Gelatin microspheres cross-linked with genipin were developed to encapsulate the probiotic Bifidobacterium lactis Bb-12 The effects of different gelatin concentrations (10-19% w/v), bloom strengths (175 and 300), surfactants, stirring rates during emulsion formation and genipin concentrations (0-10mM) on the microsphere sizes and viability of bacterial cells were investigated. Principal Component Analysis revealed microsphere size distribution differed depending on the presence or absence of surfactants as well as a trend of increasing micropshere size with increasing gelatin concentration and bloom strength. Lower stirring rates resulted in larger microspheres with higher encapsulation yields of bifidobacteria Microsphere size and cell viability were not significantly (p< 0.05) influenced by increasing genipin concentrations up to 10mM whereas microsphere stability in simulated gastric juice increased with increasing genipin concentration. The encapsulation yields were higher in 175 bloom strength gelatin microspheres than in 300. Cold-stage scanning electron microscopy showed encapsulated bacteria distributed throughout the genipin cross-linked gelatin matrix.