无标记非线性光学显微技术在脑胶质瘤精准手术切除中组织学诊断的实验研究

In the relative research of "Brain Science", "Brain Project” and “Precision Medicine", the new method and technology of the effective diagnosis and treatment of brain major diseases has become a hot research topic in the world. Glioma is the most frequent of primary brain tumor and the mortality rate is the highest except for pancreatic cancer. "Maximum resection" is the basic principle of glioma surgery. However, the so-called “Maximum resection" is still stuck in the concept of "total resection" of the image. The real histological whole resection cannot be done. The new technology which can be able to accurately identify the histological boundary and infiltrative characteristics of tumor in the precisely surgical resection of brain glioma has become a significant clinical problem and demand in the neurosurgery. This project will aim at this significant demand and use the advantages of nonlinear optical microscopy based on two-photon excited fluorescence and second harmonic generation which can provide label-free, fast, real-time pathological diagnosis. We will investigate nonlinear optical diagnostic information that can identify brain glioma, determine the grading of glioma and glioma invasion scope, distinguish normal brain tissue and brain glioma boundary. This project will provide a new real-time medical imaging technology at the molecular level which can realize the histological maximum resection of brain glioma with the precisely surgical resection.

在“脑科学”、“脑计划”和“精准医疗”相关研究中，能够对脑重大疾病进行有效诊断和治疗的新方法新技术成为国内外共同关注的前沿研究热点。脑胶质瘤在原发性脑瘤中发病率位居首位，成为除胰腺癌之外死亡率最高的癌症。“最大范围切除”是胶质瘤手术基本原则。然而，所谓“最大范围切除”还停留在影像学 “全切除” 的概念，真正意义上的组织学全切除无法做到。能够在脑胶质瘤精准手术切除中，实现肿瘤组织学边界和浸润特征实时、准确确定的技术，成为目前神经外科的重大临床问题和需求。本项目针对这一需求，利用基于双光子激发荧光和二次谐波的非线性光学显微技术具有组织病理学高分辨率、可进行无标记快速实时病理诊断的优势，通过对脑胶质瘤识别、胶质瘤分级确定、明确胶质瘤侵犯范围、正常脑组织和脑胶质瘤边界划定的非线性光学诊断信息研究，为在脑胶质瘤精准手术切除中，实现脑胶质瘤“组织学最大范围切除”提供一个全新分子水平的实时医学成像技术。

脑胶质瘤是最常见的中枢神经系统原发性肿瘤，在原发性脑瘤中发病率位居首位。胶质瘤最大的生物学特性是肿瘤细胞呈浸润性生长，边界难于确定。能够在脑胶质瘤术中提供具有组织病理学高分辨率快速病理诊断的医学影像技术，成为脑胶质瘤手术 “组织学最大范围切除”的临床迫切需求。本项目利用基于双光子激发荧光和二次谐波的非线性光学显微技术具有组织病理学高分辨率、可以进行无标记快速实时病理学诊断的优势，开展脑胶质瘤识别、胶质瘤分级确定、明确胶质瘤侵犯范围、正常脑组织和脑胶质瘤边界划定的非线性光学诊断信息研究。.利用非线性光学显微技术，无标记成像脑组织微结构，包括脑膜、大脑皮层、脉络丛、胼胝体、纹状体、海马区、上丘脑、小脑皮层和第四脑室。进一步无标记成像了小脑显微结构，包括小脑蚓体、半球、髓质、脑室、轴突束、浦肯野细胞、毛细血管和软脑膜。证明了无标记非线性光学显微技术在脑肿瘤组织上进行快速病理诊断的可行性。.利用非线性光学显微技术，开展胶质瘤的分级研究，获得可以用于识别胶质瘤分级的光学诊断特征：细胞密度、胶质瘤细胞核的异形性、血管数量增生、血管内皮细胞增生、坏死和胶原沉积。结合自主开发的算法，获得定量光学诊断参数：细胞核的周长、细胞密度、胶原含量、细胞核质比。建立光学诊断信息与脑胶质瘤发展进程之间的相关性，为在脑胶质瘤术中胶质瘤分级和手术方案选择提供数值化非线性光学诊断信息。.非线性光学成像技术不但能够区分正常脑组织的白质区和灰质区，能够识别正常脑组织的胶质细胞、神经元细胞以及微血管，也能够对胶质瘤组织学诊断要素，包括细胞密度增加、坏死、微血管增生以及肾小球样血管增生等进行清晰地成像，证明了非线性光学成像技术有能力识别正常脑组织和胶质瘤组织，准确定位胶质瘤的边界，为在脑胶质瘤精准手术切除术中，实现脑胶质瘤“组织学最大范围切除”提供一个全新分子水平的实时医学成像技术。