Infant formula (IF) containing probiotics was reconstituted at different temperatures (40, 50, 60 and 70 ℃) in order to analyze the effects of reconstitution temperature and methods (direct reconstitution in warm water and sequential reconstitution in warm and cool water) on the survival rate of probiotics. At the same time, microbial change in reconstituted IF were measured during storage at 5 ℃ for up to 24 h. The results showed that as the reconstitution increased, the survival rate of probiotics decreased. At the same reconstitution temperature, the second reconstitution method resulted in a significantly higher survival rate than did the first one. Reconstitution temperatures of 40 and 50 ℃ were beneficial to the survival of probiotics (survival rate > 60%). Storage at 5 ℃ ensured effectively the survival of the probiotics in reconstituted IF while controlling the growth of other microorganisms. In conclusion, IF containing probiotics is recommended to be reconstituted at 40–50 ℃ so as to ensure the survival of probiotics in the reconstituted IF.