as chromophores exhibit distinctive electric absorption bands within the ultraviolet–visible (UV–vis) wavelength region 13 – 18 , thereby eliciting a strong resonance enhancement of Raman scattering selectively for hemes 19 , 20 .…”

Section: Introductionmentioning

confidence: 99%

Chick sexing based on the blood analysis using Raman spectroscopy

Matsumoto,

Ogino,

Onoe

et al. 2024

Sci Rep

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Efforts are underway to develop technology for automatically determining the sex of chick embryos, aimed at establishing a stable and efficient poultry farming system while also addressing animal welfare concerns. This study investigated the possibility of chick sexing through blood analysis using Raman spectroscopy. Raman spectra were obtained from whole blood and its constituents, such as red blood cells (RBCs) and blood plasma, collected from chicks aged 1–2 days, using a 785-nm excitation wavelength. Principal component analysis (PCA) revealed statistically significant sex-dependent spectral variations in whole blood and RBCs, whereas blood plasma showed less clear dependency. These spectral differences between male and female chicks were attributed to differences in the proportion of spectral components from oxygenated (oxy-) and deoxygenated (deoxy-) RBCs, with males exhibiting a slightly stronger contribution of oxy-RBCs compared to females. This reflects the higher oxygen affinity of hemoglobin (Hb) in males compared to females. A model for discriminating chick sex was built using the ratios of certain Raman band characteristics of oxy-RBCs and deoxy-RBCs, achieving a sensitivity of 100%. This spectroscopic method holds promise for developing technology to discriminate the sex of early chicken embryos in ovo by detecting differences in oxygen saturation of RBCs based on sex.

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In situ assessment of mitochondrial respiratory activity and lipid metabolism of mouse oocytes using resonance Raman spectroscopy

Abstract: The respiratory activity and lipid metabolism depending on the degree of oocyte maturation can be assessed using resonance Raman spectroscopy in a non-invasive manner.

Cited by 6 publications

References 35 publications

Combined analytical approach empowers precise spectroscopic interpretation of subcellular components of pancreatic cancer cells

Combined analytical approach empowers precise spectroscopic interpretation of subcellular components of pancreatic cancer cells

Practices, Potential, and Perspectives for Detecting Predisease Using Raman Spectroscopy

Chick sexing based on the blood analysis using Raman spectroscopy

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