双光子显微镜在神经药理学活体研究中的应用

In Vivo Application of Two-photon Microscopy in Neuropharmacological Research

得益于生物组织双光子吸收的非线性光学效应,双光子显微镜现在已经成为活体脑功能研究中重要的研究工具。双光子成像具有较深的穿透力、更为集中的空间聚焦、较小的组织损伤性等特征。因此,一方面利用双光子显微镜能够在细胞甚至是亚细胞水平上对活体中的神经细胞结构形态、离子浓度、细胞运动、分子相互作用等生理现象和过程进行直接的成像监测,另外还能进行光裂解、光激活、光转染和光损伤等光学操纵;另一方面,双光子活体成像单细胞分辨率的大范围成像也便于高通量的系统药理学研究。在多年使用双光子显微镜进行活体研究的基础上,我们结合最新的研究进展重点阐述双光子显微镜在神经科学、神经药理学以及疾病动物模型的活体研究中的应用,并探讨其更广阔的前景。

Two-photon microscope is an useful and advanced tool for noninvasive deep fluorescence imaging in the intact brain tissue of living animals. Due to nonlinear two-photon effects,two-photon microscope enables long-term imaging in vivo with deeper detection,higher signal-to-noise ratio and lower photodamage,compared to wide-field and confocal microscopy. Two-photon microscopy can provide high-resolution images to study cellular and subcellular structure and function,including morphology,mobility and intracellular ions of cells. On the other hand,large scale two-photon imaging of cell population reveals the network construction and activity dynamics with single-cell resolution,which makes two-photon microscopy a high throughput tool in system pharmacology. Moreover,two-photon microscopy can offer some precise optical operations,such as photolysis,photoactivation,phototransfection and photodamage. Here,we give an introduction to the principles of two-photon microscopy and its in vivo applications in neuroscience and neuropharmacology researches.