This is a report on the synthesis of hexamethyl-propylene-amine oxime (HM-PAO), a new agent for regional cerebral blood flow (gamma CBF) imaging, the separation of its diastereoisomers (dl- and meso-HM-PAO), and a comparative study of three forms of Tc- 99m labelled HM-PAO (dl-, meso- and mixture-) in chromatography, mice biodistribution and rabbit imaging. The results showed that the Rf value of Tc-99m-dl-HM-PAO (0.8-1.0) on silica G thin layer chromatography with MEK as solvent was different from that of Tc-99m-meso-HM-PAO (0.6-0.8), and that the retention of dl- isomer in the brain was higher and more static than those of meso-and mixture HM-PAO but identical with Ceretec, a dl-HM-PAO kit manufactured by Amersham Inc., England. The primary clinical applications for gamma CBF SPECT imaging demonstrated a satisfactory result.