CIIMAR-Interdisciplinary Centre of Marine and Environmental Research, University of Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos, s/n, 4450-208 Matosinhos, Portugal.
Department of Biology, Faculty of Sciences of the University of Porto, Rua do Campo Alegre, s/n, 4169-007 Porto, Portugal.
Biosensors (Basel). 2024 Nov 6;14(11):538. doi: 10.3390/bios14110538.
As a highly sensitive vibrational technique, Raman spectroscopy (RS) can provide valuable chemical and molecular data useful to characterise animal cell types, tissues and organs. As a label-free, rapid detection method, RS has been considered a valuable asset in forensics, biology and medicine. The technique has been applied to zebrafish for various purposes, including physiological, biochemical or bioaccumulation analyses. The available data point out its potential for the early diagnosis of detrimental effects elicited by toxicant exposure. Nevertheless, no baseline spectra are available for zebrafish embryos and larvae that could allow for suitable planning of toxicological assessments, comparison with toxicant-elicited spectra or mechanistic understanding of biochemical and physiological responses to the exposures. With this in mind, this work carried out a baseline characterisation of Raman spectra of zebrafish embryos and larvae throughout early development. Raman spectra were recorded from the iris, forebrain, melanocytes, heart, muscle and swim bladder between 24 and 168 h post-fertilisation. A chemometrics approach, based on partial least-squares discriminant analysis (PLS-DA), was used to obtain a Raman characterisation of each tissue or organ. In total, 117 Raman bands were identified, of which 24 were well represented and, thus, retained in the data analysed. Only three bands were found to be common to all organs and tissues. The PLS-DA provided a tentative Raman spectral fingerprint typical of each tissue or organ, reflecting the ongoing developmental dynamics. The bands showed frequencies previously assigned to collagen, cholesterol, various essential amino acids, carbohydrates and nucleic acids.
作为一种高灵敏度的振动技术，拉曼光谱（RS）可以提供有价值的化学和分子数据，有助于表征动物细胞类型、组织和器官。作为一种无标记、快速检测方法，RS 已被认为是法医学、生物学和医学的宝贵资产。该技术已应用于斑马鱼，用于各种目的，包括生理、生化或生物积累分析。现有数据表明，它具有早期诊断毒物暴露引起的有害影响的潜力。然而，对于斑马鱼胚胎和幼虫，没有基线光谱可供使用，这可能会影响到毒理学评估的合适规划、与毒物诱导的光谱进行比较或对暴露引起的生化和生理反应的机制理解。考虑到这一点，本工作对斑马鱼胚胎和幼虫在整个早期发育过程中的拉曼光谱进行了基线特征描述。在受精后 24 至 168 小时之间，从虹膜、前脑、黑素细胞、心脏、肌肉和鳔记录了拉曼光谱。采用基于偏最小二乘判别分析（PLS-DA）的化学计量学方法，对每个组织或器官的拉曼光谱进行了分析。总共鉴定出 117 个拉曼带，其中 24 个带得到了很好的代表，因此保留在分析的数据中。只有三个带被发现存在于所有器官和组织中。PLS-DA 提供了每个组织或器官的暂定拉曼光谱指纹，反映了正在进行的发育动态。这些带的频率与以前分配给胶原蛋白、胆固醇、各种必需氨基酸、碳水化合物和核酸的频率相对应。
