The advance of life science research frontiers gradually increased the requirement in the performance of imaging tools. Nowadays microscopes are made to have greater resolution, higher frame rate and less photo-toxicity. This thesis will introduce some of the advanced microscopy techniques for bio-imaging. First, super-resolution technique, especially localization super-resolution technique provides nanometer resolution and enables us to build a refined understanding of the intracellular organization of cells with near-molecular resolution. Our efforts in building a user friendly localization microscope with optimized two-color performance will be introduced, together with its application in neuron science research. Second, Lightsheet microscopy using non-diffracting beams greatly shows great capability in rapid 3D imaging for live specimen with minimal photo bleaching. We will demonstrate that, using a pattern mask and an annulus, non-diffracting patterned Bessel sheets (PBS), or line Bessel sheets can be constructed easily for light sheet imaging and provide excellent performance in terms of resolution, speed and photo-toxicity.