荧光显微成像技术是开展微观生命科学研究的重要手段和工具,使用该技术可以观察生物体内的精细结构、动态追踪生物体内组织、细胞、细胞核、蛋白、小分子等不同尺度的生命活动过程。其中,研究深层组织高时空分辨率荧光显微成像技术,是当前成像领域一个前沿问题。应用自适应光学技术实时补偿经由不透明散射、非均匀生物组织传播而引入的复杂波前畸变已被证实是实现上述技术的一种有效途径。文中首先归纳了深层动态荧光显微成像的需求和特点,随后分别介绍了自适应光学技术近几年在共聚焦显微成像、随机光学重构显微成像、光激活定位显微成像、受激辐射光淬灭显微成像、双光子/多光子激发显微成像中的相关应用,并对今后的研究问题和发展方向提出展望。
Fluorescent microscopy (FM) is an essential means for the Life Sciences research in micro- scale. By use of these advanced techniques, the most finest structures of in vivo biological tissue could be observed, as well as the dynamics mechanism at different levels be traced in realtime. At present, 3D in vivo FM with high spatiotemporal resolution through the deep scattering tissue is full of challenges. Being experienced nearly one decade development, the Adaptive Optics methodologies have been proven to be an effective way to correct the wavefront aberrations in space-variant caused by the heterogeneous tissue. In this paper, the imaging requirements and characteristics of 3D in vivo FM were summarized firstly. Then many feasible At applications attaching to the Confocal Microscopy, Stochastic Optical Reconstruction Microscopy, Photoactivated Localization Microscopy, Stimulated Emission Depletion Microscopy, Two-Photon/Multi-Photon Microscopy were presented, respectively. Finally, some probable research points and further trend were given.