In this paper, a new application system with high speed photography, i.e. an observational system for the tele-micro-operation, has been proposed with a dynamic focusing system and a high-speed image processing system using the "Depth From Focus (DFF)" criteria. In micro operation, such as for the microsurgery, DNA operation and etc., the small depth of a focus on the microscope makes bad observation. For example, if the focus is on the object, the actuator cannot be seen with the microscope. On the other hand, if the focus is on the actuator, the object cannot be observed. In this sense, the "all-in-focus image," which holds the in-focused texture all over the image, is useful to observe the microenvironments on the microscope. It is also important to obtain the "depth map" which could show the 3D micro virtual environments in real-time to actuate the micro objects, intuitively. To realize the real-time micro operation with DFF criteria, which has to integrate several images to obtain "all-in-focus image" and "depth map," at least, the 240 frames par second based image capture and processing system should be required. At first, this paper briefly reviews the criteria of "depth from focus" to achieve the all-in-focus image and the 3D microenvironments' reconstruction, simultaneously. After discussing the problem in our past system, a new frame-rate system is constructed with the high-speed video camera and FPGA hardware with 240 frames par second. To apply this system in the real microscope, a new criterion "ghost filtering" technique to reconstruct the all-in-focus image is proposed. Finally, the micro observation shows the validity of this system.