Compositional and structural characterization of dorsal root ganglion neurons and co-cultured Schwann cells by confocal Raman microspectral imaging

IF 2.7 3区 化学 Q2 CHEMISTRY, ANALYTICAL Vibrational Spectroscopy Pub Date : 2024-01-01 DOI:10.1016/j.vibspec.2023.103642
Jie Li
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Abstract

Confocal Raman microspectral imaging (CRMI) is a versatile non-invasive technique that allows in vitro cell imaging without any chemical fixation, fluorescent markers or other contrast-enhancing chemicals. In this study, dorsal root ganglion (DRG) neuron and its affiliated Schwann cells (SCs) were co-cultured to unveil their underlying sub-cellular constitutional and structural nature. Both DRG neurons and SCs were derived from neonatal Sprague-Dawley rats and seeded on CaF2 subtracts for spectral analysis. After acquiring Raman hyperspectral datasets, multivariate data analyses, including K-mean cluster analysis (KCA) and principal component analysis (PCA), were successively adopted to study the subcellular structural and compositional information of the measured cells. Univariate spectral analysis was adopted to emphasize the spatial distribution of subcellular constitutions based on the acquired spectral characteristics. Results have shown Raman spectral characteristics of DRG neurons (cell membrane, cytoplasm, organelles, nucleus) and its affiliated SCs (myelin, cell membrane, cytoplasm, nucleus), as well as information on the subcellular distribution pattern of major biochemical components (proteins, cytochrome c, nucleic acids, lipids, carbohydrates). This in vitro spectral-imaging work provides a proof of principle of an analytical method for future studies on the developmental mechanisms of DRG neurons and their molecular bases for the treatment of diseases of the peripheral nervous system.

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通过共焦拉曼微光谱成像分析背根神经节神经元和共培养许旺细胞的组成和结构特征
共焦拉曼微光谱成像(CRMI)是一种多功能的非侵入性技术,无需化学固定、荧光标记或其他增强对比度的化学物质,即可进行体外细胞成像。在这项研究中,背根神经节(DRG)神经元与其附属的许旺细胞(SCs)被共同培养,以揭示其潜在的亚细胞形态和结构性质。DRG 神经元和 Schwann 细胞均来自新生 Sprague-Dawley 大鼠,并播种在 CaF2 减影剂上进行光谱分析。在获得拉曼高光谱数据集后,先后采用了包括均值聚类分析(KCA)和主成分分析(PCA)在内的多元数据分析来研究被测细胞的亚细胞结构和成分信息。根据获得的光谱特征,采用单变量光谱分析来强调亚细胞结构的空间分布。结果显示了 DRG 神经元(细胞膜、细胞质、细胞器、细胞核)及其附属 SC(髓鞘、细胞膜、细胞质、细胞核)的拉曼光谱特征,以及主要生化成分(蛋白质、细胞色素 c、核酸、脂类、碳水化合物)的亚细胞分布模式信息。这项体外光谱成像工作为今后研究 DRG 神经元的发育机制及其治疗周围神经系统疾病的分子基础提供了一种分析方法的原理证明。
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来源期刊
Vibrational Spectroscopy
Vibrational Spectroscopy 化学-分析化学
CiteScore
4.70
自引率
4.00%
发文量
103
审稿时长
52 days
期刊介绍: Vibrational Spectroscopy provides a vehicle for the publication of original research that focuses on vibrational spectroscopy. This covers infrared, near-infrared and Raman spectroscopies and publishes papers dealing with developments in applications, theory, techniques and instrumentation. The topics covered by the journal include: Sampling techniques, Vibrational spectroscopy coupled with separation techniques, Instrumentation (Fourier transform, conventional and laser based), Data manipulation, Spectra-structure correlation and group frequencies. The application areas covered include: Analytical chemistry, Bio-organic and bio-inorganic chemistry, Organic chemistry, Inorganic chemistry, Catalysis, Environmental science, Industrial chemistry, Materials science, Physical chemistry, Polymer science, Process control, Specialized problem solving.
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