An experimental system for rapid measurement of regional lung deposition of Di-2-ethylhexyl sebacate (DEHS) aerosol particles was established in the present study. The principal goal was to identify the most relevant components of the sampling train and the proper instruments to be employed. After completing the search for an optimal sampling train and instruments, a human subject test was performed. Overall, the sampling train consisted of a mouthpiece, flow meter, and particle counter. The mouthpiece was attached to a Fleisch pneumotachograph. Several TSI condensation particle counters (CPCs), a PC-LabCard and a personal computer were employed to measure and record the counts of test particles at 100 Hz. A cylinder-piston breathing machine was built to generate a series of “standard” breathing patterns. For non-human subject tests, an acrylic tube, 5 cm diameter × 60 cm length, packed with a piece of 100 ppi foam disks was used as a substitute for the human respiratory tract. The optimal sampling train was determined to be a 1TH Fleisch pneumotachograph with a CPC model 3025A because of its short response time and low flow fluctuation. A healthy non-smoking man volunteered to be the subject, and was asked to follow breathing patterns generated by a cylinder-piston breathing machine. The local deposition efficiency was calculated for 1 μm DEHS particles of each 50 cm3 volumetric region. The deposition data showed a good agreement with previous studies. Compared to the conventional bolus system, the advantage of the rapid measurement system developed in this work is its simplicity, low exposure, and high efficiency.