Transparency micro-patterned pad is used to observe fine particles by an epi-illumination microscope. The material of transparency micro-patterned pad is resin which has almost same as the refractive index of water. Therefore, it has characteristic that the micro pattern could disappear in the observation image by filling water. Epi-illumination microscope has illumination system and the detection system on the same side. In this study, we succeeded to observe the diamond particles with a diameter of 1μm on the glass substrate at 1000 frame/s in dark-field illumination. This results show that the fine particles could be observed simplicity, and also suggest the possibility that the fine particles observation on opaque substrate using the epi-illumination microscope.