In the present study, a new microbial TTI system was developed based on encapsulation techniques. TTI response (color change) was evaluated by using primarily the CIELab coordinates which were further used for the conversion into the secondary and tertiary variables and finally compared all the variables to choose the best ones for calculation of activation energy (Ea) quickly and accurately. In order to select the best variables for calculating the Ea, 95% confidence interval was also used. Primary variable $a^*$ exhibited the best linearity, narrowest 95% confidence interval and simplicity in interpretation of the color-developing reaction. Ea of the developed microbial TTIs ranged from 57.28 to 110.23 kJ/mol and maintains a consistent performance similar to that of other microbial and nonmicrobial TTIs which are commercially available. Therefore, it can be concluded that this new microbial TTI system can be used effectively in the food industry for monitoring food quality.