We studied a new method for inoculation of microorganisms to improve homogenization and their growth on plates. We named the method of Technique of Inoculation in test-Tube (TIT). It showed some advantages when compared to pour plate method such as facility and rapidity of preparation, besides easy homogenization. Using TIT, the growth of the microorganisms was reduced to 14 hours. The method was tested for counting total microorganisms in sugarcane juice using image software.