The analysis of the open loop system in the design of the closed loop control systems is very important and useful in terms of being able to learn how the closed loop system will behave. In this paper, root locus graph of fractional order transfer function is plotted with the developed application using LabVIEW graphical programming language. A user panel has been designed using the LabVIEW graphical programming language. Interactive user panel is basic and user friendly for the computation and analysis. The root locus graph of the fractional order transfer function can interactively be plotted. Matsuda method is preferred for integer order approximation. With the interactive feature of developed application, the unit step response of closed loop control system can be plotted for a selected gain on the root locus graph panel. The application which has a rich graphical interface and interactive feature can also be used in teaching of fractional order control theory.