We designed and assembled a portable 3-D miniature microscopic image system with the size of 35x35x105 mm3 . By integrating a microlens array (MLA) into the optical train of a handheld microscope, the biological specimen’s image will be captured for ease of use in a single shot. With the light field raw data and program, the focal plane can be changed digitally and the 3-D image can be reconstructed after the image was taken. To localize an object in a 3-D volume, an automated data analysis algorithm to precisely distinguish profundity position is needed. The ability to create focal stacks from a single image allows moving or specimens to be recorded. Applying light field microscope algorithm to these focal stacks, a set of cross sections will be produced, which can be visualized using 3-D rendering. Furthermore, we have developed a series of design rules in order to enhance the pixel using efficiency and reduce the crosstalk between each microlens for obtain good image quality. In this paper, we demonstrate a handheld light field microscope (HLFM) to distinguish two different color fluorescence particles separated by a cover glass in a 600um range, show its focal stacks, and 3-D position.