拉曼光谱技术能够提供与物质特定分子结构相关的光谱信息, 可用于识别生物组织微小的生化变异, 具有快速、 实时、 无损、 无需样本预处理等优点, 在临床病理诊断领域极具应用前景。 与常规组织病理学分析相比, 拉曼光谱技术能够直接检测活体组织, 简化了分析程序, 缩短了诊断时间。 人体病变组织的细胞分子组成和结构可能发生变化, 这为拉曼光谱技术在组织病理诊断中的应用提供了检测依据。 基于组织分子组成与结构的差异, 结合机器学习和化学计量学方法, 拉曼光谱技术可以提供客观的诊断信息, 实现快速、 低侵入的病理诊断。 回顾了近十年来拉曼光谱技术在组织病理诊断中的研究进展, 对取得的关键成果进行了总结, 阐述了当前离体和活体应用拉曼光谱技术的一些关键问题。 针对离体拉曼光谱检测, 重点评估福尔马林固定石蜡包埋样本、 冷冻样本和新鲜组织样本等离体样本的适用情况; 阐述拉曼光谱数据收集的关键技术, 包括适用光源、 光谱范围, 以及病理样本光谱采集的方式等。 对于活体拉曼光谱检测, 重点介绍了活体检测研究中拉曼光谱技术应用的两种形式: 结合医用内窥镜进行体内检测, 以及开放手术中的直接检测; 综述了临床适用的拉曼系统, 重点介绍了当前活体拉曼研究中应用的光纤探头。 同时, 文章也讨论了拉曼光谱数据的处理与分析方法, 通过光谱预处理, 特征提取与分类识别, 构建拉曼光谱病理诊断模型, 在小样本范围能够获得较好的诊断结果。 考虑临床实际应用, 仍需要不断优化分析方法, 实现拉曼光谱与生化信息的关联, 将样本个体差异的影响纳入分类模型中, 以提升模型性能。 文章对拉曼光谱应用于病理诊断中的关键问题进行了讨论, 为进一步开展研究提供参考。 未来需要更深入和广泛地开展离体和活体研究, 以促进拉曼光谱技术在临床中的应用。

Raman spectroscopy can provide the spectral information related to the specific molecular structure of the substance, and recognize the tiny biochemical variation of biological tissue. It is a promising diagnosis technology in clinical applications due to the advantages of fast, real-time, non-destructive detection without sample pretreatment. Compared with the routine histopathological analysis, Raman spectroscopy can detect the fresh tissue directly and it may simplify the analysis procedure and shorten the diagnosis time. Some changes may take place in molecular composition and structure of pathological tissues, which provides a theoretical basis for the application of Raman spectroscopy in histopathological diagnosis. Raman spectroscopy may provide the objective diagnosis information and achieve the rapid, low-invasive diagnosis based on the differences of molecular composition and structure between normal and pathological tissues. This paper reviewed the development of Raman spectroscopy for the pathological diagnosis of tissues in the past decade and focused on the crucial results. Some key issues in the ex vivo and in vivo studies of Raman spectroscopy in pathological diagnosis were emphasized. Firstly, for the Raman detection of ex vivo sample, the applicability of several tissue samples, such as formal infixed paraffin-embedded samples, frozen samples and fresh tissue samples, was mainly evaluated; And the emphasis of Ramandata collection in the ex vivo studies of Raman spectroscopy, including applicable laser source, spectra range, and the way of spectra collection were summarized. Meanwhile, we focused on the two application forms of Raman spectroscopy in the detection in vivo, which involved the detection of Raman spectroscopy combined with the medical endoscope, and the direct detection in open surgery. The Raman system suitable for clinical application was also taken into consideration in this review, and the fiber probes used in the current in vivo Raman studies were major. Furthermore, the analysis methods of Raman data were discussed, the pathological diagnosis models, which can provide the excellent diagnosis results in the sample set with small size, have been established through spectral preprocessing, feature extraction, classification and identification. It is necessary to optimize the analysis method to achieve the correlation between Raman spectraand biochemical information and incorporate the effects of individual differences into the classification model to improve the model. In this paper, the crucial issues of Raman spectroscopy in pathological diagnosis were discussed in order to provide a reference for further research. More extensive ex vivo and in vivo studies are required to put the Raman spectroscopy technology into clinical practice.