An X-ray particle tracking velocimetry (PTV) technique was developed to simultaneously measure the sizes and velocities of microbubbles in a fluid without optical aberration. This technique is based on a combination of in-line X-ray holography and PTV. The X-ray PTV technique uses a configuration similar to that of conventional optical imaging techniques, and is easy to implement. In the present work, microbubbles generated from a fine wire by electrical heating were used as tracer particles. The X-ray PTV technique simultaneously recorded size and velocity data for microbubbles (_b=10–60 m) moving upward in an opaque tube (inner diameter =2.7 mm). Due to the different refractive indices of water and air, phase contrast X-ray images clearly show the exact size and shape of overlapped microbubbles. In all of the working fluids tested (deionised water and 0.01 M and 0.10 M NaCl solutions), the measured terminal velocity of the microbubbles rising through the solution was proportional to the square of the bubble diameter. The proposed technique can be used to extract useful information on the behaviour of various bio/microscale fluid flows that are not amenable to analysis using conventional methods.