From the point of view of practical application of the microchannel (MC) emulsification technique, which can be used to produce super-monodispersed microspheres (MS), we fabricated a stainless steel MC and investigated the production and characterization of oil-in-water (O/W) MS using a stainless steel MC instead of a silicon MC plate. We discovered that a stainless steel MC could not be fabricated precisely at a 1-mum scale; because of its multicrystal property, it can only be processed mechanically on a 10-mum scale. O/W-MS ranging from 20 to 210 mum in average diameter were produced using a stainless steel MC with 10 to 80 mum in equivalent diameter. The MS produced were monodispersed with a coefficient of variation lower than 3% for each individual channel. This value is smaller than that of normal emulsions obtained by the conventional emulsification techniques by 1 order of magnitude. The average diameter of the MS produced at breakthrough pressure was about 2.6 times the stainless steel MC equivalent diameter. The operation pressure affects the MS formation, causing a size-stable zone, size-expanding zone, and outflow zone observed. Larger stainless steel MC demonstrated difficulties in stably producing monodispersed O/W-MS. The breakthrough pressure was approximately inversely proportional to the MC equivalent diameter.