Abstract: 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 AO 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.