Microfluidic chips such as lab-on-a-chip (LOC) include micro-channels for sample delivery, mixing, reaction, and separation. Pressure driven flow or electro-osmotic flow (EOF) has been usually employed to deliver bio-samples. Having some advantages of easy control, the flow characteristics of EOF in microchannels should be fully understood to effectively control the electro-osmotic pump for bio-sam-pie delivery. In this study, a micro PIV system with an epifluorescence inverted microscope and a cooled CCD was used to measure velocity fields of EOF in a glass microchannel and a PDMS microchannel. The EOF velocity fields were changed with respect to electric charge of seeding particles and microchannel materials used. The EOF has nearly uniform velocity distribution inside the microchannel when pressure gradient effect is negligible. The mean streamwise velocity is nearly proportional to the applied electric field. Glass microchannels give better repeatability in PIV results, compared with PDMS microchannels which are easy to fabricate and more suitable for PIV experiments.